Synpoptosis circuits for programmable cell death control

Abstract

Disclosed herein include methods, compositions, and kits suitable for use in programmable cell death control. Compositions (e.g., nucleic acid compositions, synthetic protein circuits) provided herein can comprise one or more apoptosis polypeptides; and / or one or more pyroptosis polypeptides; and / or one or more input polypeptides. In some embodiments, expression of a synthetic protein circuit of the disclosure can induce, e.g., apoptosis and / or pyroptosis in a cell.

Description

SYNPOPTOSIS CIRCUITS FOR PROGRAMMABLE CELL DEATH CONTROLRELATED APPLICATIONS

[0001] The present application claims priority to U.S. Provisional Application No. 63 / 548.335, filed November 13, 2023. The entire contents of this application is hereby expressly incorporated by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED R&D

[0002] This invention was made with government support under Grant No. EB030015 awarded by the National Institutes of Health. The government has certain rights in the invention.REFERENCE TO SEQUENCE LISTING

[0003] The present application is being filed along with a Sequence Listing in electronic format. The Sequence Listing is provided as a file entitled 30KJ-810001- WO_SequenceListing, created November 12, 2024, which is 52,267 bytes in size. The information in the electronic format of the Sequence Listing is incorporated herein by reference in its entirety.BACKGROUNDField

[0004] The present disclosure relates generally to the field of synthetic biology. Description of the Related Art

[0005] Mammalian systems use distinct cell death programs to eliminate harmful cells and shape immunity. Apoptosis is immunologically silent or “cold”. By contrast, pyroptosis is immunologically “hot” and involves substantial release of damage-associated molecular patterns (DAMPs).

[0006] Apoptosis and pyroptosis can each be advantageous depending on immunological context. The immunostimulatory nature of pyroptosis can promote cell killing. For example, inducing pyroptosis in a 15% minority of cells was sufficient to clear an entire tumor by boosting anti-tumor immunity. Consistently, expression of the gasdermin (GSDM) family of poreforming proteins, the executioners of pyroptosis, positively correlates with cancer patient survival, and cytotoxic lymphocytes upregulate GSDM expression in cancer cells. To escape pyroptosis, cancer cells generate loss-of-function GSDM mutations, silence GSDM expression, and express non-pyroptotic GSDM variants. Although beneficial in some contexts, pyroptosis can lead to pathological inflammation if triggered excessively. Therefore, it would be desirable to controllably induce apoptosis or pyroptosis and tune their relative frequencies.

[0007] Existing cell-killing approaches cannot fully direct the mode of cell death.Cytotoxic drugs are often limited to triggering apoptosis in cold tumors. CAR-T cells can effectively target cells expressing either a single antigen or multiple antigens. However, to kill target cells, CAR-T cells typically use granzymes, which may induce either apoptosis or pyroptosis. Granzyme-independent approaches have also been attempted, including engineered TRAIL-presenting cells and synthetic circuits that regulate caspases, BID, or BAX. However, these approaches are similarly restricted to induction of apoptosis.

[0008] To enable tailored control of cell death, there is a need for a set of synthetic circuits with the following features. First, the circuits should allow activation and repression of both apoptosis and pyroptosis. Second, they should steer the mode of cell death in various cell contexts. Third, they should allow the integration and computation of multiple input signals. Fourth, they should be able to selectively kill target cells. Fifth, they should support cell-cell transmission, offering the potential to engineer synthetic killer cells that use designed death programs to eliminate other cells.SUMMARY

[0009] Disclosed herein include synthetic protein circuits comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, wherein two first apoptosis polypeptides are capable of associating with each other to constitute a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0010] Disclosed herein include synthetic protein circuits comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a first partner domain; and a second apoptosis polypeptide comprising a small subunit of an apoptotic effector protein, a second partner domain capable of binding the first partner domain, a first heterologous protease cleavage site, and a first degron, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second apoptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptosispolypeptide from a second apoptosis polypeptide destabilized state to a second apoptosis polypeptide stabilized state, wherein the first apoptosis polypeptide and the second apoptosis polypeptide in the second apoptosis polypeptide stabilized state are capable of associating via binding of the first partner domain and the second partner domain to form a subunit, and wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0011] Disclosed herein include synthetic protein circuits comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, a first partner domain, a first degron, and a first heterologous protease cleavage site; and a second apoptosis polypeptide comprising a small subunit of an apoptotic effector protein and a second partner domain capable of binding the first partner domain, wherein the first apoptosis polypeptide and the second apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a subunit, wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0012] In some embodiments, the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase-4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase- 10, caspase- 14 or any variant, portion, or derivative thereof. In some embodiments, the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first heterologous protease is engineered. In some embodiments, the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2.In some embodiments, the first heterologous protease cleavage site is natural or engineered, e.g., the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site. In some embodiments, the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7. In some embodiments, the first apoptotic protein complex in the first apoptotic protein complex active state is capable of being inhibited by a small molecule inhibitor of apoptosis, e.g., Quinoline-Val- Asp-Difluorophenoxymethylketone (Q-VD-OPh), carbobenzoxy-valyl-alanyl-aspartyl-[O- methyl] -fluoromethylketone (Z-VAD-FMK), and / or emricasan.

[0013] In some embodiments, the first apoptosis polypeptide and / or the second apoptosis polypeptide comprise one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state. In some embodiments, said third partner domains and fourth partner domains are capable of multimerization. In some embodiments, the first partner domain and the second partner domain are homodimers, and / or wherein the third partner domain and the fourth partner domain are homodimers. In some embodiments, the first partner domain and the second partner domain are heterodimers and / or wherein the third partner domain and the fourth partner domain are heterodimers. In some embodiments, the binding betw een the first partner domain and the second partner domain is reversible and / or wherein the binding between the third partner domain and the fourth partner domain is reversible.

[0014] In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise SYNZIP1. SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18, SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS. ATF4. BACH1, JUND, NFE2L3, AZip, BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13 1 :234 heterodimer a, DHD 15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a. DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a. DHD33 heterodimer a,DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a. DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a, DHD37_3: 124 heterodimer a, DHD37_1:234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a. DHD39 heterodimer a, DHD40 heterodimer a. DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a, DHD94_2: 143 heterodimer a, DHD95 heterodimer a. DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a. DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103_l:423 heterodimer a, DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a. DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a, DHD122 heterodimer a, DHD123 heterodimer a. DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a. DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a, DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a, DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a. DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a, DHD159 heterodimer a, DHD160 heterodimer a. DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a, DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a, DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD 13 2: 341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13 4: 123 heterodimer b, DHD13 1 :234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b. DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1:234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b. DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3:214 heterodimer b, DHD94_2:143 heterodimer b. DHD95 heterodimer b. DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l:243 heterodimer b, DHD103 heterodimer b, DHD103_l:423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b. DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD111 heterodimer b, DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD1 15 heterodimer b, DHD116 heterodimer b, DHD1 17 heterodimer b. DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b, DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b, DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b, DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b. DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b. DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD148 heterodimer b, DHD 149 heterodimer b, DHD150 heterodimer b. DHD151 heterodimer b, DHD 152 heterodimer b, DHD 153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b. DHD 162 heterodimer b, DHD 163 heterodimer b, DHD 164 heterodimer b,DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b, DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, Pl peptide, P2 peptide, P3 peptide, P4 peptide, P5 peptide, P6 peptide, P7 peptide, P8 peptide, P9 peptide, PIO peptide, Pl l peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer, N8 heterodimer, P8A heterodimer, an Sih peptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0015] In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3-3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine- phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, a Dbl homology domain, a gelsolin homology domain, a PB1 domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof. In some embodiments, the first partner domain and the second partner domain and / or the third partner domain and the fourth partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (f) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise nHalo, cHalo, or any combination thereof.

[0016] Disclosed herein include synthetic protein circuits comprising: a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domainseparated by a first heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0017] In some embodiments, the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, e.g., a bulk}' domain. In some embodiments, the first inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein. In some embodiments, the cell does not express an endogenous protein comprising a pyroptosis effector domain.

[0018] Disclosed herein include synthetic protein circuits comprising: a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first degron separated by a first heterologous protease cleavage site, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from a first pyroptosis polypeptide destabilized state to a first pyroptosis polypeptide stabilized state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide stabilized state is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0019] Disclosed herein include synthetic protein circuits comprising: a first pyroptosis polypeptide comprising a pyroptosis effector domain, a first degron, and a first heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, and wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the firstpyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0020] In some embodiments, the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is from the gasdermin (GSDM) family, including GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59. In some embodiments, the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukary otic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first heterologous protease is engineered. In some embodiments, the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP). derivatives thereof, or any combination thereof. In some embodiments, the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the first heterologous protease cleavage site is natural or engineered, e.g., the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site. In some embodiments, the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7.

[0021] In some embodiments, the first pyroptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to the first pyroptosis polypeptide active state in the absence of the first heterologous protease in the first heterologous protease active state. In some embodiments, said third partner domains and fourth partner domains are capable of multimerization . In some embodiments, the third partner domain and the fourth partner domain are homodimers. In some embodiments, the third partner domain and the fourth partner domain are heterodimers. In some embodiments, the binding between the third partner domain and the fourth partner domain is reversible.

[0022] In some embodiments, the third partner domain and / or the fourth partner domain comprise SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18. SYNZIP19, SYNZIP20, SYNZIP21. SYNZIP22,SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip, BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13_1:234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a. DHD21 heterodimer a, DHD25 heterodimer a. DHD27 heterodimer a, DHD30 heterodimer a, DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a. DHD37 3: 124 heterodimer a, DHD37 1 :234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a. DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103 E423 heterodimer a. DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD1 14 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a. DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a. DHD16 heterodimer a, DHD18 heterodimer a. DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a. DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a. DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a. DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a,DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a. DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a. DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a. DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD13_2:341 heterodimer b, DHD13 AAAA heterodimer b. DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b. DHD13 1:234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1 :234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b. DHD94 3:214 heterodimer b, DHD94 2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b,DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l:243 heterodimer b, DHD103 heterodimer b, DHD103_l:423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD1 11 heterodimer b. DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD115 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b. DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b. DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b. DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b. DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67heterodimer b, DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD151 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b. DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b, DHD163 heterodimer b, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b. DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, Pl peptide. P2 peptide, P3 peptide, P4 peptide, P5 peptide, P6 peptide, P7 peptide, P8 peptide, P9 peptide, PIO peptide, Pl l peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer, N8 heterodimer, P8A heterodimer, an S ih peptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0023] In some embodiments, the third partner domain and / or the fourth partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, a Dbl homology domain, a gelsolin homology domain, aPBl domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof. In some embodiments, the third partner domain and the fourth partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (I) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the third partner domain and / or the fourth partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, the third partner domain and / or the fourth partner domain comprise nHalo, cHalo, or any combinationthereof. In some embodiments, inducing pyroptosis in the cell causes the cell to induce key signatures of pyroptosis: chromatin condensation and DNA fragmentation, pore formation, cell swelling, and osmotic lysis, followed by release of one or more inflammatory cytokines. In some embodiments, the one or more inflammatory cytokines comprise IL-18, IL-1(3, IL-6. IL-8, interferon gamma (IFN-y). and / or tumor necrosis factor-alpha (TNF-a).

[0024] Disclosed herein include synthetic protein circuits comprising: one or more apoptosis polypeptides; and / or one or more pyroptosis polypeptides; and / or one or more input polypeptides.

[0025] Disclosed herein include synthetic protein circuits comprising: (i) one or more apoptosis polypeptides or one or more pyroptosis polypeptides and (ii) one or more input polypeptides, configured to form one or more logic gates selected from the group comprising an OR logic gate, AND logic gate, NOR logic gate, NAND logic gate, IMPLY logic gate, NIMPLY logic gate, XOR logic gate, and an XNOR logic gate.

[0026] The synthetic protein circuit can comprise: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, a second heterologous protease cleavage site, and a first degron, wherein the first apoptosis polypeptide is capable of being in a first apoptosis polypeptide destabilized state; (ii) a first input polypeptide comprising a first heterologous protease; and / or (iii) a second input polypeptide comprising a second heterologous protease, wherein the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide, wherein the second heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptosis polypeptide from the first apoptosis polypeptide destabilized state to a first apoptosis polypeptide stabilized state; wherein two of the first apoptosis polypeptides in the first apoptosis polypeptide stabilized state are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, and wherein the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, and wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell.

[0027] The synthetic protein circuit can comprise: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site and a second heterologousprotease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state; (ii) a first input polypeptide comprising a first heterologous protease; and / or (iii) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state; (II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the second heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state; or (III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state, and / or the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the second heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell.

[0028] The synthetic protein circuit can comprise: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, a first partner domain, a first degron, and a first heterologous protease cleavage site; (ii) a second apoptosis polypeptide comprising a small subunit of an apoptotic effector protein, a second partner domain capable of binding the first partner domain, a second heterologous protease cleavage site, and a second degron, (iii) a first input polypeptide comprising a first heterologous protease; and / or (iv) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state; (II) the second heterologous protease in a second heterologous protease active state is capable of cuttingthe second heterologous protease cleavage site of the second apoptosis polypeptide and thereby releasing the second degron, and wherein the second heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptosis polypeptide from a second apoptosis polypeptide destabilized state to a second apoptosis polypeptide stabilized state, wherein the first apoptosis polypeptide and the second apoptosis polypeptide in the second apoptosis polypeptide stabilized state are capable of associating via binding of the first partner domain and the second partner domain to form a subunit, and wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; or (III) wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from the first apoptosis polypeptide stabilized state to the first apoptosis polypeptide destabilized state, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide and thereby releasing the second degron, and wherein the second heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptosis polypeptide from the second apoptosis polypeptide destabilized state to the second apoptosis polypeptide stabilized state.

[0029] The synthetic protein circuit can comprise: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, a second heterologous protease cleavage site, and a first degron; (ii) a second apoptosis polypeptide comprising the large subunit of the apoptotic effector protein and the small subunit of the apoptotic effector protein separated by the second heterologous protease cleavage site, the first heterologous protease cleavage site, and a second degron. wherein two of the first apoptosis polypeptides are capable forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, two of the second apoptosis polypeptides are capable forming a second apoptotic protein complex in a second apoptotic protein complex inactive state, and / or one of the first apoptosis polypeptide and one of the second apoptosis polypeptide are capable forming a third apoptotic protein complex in a third apoptotic protein complex inactive state; (iii) a first input polypeptide comprising a first heterologous protease; and / or (iv) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide,and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, and the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second apoptosis polypeptide to expose the second degron, and wherein the second degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from a second apoptosis polypeptide stabilized state to a second apoptosis polypeptide destabilized state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; (II) wherein the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide, and wherein the second heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptotic protein complex from the second apoptotic protein complex inactive state to a second apoptotic protein complex active state, and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state, wherein the second apoptotic protein complex in the second apoptotic protein complex active state is capable of inducing apoptosis in the cell; or (III) wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second apoptosis polypeptide to expose the first degron, and wherein the first degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from the second apoptosis polypeptide stabilized state to the second apoptosis polypeptide destabilized state; and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron. and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from the first apoptosis polypeptide stabilized state to the first apoptosis polypeptide destabilized state.

[0030] The synthetic protein circuit can comprise: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, and a first partner domain and a first degron separated by a first heterologous protease cleavage site, and a second heterologous protease cleavage site; (ii) a second apoptosis polypeptide comprising a small subunit of an apoptotic effector protein and a second partner domain capable of binding the first partner domain,wherein the first apoptosis polypeptide and the second apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a subunit; wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; (iii) a first input polypeptide comprising a first heterologous protease; and / or (iv) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, thereby releasing the first degron; (II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state.

[0031] The synthetic protein circuit can comprise: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, a first partner domain, a first degron, and a first heterologous protease cleavage site; (ii) a second apoptosis polypeptide comprising the large subunit of the apoptotic effector protein, the first partner domain, a second degron. and a second heterologous protease cleavage site; (iii) a third apoptosis polypeptide comprising a small subunit of an apoptotic effector protein and a second partner domain capable of binding the first partner domain, wherein the first apoptosis polypeptide and the third apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a first subunit, and / or the second apoptosis polypeptide and the third apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a second subunit, wherein two first subunits, two second subunits, and / or one first subunit and one second subunit are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; (iv) a first input polypeptide comprising a first heterologous protease; and / or (v) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state; (II) the second heterologous protease in a second heterologousprotease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide to expose the second degron, and wherein the second degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from a second apoptosis polypeptide stabilized state to a second apoptosis polypeptide destabilized state; or (III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from the first apoptosis polypeptide stabilized state to the first apoptosis polypeptide destabilized state, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide to expose the second degron, and wherein the second degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from the second apoptosis polypeptide stabilized state to the second apoptosis polypeptide destabilized state.

[0032] In some embodiments, the apoptotic effector protein comprises caspase- 1, caspase-2, caspase-3, caspase-4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase- 10, caspase- 14 or any variant, portion, or derivative thereof. In some embodiments, the first and / or second heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first and / or second heterologous protease is engineered. In some embodiments, wherein the first and / or second heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the first and / or second heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the first and / or second heterologous protease cleavage site is natural or engineered, e.g., a cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the first and / or the second heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first and / or second heterologous protease cleavage site. In some embodiments, the first and / or second heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7. In some embodiments, the first and second heterologous protease are different from each other. In some embodiments, the first, second, and / or third apoptotic protein complex in the first, second, and / or third apoptotic protein complex active state is capable of being inhibited by a smallmolecule inhibitor of apoptosis, e.g., Quinoline-Val-Asp-Difluorophenoxymethylketone (Q-VD- OPh), carbobenzoxy-valyl-alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK), and / or emricasan

[0033] In some embodiments, the first apoptosis polypeptide, the second apoptosis polypeptide and / or the third apoptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first, second, and / or third apoptotic protein complex in the first, second, and / or third apoptotic protein complex inactive state to the first, second, and / or third apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state and / or the second heterologous protease in the second heterologous protease in the second heterologous protease active state. In some embodiments, said third partner domains and fourth partner domains are capable of multimerization. In some embodiments, the first partner domain and the second partner domain are homodimers and / or wherein the third partner domain and the fourth partner domain are homodimers. In some embodiments, the first partner domain and the second partner domain are heterodimers and / or wherein the third partner domain and the fourth partner domain are heterodimers. In some embodiments, the binding between the first partner domain and the second partner domain is reversible and / or wherein the binding between the third partner domain and the fourth partner domain is reversible.

[0034] In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6. SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18, SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip, BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13 2 341 heterodimer a, DHD13 AAAA heterodimer a. DHD13 BAAA heterodimer a. DHD13_4: 123 heterodimer a, DHD13 1 :234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a, DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34_XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a, DHD37_3: 124 heterodimer a, DHD37_1:234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a. DHD89 heterodimer a, DHD90heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a, DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l:243 heterodimer a, DHD103 heterodimer a, DHD103_l:423 heterodimer a, DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a. DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a, DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a. DHD145 heterodimer a. DHD146 heterodimer a. DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a. DHD29 heterodimer a, DHD31 heterodimer a. DHD32 heterodimer a, DHD38 heterodimer a. DHD60 heterodimer a, DHD63 heterodimer a, DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a, DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a, DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a, DHS17 heterodimer a, DHD17 heterodimer a. DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a. DHD135 heterodimer a, DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13_XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13_XAAX heterodimer b. DHD 13 2:341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b, DHD13_1 :234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b,DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1:234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b. DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3:214 heterodimer b, DHD94_2:143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD 102 1:243 heterodimer b, DHD103 heterodimer b, DHD103_l:423 heterodimer b, DHD 104 heterodimer b, DHD 105 heterodimer b, DHD 106 heterodimer b, DHD 107 heterodimer b, DHD 108 heterodimer b, DHD 109 heterodimer b, DHD110 heterodimer b, DHD111 heterodimer b, DHD112 heterodimer b, DHD113 heterodimer b, DHD 114 heterodimer b, DHD115 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD 120 heterodimer b, DHD121 heterodimer b, DHD 122 heterodimer b, DHD 123 heterodimer b, DHD 124 heterodimer b, DHD 125 heterodimer b, DHD 126 heterodimer b, DHD 127 heterodimer b, DHD 128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD 146 heterodimer b. DHD 147 heterodimer b, DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b, DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD 148 heterodimer b, DHD 149 heterodimer b, DHD 150 heterodimer b, DHD 151 heterodimer b, DHD 152 heterodimer b, DHD 153 heterodimer b, DHD154 heterodimer b. DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD 162 heterodimer b, DHD 163 heterodimer b, DHD 164 heterodimer b, DHD 165 heterodimer b, DHD 166 heterodimer b, DHS17 heterodimer b, DHD 17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b, DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD 139 heterodimer b, DHD 140 heterodimer b, DHD 141 heterodimer b, DHD 142 heterodimer b, DHD 143 heterodimer b, DHD 144 heterodimer b, P 1 peptide, P2 peptide, P3 peptide. P4 peptide, P5 peptide, P6 peptide, P7 peptide. P8 peptide, P9 peptide, P10 peptide,P 11 peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer, N8 heterodimer, P8A heterodimer, an Sih peptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0035] In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3-3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine- phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain. aDbl homology domain, a gelsolin homology domain, a PB1 domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof. In some embodiments, the first partner domain and the second partner domain and / or the third partner domain and the fourth partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (1) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, the first partner domain, the second partner domain, the third partner domain, and / or the fourth partner domain comprise nHalo. cHalo. or any combination thereof.

[0036] The synthetic protein circuit can comprise: (i) a first pyroptosis polypeptide comprising a pyroptosis effector domain, a first inhibitory' domain, and a second inhibitory' domain, wherein the pyroptosis effector domain and the first inhibitory' domain are separated by a first heterologous protease cleavage site, and the pyroptosis effector domain and the second inhibitory domain are separated by a second heterologous cleavage site, wherein the first and / or second inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state; (ii) a first input polypeptide comprising a first heterologous protease; and / or (iii) a second input polypeptidecomprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut releases the first inhibitory domain from the first pyroptosis polypeptide, thereby the second inhibitory domain is capable of inhibiting the activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in the first pyroptosis polypeptide inactive state; (II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut releases the second inhibitory domain from the first pyroptosis polypeptide, thereby the first inhibitory domain is capable of inhibiting the activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in the first pyroptosis polypeptide inactive state; or (III) wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut releases the first inhibitory7domain from the first pyroptosis polypeptide, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut releases the second inhibitory- domain from the first pyroptosis polypeptide, wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut and the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, and wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell.

[0037] In some embodiments, the first and / or second inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, e.g., a bulky domain. In some embodiments, the first and / or second inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein. In some embodiments, the cell does not express an endogenous protein comprising a pyroptosis effector domain.

[0038] The synthetic protein circuit can comprise: (i) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domain separated by a first heterologous protease cleavage site and a second heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, therebythe first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state; (ii) a first input polypeptide comprising a first heterologous protease; and / or (iii) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell; (II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell; or (III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, and / or the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in the cell.

[0039] In some embodiments, the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, e.g., a bulky domain. In some embodiments, the first inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein. In some embodiments, the cell does not express an endogenous protein comprising a pyroptosis effector domain.

[0040] The synthetic protein circuit can comprise: (i) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first degron separated by a first heterologous protease cleavage site, a second degron, and a second heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide destabilized state; (ii) a first input polypeptide comprising a first heterologous protease; and / or (iii) a secondinput polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide destabilized state to a first pyroptosis polypeptide stabilized state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide stabilized state is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell; or (II) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide destabilized state to the first pyroptosis polypeptide stabilized state, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide in the first pyroptosis polypeptide destabilized state to expose the second degron. and wherein the second degron of the first pyroptosis polypeptide in the first pyroptosis polypeptide stabilized state being exposed changes the first pyroptosis polypeptide from the first pyroptosis polypeptide stabilized state to the first pyroptosis polypeptide destabilized state.

[0041] The synthetic protein circuit can comprise: (i) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domain separated by a first heterologous protease cleavage site, a second heterologous cleavage site, and a first degron, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain of the first pyroptosis polypeptide, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state; (ii) a second pyroptosis polypeptide comprising a pyroptosis effector domain and a second inhibitory domain separated by the second heterologous protease cleavage site, the first heterologous cleavage site, and a second degron, wherein the second inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain of the second pyroptosis polypeptide, thereby the second pyroptosis polypeptide is in a second pyroptosis polypeptide inactive state; (iii) a first input polypeptide comprising a first heterologous protease; and / or (iv) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the firstpyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, and wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron, and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from a second pyroptosis polypeptide stabilized state to a second pyroptosis polypeptide destabilized state: (II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the second pyroptosis polypeptide being cut changes the second pyroptosis polypeptide from the second pyroptosis polypeptide inactive state to a second pyroptosis polypeptide active state, wherein the second pyroptosis polypeptide in the second pyroptosis polypeptide active state is capable of inducing pyroptosis in the cell, and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state; or (III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron, and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from the second pyroptosis polypeptide stabilized state to the second pyroptosis polypeptide destabilized state, and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from the first pyroptosis polypeptide stabilized state to the first pyroptosis polypeptide destabilized state.

[0042] In some embodiments, the first and / or second inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, e.g., a bulky domain. In some embodiments, the first and / or second inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein. In some embodiments, the cell does not express an endogenous protein comprising a pyroptosis effector domain.

[0043] The synthetic protein circuit can comprise: (i) a first pyroptosis polypeptidecomprising a pyroptosis effector domain and a first degron separated by a first heterologous protease cleavage site, and a second heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell ; (ii) a first input polypeptide comprising a first heterologous protease; and / or (iii) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron from the first pyroptosis polypeptide; or (II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state.

[0044] The synthetic protein circuit can comprise: (i) a first pyroptosis polypeptide comprising a pyroptosis effector domain, a first degron, and a first heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell; (ii) a second pyroptosis polypeptide comprising a pyroptosis effector domain, a second degron, and a second heterologous protease cleavage site, wherein the second pyroptosis polypeptide is capable of being in a second pyroptosis polypeptide active state, wherein the second pyroptosis polypeptide in the second pyroptosis polypeptide active state is capable of inducing pyroptosis in the cell; (iii) a first input polypeptide comprising a first heterologous protease; and / or (iv) a second input polypeptide comprising a second heterologous protease, wherein: (I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state; (II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron, and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from a second pyroptosis polypeptide stabilized state to a second pyroptosis polypeptide destabilized state; or (III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologousprotease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from the first pyroptosis polypeptide stabilized state to the first pyroptosis polypeptide destabilized state, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron, and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from the second pyroptosis polypeptide stabilized state to the second pyroptosis polypeptide destabilized state.

[0045] In some embodiments, the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is from the gasdermin (GSDM) family, including GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59. In some embodiments, the first and / or second heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first and / or second heterologous protease is engineered. In some embodiments, the first and / or second heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the first and / or second heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the first and / or second heterologous protease cleavage site is natural or engineered, e.g., the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the first and / or second heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first and / or second heterologous protease cleavage site. In some embodiments, the first and / or second heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7. In some embodiments, the first and / or second heterologous protease are different from each other.

[0046] In some embodiments, the first pyroptosis polypeptide and / or the second pyroptosis polypeptide comprise one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of, inhibiting change of the first and / or second pyroptosis polypeptide from the first and / or second pyroptosis polypeptide inactive state to the first and / or second pyroptosis polypeptide active state, in the absence of the first and / or second heterologous protease in the first and / or second heterologous protease active state. In some embodiments, saidthird partner domains and fourth partner domains are capable of multimerization. In some embodiments, the third partner domain and the fourth partner domain are homodimers. In some embodiments, the third partner domain and the fourth partner domain are heterodimers. In some embodiments, the binding between the third partner domain and the fourth partner domain is reversible.

[0047] In some embodiments, the third partner domain and / or the fourth partner domain comprise SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18. SYNZIP19, SYNZIP20, SYNZIP21. SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip, BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD I 3 I 234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a, DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a. DHD37 3: 124 heterodimer a, DHD37 1 :234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a. DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103 E423 heterodimer a. DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a. DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a. DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8heterodimer a, DHD16 heterodimer a, DHD 18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a. DHD66 heterodimer a, DHD67 heterodimer a. DHD69 heterodimer a, DHD71 heterodimer a. DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a. DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a. DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD 165 heterodimer a, DHD166 heterodimer a. DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a. DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD 13 2: 341 heterodimer b, DHD13 AAAA heterodimer b. DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b. DHD 13 1:234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1 :234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b. DHD94 heterodimer b. DHD94_3:214 heterodimer b. DHD94 2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l:243 heterodimer b, DHD 103 heterodimer b, DHD103_l:423 heterodimer b, DHD 104 heterodimer b, DHD 105 heterodimer b, DHD 106 heterodimer b, DHD 107 heterodimer b, DHD 108 heterodimer b, DHD 109 heterodimer b, DHD 110 heterodimer b, DHD1 11 heterodimer b. DHD112 heterodimer b, DHD113 heterodimer b, DHD 114 heterodimer b, DHD115 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD 120 heterodimer b, DHD 121 heterodimer b, DHD 122heterodimer b, DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b. DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b. DHD5 heterodimer b. DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b. DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b. DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD151 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b, DHD163 heterodimer b, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b. DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, Pl peptide. P2 peptide, P3 peptide, P4 peptide, P5 peptide, P6 peptide, P7 peptide, P8 peptide, P9 peptide, PIO peptide, Pl 1 peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer, N8 heterodimer, P8A heterodimer, an S ih peptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0048] In some embodiments, the third partner domain and / or the fourth partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology7domain, aDbl homology7domain, a gelsolin homology7domain, aPBl domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof,variants thereof, or any combination thereof. In some embodiments, the third partner domain and the fourth partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (f) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the third partner domain and / or the fourth partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, the third partner domain and / or the fourth partner domain comprise nHalo, cHalo, or any combination thereof. In some embodiments, inducing pyroptosis in the cell causes the cell to release one or more inflammatory cytokines. In some embodiments, the one or more inflammatory cytokines comprise IL-18, IL-1 (3, IL-6, IL-8, interferon gamma (IFN-y), and / or tumor necrosis factor-alpha (TNF-a).

[0049] In some embodiments, the first apoptosis polypeptide, the second apoptosis polypeptide, the third apoptosis polypeptide, the first pyroptosis polypeptide, the second pyroptosis polypeptide, the first input polypeptide, and / or the second input polypeptide comprise one or more linkers. In some embodiments, the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, e.g., about 1 to about 18 flexible amino acid residues. In some embodiments, the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 2 repeating amino acid subunits or more.

[0050] In some embodiments: (i) the first, second, and / or third apoptosis polypeptide; (ii) the first and / or second pyroptosis polypeptide; and / or (iii) the first and / or second input polypeptide, are configured to be in a first localized state. In some embodiments, the first localized state comprises a state created by phase separation, a state defined by the proximity7to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome. microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome. ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof. In some embodiments, the first, second, and / or third apoptosis polypeptide, the first and / or second pyroptosis polypeptide, and / or the first and / or second input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state. In some embodiments:(i) the first, second, and / or third apoptosis polypeptide; (ii) the first and / or second pyroptosis polypeptide; and / or (iii) the first and / or second input polypeptide, are configured to be in second localized state(s), wherein the first localized state and the second localized state(s) are different. In some embodiments, the second localized state(s) comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof. In some embodiments, the first, second, and / or third apoptosis polypeptide, the first and / or second pyroptosis polypeptide, and / or the first and / or second input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state. In some embodiments: (i) the first, second, and / or third apoptosis polypeptide; (ii) the first and / or second pyroptosis polypeptide; and / or (iii) the first and / or second input polypeptide comprise a first localization signal. In some embodiments, the first localization signal is adjacent to a third degron and / or a third heterologous cleavage site. In some embodiments: (i) the first, second, and / or third apoptosis polypeptide; (ii) the first and / or second pyroptosis polypeptide; and / or (iii) the first and / or second input polypeptide comprise second localization signal(s). In some embodiments, the second localization signal is adjacent to a third degron and / or a third heterologous protease cleavage site. In some embodiments, the presence of the third degron and / or wherein the third heterologous cleavage site being cut changes: (i) the first, second, and / or third apoptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or to a non-localized state; (ii) the first and / or second pyroptosis polypeptide from a nonlocalized state or the first localized state to the second localized state(s) or to a non-localized state; and / or (iii) the first and / or second input polypeptide from a non-localized state or the first localized state to the second localized state(s) or to a non-localized state. In some embodiments, the synthetic protein circuit further comprises a third input polypeptide comprising a third heterologous protease capable of cutting the third heterologous protease cleavage site. In some embodiments, the third heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the third heterologous protease is engineered. In some embodiments, the third heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease,hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the third heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the third heterologous protease cleavage site is natural or engineered, e.g., the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the third heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild ty pe third heterologous protease cleavage site. In some embodiments, the third heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7. In some embodiments, the first localization signal and / or second localization signal(s) is selected from the group comprising a nuclear localization signal (NLS), a nuclear export signal (NES), a peroxisomal targeting signal (PTS), a mitochondrial targeting sequence (MTS), a ER signal peptide, CAAX box, a peroxisomal targeting signal (PTS). PTS1, PTS2, a dileucine motif, YxxO motif, a palmitoylation motif, a GPI anchor signal, myristoylation signal, HDEL, KDEL, KKXX motif, RXR motif, SV40 NLS, SV40 NES, CAAX membrane tether, ER recruitment, portions thereof, derivatives thereof, or any combination thereof. In some embodiments: (a) the first, second, and / or third apoptosis polypeptide in the first localized state; (b) the first, second, and / or third apoptosis polypeptide in the second localized state(s); (c) the first and / or second pyroptosis in the first localized state; (d) the first and / or second pyroptosis polypeptide in the second localized state(s); (e) the first and / or second input polypeptide in the first localized state; and / or (f) the first and / or second input polypeptide in the second localized state(s); is capable of modulating an activation threshold and / or sensitivity’ of the synthetic protein circuit.

[0051] In some embodiments, the synthetic protein circuit is present in a cell. In some embodiments, the cell is: a cell of a subject, e.g., a subject suffering from a disease or disorder. In some embodiments, the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof; a cell derived from a donor; and / or an in vivo cell, an ex vivo cell, an in vitro cell or an in situ cell. In some embodiments, the cell is a eukaryotic cell, e.g.. a mammalian cell. In some embodiments, the mammalian cell comprises an antigen-presenting cell, a dendritic cell, a macrophage, a neural cell, a brain cell, an astrocyte, a microglial cell, and a neuron, a spleen cell, a lymphoid cell, a lung cell, a lung epithelial cell, a skin cell, a keratinocyte, an endothelial cell, an alveolar cell, an alveolar macrophage, an alveolar pneumocyte, a vascular endothelial cell, a mesenchymal cell, anepithelial cell, a colonic epithelial cell, a hematopoietic cell, a bone marrow cell, a Claudius cell, Hensen cell, Merkel cell, Muller cell, Paneth cell, Purkinje cell, Schwann cell, Sertoli cell, acidophil cell, acinar cell, adipoblast, adipocyte, brown or white alpha cell, amacrine cell, beta cell, capsular cell, cementocyte, chief cell, chondroblast, chondrocyte, chromaffin cell, chromophobic cell, corticotroph, delta cell, Langerhans cell, follicular dendritic cell, enterochromaffin cell, ependymocyte, epithelial cell, basal cell, squamous cell, endothelial cell, transitional cell, erythroblast, erythrocyte, fibroblast, fibrocyte, follicular cell, germ cell, gamete, ovum, spermatozoon, oocyte, primary oocyte, secondary oocyte, spermatid, spermatocyte, primary spermatocyte, secondary’ spermatocyte, germinal epithelium, giant cell, glial cell, astroblast, astrocyte, oligodendroblast, oligodendrocyte, glioblast, goblet cell, gonadotroph, granulosa cell, haemocytoblast, hair cell, hepatoblast, hepatocyte, hyalocyte, interstitial cell, juxtaglomerular cell, keratinocyte, keratocyte, lemmal cell, leukocyte, granulocyte, basophil, eosinophil, neutrophil, lymphoblast, B-lymphoblast, T-lymphoblast, lymphocyte, B-lymphocyte, T-lymphocyte, helper induced T-lymphocyte, Thl T-lymphocyte, Th2 T-lymphocyte, natural killer cell, thymocyte, macrophage, Kupffer cell, alveolar macrophage, foam cell, histiocyte, luteal cell, lymphocytic stem cell, lymphoid cell, lymphoid stem cell, macroglial cell, mammotroph, mast cell, medulloblast, megakaryoblast, megakaryocyte, melanoblast, melanocyte, mesangial cell, mesothelial cell, metamyelocyte, monoblast, monocyte, mucous neck cell, myoblast, myocyte, muscle cell, cardiac muscle cell, skeletal muscle cell, smooth muscle cell, myelocyte, myeloid cell, myeloid stem cell, myoblast, myoepithelial cell, myofibrobast, neuroblast, neuroepithelial cell, neuron, odontoblast, osteoblast, osteoclast, osteocyte, oxyntic cell, parafollicular cell, paraluteal cell, peptic cell, pericyte, peripheral blood mononuclear cell, phaeochromocyte, phalangeal cell, pinealocyte, pituicyte, plasma cell, platelet, podocyte, proerythroblast, promonocyte, promyeloblast, promyelocyte, pronormoblast, reticulocyte, retinal pigment epithelial cell, retinoblast, small cell, somatotroph, stem cell, sustentacular cell, teloglial cell, a zymogenic cell, or any combination thereof. In some embodiments, the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell (iPSC), a hematopoietic stem / progenitor cell (HSPC), or any combination thereof.

[0052] Disclosed herein include synthetic protein circuits comprising: one or more first, second, or third apoptosis polypeptides; one or more first or second pyroptosis polypeptides; one or more input polypeptides; one or more pyroptosis effector proteins; and / or one or more mutant pyroptosis effector proteins, wherein the synthetic protein circuit is capable of inducing in a cell: apoptosis via a first apoptotic protein complex in first apoptotic protein complex active state; and / or pyroptosis via a first and / or second pyroptosis polypeptide in a first and / or second pyroptosis polypeptide active state or a pyroptosis effector protein in a pyroptosis effector proteinactive state.

[0053] Disclosed herein include synthetic protein circuits comprising: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, and wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; and (ii) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domain separated by the first heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide inactive state, wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first of the first pyroptosis polypeptide, wherein the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, and wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of: (a) inducing pyroptosis in the cell; and (b) inhibiting the induction of apoptosis in the cell by the first apoptotic protein complex in the first apoptotic protein complex active state. In some embodiments, said inhibition is at least 10%. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0054] In some embodiments, the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase-4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase- 10, caspase-14 or any variant, portion, or derivative thereof. In some embodiments, the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59. In some embodiments, the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first heterologous protease is engineered. In some embodiments, the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease,hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the first heterologous protease cleavage site is natural or engineered, e.g., the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site. In some embodiments, the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7.

[0055] In some embodiments, the first apoptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state. In some embodiments, said third partner domains and fourth partner domains are capable of multimerization. In some embodiments, the first pyroptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to the first pyroptosis polypeptide active state in the absence of the first heterologous protease in the first heterologous protease active state. In some embodiments, said third partner domains and fourth partner domains are capable of multimerization. In some embodiments, the third partner domain and the fourth partner domain are homodimers. In some embodiments, the third partner domain and the fourth partner domain are heterodimers. In some embodiments, the binding between the third partner domain and the fourth partner domain is reversible.

[0056] In some embodiments, the third partner domain and / or the fourth partner domain comprise SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18. SYNZIP19, SYNZIP20, SYNZIP21. SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip, BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13 E234 heterodimer a, DHD15 heterodimer a, DHD20heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a, DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a. DHD37_3: 124 heterodimer a, DHD37_1 :234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94 3:214 heterodimer a. DHD94 2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103_l:423 heterodimer a. DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a. DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a. DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a. DHD66 heterodimer a, DHD67 heterodimer a. DHD69 heterodimer a, DHD71 heterodimer a. DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a. DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a. DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a. DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a. DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a,DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD13_2:341 heterodimer b, DHD13 AAAA heterodimer b. DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b. DHD13_1:234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1 :234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3:214 heterodimer b, DHD94 2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l:243 heterodimer b, DHD103 heterodimer b, DHD103_l:423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD1 11 heterodimer b. DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD115 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b, DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b. DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b. DHD5 heterodimer b. DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b. DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b. DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b. DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD151 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b, DHD163 heterodimer b, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b. DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, Pl peptide. P2 peptide, P3 peptide, P4 peptide, P5 peptide, P6 peptide, P7 peptide, P8 peptide, P9 peptide, PIO peptide, Pl 1 peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer, N8 heterodimer, P8A heterodimer, an Sih peptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0057] In some embodiments, the third partner domain and / or the fourth partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology7domain, aDbl homology7domain, a gelsolin homology7domain, aPBl domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof. In some embodiments, the third partner domain and the fourth partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (1) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the third partner domain and / or the fourth partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, the third partner domain and / or the fourth partner domain comprise nHalo, cHalo, or any combination thereof.

[0058] In some embodiments, the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity7of the pyroptosis effector domain, e.g., a bulky7domain In some embodiments, the first inhibitory domain comprises an auto-inhibitory domain ofa gasdermin family of pore-forming proteins or maltose-binding protein. In some embodiments, the cell does not express an endogenous protein comprising a pyroptosis effector domain.

[0059] Disclosed herein include sy nthetic protein circuits comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein, separated by a first heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptosis protease complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of: (a) inducing apoptosis in a cell expressing a pyroptosis effector protein; and / or (b) cutting the pyroptosis effector protein, wherein the pyroptosis effector protein being cut changes the pyroptosis effector protein from a pyroptosis effector protein inactive state to a pyroptosis effector protein active state, wherein the pyroptosis effector protein in the pyroptosis effector protein active state is capable of: (c) inducing pyroptosis in the cell; and (d) inhibiting the induction of apoptosis in the cell by the first apoptotic protein complex in the first apoptotic protein complex active state. In some embodiments, the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

[0060] Disclosed herein include synthetic protein circuits comprising: (i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein, separated by a first heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of: (a) inducing apoptosis in a cell expressing a pyroptosis effector protein; and (b) cutting the pyroptosis effector protein, wherein the pyroptosis effector protein being cut changes the pyroptosis effector protein from a pyroptosis effector protein inactive state to a pyroptosis effector protein active state, wherein the pyroptosis effector protein in the pyroptosis effector protein active state is capable of: (c) inducing pyroptosis in the cell; and (d) inhibiting theinduction of apoptosis in the cell by the apoptotic protein complex in the first apoptotic protein complex active state; and (ii) a first pyroptosis polypeptide comprising a mutant pyroptosis effector domain comprising a mutation, capable of inhibiting the pyroptosis effector protein in the pyroptosis effector protein active state, thereby changing the pyroptosis effector protein from the pyroptosis effector protein active state to a pyroptosis effector protein inactive state; thereby the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in the cell. In some embodiments, the synthetic protein circuit comprises a first input polypeptide comprising the first heterologous protease. In some embodiments, the apoptotic effector protein comprises caspase- 1, caspase-2, caspase-3, caspase-4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, caspase-14 or any variant, portion, or derivative thereof.

[0061] In some embodiments, the first apoptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state, in the absence of the first heterologous protease in the first heterologous protease active state. In some embodiments, said third partner domains and fourth partner domains are capable of multimerization. In some embodiments, the third partner domain and the fourth partner domain are homodimers. In some embodiments, the third partner domain and the fourth partner domain are heterodimers. In some embodiments, the binding between the third partner domain and the fourth partner domain is reversible.

[0062] In some embodiments, the third partner domain and / or the fourth partner domain comprise SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18. SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip. BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13_1:234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a. DHD27 heterodimer a, DHD30 heterodimer a, DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a. DHD37_3: 124 heterodimer a, DHD37_1 :234 heterodimer a, DHD37 AXBBheterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a. DHD94_2: 143 heterodimer a. DHD95 heterodimer a, DHD96 heterodimer a. DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103_l:423 heterodimer a. DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a, DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD1 14 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a. DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a. DHD16 heterodimer a, DHD18 heterodimer a. DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a. DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a. DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a. DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a. DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a. DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a. DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a, DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD13_2:341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b. DHD13_1:234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b,DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3: 124 heterodimer b, DHD37_1 :234 heterodimer b. DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b. DHD94 3:214 heterodimer b, DHD94 2: 143 heterodimer b, DHD95 heterodimer b. DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l:243 heterodimer b, DHD103 heterodimer b, DHD103_l:423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD1 11 heterodimer b. DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD115 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b. DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b. DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b. DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b. DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b. DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD151 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b. DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b, DHD163 heterodimer b, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b. DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b,DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, Pl peptide. P2 peptide, P3 peptide, P4 peptide, P5 peptide, P6 peptide, P7 peptide, P8 peptide, P9 peptide, PIO peptide, Pl l peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer. N8 heterodimer, P8A heterodimer, an Sih peptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0063] In some embodiments, the third partner domain and / or the fourth partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, a Dbl homology domain, a gelsolin homology domain, a PB1 domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof. In some embodiments, the third partner domain and the fourth partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (1) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the third partner domain and / or the fourth partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, the third partner domain and / or the fourth partner domain comprise nHalo, cHalo, or any combination thereof.

[0064] In some embodiments, the pyroptosis effector protein comprises a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59.

[0065] In some embodiments, the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first heterologous protease is engineered. In some embodiments, the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus(TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the first heterologous protease cleavage site is natural or engineered, e.g.. the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site. In some embodiments, the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7.

[0066] In some embodiments, the mutant pyroptosis effector domain is derived from an N-terminal domain of a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59. In some embodiments, the one or more mutations comprises a V99N, L101N, L103N, V193E, A195E, G199E, and / or I217N mutation in GSDME. In some embodiments, the mutation is an I217N mutation in GSDME.

[0067] In some embodiments, inducing pyroptosis in the cell causes the cell to release one or more inflammatory cytokines. In some embodiments, the one or more inflammatory cytokines comprise IL- 18. IL-10, IL-6. IL-8, interferon gamma (IFN-y), and / or tumor necrosis factor-alpha (TNF-a). In some embodiments, inhibition of the pyroptosis effector protein by the first pyroptosis polypeptide is dose-dependent, thereby the induction of apoptosis in the cell is dose-dependent. In some embodiments, a concentration of the first pyroptosis peptide is at least two-folder higher than a concentration of the pyroptosis effector protein in the cell, thereby the inhibition of the pyroptosis effector protein by the first pyroptosis polypeptide is increased relative to a cell wherein the concentration of the first pyroptosis polypeptide is not at least two-fold higher than the concentration of the pyroptosis effector protein.

[0068] In some embodiments, the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide comprise one or more linkers. In some embodiments, the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, e.g., about 1 to about 18 flexible amino acid residues. In some embodiments, the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 2 repeating amino acid subunits or more.

[0069] In some embodiments: (i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide, are configured to be in a first localized state. In some embodiments, the first localized state comprises a state created by phaseseparation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome. microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof. In some embodiments, the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state. In some embodiments: (i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide, are configured to be in second localized state(s), wherein the first localized state and the second localized state(s) are different. In some embodiments, the second localized state(s) comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory' vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome. microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof. In some embodiments, the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state. In some embodiments: (i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide comprise a first localization signal. In some embodiments, the first localization signal is adjacent to a third degron and / or a third heterologous cleavage site. In some embodiments: (i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide comprise second localization signal(s). In some embodiments, the second localization signal is adjacent to a third degron and / or a third heterologous protease cleavage site. In some embodiments, the presence of the third degron and / or wherein the third heterologous cleavage site being cut changes: (i) the first apoptosis polypeptide from a nonlocalized state or the first localized state to the second localized state(s) or a non-localized state; (ii) the first pyroptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state; and / or (iii) the first input polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localizedstate. In some embodiments, the synthetic protein circuit further comprises a third input polypeptide comprising a third heterologous protease capable of cutting the third heterologous protease cleavage site. In some embodiments, the third heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the third heterologous protease is engineered. In some embodiments, the third heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the third heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the third heterologous protease cleavage site is natural or engineered, e.g., the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the third heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type third heterologous protease cleavage site. In some embodiments, the third heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7. In some embodiments: (a) the first apoptosis polypeptide in the first localized state; (b) the first apoptosis polypeptide in the second localized state(s); (c) the first pyroptosis in the first localized state; (d) the first pyroptosis polypeptide in the second localized state(s); (e) the first input polypeptide in the first localized state; and / or (I the first second input polypeptide in the second localized state(s), is capable of modulating an activation threshold and / or sensitivity’ of the synthetic protein circuit. In some embodiments, the first localization signal and / or second localization signal(s) is selected from the group comprising a nuclear localization signal (NLS), a nuclear export signal (NES), a peroxisomal targeting signal (PTS), a mitochondrial targeting sequence (MTS), a ER signal peptide, CAAX box, a peroxisomal targeting signal (PTS), PTS1, PTS2, a dileucine motif, YxxQ motif, a palmitoylation motif, a GPI anchor signal, myristoylation signal. HDEL. KDEL. KKXX motif, RXR motif. SV40 NLS. SV40 NES, CAAX membrane tether, ER recruitment, portions thereof, derivatives thereof, or any combination thereof.

[0070] In some embodiments, the synthetic protein circuit is present in a cell. In some embodiments, the cell is: a cell of a subject, e.g., a subject suffering from a disease or disorder. In some embodiments, the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof; a cell derived from a donor;and / or an in vivo cell, an ex vivo cell, an in vitro cell or an in situ cell. In some embodiments, the cell is a eukaryotic cell, e.g., a mammalian cell. In some embodiments, the mammalian cell comprises an antigen-presenting cell, a dendritic cell, a macrophage, a neural cell, a brain cell, an astrocyte, a microglial cell, and a neuron, a spleen cell, a lymphoid cell, a lung cell, a lung epithelial cell, a skin cell, a keratinocyte, an endothelial cell, an alveolar cell, an alveolar macrophage, an alveolar pneumocyte, a vascular endothelial cell, a mesenchymal cell, an epithelial cell, a colonic epithelial cell, a hematopoietic cell, a bone marrow cell, a Claudius cell, Hensen cell, Merkel cell, Muller cell, Paneth cell, Purkinje cell, Schwann cell, Sertoli cell, acidophil cell, acinar cell, adipoblast, adipocyte, brown or white alpha cell, amacrine cell, beta cell, capsular cell, cementocyte, chief cell, chondroblast, chondrocyte, chromaffin cell, chromophobic cell, corticotroph, delta cell, Langerhans cell, follicular dendritic cell, enterochromaffin cell, ependymocyte, epithelial cell, basal cell, squamous cell, endothelial cell, transitional cell, erythroblast, erythrocyte, fibroblast, fibrocyte, follicular cell, germ cell, gamete, ovum, spermatozoon, oocyte, primary oocyte, secondary oocyte, spermatid, spermatocyte, primary spermatocyte, secondary spermatocyte, germinal epithelium, giant cell, glial cell, astroblast, astrocyte, oligodendroblast, oligodendrocyte, glioblast, goblet cell, gonadotroph, granulosa cell, haemocytoblast. hair cell, hepatoblast, hepatocyte, hyalocyte, interstitial cell, juxtaglomerular cell, keratinocyte, keratocyte. lemmal cell, leukocyte, granulocyte, basophil, eosinophil, neutrophil, lymphoblast, B-lymphoblast, T-lymphoblast, lymphocyte, B-lymphocyte, T-lymphocyte, helper induced T-lymphocyte, Thl T-lymphocyte, Th2 T-lymphocyte, natural killer cell, thymocyte, macrophage, Kupffer cell, alveolar macrophage, foam cell, histiocyte, luteal cell, lymphocytic stem cell, lymphoid cell, lymphoid stem cell, macroglial cell, mammotroph, mast cell, medulloblast, megakaryoblast, megakaryocyte, melanoblast, melanocyte, mesangial cell, mesothelial cell, metamyelocyte, monoblast, monocyte, mucous neck cell, myoblast, myocyte, muscle cell, cardiac muscle cell, skeletal muscle cell, smooth muscle cell, myelocyte, myeloid cell, myeloid stem cell, myoblast, myoepithelial cell, myofibrobast, neuroblast, neuroepithelial cell, neuron, odontoblast, osteoblast, osteoclast, osteocyte, oxyntic cell, parafollicular cell, paraluteal cell, peptic cell, pericyte, peripheral blood mononuclear cell, phaeochromocyte, phalangeal cell, pinealocyte, pituicyte, plasma cell, platelet, podocyte, proerythroblast, promonocyte, promyeloblast, promyelocyte, pronormoblast, reticulocyte, retinal pigment epithelial cell, retinoblast, small cell, somatotroph, stem cell, sustentacular cell, teloglial cell, a zymogenic cell, or any combination thereof. In some embodiments, the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell (iPSC), a hematopoietic stem / progenitor cell (HSPC), or any combination thereof.

[0071] Disclosed herein include synthetic protein circuits comprising: a firstpolypeptide comprising a first signal transducer binding domain and a first part of a first cell death executioner, wherein the first signal transducer binding domain is capable of binding a first signal transducer to form a first signal transducer-bound polypeptide; a second polypeptide comprising a second signal transducer binding domain and a second part of the first cell death executioner, wherein the second signal transducer binding domain is capable of binding a second signal transducer to form a second signal transducer-bound polypeptide, and wherein the first part of the first cell death executioner and the second part of the first cell death executioner are capable of associating with each other to constitute a first cell death executioner capable of being in a first cell death executioner active state when the first signal transducer and the second signal transducer are in close proximity at an association location; and wherein the first cell death executioner in the first cell death executioner active state is capable of inducing apoptosis or pyroptosis in a cell.

[0072] In some embodiments, the first signal transducer binding domain of the first polypeptide and the second signal transducer binding domain of the second polypeptide are identical. In some embodiments, the first transducer and the second transducer are identical and / or are the same protein. In some embodiments, the first cell death executioner comprises an apoptosis effector protein or a pyroptosis effector protein. In some embodiments, the first part of the first cell death executioner comprises a large subunit of the apoptotic effector protein and / or wherein the second part of second first cell death executioner comprises a small subunit of the apoptotic effector protein. In some embodiments, the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase-4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase- 10, caspase-14 or any variant, portion, or derivative thereof. In some embodiments, the pyroptosis effector protein comprises a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59.

[0073] In some embodiments, the first signal transducer, the second signal transducer, or both, are capable of being localized at the association location. In some embodiments, the first signal transducer when in a first signal transducer active state, the second signal transducer when in a second signal transducer active state, or both, are capable of being localized at the association location. In some embodiments, the first signal transducer when in a first inactive state, the second signal transducer when in a second inactive state, or both, are capable of being localized at the association location. In some embodiments, the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer, or both. In some embodiments, the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first signal transducer active state, wherein the second signal transducer binding domain of the second polypeptide is capableof binding to the second signal transducer in a second signal transducer active state, or both. In some embodiments, the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first inactive state, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer in a second inactive state, or both. In some embodiments, the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, or both. In some embodiments, the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at a first cellular location other than the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducerbound polypeptide at a second cellular location other than the association location, or both. In some embodiments, the first cellular location, the second cellular location, or both comprise one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof In some embodiments, the association location comprises one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory' vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium. sarcomere, focal contact, podosome. ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof.

[0074] In some embodiments, a first concentration of the first signal transducer-bound polypeptide is at least two-fold higher at the association location as compared to a first cellular location other than the association location when the first signal transducer is in a first signal transducer active state, and / or wherein a second concentration of the second signal transducerbound polypeptide is at least two-fold higher at the association location as compared to a second cellular location other than the association location when the second signal transducer is in a second signal transducer active state. In some embodiments, a first concentration of the first celldeath executioner in the first cell death executioner active state is at least two-fold higher at the association location as compared to a cellular location other than the association location when the first signal transducer is in a first signal transducer active state and / or when the second signal transducer is in a second signal transducer active state. In some embodiments, the first part of the first cell death executioner and the second part of the first cell death executioner have the weak association affinity when the first signal transducer is in a first signal transducer inactive state and / or the second signal transducer is in a second signal transducer inactive state. In some embodiments, the first part of the first cell death executioner and the second part of the first cell death executioner are incapable of associating to form the first cell death executioner in the first cell death executioner active state when the first signal transducer is in a first signal transducer inactive state and / or the second signal transducer is in a second signal transducer inactive state. In some embodiments, a first concentration of the first signal transducer-bound polypeptide and a second concentration of the second signal transducer-bound polypeptide at the association location are insufficient for the first part of the first cell death executioner and the second part of the first cell death executioner to form an active first cell death executioner when the first signal transducer is in a first signal transducer inactive state and / or the second signal transducer is in a second signal transducer inactive state. In some embodiments, a first concentration of the first signal transducer-bound polypeptide at the association location is comparable to a first cellular location other than the association location when the first signal transducer is in a first signal transducer inactive state, and / or wherein a second concentration of the second signal transducerbound polypeptide at the association location is comparable to a second cellular location other than the association location when the second signal transducer is in a second signal transducer inactive state. In some embodiments, the first part of the first cell death executioner and the second part of the first cell death executioner are capable of associating with each other to form the first cell death executioner in the first cell death executioner active state at a threshold first polypeptide concentration and a threshold second polypeptide concentration at the association location. In some embodiments, the threshold first polypeptide concentration and the threshold second polypeptide concentration at the association location are reached at a threshold signal transducer activation level of the signal transducer.

[0075] In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain are identical. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain are different. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain each is capable of binding molecules of the first signal transducer and / or the second signal transducer. In some embodiments, the first signal transducer and / or the secondsignal transducer belong to a signal transduction pathway. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain comprise a RAS binding domain (RBD) and / or RAS association domain (RAD). In some embodiments, the RAS binding domain comprises or is derived from a RAS interacting protein, optionally selected from the group compnsing AG02, APBB1IP, APPL1, ARAF, ARL1. ARL2. ARRB1, ARRB2, BAIAP2, BCL2, BCL2L1, BRAF, BRAP, BSG, CALM1, CALM3, CALML3, CALML4, CALML5, CALML6, CNKSR1, CNKSR2, CSK, DAB2IP, EGFR, ERBIN, FGA, FGB, FGG, FN1, GRB2, HK1, IFNGR1, IL6, IQGAP1, ITGA2B, ITGB3, KSR1, KSR2, LGALS3, LYN, LZTR1, MAP2K1, MAP2K2, MAPK1, MAPK14, MAPK3, MAPKAP1, MARK2. MARK3, MBP, MSI2, MTOR, NCBP2AS2, NF1, NIBAN2, PDE4DIP, PDE6D, PDPK1, PEBP1, PIK3CA, PIK3CB, PIK3CD, PIK3R1, PIK3R2, PIP5K1A, PLCE1, PPIA, PRKCZ, PTGS2, RAFI, RALB, RALGDS, RAP1A, RAP1B, RAP1GDS1, RASA1, RASA2, RASA3, RASA4, RASAL1, RASAL2, RASAL3. RASSF1, RASSF2, RASSF5, RGL1, RGL3, RIN1, SHOC2, SOS 1, SOS2, SPRED1, SPRED2, SPRED3, SRC, SYNGAP1, TIAM1, TLN1, VCL, VWF, YWHAB, or any combination thereof. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain comprises a lipid binding domain. In some embodiments, the lipid binding domain comprises a Pleckstrin homology (PH) domain. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain comprises an antibody, an antibody fragment, a binding domain derived from a natural protein, an scFv, a Fv, a Fab, a (Fab')2, a single domain antibody (SDAB), a VH or VL domain, a camelid VHH domain, a Fab, a Fab1, a F(ab')2, a Fv, a scFv, a dsFv, a diabody, a triabody, a tetrabody, a multispecific antibody formed from antibody fragments, a singledomain antibody (sdAb), a single chain comprising cantiomplementary scFvs (tandem scFvs) or bispecific tandem scFvs, an Fv construct, a disulfide-linked Fv, a dual variable domain immunoglobulin (DVD-Ig) binding protein or a nanobody, an aptamer, an affibody, an affilin, an affitin, an affimer, an alphabody, an anticalin, an avimer, a DARPin, a Fynomer, a Kunitz domain peptide, a monobody, or any combination thereof.

[0076] In some embodiments, the first signal transducer is capable of binding the first signal transducer binding domain and / or the second signal transducer is capable of binding the second signal transducer binding domain following a modification selected from the group comprising phosphorylation, dephosphorylation, acetylation, methylation, acylation, glycosylation, glycosylphosphatidylinositol (GPI) anchoring, sulfation, disulfide bond formation, deamidation, ubiquitination, sumoylation, nitration of tyrosine, hydrolysis of ATP or GTP, binding of ATP or GTP, cleavage, or any combination thereof. In some embodiments, the first signal transducer, the second signal transducer, or both are endogenous proteins. In someembodiments, the first signal transducer, the second signal transducer, or both comprise AKT, PI3K, MAPK, p44 / 42 MAP kinase, TYK2, p38 MAP kinase, PKC, PKA, SAPK, ELK, JNK, eJun, RAS, Raf, MEK 1 / 2, MEK 3 / 6, MEK 4 / 7, ZAP-70, LAT, SRC, LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX, PLCy, PLCy, NF-kB, FAK, CREB, aIII|33, FceRI, BAD, p70S6K, STAT1, STAT2, STAT3. STAT5, STAT6. or any combination thereof. In some embodiments, the first signal transducer and / or the second signal transducer are capable of regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof. In some embodiments, the first signal transducer, the second signal transducer, or both comprise a RAS protein. In some embodiments, the RAS protein is KRAS, NRAS, HRAS, or any combination thereof. In some embodiments, the first signal transducer, the second signal transducer, or both are exogenous proteins. In some embodiments, the synthetic protein circuit comprises the first signal transducer, the second signal transducer, or both. In some embodiments, the first signal transducer, the second signal transducer, or both comprise a lipid. In some embodiments, the lipid comprises a phospholipid. In some embodiments, the phospholipid is phosphatidylinositol 3- phosphate.

[0077] In some embodiments, the synthetic protein circuit is capable of detecting an activity of the first signal transducer and an activity of the second signal transducer. In some embodiments, an activity of the first cell death executioner correlates with an activity of the first signal transducer and / or an activity of the second signal transducer. In some embodiments, the synthetic protein circuit is capable of detecting activities of the first signal transducer and activities of the second signal transducer over a period of time. In some embodiments, activities of the first cell death executioner correlate with activities of the first signal transducer and activities of the second signal transducer over a period of time. In some embodiments, the synthetic protein circuit is capable of detecting an aberrant signaling. In some embodiments, aberrant signaling involves an active signal transducer. In some embodiments, the aberrant signaling involves an overactive signal transducer. In some embodiments, the aberrant signaling involves a constitutively active signal transducer over a period of time. In some embodiments, the synthetic protein circuit is capable of detecting an activity of a signal transducer activator and / or an activity of a signal transducer repressor. In some embodiments, the aberrant signaling involves an active signal transducer repressor and an active signal transducer. In some embodiments, the aberrant signaling involves an inactive signal transducer activator and an active signal transducer. In some embodiments, the aberrant signaling involves an inactive signal transducer. In some embodiments, the aberrant signaling involves an underactive signal transducer. In some embodiments, the aberrant signaling involves a constitutively inactive signal transducer over a period of time. Insome embodiments, the aberrant signaling involves an inactive signal transducer repressor and an inactive signal transducer. In some embodiments, the aberrant signaling involves an active signal transducer activator and an inactive signal transducer. In some embodiments, the aberrant signaling involves an active signal transducer, and wherein the aberrant signaling comprises an aberrant signal of at least one signal transduction pathway regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof. In some embodiments, the synthetic protein circuit is capable of directly or indirectly inducing cell death in the presence of the aberrant signaling. In some embodiments, the first cell death executioner is capable of directly or indirectly inducing cell death in the presence of aberrant signaling. In some embodiments, the synthetic protein circuit is capable of directly or indirectly inducing cell death when a first level of activation of the first signal transducer is above a first signal transducer activation threshold and / or a second level of activation of the second signal transducer is below a second signal transducer activation threshold. In some embodiments, the effector protein is capable of directly or indirectly inducing cell death when a first level of activation of the first signal transducer is above a first signal transducer activation threshold and / or a second level of activation of the second signal transducer is below a second signal transducer activation threshold.

[0078] In some embodiments, the synthetic protein circuit is present in a cell. In some embodiments, the cell is: a cell of a subject, e.g., a subject suffering from a disease or disorder. In some embodiments, the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof; a cell derived from a donor; and / or an in vivo cell, an ex vivo cell, an in vitro cell or an in situ cell. In some embodiments, the cell is a eukary otic cell, e.g., a mammalian cell. In some embodiments, the mammalian cell comprises an antigen-presenting cell, a dendritic cell, a macrophage, a neural cell, a brain cell, an astrocyte, a microglial cell, and a neuron, a spleen cell, a lymphoid cell, a lung cell, a lung epithelial cell, a skin cell, a keratinocyte, an endothelial cell, an alveolar cell, an alveolar macrophage, an alveolar pneumocyte, a vascular endothelial cell, a mesenchymal cell, an epithelial cell, a colonic epithelial cell, a hematopoietic cell, a bone marrow cell, a Claudius cell, Hensen cell, Merkel cell. Muller cell, Paneth cell, Purkinje cell, Schwann cell, Sertoli cell, acidophil cell, acinar cell, adipoblast, adipocyte, brown or white alpha cell, amacrine cell, beta cell, capsular cell, cementocyte, chief cell, chondroblast, chondrocyte, chromaffin cell, chromophobic cell, corticotroph, delta cell, Langerhans cell, follicular dendritic cell, enterochromaffin cell, ependymocyte. epithelial cell, basal cell, squamous cell, endothelial cell,transitional cell, erythroblast, erythrocyte, fibroblast, fibrocyte, follicular cell, germ cell, gamete, ovum, spermatozoon, oocyte, primary oocyte, secondary oocyte, spermatid, spermatocyte, primary spermatocyte, secondary' spermatocyte, germinal epithelium, giant cell, glial cell, astroblast, astrocyte, oligodendroblast, oligodendrocyte, glioblast, goblet cell, gonadotroph, granulosa cell, haemocytoblast. hair cell, hepatoblast, hepatocyte, hyalocyte, interstitial cell, juxtaglomerular cell, keratinocyte, keratocyte, lemmal cell, leukocyte, granulocyte, basophil, eosinophil, neutrophil, lymphoblast, B-lymphoblast, T-lymphoblast, lymphocyte, B-lymphocyte, T-lymphocyte, helper induced T-lymphocyte, Thl T-lymphocyte, Th2 T-lymphocyte, natural killer cell, thymocyte, macrophage, Kupffer cell, alveolar macrophage, foam cell, histiocyte, luteal cell, lymphocytic stem cell, lymphoid cell, lymphoid stem cell, macroglial cell, mammotroph, mast cell, medulloblast, megakaryoblast, megakaryocyte, melanoblast, melanocyte, mesangial cell, mesothelial cell, metamyelocyte, monoblast, monocyte, mucous neck cell, myoblast, myocyte, muscle cell, cardiac muscle cell, skeletal muscle cell, smooth muscle cell, myelocyte, myeloid cell, myeloid stem cell, myoblast, myoepithelial cell, myofibrobast, neuroblast, neuroepithelial cell, neuron, odontoblast, osteoblast, osteoclast, osteocyte, oxyntic cell, parafollicular cell, paraluteal cell, peptic cell, pericyte, peripheral blood mononuclear cell, phaeochromocyte, phalangeal cell, pinealocyte, pituicyte, plasma cell, platelet, podocyte, proerythroblast, promonocyte, promyeloblast, promyelocyte, pronormoblast, reticulocyte, retinal pigment epithelial cell, retinoblast, small cell, somatotroph, stem cell, sustentacular cell, teloglial cell, a zymogenic cell, or any combination thereof. In some embodiments, the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell (iPS C), a hematopoietic stem / progenitor cell (HSPC), or any combination thereof.

[0079] Disclosed herein include nucleic acid compositions. In some embodiments, the composition comprises: one or more polynucleotides encoding any of the synthetic protein circuits of the disclosure. In some embodiments, the one or more polynucleotides comprise: one or more first polynucleotides encoding a first apoptosis polypeptide, a first pyroptosis polypeptide, or a first polypeptide: one or more second polynucleotides encoding a second apoptosis polypeptide, a second pyroptosis polypeptide, or a second polypeptide: and / or one or more third polynucleotides encoding a third apoptosis polypeptide. In some embodiments, the nucleic acid composition comprises one or more polynucleotides encoding a first and / or second input polypeptide.

[0080] In some embodiments, at least two of the one or more polynucleotides are operably linked to a tandem gene expression element. In some embodiments, the one or more polynucleotides comprise: a 5'UTR and / or a 3’UTR; a tandem gene expression element selected from the group an internal ribosomal entry site (IRES), foot-and-mouth disease vims 2A peptide(F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2 A peptide (T2A), or any combination thereof; and / or a transcript stabilization element. In some embodiments, the transcript stabilization element comprises woodchuck hepatitis post-translational regulatory element (WPRE), bovine growth hormone polyadenylation (bGH-polyA) signal sequence, human growth hormone polyadenylation (hGH-polyA) signal sequence, or any combination thereof.

[0081] In some embodiments, the one or more polynucleotides are operably connected to a promoter selected from the group comprising: a minimal promoter, e.g., TATA, miniCMV, and / or miniPromo; a tissue-specific promoter and / or a lineage-specific promoter; and / or a ubiquitous promoter, e.g., a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter. H5, P7.5, and Pl l promoters from vaccinia virus, an elongation factor 1 -alpha (EFla) promoter, early growth response 1 (EGR1), ferritin H (FerH), ferritin L (FerL), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), heat shock 70 kDa protein 5 (HSPA5), heat shock protein 90 kDa beta, member 1 (HSP90B1), heat shock protein 70 kDa (HSP70), (3-kinesin ( -KIN), the human ROSA 26 locus, a Ubiquitin C promoter (UBC). a phosphoglycerate kinase- 1 (PGK) promoter. 3- phosphogly cerate kinase promoter, a cytomegalovirus enhancer, human P-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CASI promoter, a CBH promoter, or any combination thereof.

[0082] In some embodiments, the nucleic acid composition is configured to enhance stability, durability, and / or expression level, optionally a 5' untranslated region (UTR), a 3' UTR, and / or a 5’ cap; optionally one or more modified nucleotides, further optionally selected from the group comprising pseudouridine, N-l-methyl-pseudouridine, 2-aminoadenosine, 2- thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C-5 propynyl- cytidine, C-5 propynyl-undine, 2-aminoadenosine, C5-bromouridine. C5-fluorouridine, C5- iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8 -oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and 2-thiocytidine; and / or optionally a modified nucleotide in place of one or more uridines. In some embodiments, the modified nucleoside is selected from pseudouridine (\| / ). N 1-methyl- pseudouridine (m I ), and 5-methyl-uridine (m5U).

[0083] In some embodiments, the nucleic acid composition is complexed or associated with one or more lipids or lipid-based carriers, thereby forming liposomes, lipid nanoparticles (LNPs), lipoplexes, and / or nanoliposomes, e.g., encapsulating the nucleic acid composition. Insome embodiments, the nucleic acid composition is, comprises, or further comprises, one or more vectors. In some embodiments, at least one of the one or more vectors is a viral vector, a plasmid, a transposable element, a naked DNA vector, a lipid nanoparticle (LNP), a bacterial cell, a bacteriophage, or any combination thereof. In some embodiments, the viral vector is an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MV A vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, a Sindbis virus vector, a VEE vector, a Measles virus vector, an influenza virus vector, a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector, or any combination thereof. In some embodiments, the transposable element is piggybac transposon or sleeping beauty transposon. In some embodiments, the one or more polynucleotides are comprised in the one or more vectors. In some embodiments, the one or more polynucleotides are comprised in the same vector and / or different vectors. In some embodiments, the one or more polynucleotides are situated on the same nucleic acid and / or different nucleic acids. In some embodiments, the nucleic acid composition comprises circular mRNA, circular DNA, self-amplifying RNA, selfamplifying RNA, and / or mRNA.

[0084] In some embodiments, the nucleic acid composition is configured to achieve relative levels of the first apoptotic polypeptide, the second apoptotic polypeptide, the third apoptotic polypeptide, the first input polypeptide, the second input polypeptide, the first polypeptide and / or the second polypeptide desired by a user. In some embodiments, the nucleic acid composition is configured to achieve relative levels of the first pyroptotic polypeptide, the second pyroptotic polypeptide, the first input polypeptide, the second input polypeptide, the first polypeptide and / or the second polypeptide desired by a user. In some embodiments, the expression of one or more of the first apoptotic polypeptide, the second apoptotic polypeptide, the third apoptotic polypeptide, first pyroptotic polypeptide, the second pyroptotic polypeptide, the first input polypeptide, and / or the second polypeptide is configured to be dosage invariant and / or robust to tissue tropism and stochastic expression. In some embodiments, the induction of apoptosis or the induction of pyroptosis can be tuned by adjusting the relative levels of the first apoptotic polypeptide, the second apoptotic polypeptide, the third apoptotic polypeptide, first pyroptotic polypeptide, the second pyroptotic polypeptide, the first input polypeptide, and / or the second polypeptide.

[0085] Disclosed herein include compositions. In some embodiments, the composition comprises: a first population of sender cells comprising: (i) one or more first polynucleotide(s) encoding a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous proteasecleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, and wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; and (ii) a first vector; and a second population of sender cells comprising: (i) one or more second polynucleotide(s) encoding a first input polypeptide; and (ii) a second vector.

[0086] In some embodiments, the first vector of the first population of sender cells and the second vector of the second population of sender cells are capable of delivering the one or more first polynucleotide(s) and the one or more second polynucleotide(s) to receiver cells. In some embodiments, the first apoptosis polypeptide and the first input polypeptide are expressed in the receiver cells, thereby the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in the receiver cells. In some embodiments, the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase-4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase- 10, caspase- 14 or any variant, portion, or derivative thereof. In some embodiments, the first heterologous protease comprises or is derived from one or more of a prokary otic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first heterologous protease is engineered. In some embodiments, the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C vims protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the first heterologous protease cleavage site is natural or engineered, e.g.. the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site. In some embodiments, the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7. In some embodiments, the first apoptosis polypeptide, the first input polypeptide, or both comprise a membrane-localization domain. In some embodiments, the membrane-localization domain comprises a CAAX domain.

[0087] Disclosed herein include compositions. In some embodiments, the composition comprises: a first population of sender cells comprising: (i) one or more first polynucleotide(s) encoding a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domain separated by a first heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, and (ii) a first vector; and a second population of sender cells comprising: (i) one or more second polynucleotide(s) encoding a first input polypeptide comprising the first heterologous protease; and (ii) a second vector.

[0088] In some embodiments, the first vector of the first population of sender cells and the second vector of the second population of sender cells are capable of delivering the one or more first polynucleotide(s) and the one or more second polynucleotide(s) to receiver cells. In some embodiments, the first pyroptosis polypeptide and the first input polypeptide are expressed in the receiver cells, thereby the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in the receiver cells. In some embodiments, the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59. In some embodiments, the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the first heterologous protease is engineered. In some embodiments, wherein the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof. In some embodiments, the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the first heterologous protease cleavage site is natural or engineered, wherein the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild ty pe first heterologous protease cleavage site. In some embodiments, the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7.

[0089] In some embodiments, the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, e.g., a bulky domain. In some embodiments, the first inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein. In some embodiments, the cell does not express an endogenous protein comprising a pyroptosis effector domain.

[0090] In some embodiments, the first apoptosis polypeptide or the first pyroptosis polypeptide comprise one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain of the first apoptosis polypeptide is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state, or wherein binding of the third partner domain and the fourth partner domain of the first pyroptosis polypeptide is capable of inhibiting change of the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to the first pyroptosis polypeptide active state in the absence of the first heterologous protease in the first heterologous protease active state. In some embodiments, said third partner domains and fourth partner domains are capable of multimerization. In some embodiments, the third partner domain and the fourth partner domain are homodimers. In some embodiments, the third partner domain and the fourth partner domain are heterodimers. In some embodiments, the binding between the third partner domain and the fourth partner domain is reversible.

[0091] In some embodiments, the third partner domain and / or the fourth partner domain comprise SYNZIP1. SYNZIP2. SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18, SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip, BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a. DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4: 123 heterodimer a, DHD13_1 :234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a, DHD27 heterodimer a, DHD30 heterodimer a. DHD33 heterodimer a, DHD34_XAAXA heterodimer a, DHD34_XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a, DHD37_3: 124 heterodimer a, DHD37_1 :234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a. DHD70 heterodimer a, DHD88 heterodimer a,DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a, DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a, DHD97 heterodimer a, DHD98 heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a. DHD103_l:423 heterodimer a, DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a. DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a. DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a, DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a, DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a, DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a. DHD26 heterodimer a, DHD28 heterodimer a. DHD29 heterodimer a, DHD31 heterodimer a. DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a, DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a, DHD72 heterodimer a, DHD73 heterodimer a, DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a, DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a. DHD166 heterodimer a, DHS17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a. DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a, DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a. DHD143 heterodimer a, DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b. DHD13 XAXA heterodimer b. DHD13 XAAX heterodimer b, DHD 13 2:341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b, DHD13_1:234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b, DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b,DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3:124 heterodimer b, DHD37_1 :234 heterodimer b, DHD37 AXBB heterodimer b, DHD37 XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b. DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3:214 heterodimer b, DHD94 2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b, DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l :243 heterodimer b, DHD103 heterodimer b, DHD103_l:423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD111 heterodimer b. DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD 115 heterodimer b, DHD 116 heterodimer b, DHD 117 heterodimer b, DHD 118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b, DHD 123 heterodimer b, DHD 124 heterodimer b, DHD 125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD 130 heterodimer b, DHD 145 heterodimer b, DHD 146 heterodimer b, DHD 147 heterodimer b, DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b, DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b, DHD 16 heterodimer b, DHD 18 heterodimer b, DHD 19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD 148 heterodimer b, DHD 149 heterodimer b, DHD 150 heterodimer b, DHD151 heterodimer b. DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD 159 heterodimer b, DHD 160 heterodimer b, DHD 161 heterodimer b, DHD 162 heterodimer b, DHD 163 heterodimer b, DHD 164 heterodimer b, DHD 165 heterodimer b, DHD 166 heterodimer b, DHS17 heterodimer b, DHD 17 heterodimer b, DHD 131 heterodimer b, DHD 132 heterodimer b, DHD 133 heterodimer b, DHD 134 heterodimer b, DHD 135 heterodimer b, DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD 140 heterodimer b, DHD 141 heterodimer b, DHD 142 heterodimer b, DHD 143 heterodimer b, DHD 144 heterodimer b, Pl peptide, P2 peptide, P3 peptide, P4peptide, P5 peptide, P6 peptide, P7 peptide, P8 peptide, P9 peptide, PIO peptide, Pl 1 peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer, N8 heterodimer, P8A heterodimer, an Sih peptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0092] In some embodiments, the third partner domain and / or the fourth partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosme-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, aDbl homology domain, a gelsolin homology domain, aPBl domain, a SOCS box, an RGS domain, a Toll / IL- 1 receptor domain, a tetratncopeptide repeat, a TRAF domain, a Bcl-2 homology' domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof. In some embodiments, the third partner domain and the fourth partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (1) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the third partner domain and / or the fourth partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, the third partner domain and / or the fourth partner domain comprise nHalo, cHalo, or any combination thereof.

[0093] Disclosed herein include compositions. In some embodiments, the composition comprises: a first population of sender cells comprising: (i) one or more first polynucleotide(s) encoding a first pyroptosis polypeptide comprising a pyroptosis effector domain and and a first partner domain; and (ii) a first vector, wherein the first population of sender cells express a silencer polypeptide comprising a first inhibitory domain and a second partner domain capable of binding the first partner domain, and wherein the inhibitor domain of the silencer polypeptide is capable of inhibiting the first pyroptosis polypeptide when the first pyroptosis polypeptide associates with the silencer polypeptide via binding of the first partner domain and the second partner domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state.

[0094] In some embodiments, the first vector is capable of delivering the one or morefirst polynucleotide(s) to receiver cells. In some embodiments, the first pyroptosis polypeptide is expressed in the receiver cells, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in the receiver cells. In some embodiments, the receiver cells do not express the silencer polypeptide. In some embodiments, the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, e.g., a bulky domain. In some embodiments, the first inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein. In some embodiments, the cell does not express an endogenous protein comprising a pyroptosis effector domain. In some embodiments, the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein. In some embodiments, the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59.

[0095] In some embodiments, the first partner domain and the second partner domain are homodimers. In some embodiments, the first partner domain and the second partner domain are heterodimers. In some embodiments, the binding between the first partner domain and the second partner domain is reversible.

[0096] In some embodiments, the first partner domain and / or the second partner domain comprise SYNZIP1, SYNZIP2, SYNZIP3, SYNZIP4, SYNZIP5, SYNZIP6, SYNZIP7, SYNZIP8, SYNZIP9, SYNZIP10, SYNZIP11, SYNZIP12, SYNZIP13, SYNZIP14, SYNZIP15, SYNZIP16, SYNZIP17, SYNZIP18, SYNZIP19, SYNZIP20, SYNZIP21, SYNZIP22, SYNZIP23, BATF, FOS, ATF4, BACH1, JUND, NFE2L3, AZip. BZip, DHD9 heterodimer a, DHD13 XAAA heterodimer a, DHD13 XAXA heterodimer a, DHD13 XAAX heterodimer a, DHD13_2:341 heterodimer a, DHD13 AAAA heterodimer a, DHD13 BAAA heterodimer a, DHD13_4:123 heterodimer a, DHD13 E234 heterodimer a, DHD15 heterodimer a, DHD20 heterodimer a, DHD21 heterodimer a, DHD25 heterodimer a. DHD27 heterodimer a, DHD30 heterodimer a. DHD33 heterodimer a, DHD34 XAAXA heterodimer a, DHD34 XAXXA heterodimer a, DHD34 XAAAA heterodimer a, DHD36 heterodimer a, DHD37 ABXB heterodimer a, DHD37 BBBB heterodimer a, DHD37 XBXB heterodimer a, DHD37 AXXB heterodimer a, DHD37_3: 124 heterodimer a, DHD37_1 :234 heterodimer a, DHD37 AXBB heterodimer a, DHD37 XBBA heterodimer a, DHD39 heterodimer a, DHD40 heterodimer a, DHD43 heterodimer a, DHD65 heterodimer a, DHD70 heterodimer a, DHD88 heterodimer a, DHD89 heterodimer a, DHD90 heterodimer a, DHD91 heterodimer a, DHD92 heterodimer a, DHD93 heterodimer a, DHD94 heterodimer a, DHD94_3:214 heterodimer a, DHD94_2: 143 heterodimer a, DHD95 heterodimer a, DHD96 heterodimer a. DHD97 heterodimer a, DHD98heterodimer a, DHD99 heterodimer a, DHD100 heterodimer a, DHD101 heterodimer a, DHD102 heterodimer a, DHD102_l :243 heterodimer a, DHD103 heterodimer a, DHD103_l:423 heterodimer a, DHD104 heterodimer a, DHD105 heterodimer a, DHD106 heterodimer a, DHD107 heterodimer a, DHD108 heterodimer a, DHD109 heterodimer a, DHD110 heterodimer a. DHD111 heterodimer a, DHD112 heterodimer a, DHD113 heterodimer a, DHD114 heterodimer a, DHD115 heterodimer a, DHD116 heterodimer a, DHD117 heterodimer a, DHD118 heterodimer a, DHD119 heterodimer a, DHD120 heterodimer a, DHD121 heterodimer a. DHD122 heterodimer a, DHD123 heterodimer a, DHD124 heterodimer a, DHD125 heterodimer a. DHD126 heterodimer a, DHD127 heterodimer a, DHD128 heterodimer a, DHD129 heterodimer a, DHD130 heterodimer a, DHD145 heterodimer a, DHD146 heterodimer a, DHD147 heterodimer a, DHD1 heterodimer a, DHD2 heterodimer a, DHD3 heterodimer a, DHD4 heterodimer a, DHD5 heterodimer a, DHD6 heterodimer a, DHD7 heterodimer a, DHD8 heterodimer a, DHD16 heterodimer a, DHD18 heterodimer a. DHD19 heterodimer a, DHD22 heterodimer a, DHD23 heterodimer a, DHD24 heterodimer a, DHD26 heterodimer a, DHD28 heterodimer a, DHD29 heterodimer a, DHD31 heterodimer a, DHD32 heterodimer a, DHD38 heterodimer a, DHD60 heterodimer a, DHD63 heterodimer a, DHD66 heterodimer a, DHD67 heterodimer a, DHD69 heterodimer a, DHD71 heterodimer a. DHD72 heterodimer a, DHD73 heterodimer a. DHD148 heterodimer a, DHD149 heterodimer a, DHD150 heterodimer a, DHD151 heterodimer a, DHD152 heterodimer a, DHD153 heterodimer a, DHD154 heterodimer a, DHD155 heterodimer a, DHD156 heterodimer a, DHD157 heterodimer a, DHD158 heterodimer a, DHD159 heterodimer a, DHD160 heterodimer a, DHD161 heterodimer a, DHD162 heterodimer a, DHD163 heterodimer a, DHD164 heterodimer a, DHD165 heterodimer a, DHD166 heterodimer a, DHS 17 heterodimer a, DHD17 heterodimer a, DHD131 heterodimer a, DHD132 heterodimer a, DHD133 heterodimer a, DHD134 heterodimer a, DHD135 heterodimer a, DHD136 heterodimer a, DHD137 heterodimer a, DHD138 heterodimer a, DHD139 heterodimer a, DHD140 heterodimer a, DHD141 heterodimer a, DHD142 heterodimer a. DHD143 heterodimer a. DHD144 heterodimer a, DHD9 heterodimer b, DHD13 XAAA heterodimer b, DHD13 XAXA heterodimer b, DHD13 XAAX heterodimer b, DHD13_2:341 heterodimer b, DHD13 AAAA heterodimer b, DHD13 BAAA heterodimer b, DHD13_4: 123 heterodimer b, DHD13_1:234 heterodimer b, DHD15 heterodimer b, DHD20 heterodimer b, DHD21 heterodimer b, DHD25 heterodimer b. DHD27 heterodimer b, DHD30 heterodimer b, DHD33 heterodimer b, DHD34 XAAXA heterodimer b, DHD34 XAXXA heterodimer b, DHD34 XAAAA heterodimer b, DHD36 heterodimer b, DHD37 ABXB heterodimer b, DHD37 BBBB heterodimer b, DHD37 XBXB heterodimer b, DHD37 AXXB heterodimer b, DHD37_3:124 heterodimer b, DHD37_1 :234 heterodimer b. DHD37 AXBB heterodimer b,DHD37_XBBA heterodimer b, DHD39 heterodimer b, DHD40 heterodimer b, DHD43 heterodimer b, DHD65 heterodimer b, DHD70 heterodimer b, DHD88 heterodimer b, DHD89 heterodimer b, DHD90 heterodimer b, DHD91 heterodimer b, DHD92 heterodimer b, DHD93 heterodimer b, DHD94 heterodimer b, DHD94_3:214 heterodimer b, DHD94 2: 143 heterodimer b, DHD95 heterodimer b, DHD96 heterodimer b, DHD97 heterodimer b. DHD98 heterodimer b, DHD99 heterodimer b, DHD100 heterodimer b, DHD101 heterodimer b, DHD102 heterodimer b, DHD102_l:243 heterodimer b, DHD103 heterodimer b, DHD103_l:423 heterodimer b, DHD104 heterodimer b, DHD105 heterodimer b, DHD106 heterodimer b, DHD107 heterodimer b, DHD108 heterodimer b, DHD109 heterodimer b, DHD110 heterodimer b, DHD111 heterodimer b. DHD112 heterodimer b, DHD113 heterodimer b, DHD114 heterodimer b, DHD115 heterodimer b, DHD116 heterodimer b, DHD117 heterodimer b, DHD118 heterodimer b, DHD119 heterodimer b, DHD120 heterodimer b, DHD121 heterodimer b, DHD122 heterodimer b. DHD123 heterodimer b, DHD124 heterodimer b, DHD125 heterodimer b, DHD126 heterodimer b, DHD127 heterodimer b, DHD128 heterodimer b, DHD129 heterodimer b, DHD130 heterodimer b, DHD145 heterodimer b, DHD146 heterodimer b, DHD147 heterodimer b, DHD1 heterodimer b, DHD2 heterodimer b, DHD3 heterodimer b, DHD4 heterodimer b. DHD5 heterodimer b, DHD6 heterodimer b, DHD7 heterodimer b, DHD8 heterodimer b. DHD16 heterodimer b, DHD18 heterodimer b, DHD19 heterodimer b, DHD22 heterodimer b, DHD23 heterodimer b, DHD24 heterodimer b, DHD26 heterodimer b, DHD28 heterodimer b, DHD29 heterodimer b, DHD31 heterodimer b, DHD32 heterodimer b, DHD38 heterodimer b, DHD60 heterodimer b, DHD63 heterodimer b, DHD66 heterodimer b, DHD67 heterodimer b, DHD69 heterodimer b, DHD71 heterodimer b, DHD72 heterodimer b, DHD73 heterodimer b, DHD148 heterodimer b, DHD149 heterodimer b, DHD150 heterodimer b, DHD151 heterodimer b, DHD152 heterodimer b, DHD153 heterodimer b, DHD154 heterodimer b, DHD155 heterodimer b, DHD156 heterodimer b, DHD157 heterodimer b, DHD158 heterodimer b, DHD159 heterodimer b, DHD160 heterodimer b, DHD161 heterodimer b, DHD162 heterodimer b. DHD163 heterodimer b, DHD164 heterodimer b, DHD165 heterodimer b, DHD166 heterodimer b, DHS17 heterodimer b, DHD17 heterodimer b, DHD131 heterodimer b, DHD132 heterodimer b, DHD133 heterodimer b, DHD134 heterodimer b, DHD135 heterodimer b, DHD136 heterodimer b, DHD137 heterodimer b, DHD138 heterodimer b, DHD139 heterodimer b, DHD140 heterodimer b, DHD141 heterodimer b, DHD142 heterodimer b, DHD143 heterodimer b, DHD144 heterodimer b, Pl peptide, P2 peptide, P3 peptide, P4 peptide, P5 peptide, P6 peptide, P7 peptide, P8 peptide, P9 peptide, PIO peptide, Pl 1 peptide, P12 peptide, N5 heterodimer, P5A heterodimer, N6 heterodimer, P6A heterodimer, N7 heterodimer, P7A heterodimer, N7 heterodimer, P7A heterodimer, N8 heterodimer, P8A heterodimer, an Sihpeptide, an S2h peptide, an S3h peptide, an S4h peptide, a P5f peptide, a P6f peptide, a P13f peptide, a P14f peptide, portions thereof, derivatives thereof, or any combination thereof.

[0097] In some embodiments, the first partner domain and / or the second partner domain comprise an SH3 domain, a PDZ domain, a GTPase binding domain, a leucine zipper domain, an SH2 domain, a PTB domain, an FHA domain, a WW domain, a 14-3- 3 domain, a death domain, a caspase recruitment domain, a bromodomain, a chromatin organization modifier, a shadow chromo domain, an F-box domain, a HECT domain, a RING finger domain, a sterile alpha motif domain, a glycine-tyrosine-phenylalanine domain, a SNAP domain, a VHS domain, an ANK repeat, an armadillo repeat, a WD40 repeat, an MH2 domain, a calponin homology domain, a Dbl homology domain, a gelsolin homology domain, a PB1 domain, a SOCS box, an RGS domain, a Toll / IL-1 receptor domain, a tetratricopeptide repeat, a TRAF domain, a Bcl-2 homology domain, a coiled-coil domain, a bZIP domain, portions thereof, variants thereof, or any combination thereof. In some embodiments, the first partner domain and the second partner domain are a pair of constitutive protein partner domains selected from the group consisting of (a) cognate leucine zipper domains, (b) cognate PSD95-Dlgl-Zo-1 (PDZ) domains, (c) a streptavidin domain and cognate streptavidin binding protein (SBP) domain, (d) a PYL domain and cognate ABI domain, (e) a pair of cognate zinc finger domains, (!) a pair of cognate SH3 domains, and (g) a peptide and antibody or antigen-binding fragment thereof that specifically binds to the peptide. In some embodiments, the first partner domain and / or the second partner domain comprise CZp, NZp, or any combination thereof. In some embodiments, inducing pyroptosis in a receiver cell causes the receiver cell to release one or more inflammatory cytokines. In some embodiments, the one or more inflammatory cytokines comprise IL- 18, IL-10, IL-6, IL- 8, interferon gamma (IFN-y), and / or tumor necrosis factor-alpha (TNF-a).

[0098] In some embodiments, the first apoptosis polypeptide, the first pyroptosis polypeptide, the first input polypeptide, and / or the silencer polypeptide comprises one or more linkers. In some embodiments, the linker: is a flexible linker, a rigid linker, or a hybrid linker; is hydrophilic or hydrophobic; is between 1 and 250 amino acids; comprises one or more flexible amino acid residues, e.g., about 1 to about 18 flexible amino acid residues. In some embodiments, the flexible amino acid residues comprise glycine, serine, or a combination thereof; and / or comprises 2 repeating amino acid subunits or more. In some embodiments: (i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide, are configured to be in a first localized state. In some embodiments, the first localized state comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, innermitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof. In some embodiments, first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state. In some embodiments:(i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide, are configured to be in second localized state(s), wherein the first localized state and the second localized state(s) are different. In some embodiments, the second localized state(s) comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof. In some embodiments, the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state. In some embodiments: (i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide comprise a first localization signal. In some embodiments, the first localization signal is adjacent to a third degron and / or a third heterologous cleavage site. In some embodiments: (i) the first apoptosis polypeptide; (ii) the first pyroptosis polypeptide, and / or (iii) the first input polypeptide comprise second localization signal(s). In some embodiments, the second localization signal is adjacent to a third degron and / or a third heterologous protease cleavage site. In some embodiments, the presence of the third degron and / or wherein the third heterologous cleavage site being cut changes: (i) the first apoptosis polypeptide from a nonlocalized state or the first localized state to the second localized state(s) or a non-localized state;(ii) the first pyroptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state; and / or (iii) the first input polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state. In some embodiments, the synthetic protein circuit further comprises a third input polypeptide comprising a third heterologous protease capable of cutting the third heterologous protease cleavage site. In some embodiments, the third heterologous protease comprises or isderived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease. In some embodiments, the third heterologous protease is engineered. In some embodiments, the third heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP). derivatives thereof, or any combination thereof. In some embodiments, the third heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2. In some embodiments, the third heterologous protease cleavage site is natural or engineered, e.g., the cleavage sequence of a human apoptotic effector protease, e.g., a caspase cleavage sequence. In some embodiments, the third heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit. In some embodiments, said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type third heterologous protease cleavage site. In some embodiments, the third heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7. In some embodiments: (a) the first apoptosis polypeptide in the first localized state: (b) the first apoptosis polypeptide in the second localized state(s); (c) the first pyroptosis in the first localized state; (d) the first pyroptosis polypeptide in the second localized state(s); (e) the first input polypeptide in the first localized state; and / or (f) the first second input polypeptide in the second localized state(s), is capable of modulating an activation threshold and / or sensitivity of the synthetic protein circuit. In some embodiments, the first localization signal and / or second localization signal(s) is selected from the group comprising a nuclear localization signal (NLS), a nuclear export signal (NES), a peroxisomal targeting signal (PTS), a mitochondrial targeting sequence (MTS), a ER signal peptide, CAAX box. a peroxisomal targeting signal (PTS), PTS1, PTS2, a dileucine motif, YxxO motif, a palmitoylation motif, a GPI anchor signal, myristoylation signal, HDEL, KDEL, KKXX motif, RXR motif, SV40 NLS, SV40 NES, CAAX membrane tether, ER recruitment, portions thereof, derivatives thereof, or any combination thereof.

[0099] In some embodiments, the first and / or second vector is a viral vector, a plasmid, a naked DNA vector, a naked RNA vector, a lipid nanoparticle, or any combination thereof. In some embodiments, the viral vector is an AAV vector, a lentivirus vector, a retrovirus vector, an integration-deficient lentivirus (IDLV) vector. In some embodiments, the AAV vector comprises single-stranded AAV (ssAAV) vector or a self-complementary AAV (scAAV) vector. In some embodiments, the receiver cell or the sender cell is: a cell of a subject, e.g., a subject suffering from a disease or disorder. In some embodiments, the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused byaberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof; a cell derived from a donor; and / or an in vivo cell, an ex vivo cell, or an in situ cell.

[0100] In some embodiments, the first and / or the second vector is capable of delivering the one or more first polynucleotides and / or the one or more second polynucleotides to one or more tissues of a subject. In some embodiments, the receiver cells are situated within one or more tissues of a subject; and / or the one or more tissues comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue, hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof.

[0101] In some embodiments, the sender cell and / or the receiver cell is a eukaryotic cell, e.g., a mammalian cell. In some embodiments, the mammalian cell comprises an antigen- presenting cell, a dendritic cell, a macrophage, a neural cell, a brain cell, an astrocyte, a microglial cell, and a neuron, a spleen cell, a lymphoid cell, a lung cell, a lung epithelial cell, a skin cell, a keratinocyte, an endothelial cell, an alveolar cell, an alveolar macrophage, an alveolar pneumocyte, a vascular endothelial cell, a mesenchymal cell, an epithelial cell, a colonic epithelial cell, a hematopoietic cell, a bone marrow cell, a Claudius cell, Hensen cell, Merkel cell, Muller cell, Paneth cell, Purkinje cell, Schwann cell. Sertoli cell, acidophil cell, acinar cell, adipoblast, adipocyte, brown or white alpha cell, amacrine cell, beta cell, capsular cell, cementocyte, chief cell, chondroblast, chondrocyte, chromaffin cell, chromophobic cell, corticotroph, delta cell, Langerhans cell, follicular dendritic cell, enterochromaffin cell, ependymocyte, epithelial cell, basal cell, squamous cell, endothelial cell, transitional cell, erythroblast, erythrocyte, fibroblast, fibrocyte, follicular cell, germ cell, gamete, ovum, spermatozoon, oocyte, primary oocyte, secondary oocyte, spermatid, spermatocyte, primary spermatocyte, secondary spermatocyte, germinal epithelium, giant cell, glial cell, astroblast, astrocyte, oligodendroblast, oligodendrocyte, glioblast, goblet cell, gonadotroph, granulosa cell, haemocytoblast, hair cell, hepatoblast, hepatocyte, hyalocyte, interstitial cell, juxtaglomerular cell, keratinocyte, keratocyte, lemmal cell, leukocyte, granulocyte, basophil, eosinophil, neutrophil, lymphoblast, B-lymphoblast, T- lymphoblast, lymphocyte, B-lymphocyte, T-lymphocyte, helper induced T-lymphocyte, Thl T- lymphocyte, Th2 T-lymphocyte, natural killer cell, thymocyte, macrophage, Kupffer cell, alveolar macrophage, foam cell, histiocyte, luteal cell, lymphocytic stem cell, lymphoid cell, lymphoidstem cell, macroglial cell, mammotroph, mast cell, medulloblast, megakaryoblast, megakaryocyte, melanoblast, melanocyte, mesangial cell, mesothelial cell, metamyelocyte, monoblast, monocyte, mucous neck cell, myoblast, myocyte, muscle cell, cardiac muscle cell, skeletal muscle cell, smooth muscle cell, myelocyte, myeloid cell, myeloid stem cell, myoblast, myoepithelial cell, myofibrobast, neuroblast, neuroepithelial cell, neuron, odontoblast, osteoblast, osteoclast, osteocyte, oxyntic cell, parafollicular cell, paraluteal cell, peptic cell, pericyte, peripheral blood mononuclear cell, phaeochromocyte, phalangeal cell, pinealocyte, pituicyte, plasma cell, platelet, podocyte, proerythroblast, promonocyte, promyeloblast, promyelocyte, pronormoblast, reticulocyte, retinal pigment epithelial cell, retinoblast, small cell, somatotroph, stem cell, sustentacular cell, teloglial cell, a zymogenic cell, or any combination thereof. In some embodiments, the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell (iPSC), a hematopoietic stem / progenitor cell (HSPC), or any combination thereof.

[0102] In some embodiments, the receiver cell comprises a unique cell type and / or a unique cell state. In some embodiments, the unique cell type and / or the unique cell state comprises a unique gene expression pattern. In some embodiments, the unique cell type and / or unique cell state comprises (i) one or more mutations of a protein, (ii) structural variants and / or copy -number alternations of one or more protein-coding genes, (iii) epigenetic signature(s), and / or (iv) a unique anatomic location. In some embodiments, the unique cell type and / or the unique cell state comprises anatomically locally unique gene expression; wherein the unique cell type and / or the unique cell state is caused by hereditable, environmental, and / or idiopathic factors; wherein the unique cell ty pe and / or the cell in the unique cell state (i) causes and / or aggravates a disease or disorder and / or (ii) is associated with the pathology of a disease or disorder; and / or wherein the unique cell state comprises a senescent cell state induced by a tumor microenvironment. In some embodiments, the senescent cell state induced by atumor microenvironment comprises expression of CD57, KRLG1, TIGIT, p21, p53, phospho-p53, DECI, PPP1A, yH2AX, 53BPI, Radl7. ATR, ATM, MDC1, TIF, IL-6, IL-8, CXCR2, IGF2, IGFBP3, IGFBP5, IGFBP7, STC1, GDF15, SERPIN. ICAM-1, DEP1 , B2MG, NOTCH3, DcR2, or any combination thereof. In some embodiments, the unique cell state and / or unique cell type is characterized by one or more of: aberrant signaling of one or more signal trans ducer(s); cell proliferation, stress pathways, oxidative stress, stress kinase activation, DNA damage, lipid metabolism, carbohydrate regulation, metabolic activation including Phase I and Phase II reactions, Cytochrome P-450 induction or inhibition, ammonia detoxification, mitochondrial function, peroxisome proliferation, organelle function, cell cycle state, morphology7, apoptosis, DNA damage, metabolism, signal transduction, cell differentiation, cell-cell interaction and cell to non-cellular compartment; acute phase stress, cell adhesion, AH-response. anti-apoptosis and apoptosis,antimetabolism, anti- proliferation, arachidonic acid release, ATP depletion, cell cycle disruption, cell matrix disruption, cell migration, cell proliferation, cell regeneration, cell-cell communication, cholestasis, differentiation, DNA damage, DNA replication, early response genes, endoplasmic reticulum stress, estogenicity. fatty liver, fibrosis, general cell stress, glucose deprivation, growth arrest, heat shock, hepatotoxicity, hypercholesterolemia, hypoxia, immunotox, inflammation, invasion, ion transport, liver regeneration, cell migration, mitochondrial function, mitogenesis, multidrug resistance, nephrotoxicity, oxidative stress, peroxisome damage, recombination, ribotoxic stress, sclerosis, steatosis, teratogenesis, transformation, disrupted translation, transport, and tumor suppression; and nutrient deprivation, hypoxia, oxidative stress, hyperproliferative signals, oncogenic stress, DNA damage, ribonucleotide depletion, replicative stress, and telomere attrition, promotion of cell cycle arrest, promotion of DNA-repair, promotion of apoptosis, promotion of genomic stability, promotion of senescence, and promotion of autophagy’, regulation of cell metabolic reprogramming, regulation of tumor microenvironment signaling, inhibition of cell sternness, survival, and invasion. In some embodiments, the unique cell state comprises: a physiological state, e.g., a cell cycle state, a differentiation state, a development state a metabolic state, or a combination thereof; and / or a pathological state, e.g.. a disease state, a human disease state, a diabetic state, an immune disorder state, a neurodegenerative disorder state, an oncogenic state, or a combination thereof. In some embodiments, the unique cell type is: an antigen-presenting cell, a dendritic cell, a macrophage, a neural cell, a brain cell, an astrocyte, a microglial cell, and a neuron, a spleen cell, a lymphoid cell, a lung cell, a lung epithelial cell, a skin cell, a keratinocyte, an endothelial cell, an alveolar cell, an alveolar macrophage, an alveolar pneumocyte, a vascular endothelial cell, a mesenchymal cell, an epithelial cell, a colonic epithelial cell, a hematopoietic cell, a bone marrow cell, a Claudius cell, Hensen cell, Merkel cell, Muller cell, Paneth cell, Purkinje cell, Schwann cell, Sertoli cell, acidophil cell, acinar cell, adipoblast, adipocyte, brown or white alpha cell, amacrine cell, beta cell, capsular cell, cementocyte, chief cell, chondroblast, chondrocyte, chromaffin cell, chromophobic cell, corticotroph, delta cell, Langerhans cell, follicular dendritic cell, enterochromaffin cell, ependymocyte, epithelial cell, basal cell, squamous cell, endothelial cell, transitional cell, erythroblast, erythrocyte, fibroblast, fibrocyte, follicular cell, germ cell, gamete, ovum, spermatozoon, oocyte, primary oocyte, secondary oocyte, spermatid, spermatocyte, primary spermatocyte, secondary’ spermatocyte, germinal epithelium, giant cell, glial cell, astroblast, astrocyte, oligodendroblast, oligodendrocyte, glioblast, goblet cell, gonadotroph, granulosa cell, haemocytoblast, hair cell, hepatoblast, hepatocyte, hyalocyte, interstitial cell, juxtaglomerular cell, keratinocyte, keratocyte, lemmal cell, leukocyte, granulocyte, basophil, eosinophil, neutrophil, lymphoblast, B-lymphoblast, T-lymphoblast, lymphocyte, B-lymphocyte,T-lymphocyte, helper induced T-lymphocyte, Thl T-lymphocyte, Th2 T-lymphocyte, natural killer cell, thymocyte, macrophage, Kupffer cell, alveolar macrophage, foam cell, histiocyte, luteal cell, lymphocytic stem cell, lymphoid cell, lymphoid stem cell, macroglial cell, mammotroph, mast cell, medulloblast, megakaryoblast, megakaryocyte, melanoblast, melanocyte, mesangial cell, mesothelial cell, metamyelocyte, monoblast, monocyte, mucous neck cell, myoblast, myocyte, muscle cell, cardiac muscle cell, skeletal muscle cell, smooth muscle cell, myelocyte, myeloid cell, myeloid stem cell, myoblast, myoepithelial cell, myofibrobast, neuroblast, neuroepithelial cell, neuron, odontoblast, osteoblast, osteoclast, osteocyte, oxyntic cell, parafollicular cell, paraluteal cell, peptic cell, pericyte, peripheral blood mononuclear cell, phaeochromocyte, phalangeal cell, pinealocyte, pituicyte, plasma cell, platelet, podocyte, proerythroblast, promonocyte, promyeloblast, promyelocyte, pronormoblast, reticulocyte, retinal pigment epithelial cell, retinoblast, small cell, somatotroph, stem cell, sustentacular cell, teloglial cell, a zymogenic cell, or any combination thereof. In some embodiments, the stem cell comprises an embryonic stem cell, an induced pluripotent stem cell (iPSC), a hematopoietic stem / progenitor cell (HSPC), or any combination thereof. In some embodiments, the receiver cell is characterized by aberrant signaling of one or more signal transducers, and wherein the aberrant signaling involves: an overactive signal transducer; a constitutively active signal transducer over a period of time; an active signal transducer repressor and an active signal transducer; an inactive signal transducer activator and an active signal transducer; an inactive signal transducer; an underactive signal transducer; a constitutively inactive signal transducer over a period of time; an inactive signal transducer repressor and an inactive signal transducer; and / or an active signal transducer activator and an inactive signal transducer. In some embodiments, the aberrant signaling comprises an aberrant signal of at least one signal transduction pathway regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof. In some embodiments, the signal transduscer(s) is AKT, PI3K, MAPK, p44 / 42 MAP kinase, TYK2, p38 MAP kinase, PKC. PKA, SAPK. ELK. JNK, eJun, RAS, Raf. MEK 1 / 2, MEK 3 / 6, MEK 4 / 7. ZAP-70, LAT. SRC. LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX, PLCy, PLCy, NF-kB, FAK, CREB, aIII 3, FceRI, BAD, p70S6K, STAT1, STAT2, STAT3, STAT5, STAT6, or any combination thereof.

[0103] Provided herein are methods of selectively killing a target cell. In some embodiments, the method comprises: expressing any of the synthetic protein circuits or any of the nucleic acid compositions of the disclosure in the target cell, wherein the synthetic protein circuit is configured to be responsive to a unique cell type and / or unique cell state of the target cell. In some embodiments, the first and / or second heterologous protease is configured to be in the firstand / or second heterologous protease active state in response to the unique cell type and / or unique cell state of the target cell.

[0104] In some embodiments, the unique cell type and / or the unique cell state comprises a unique gene expression pattern. In some embodiments, the unique cell t pe and / or unique cell state comprises: (i) one or more mutations of a protein, (ii) structural variants and / or copy -number alternations of one or more protein-coding genes, (iii) epigenetic signature(s), and / or (iv) a unique anatomic location. In some embodiments, the unique cell type and / or the unique cell state comprises anatomically locally unique gene expression. In some embodiments, the unique cell type and / or the unique cell state is caused by hereditable, environmental, and / or idiopathic factors. In some embodiments, the unique cell type and / or the cell in the unique cell state (i) causes and / or aggravates a disease or disorder and / or (ii) is associated with the pathology of a disease or disorder. In some embodiments, the unique cell state comprises a senescent cell state induced by a tumor microenvironment. In some embodiments, the senescent cell state induced by a tumor microenvironment comprises expression of CD57. KRLG1, TIGIT. p21, p53. phospho-p53, DECI, PPP1A. yH2AX, 53BPL Radi 7, ATR, ATM, MDC1, TIF, IL-6, IL-8, CXCR2, IGF2. IGFBP3, IGFBP5, IGFBP7, STC1, GDF15, SERPIN, ICAM-1 , DEP1 , B2MG, NOTCH3, DcR2, or any combination thereof. In some embodiments, the unique cell state and / or unique cell type is characterized by aberrant signaling of one or more signal transducer(s). In some embodiments, the unique cell state comprises: a physiological state, e.g., a cell cycle state, a differentiation state, a development state a metabolic state, or a combination thereof; and / or a pathological state, e.g., a disease state, a human disease state, a diabetic state, an immune disorder state, a neurodegenerative disorder state, an oncogenic state, or a combination thereof. In some embodiments, the unique cell state and / or unique cell type is characterized by one or more of cell proliferation, stress pathways, oxidative stress, stress kinase activation, DNA damage, lipid metabolism, carbohydrate regulation, metabolic activation including Phase I and Phase II reactions, Cytochrome P-450 induction or inhibition, ammonia detoxification, mitochondrial function, peroxisome proliferation, organelle function, cell cycle state, morphology, apoptosis, DNA damage, metabolism, signal transduction, cell differentiation, cell-cell interaction and cell to non-cellular compartment. In some embodiments, the unique cell state and / or unique cell type is characterized by one or more of acute phase stress, cell adhesion, AH-response, anti-apoptosis and apoptosis, antimetabolism, anti- proliferation, arachidonic acid release, ATP depletion, cell cycle disruption, cell matrix disruption, cell migration, cell proliferation, cell regeneration, cellcell communication, cholestasis, differentiation, DNA damage, DNA replication, early response genes, endoplasmic reticulum stress, estogenicity, fatty liver, fibrosis, general cell stress, glucose deprivation, growth arrest, heat shock, hepatotoxicity, hypercholesterolemia, hypoxia,immunotox, inflammation, invasion, ion transport, liver regeneration, cell migration, mitochondrial function, mitogenesis, multidrug resistance, nephrotoxicity, oxidative stress, peroxisome damage, recombination, ribotoxic stress, sclerosis, steatosis, teratogenesis, transformation, disrupted translation, transport, and tumor suppression. In some embodiments, the unique cell state and / or unique cell type is characterized by one or more of nutrient deprivation, hypoxia, oxidative stress, hyperproliferative signals, oncogenic stress, DNA damage, ribonucleotide depletion, replicative stress, and telomere attrition, promotion of cell cycle arrest, promotion of DNA-repair, promotion of apoptosis, promotion of genomic stability, promotion of senescence, and promotion of autophagy’, regulation of cell metabolic reprogramming, regulation of tumor microenvironment signaling, inhibition of cell sternness, survival, and invasion. In some embodiments, the unique cell type is: an antigen-presenting cell, a dendritic cell, a macrophage, a neural cell, a brain cell, an astrocyte, a microglial cell, and a neuron, a spleen cell, a lymphoid cell, a lung cell, a lung epithelial cell, a skin cell, a keratinocyte, an endothelial cell, an alveolar cell, an alveolar macrophage, an alveolar pneumocyte, a vascular endothelial cell, a mesenchymal cell, an epithelial cell, a colonic epithelial cell, a hematopoietic cell, a bone marrow cell, a Claudius cell, Hensen cell, Merkel cell, Muller cell, Paneth cell, Purkinje cell, Schwann cell, Sertoli cell, acidophil cell, acinar cell, adipoblast, adipocyte, brown or white alpha cell, amacrine cell, beta cell, capsular cell, cementocyte, chief cell, chondroblast, chondrocyte, chromaffin cell, chromophobic cell, corticotroph, delta cell, Langerhans cell, follicular dendritic cell, enterochromaffin cell, ependymocyte, epithelial cell, basal cell, squamous cell, endothelial cell, transitional cell, ery throblast, ery throcyte, fibroblast, fibrocyte, follicular cell, germ cell, gamete, ovum, spermatozoon, oocyte, primary oocyte, secondary oocyte, spermatid, spermatocyte, primary' spermatocyte, secondary' spermatocyte, germinal epithelium, giant cell, glial cell, astroblast, astrocyte, oligodendroblast, oligodendrocyte, glioblast, goblet cell, gonadotroph, granulosa cell, haemocytoblast, hair cell, hepatoblast, hepatocyte, hyalocyte, interstitial cell, juxtaglomerular cell, keratinocyte, keratocyte, lemmal cell, leukocyte, granulocyte, basophil, eosinophil, neutrophil, lymphoblast. B-lymphoblast, T-lymphoblast, lymphocyte, B-lymphocyte, T-lymphocyte, helper induced T-lymphocyte, Thl T-lymphocyte, Th2 T-lymphocyte, natural killer cell, thymocyte, macrophage, Kupffer cell, alveolar macrophage, foam cell, histiocyte, luteal cell, lymphocytic stem cell, lymphoid cell, lymphoid stem cell, macroglial cell, mammotroph, mast cell, medulloblast. megakaryoblast. megakaryocyte, melanoblast, melanocyte, mesangial cell, mesothelial cell, metamyelocyte, monoblast, monocyte, mucous neck cell, myoblast, myocyte, muscle cell, cardiac muscle cell, skeletal muscle cell, smooth muscle cell, myelocyte, myeloid cell, myeloid stem cell, myoblast, myoepithelial cell, myofibrobast, neuroblast, neuroepithelial cell, neuron, odontoblast, osteoblast, osteoclast, osteocyte, oxyntic cell,parafollicular cell, paraluteal cell, peptic cell, pericyte, peripheral blood mononuclear cell, phaeochromocyte, phalangeal cell, pinealocyte, pituicyte, plasma cell, platelet, podocyte, proery throblast, promonocyte, promyeloblast, promyelocyte, pronormoblast, reticulocyte, retinal pigment epithelial cell, retinoblast, small cell, somatotroph, stem cell, sustentacular cell, teloglial cell, a zymogenic cell, or any combination thereof. In some embodiments, the stem cell comprises an ernbry onic stem cell, an induced pluripotent stem cell (iPSC), a hematopoietic stem / progenitor cell (HSPC), or any combination thereof. In some embodiments, the aberrant signaling involves: an overactive signal transducer; a constitutively active signal transducer over a period of time; an active signal transducer repressor and an active signal transducer; an inactive signal transducer activator and an active signal transducer; an inactive signal transducer; an underactive signal transducer; a constitutively inactive signal transducer over a period of time; an inactive signal transducer repressor and an inactive signal transducer; and / or an active signal transducer activator and an inactive signal transducer. In some embodiments, the aberrant signaling comprises an aberrant signal of at least one signal transduction pathway regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology', cell differentiation, apoptosis, or any combination thereof. In some embodiments, the signal transducer(s) is AKT, PI3K, MAPK, p44 / 42 MAP kinase, TYK2, p38 MAP kinase, PKC. PKA, SAPK. ELK. JNK, eJun, RAS, Raf. MEK 1 / 2, MEK 3 / 6, MEK 4 / 7, ZAP-70, LAT. SRC. LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX, PLCy, PLCy, NF-kB, FAK, CREB, aIII 3, FcsRI, BAD, p70S6K, STAT1, STAT2, STAT3, STAT5, STAT6.

[0105] In some embodiments, configuring the first and / or the second heterologous protease to be in the first and / or the second heterologous protease active state in response to the unique cell type and / or the unique cell state of the target cell comprises: expressing a second synthetic protein circuit in the target cell, wherein the second synthetic protein circuit comprises: a first polypeptide comprising a first signal transducer binding domain and a first part of a first protease domain of the first or second heterologous protease, wherein the first signal transducer binding domain is capable of binding a first signal transducer to form a first signal transducerbound polypeptide; a second polypeptide comprising a second signal transducer binding domain and a second part of the first protease domain of the first or second heterologous protease, wherein the second signal transducer binding domain is capable of binding a second signal transducer to form a second signal transducer-bound polypeptide, wherein the first part of the first protease domain and the second part of the first protease domain have weak association affinity, and wherein the first part of the first protease domain and the second part of the first protease domain are capable of associating with each other to constitute the first or second heterologous protease capable in a first or second heterologous protease active state capable of cutting: (i) the first,-n-second, or third apoptosis polypeptide at the first or second heterologous protease cleavage site when the first signal transducer and the second signal transducer are in close proximity at an association location; or (ii) the first or second pyroptosis polypeptide at the first or second heterologous protease cleavage site when the first signal transducer and the second signal transducer are in close proximity at an association location.

[0106] In some embodiments, the first signal transducer binding domain of the first polypeptide and the second signal transducer binding domain of the second polypeptide are identical. In some embodiments, the first signal transducer and the second signal transducer are identical and / or are the same protein. In some embodiments, the first signal transducer, the second signal transducer, or both, are capable of being localized at the association location. In some embodiments, the first signal transducer when in a first signal transducer active state, the second signal transducer when in a second signal transducer active state, or both, are capable of being localized at the association location. In some embodiments, the first signal transducer when in a first inactive state, the second signal transducer when in a second inactive state, or both, are capable of being localized at the association location. In some embodiments, the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer, or both. In some embodiments, the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first signal transducer active state, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer in a second signal transducer active state, or both. In some embodiments, the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first inactive state, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer in a second inactive state, or both. In some embodiments, the first signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, or both. In some embodiments, the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at a first cellular location other than the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at a second cellular location other than the association location, or both.

[0107] In some embodiments, the first cellular location, the second cellular location, or both comprise one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory’ vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome. microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof In some embodiments, the association location comprises one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof.

[0108] In some embodiments, the first signal transducer binding domain and the second signal transducer binding domain are different. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain each is capable of binding molecules of the first signal transducer and / or the second signal transducer. In some embodiments, the first signal transducer and / or the second signal transducer belong to a signal transduction pathway. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain comprise a RAS binding domain (RBD) and / or RAS association domain (RAD). In some embodiments, the RAS binding domain comprises or is derived from a RAS interacting protein, optionally selected from the group comprising AGO2, APBB1IP, APPL1, ARAF, ARL1, ARL2, ARRB1, ARRB2, BAIAP2, BCL2, BCL2L1, BRAF, BRAP, BSG, CALM1, CALM3, CALML3. CALML4, CALML5, CALML6, CNKSR1, CNKSR2, CSK, DAB2IP. EGFR. ERBIN, FGA. FGB. FGG, FN1, GRB2, HK1, IFNGR1, IL6, IQGAP1, ITGA2B, ITGB3, KSR1, KSR2, LGALS3, LYN, LZTR1, MAP2K1, MAP2K2, MAPK1, MAPK14, MAPK3, MAPKAP1, MARK2, MARK3, MBP, MSI2, MTOR, NCBP2AS2, NF1, NIBAN2, PDE4DIP, PDE6D, PDPK1, PEBP1, PIK3CA, PIK3CB, PIK3CD, PIK3R1, PIK3R2, PIP5K1A, PLCE1, PPIA, PRKCZ. PTGS2, RAFI, RALB, RALGDS. RAP1A, RAP1B, RAP1GDS1, RASA1, RASA2, RASA3, RASA4, RASAL1, RASAL2, RASAL3, RASSF1, RASSF2, RASSF5, RGL1, RGL3, RIN1, SHOC2, S0S1, S0S2, SPRED1, SPRED2, SPRED3, SRC, SYNGAP1, TIAM1, TLN1, VCL, VWF, YWHAB, or any combination thereof. In some embodiments, the first signal transducer binding domain and / or the second signal transducerbinding domain comprises a lipid binding domain. In some embodiments, the lipid binding domain comprises a Pleckstrin homology (PH) domain. In some embodiments, the first signal transducer binding domain and / or the second signal transducer binding domain comprises an antibody, an antibody fragment, a binding domain derived from a natural protein, an scFv, a Fv, a Fab, a (Fab')2, a single domain antibody (SDAB), a VH or VL domain, a camelid VHH domain, a Fab, a Fab1, a F(ab’)2, a Fv, a scFv, a dsFv, a diabody, a triabody, a tetrabody, a multispecific antibody formed from antibody fragments, a single-domain antibody (sdAb), a single chain comprising cantiomplementary scFvs (tandem scFvs) or bispecific tandem scFvs, an Fv construct, a disulfide-linked Fv, a dual variable domain immunoglobulin (DVD-Ig) binding protein or a nanobody, an aptamer, an affibody, an affilin, an affitin, an affimer, an alphabody, an anticalin, an avimer, a DARPin, a Fynomer, a Kunitz domain peptide, a monobody, or any combination thereof. In some embodiments, the first signal transducer is capable of binding the first signal transducer binding domain and / or the second signal transducer is capable of binding the second signal transducer binding domain following a modification selected from the group comprising phosphorylation, dephosphorylation, acetylation, methylation, acylation, glycosylation, glycosylphosphatidylinositol (GPI) anchoring, sulfation, disulfide bond formation, deamidation, ubiquitination, sumoylation, nitration of tyrosine, hydrolysis of ATP or GTP, binding of ATP or GTP, cleavage, or any combination thereof.

[0109] In some embodiments, the first signal transducer, the second signal transducer, or both are endogenous proteins. In some embodiments, the first signal transducer, the second signal transducer, or both comprise AKT, PI3K, MAPK, p44 / 42 MAP kinase, TYK2, p38 MAP kinase, PKC. PKA, SAPK, ELK, JNK, eJun, RAS, Raf. MEK 1 / 2, MEK 3 / 6. MEK 4 / 7, ZAP-70, LAT, SRC, LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX, PLCy, PLCy, NF- kB, FAK, CREB, aHI|33, FccRI, BAD, p70S6K, STAT1, STAT2, STAT3, STAT5, STAT6, or any combination thereof. In some embodiments, the first signal transducer and / or the second signal transducer are capable of regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof. In some embodiments, the first signal transducer, the second signal transducer, or both comprise a RAS protein, a CTNNB1 protein, or a TP53 protein. In some embodiments, the RAS protein is KRAS. NHAS, HRAS, or any combination thereof. In some embodiments, the RAS protein comprises a G12 mutation, G13 mutation, a Q61 mutation, and / or an A146 mutation. In some embodiments, the G12 mutation is selected from the group comprising G12A, G12C, G12D, G12R, G12S, G12V, and any combination thereof. In some embodiments, the G13 mutation is selected from the group comprising G13C, G13D, G13dup, G13R, G13S, G13V, and any combination thereof. In some embodiments, the Q61 mutation is selected from thegroup comprising Q61H, Q61K, Q61L, Q61R, Q61E, Q61P, Q61*, and any combination thereof. In some embodiments, the Al 46 mutation is selected from the group comprising A146P, A146T, Al 46V, and any combination thereof. In some embodiments, the signature detected by the input polypeptide(s) is correlated with any other cellular signature, protein state, cell type, and / or cell state capable of being read out as a biomarker. In some embodiments, the signal transducer is CTNNB1, and wherein the cell state is defined by CTNNB1 mutation(s) and / or localization and / or concentration and / or protein turnover and / or multimerization and / or PTM(s). In some embodiments, the signal transducer is TP53, and wherein the cell state is defined by TP53 mutation(s) and / or elevated TP53 concentration and / or altered TP53 oligomerization / multimerization state and / or TP53 localization pattern and / or PTM(s) and / or turnover. In some embodiments, the signal transducer(s) are associated with disease, e.g., cancer. In some embodiments, the first signal transducer, the second signal transducer, or both are exogenous proteins. In some embodiments, the second synthetic protein circuit comprises the first signal transducer, the second signal transducer, or both. In some embodiments, the first signal transducer, the second signal transducer, or both comprise a lipid. In some embodiments, the lipid comprises a phospholipid. In some embodiments, the phospholipid is phosphatidylinositol 3- phosphate.

[0110] Disclosed herein include methods of treating or preventing a disease or disorder in a subject in need thereof. In some embodiments, the method comprises: expressing any of the synthetic protein circuit, the synthetic protein circuit and / or the second synthetic protein circuit of the disclosure, in a cell of the subject.[OHl] In some embodiments, the method comprises: administering to the subject an effective amount of a nucleic acid composition or a composition disclosed herein, thereby treating or preventing the disease or disorder in the subject. In some embodiments, administering comprises: (i) isolating one or more cells from the subject; (ii) contacting said one or more cells with any of the nucleic acid compositions disclosed herein, thereby generating engineered cells, optionally the contacting comprises transfection; and (iii) administering the one or more engineered cells into a subject after the contacting step.

[0112] In some embodiments, the disease or disorder is a blood disease, an immune disease, a neurological disease or disorder, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA. a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof, e.g., a solid tumor.

[0113] In some embodiments, the disease or disorder is an infectious disease selected from the group consisting of an Acute Flaccid Myelitis (AFM), Anaplasmosis. Anthrax,Babesiosis, Botulism, Brucellosis, Campylobacteriosis, Carbapenem-resistant Infection, Chancroid, Chikungunya Virus Infection, Chlamydia, Ciguatera, Difficile Infection, Perfringens, Coccidioidomycosis fungal infection, coronavirus infection, Covid- 19 (SARS-CoV-2), Creutzfeldt-Jacob Disease / transmissible spongiform encephalopathy, Cryptosporidiosis (Crypto), Cyclosporiasis, Dengue 1.2.3 or 4, Diphtheria, E. coli infection / Shiga toxin-producing (STEC), Eastern Equine Encephalitis, Hemorrhagic Fever (Ebola), Ehrlichiosis, Encephalitis, Arboviral or parainfectious, Non-Polio Enterovirus, D68 Entero viru(EV-D68), Giardiasis, Glanders, Gonococcal Infection, Granuloma inguinale, Haemophilus Influenza disease Type B (Hib or H- flu), Hantavirus Pulmonary Syndrome (HPS). Hemolytic Uremic Syndrome (HUS). Hepatitis A (Hep A), Hepatitis B (Hep B), Hepatitis C (Hep C), Hepatitis D (Hep D), Hepatitis E (Hep E), Herpes, Herpes Zoster (Shingles), Histoplasmosis infection, Human Immunodeficiency Virus / AIDS (HIV / AIDS), Human Papillomavirus (HPV), Influenza (Flu), Legionellosis (Legionnaires Disease). Leprosy (Hansens Disease), Leptospirosis, Listeriosis (Listeria), Lyme Disease, Lymphogranuloma venereum infection (LGV), Malaria, Measles, Melioidosis, Meningitis (Viral), Meningococcal Disease (Meningitis (Bacterial)), Middle East Respirator}' Syndrome Coronavirus (MERS-CoV), Mumps, Norovirus, Pediculosis, Pelvic Inflammatory' Disease (PID), Pertussis (Whooping Cough), Plague (Bubonic, Septicemic, Pneumonic), Pneumococcal Disease (Pneumonia), Poliomyelitis (Polio), Powassan, Psittacosis, Pthiriasis, Pustular Rash diseases (Small pox, monkeypox, cowpox), Q-Fever, Rabies, Rickettsiosis (Rocky’ Mountain Spotted Fever), Rubella (German Measles), Salmonellosis gastroenteritis (Salmonella), Scabies, Scombroid, Sepsis, Severe Acute Respiratory Syndrome (SARS), Shigellosis gastroenteritis (Shigella), Smallpox, Staphyloccal Infection Methicillin-resistant (MRSA), Staphylococcal Food Poisoning Enterotoxin B Poisoning (Staph Food Poisoning), Saphylococcal Infection Vancomycin Intermediate (VISA), Staphylococcal Infection Vancomycin Resistant (VRSA), Streptococcal Disease Group A (invasive) (Strep A (invasive), Streptococcal Disease, Group B (Strep-B), Streptococcal Toxic-Shock Syndrome STSS Toxic Shock, Syphilis (primary, secondary, early latent, late latent, congenital). Tetanus Infection. Trichomoniasis, Trichonosis Infection, Tuberculosis (TB), Tuberculosis Latent (LTBI), Tularemia, Typhoid Fever Group D, Vaginosis, Varicella (Chickenpox), Vibrio cholerae (Cholera), Vibriosis (Vibrio), Ebola Virus Hemorrhagic Fever, Lasa Virus Hemorrhagic Fever, Marburg Virus Hemorrhagic Fever, West Nile Virus. Yellow Fever, Yersenia. and Zika Virus Infection.

[0114] In some embodiments, the disease is associated with expression of a tumor- associated antigen. In some embodiments, the disease associated with expression of a tumor antigen-associated is selected from the group consisting of a proliferative disease, a precancerous condition, a cancer, and a non-cancer related indication associated with expression of the tumorantigen.

[0115] In some embodiments, the cancer is selected from the group consisting of colon cancer, rectal cancer, renal-cell carcinoma, liver cancer, non-small cell carcinoma of the lung, cancer of the small intestine, cancer of the esophagus, melanoma, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin lymphoma, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, solid tumors of childhood, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers, combinations of said cancers, and metastatic lesions of said cancers. In some embodiments, the cancer is non-resectable.

[0116] In some embodiments, the cancer comprises one or more mutations in KRAS, NRAS. HRAS. or any combination thereof, e.g., a G12 mutation. G13 mutation, a Q61 mutation, and / or an A146 mutation. In some embodiments, the G12 mutation is selected from the group comprising G12A, G12C, G12D, G12R, G12S, G12V, and any combination thereof. In some embodiments, the G13 mutation is selected from the group comprising G13C, G13D, G13dup, G13R, G13S, G13V, and any combination thereof. In some embodiments, the Q61 mutation is selected from the group comprising Q61H, Q61K, Q61L, Q61R, Q61E, Q61P, Q61*, and any combination thereof. In some embodiments, the A146 mutation is selected from the group comprising A146P, A146T, A146V, and any combination thereof

[0117] In some embodiments, the subject was previously treated with approved medication(s) and / or surgery. In some embodiments, the cancer comprises metastasis to liver and / or lung, e.g., (i) metastasis of colorectal cancer to liver and / or lung or (ii) metastasis of pancreatic cancer to liver and / or lung. In some embodiments, said metastasis is non-resectable. In some embodiments, the cancer is a hematologic cancer chosen from one or more of chronic lymphocytic leukemia (CLL). acute leukemias, acute lymphoid leukemia (ALL), B-cell acute lymphoid leukemia (B-ALL), T-cell acute lymphoid leukemia (T-ALL), chronic myelogenous leukemia (CML), B cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt's lymphoma, diffuse large B cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell- or a large cell-follicular lymphoma, malignant lymphoproliferative conditions, MALTlymphoma, mantle cell lymphoma, marginal zone lymphoma, multiple myeloma, myelodysplasia and myelodysplastic syndrome, non-Hodgkin's lymphoma, Hodgkin's lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, or preleukemia.

[0118] In some embodiments, administering comprises one or more administrations, e.g., via aerosol delivery, nasal delivery, vaginal delivery, rectal delivery, buccal delivery, ocular delivery, local delivery, topical delivery, intracistemal delivery', intraperitoneal delivery', oral delivery', intramuscular injection, intravenous injection, subcutaneous injection, intranodal injection, intratumoral injection, intraperitoneal injection, intradermal injection, or any combination thereof. In some embodiments, the administering comprises continuous administration. In some embodiments, the administering comprises a first administration and a second administration. In some embodiments, separated by a period of time of at least one hour, at least one day, at least one week, or at least one month. In some embodiments, administering comprises one or more repeated administrations. In some embodiments, repeated hourly, every 2 hours, every 4 hours, every' 6 hours, every 8 hours, every' 10 hours, every' 12 hours, every 16 hours, every' 20 hours, every' 24 hours, every' 2 days, every' 3 days, every' 4 days, every' 5 days, every' 6 days, every 7 days, every' 10 days, or every 14 days.

[0119] Any of the synthetic protein circuits, nucleic acid compositions, or methods can comprise a supplementary' protein circuit. In some embodiments, the supplementary' protein circuit comprises: a first polypeptide comprising an optional first supplementary domain and a first part of a first protease domain of a supplementary heterologous protease; a second polypeptide comprising a second supplementary domain and a second part of the first protease domain of the supplementary protease, wherein the first part of the first protease domain and the second part of the first protease domain have weak association affinity, and wherein the first part of the first protease domain and the second part of the first protease domain are capable of associating with each other to constitute the supplementary heterologous protease, optionally the first and / or second supplementary domain is a signal transducer binding domain, optionally the supplementary heterologous protease in a supplementary heterologous protease active state is capable of cutting (i) the first, second, or third apoptosis polypeptide at the first or second heterologous protease cleavage site, and / or (ii) the first or second pyroptosis polypeptide at the first or second heterologous protease cleavage site, further optionally when a first signal transducer and a second signal transducer are in close proximity at an association location the supplementary heterologous protease is the first or second heterologous protease.

[0120] In some embodiments, the induction of apoptosis and / or pyroptosis is dependent on the dose of one or more synthetic protein circuit components. In some embodiments,cell death is triggered when a threshold amount of one or more of the following is reached: (i) the first, second, and / or third apoptotic protein complex in the first, second, and / or third apoptotic protein complex active state; (ii) the first and / or second pyroptosis polypeptide in a first and / or second pyroptosis polypeptide active state; and / or (iii) a pyroptosis effector protein in a pyroptosis effector protein active state.BRIEF DESCRIPTION OF THE DRAWINGS

[0121] FIG. 1A-FIG. IE display non-limiting exemplar}' data showing synpoptosis circuits control user-selectable cell death programs. The schematic in FIG. 1A shows synthetic cell death “synpoptosis’' circuits steer the mode of cell death by operating orthogonally to cell- intrinsic death programs. Shown in FIG. IB are schematics of exemplary molecular building blocks of synpoptosis circuits of the disclosure, include caspase-3 subunits, gasdermin domains, viral proteases such as TEVP, degrons, maltose-binding protein, and leucine zippers. As shown in FIG. 1C, synthetic apoptosis modules use viral proteases, such as TEVP, to activate or repress engineered variants of caspase-3. HEK cells were transiently transfected with plasmid DNA encoding the synthetic modules and then cell death was quantified by staining and flow cytometry. Throughout the figure, the gray window indicates the fractions established by negative and positive transient DNA transfection controls; dots represent biological replicates (distinct culture wells). As shown in FIG. ID, synthetic pyroptosis modules similarly use TEVP to regulate engineered GSDMA. FIG. IE displays exemplary data showing apoptosis and pyroptosis exhibit different staining patterns with Annexin and Sytox. Annexin stains both apoptotic and pyroptotic cells, while Sytox primarily stains pyroptotic cells. We calculated the fraction of cells that stained positive for each dye after gating on the transfected cells based on fluorescence of a co-transfected marker. Data represent three independent experiments.

[0122] FIG. 2A-FIG. 2H display non-limiting exemplar}’ data showing synpoptosis circuits lead to canonical features of cell death. FIG. 2A displays data showing transient transfection of HEK cells with plasmid DNA encoding the TEVP-activated caspase-3 circuit triggered asynchronous apoptosis, shown by flow cytometry. Between 16 and 24 hours after transfection (gray window). Sytox remains low in apoptotic cells and therefore can reliably distinguish between apoptotic and pyroptotic cells. Throughout the disclosure, dots represent biological replicates (distinct culture wells). Similarly, the TEVP-activated GSDMA circuits triggered asynchronous pyroptosis (FIG. 2B). FIG. 2C displays graphs showing Q-VD-OPh suppressed circuit-induced apoptosis but not pyroptosis, shown by flow cytometry. The gray window indicates the fractions established by negative and positive transient DNA transfection controls. As shown in FIG. 2D, TO-PRO-3 stains cells killed by the apoptosis and pyroptosis circuits, shown by flow’ cytometry'. FIG. 2E displays exemplary' data showing the apoptosis andpyroptosis circuits differently triggered the release of IL-1 P and IL-18 from engineered HEK cells that stably express these cytokines. As shown in FIG. 2F, transient transfection using in vitro transcribed mRNA transcripts of the synpoptosis circuits led to loss of ATP-based cell viability. FIG. 2G displays exemplary data showing the mRNA version of the pyroptosis circuit triggered more LDH release than the apoptosis circuit. The mRNA version of the pyroptosis circuit triggered more ATP release than the apoptosis circuit. The same wells were repeatedly captured across a time course (FIG. 2H).

[0123] FIG. 3A-FIG. 3F display non-limiting exemplary data showing synpoptosis circuits direct the mode of cell death. FIG. 3A displays exemplary data showing transient transfection of plasmid DNA encoding TEVP and engineered TEVP-activatable caspase-3 induced apoptosis in HEK cells, shown by flow cytometry. TEVP-activatable GSDMA overrode the apoptotic program, leading to pyroptosis. Throughout the figure, horizontal lines indicate the fractions established by negative and positive transient DNA transfection controls, established separately for Annexin and Sytox; dots represent biological replicates (distinct culture wells). As shown in FIG. 3B, cells transfected with plasmid DNA encoding wildtype GSDME, a natural substrate of caspase-3, underw ent pyroptosis in response to TEVP-mediated caspase-3 activation. GSDME N(mut) overcame the tendency of wildtype GSDME-expressing cells to undergo pyroptosis downstream of caspase-3 activation and promoted apoptosis (FIG. 3C). FIG. 3D displays exemplary data showing the ratio between the two forms of cell death is tunable by adjusting the plasmid DNA amount of GSDME N(mut) relative to wildtype GSDME (low, 1 : 1; medium, 2: 1; high, 4:1). The heights of the stacked bars indicate means of three biological replicates (distinct culture wells) measured by flow cytometry. FIG. 3E displays exemplary data showing in vitro transcribed mRNA versions of synpoptosis circuits drove apoptosis and pyroptosis in Jurkat and THP-1 cells, which have more sophisticated endogenous cell death circuitry than HEK cells. As shown in FIG. 3F, in response to natural death inducers, cells execute their default death programs. GSDM- negative and GSDM-positive cells tend to die by apoptosis and pyroptosis, respectively. Described herein are synpoptosis circuits that dictate death mode by overriding these cell -intrinsic preferences.

[0124] FIG. 4A-FIG. 4F display non-limiting exemplary data showing synpoptosis circuits perform combinatorial computation. As shown in FIG. 4A, synpoptosis circuits enabled context-dependent apoptosis by responding to logical combinations of protease inputs (TEVP as Input 1 and TVMVP as Input 2). In the first demonstration, transient transfection of plasmid DNA encoding both protease inputs must be present to activate the engineered caspase-3 and cause apoptosis, shown by flow cytometry7. In the second demonstration, either input is sufficient to trigger apoptosis. In the third demonstration, apoptosis occurs only when the first input is presentand the second input is absent. Throughout the figure, the gray window indicates the fractions established by negative and positive transient DNA transfection controls; dots represent biological replicates (distinct culture wells). Similar design principles were applied to engineer the same gating functions for pyroptosis (FIG. 4B). FIG. 4C displays exemplary data showing that using these synpoptosis gates, one can eliminate specific cells within mixed populations that exhibit distinct intracellular states defined by the expression profiles of input proteases and their fluorescence reporters, shown by flow cytometry. The cell states were established by stable protein expression using lentiviral transduction. Data represent three independent experiments. Quantification of Annexin staining revealed the efficacy of the synpoptosis gates (FIG. 4D). In FIG. 4D, each quadrant is shown from left to right for the indicated circuit. Horizontal lines indicate quadrant-specific negative and positive controls of the Annexin signal. Bars indicate means of three biological replicates (distinct culture wells). FIG. 4E displays a schematic showing synpoptosis circuits can achieve context-dependent killing by responding to logical combinations of protease inputs. As shown in FIG. 4F, using these synpoptosis gates, we can eliminate specific cells within mixed populations that exhibit distinct intracellular states.

[0125] FIG. 5A-FIG. 5D display non-limiting exemplary data showing synpoptosis circuits selectively eliminate target cells. As shown in FIG. 5A, synpoptosis circuits can selectively kill target cells by incorporating a sensor module that identifies the target cells. A synthetic sensor of active Ras was used. The sensor is a split TEVP with each half - nTEVP and cTEVP - tethered to a Ras-binding domain (RBD). In wildtype (WT) HEK cells with inactive Ras, the sensor is catalytically inactive. In cells stably expressing active Ras (RasGC cells), TEVP is reconstituted by proximity. FIG. 5B displays exemplary data showing that transient DNA transfection of a caspase-3-based synpoptosis circuit including the synthetic Ras sensor module enabled selective apoptosis of RasGC cells, but not WT cells. Throughout the figure, horizontal lines in the bar plots indicate the fractions established by negative and positive transient DNA transfection controls, separately for each dye and each cell line; dots represent biological replicates (distinct culture wells); bars indicate means; histograms represent three independent experiments. Similarly, a Ras-sensing synpoptosis circuit using GSDMA as the output triggered selective pyroptosis of RasGC cells (FIG. 5C). As shown in FIG. 5D, synpoptosis circuits can selectively kill target cells by incorporating a sensor module that identifies the targets. This feature is desired if we need to restrict synpoptosis to harmful cells within a mixture among healthy cells.

[0126] FIG. 6A-FIG. 6H display non-limiting exemplary data showing that synpoptosis circuits support intercellular operations. FIG. 6A displays an exemplary schematic showing synpoptosis circuits can be transmitted intercellularly by virus-like particles (VLPs). To address the key challenge of sender cell death, a simple strategy is to use a split-sender system, inwhich an inactive executioner and an activating protease are packaged separately. As shown in FIG. 6B, a model VLP transiently delivered nucleic acid encoding Cherry from sender HEK cells to receiver HEK cells, where the cargo was expressed. Experiments were performed by supernatant transfer from sender to receiver cells after transient DNA transfection of sender cells. Line segments connect means at different time points. Dots represent biological replicates (distinct culture wells). In FIG. 6C, two model cargoes, Cherry and Citrine, were separately packaged by two sender populations and co-delivered to the same receiver population, supporting the splitsender system. Scatter plots represent three independent experiments. FIG. 6D displays exemplary' data showing a synthetic pyroptosis circuit, consisting of inactive GSDMA and the activating protease TEVP, was delivered by VLP using the split-sender system. Throughout the figure, the gray window indicates the fractions established by negative and positive transient DNA transfection controls; bars indicate means; dots in the two-dimensional sender death-receiver death plots indicate means of three biological replicates (distinct culture wells). Analogously, the split-sender system enabled VLP delivery of a synthetic apoptosis circuit, by separately packaging inactive caspase-3 and the activating protease TEVP (FIG. 6E). Appending CAAX tails to both circuit components enhanced apoptosis. A more elegant, compact, single-sender system directly delivers an active executioner. This strategy requires sender-specific silencing of the active executioner (FIG. 6F). As shown in FIG. 6G, a synthetic protein-level silencer (GSDMA Z-C) potently inhibited an engineered active executioner (GSDMA N-Z), likely by using leucine zippers (Z) to mask a loop critical for the executioner’s activity, shown by AlphaFold models. The inhibitory curve was obtained by transient transfection of HEK cells with plasmid DNA encoding the active executioner and the silencer at indicated plasmid mass ratios. Senders transfected with plasmid DNA encoding the silencer, but not wildtype senders, could directly package VLPs that express the active executioner (FIG. 6H).

[0127] FIG. 7A-FIG. 7E display non-limiting exemplary' data showing synpoptosis circuits orthogonally control cell death. FIG. 7A displays a graph showing that HEK cells express negligible levels of endogenous GSDMs. Expression data were obtained from the Human Protein Atlas. As shown in FIG. 7B, the majority of experiments in instant disclosure were performed using the transient transfection method, as show n in the schematic with mock data. The cells were co-transfected with plasmid DNA encoding the synthetic circuits and a fluorescent protein marker. After 16-24 hours post-transfection, both floating and attached cells were collected for staining and flow cytometry. As shown in FIG. 7C, there are several possible sources of cells that stain low for the fluorescent co-transfection marker (Cherry ) and high for the death dye (Annexin). First, the transfection reagent itself causes some toxicity; second, the cell collection procedure involving pelleting and resuspending kills some cells; and third, when the circuits kill the cells,they also reduce the level of the fluorescent protein in the cells, making some transfected cells appear Cherry-low. Given the uncertain origins, Cherry-high cells were focused on by gating, which allowed definitive attribution of observed cell death to the transfected synthetic circuits. Data represent three independent experiments. FIG. 7D displays exemplary data showing three viral proteases, TEVP, TVMVP, and HCVP, orthogonally activated their cognate engineered GSDMs containing the cleavage sites (tev, tvmv, and hcv, respectively). Data represent three independent experiments. Colors indicate means. As shown in FIG. 7E, naturally occurring GSDMD inactivation by caspase-3 prompted us to insert a TEVP cleavage site (tev) at an equivalent location in GSDMA. The GSDMD structure was from the Protein Data Bank (PDB 6N9O). The engineered GSDMA model was predicted by AlphaFold.

[0128] FIG. 8A-FIG. 8D display non-limiting exemplary data related to synpoptosis circuits show ty pical features of natural death programs. As shown in FIG. 8A, in transient DNA transfection experiments, engineered auto-inhibited caspase-3 or GSDMA induced modest cell death when they were highly expressed, without the activating TEVP. Within each bin on the x- axis, dots represent biological replicates (distinct culture wells). FIG. 8B displays exemplary data showing titrating the DNA amounts of the circuit plasmids enabled dose-dependent control over the fraction of cell killing. Throughout the figure, the gray window indicates the y-ranges established by positive and negative transient DNA transfection controls; dots represent biological replicates (distinct culture wells). As shown in FIG. 8C, at a constant amount of the plasmid DNA, the penetrance of synpoptosis circuits could be tuned by modulating protein expression at the mRNA level, using a synthetic miRNA-based incoherent feedforward loop (IFFL). The strength of mRNA inhibition by the miRNA is tunable by adjusting base complementarity between mRNA and miRNA. In the GFP panel, dots mark median fluorescence values. FIG. 8D displays exemplary images showing cells killed by synpoptosis circuits displayed canonical morphological characteristics of apoptosis and pyroptosis. Images are representative of three independent experiments and on the same brightness and color scales. Scale bars: 200 pm (left) and 20 pm (right).

[0129] FIG. 9A-FIG. 9D display non-limiting exemplary- data showing synpoptosis circuits modulate cell death. As shoyvn in FIG. 9A, GSDME mutants displayed various pyroptotic activity compared to wildt pe GSDME N-domain (N), shoyvn by transient DNA transfection and flow cytometry. The last mutant was defective and later referred to as GSDME N(mut). GSDME N(mut) inhibited wildtype GSDME N-induced pyroptosis. Ratio of mutant to wildtype GSDME (mass of DNA plasmids) from left to right: 1-to-l (low), 2-to-l (medium), and 4-to-l (high). The gray window indicates the fractions established by negative and positive transient DNA transfection controls. Throughout the figure, dots represent biological replicates (distinct culturewells). Time-course flow cytometry experiments suggested that GSDME N(mut) promoted apoptosis to various degrees, shown by different Sytox levels and similar Annexin levels, in cells expressing wildtype GSDME (FIG. 9B). The gray window indicates that Sytox remains low in apoptotic cells between 16 and 24 hours post DNA transfection. Using GSDMD-KO THP-1 cells, we demonstrated that the mRNA-encoded synpoptosis circuits trigger the expected mode of cell death, independently of endogenous death circuitry (FIG. 9C). Horizontal lines indicate the fractions established by negative and positive transient mRNA transfection controls. Shown in FIG. 9D is a schematic of natural mechanism the cell uses to attenuate pyroptosis is the trans inhibition of pyroptotic GSDMB (variants 3 and 4) by non-pyroptotic GSDMB (variants 1. 2, and 5). This architecture inspired the design of synpoptosis circuits that force apoptosis in GSDM- positive cells.

[0130] FIG. 10A-FIG. 10E display non-limiting exemplary data showing synpoptosis circuits enable complex functions. As shown in FIG. 10A, more gates were built that synthetically control apoptosis based on different protease input combinations, shown by transient DNA transfection and flow cytometry experiments. Throughout the figure, the gray window indicates the fractions established by negative and positive transient DNA transfection controls; dots represent biological replicates (distinct culture wells). The corresponding logic gates for synthetic control of pyroptosis were constructed similarly (FIG. 10B). As shown in FIG. 10C, cell death programs naturally respond to logical input combinations. GSDMD is proteolytically activated by either caspase-1 or caspase-11, like an OR gate. On the other hand, cleavage and lipidation of GSDMD by the zDHHC enzy mes are both required for GSDMD activity, like an AND gate. Besides AND. OR, and NIMPLY. we built the other five binary logic gating functions for synthetic apoptosis (FIG. 10D). Similar principles were transferable to the design of synthetic pyroptosis gates (FIG. 10E).

[0131] FIG. 11A-FIG. 11C display non-limiting exemplary data showing synpoptosis circuits exhibit target cell selectivity. FIG. 11A shows a schematic of Proximity-reconstituted TEVP in RasGC cells should protect a degron-tagged Citrine reporter (Citrine-tev-Deg) from degradation, enhancing Citrine fluorescence. As shown in FIG. 11B, the synthetic Ras sensor enabled selective targeting of RasGC cells, shown by RasGC-specific enhancement of Citrine fluorescence, measured by flow cytometry after transient DNA transfection. Light and dark gray colors indicate WT and RasGC cells, respectively. The Citrine-tev-Deg reporter was stably integrated into RasGC and WT cells by lentiviral transduction. Data represent three independent experiments. FIG. 11C displays exemplary data showing the combination of nTEVP-RBD and cTEVP-RBD enhanced Citrine fluorescence in RasGC cells, shown on both linear and log scales. Horizontal lines indicate the median fluorescence values established by negative and positivetransient DNA transfection controls, established separately for each cell line; bars indicate the means of median fluorescence values; dots represent biological replicates (distinct culture wells).

[0132] FIG. 12A-FIG. 12E display non-limiting exemplary' data showing synpoptosis circuits allow cell-cell transmission. As shown in FIG. 12A, VLPs expressing a circuit component could complement the missing component in the cell, shown by flow cytometry after supernatant transfer. The gray window indicates the fractions established by negative and positive transient DNA transfection controls. Throughout the figure, dots represent biological replicates (distinct culture wells), and bars indicate means. FIG. 12B displays a graph showing inhibition of GSDMA N by GSDMA C requires a zipper (Z) on each of the two molecules, shown by transient DNA transfection and flow cytometry experiments. The gray window indicates the fractions established by negative and positive transient DNA transfection controls. FIG. 12C displays a graph of data showing senders expressing GSDMA Z-C (the silencer) after transient DNA transfection were protected from pyroptosis when treated with supernatants containing VLPs that express GSDMA N-Z (the engineered active executioner). The gray window indicates the fractions established by wild type senders treated with empty VLPs and GSDMA N-Z VLPs, respectively. FIG. 12D displays exemplary' images showing GSDMA Z-C (the silencer) suppressed cytotoxicity' caused by GSDMA N-Z (the engineered active executioner). Images are representative of three independent experiments and on the same brightness and color scales. Scale bar: 200 pm. FIG. 12E displays exemplary data related to the single-sender system functions in a co-culture context. Sender cells were transiently transfected with DNA to express the silencer and package GSDMA N-Z VLPs or empty' VLPs as a control. The senders, but not receivers, were stained with Vybrant DiD. To show that Vybrant DiD persists in dead cells HEK cells pre-stained by Vybrant DiD were transfected with plasmid DNA encoding GSDMA N-Z, followed by Sytox staining and flow cytometry.

[0133] FIG. 13 display s a schematic of the exemplary7methods and compositions of the disclosure.

[0134] FIG. 14 displays a diagrams of the motivation for fine-tuning of cell death executioner activation thresholds.

[0135] FIG. 15A-FIG. 15B show non-limiting exemplary' data related to the finetuning of cell death executioner activation thresholds. Graphs in FIG. 15 A show mutation of protease cut site or addition of protease-cleavable degrons enables background suppression (cf. N.C. panel) and fine-tuning of the caspase activation threshold. Cell death quantified with Annexin V (APC channel). Caspase co-transfected with GFP to quantify caspase dosage. N.C. = nTEVP + cTEVP (no active TEVP reconstitution); P.C. = P3-nTEVP + P4-cTEVP (P3, P4 zipper domains trigger TEVP reconstitution); NeoR = no TEVP transfected. As shown in FIG. 15B, fine-tuning with select protease cleavage site modifications does not affect maximal killing. Cell death quantified with Annexin V.

[0136] FIG. 16 displays non-limiting exemplary data showing caspase localization can boost input sensitivity or reduce background killing. On the left is shown a schematic of design strategy. As shown in the graph on the right, when membrane TEVP is co-localized with membrane Casp3, the amount of induced cell death is higher than that of membrane TEVP with cytoplasmic Casp3.

[0137] FIG. 17 displays exemplary data showing gasdermin localization can boost input sensitivity or reduce background killing.

[0138] FIG. 18 shows data related to rational protein design can reduce background and boost sensitivity.

[0139] FIG. 19A-FIG. 19B display non-limiting exemplary7data showing that split caspases can sense Ras and conditionally trigger cell death without requiring protease module. FIG. 19A depicts an exemplary schematic of a split-caspase circuit of the disclosure. FIG. 19B show s a graph of apoptosis induction in cells using an exemplary split caspase circuit.DETAILED DESCRIPTION

[0140] In the following detailed description, reference is made to the accompanying drawings, which form a part hereof. In the drawings, similar symbols typically identify similar components, unless context dictates otherwise. The illustrative embodiments described in the detailed description, drawings, and claims are not meant to be limiting. Other embodiments may be utilized, and other changes may be made, without departing from the spirit or scope of the subject matter presented herein. It will be readily understood that the aspects of the present disclosure, as generally described herein, and illustrated in the Figures, can be arranged, substituted, combined, separated, and designed in a wide variety of different configurations, all of which are explicitly contemplated herein and made part of the disclosure herein.

[0141] All patents, published patent applications, other publications, and sequences from GenBank, and other databases referred to herein are incorporated by reference in their entirety with respect to the related technology.Definitions

[0142] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary7skill in the art to which the present disclosure belongs. See, e.g. Singleton et al., Dictionary7of Microbiology and Molecular Biology72nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al.. Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Press (Cold Spring Harbor, NY 1989). For purposes of the present disclosure, the follow ing terms are defined below.

[0143] As used herein, the term “vector” refers to a polynucleotide construct, typically a plasmid or a virus, used to transmit genetic material to a host cell (e.g., a target cell). Vectors can be, for example, viruses, plasmids, cosmids, or phage. A vector can be a viral vector. A vector as used herein can be composed of either DNA or RNA. In some embodiments, a vector is composed of DNA. An “expression vector” is a vector that is capable of directing the expression of a protein encoded by one or more genes carried by the vector when it is present in the appropriate environment. Vectors are preferably capable of autonomous replication. Typically, an expression vector comprises a transcription promoter, a gene, and a transcription terminator. Gene expression is usually placed under the control of a promoter, and a gene is said to be “operably linked to” the promoter.

[0144] As used herein, the term “operably linked” is used to describe the connection between regulatory elements and a gene or its coding region. Typically, gene expression is placed under the control of one or more regulatory’ elements, for example, without limitation, constitutive or inducible promoters, tissue-specific regulatory elements, and enhancers. A gene or coding region is said to be “operably linked to” or “operatively linked to” or “operably associated with” the regulatory’ elements, meaning that the gene or coding region is controlled or influenced by the regulatory element. For instance, a promoter is operably linked to a coding sequence if the promoter effects transcription or expression of the coding sequence.

[0145] The term “construct,” as used herein, refers to a recombinant nucleic acid that has been generated for the purpose of the expression of a specific nucleotide sequence(s), or that is to be used in the construction of other recombinant nucleotide sequences.

[0146] As used herein, the terms “nucleic acid” and “polynucleotide” are interchangeable and refer to any nucleic acid, whether composed of phosphodiester linkages or modified linkages such as phosphotriester, phosphoramidate, siloxane, carbonate, carboxymethylester, acetamidate, carbamate, thioether, bridged phosphoramidate, bridged methylene phosphonate, bridged phosphoramidate, bridged phosphoramidate, bridged methylene phosphonate. phosphorothioate, methylphosphonate, phosphorodithioate. bridged phosphorothioate or sultone linkages, and combinations of such linkages. The terms “nucleic acid” and “polynucleotide” also specifically include nucleic acids composed of bases other than the five biologically occurring bases (adenine, guanine, thymine, cytosine and uracil).

[0147] The term “regulatory element” and “expression control element” are used interchangeably and refer to nucleic acid molecules that can influence the expression of an operably linked coding sequence in a particular host organism. These terms are used broadly to and cover all elements that promote or regulate transcription, including promoters, core elements required for basic interaction of RNA polymerase and transcription factors, upstream elements,enhancers, and response elements (see, e.g. , Lewin, ’‘Genes V” (Oxford University Press, Oxford) pages 847-873). Exemplary regulatory elements in prokaryotes include promoters, operator sequences and a ribosome binding sites. Regulatory elements that are used in eukary otic cells can include, without limitation, transcriptional and translational control sequences, such as promoters, enhancers, splicing signals, polyadenylation signals, terminators, protein degradation signals, internal ribosome-entry element (IRES), 2A sequences, and the like, that provide for and / or regulate expression of a coding sequence and / or production of an encoded polypeptide in a host cell.

[0148] As used herein, 2A sequences or elements refer to small peptides introduced as a linker between two proteins, allowing autonomous intraribosomal self- processing of polyproteins (See e.g. , de Felipe. Genetic Vaccines and Then 2: 13 (2004); deFelipe et al. Traffic 5:616-626 (2004)). These short peptides allow co-expression of multiple proteins from a single vector. Many 2A elements are know n in the art. Examples of 2A sequences that can be used in the methods and system disclosed herein, without limitation, include 2A sequences from the foot- and-mouth disease virus (F2A), equine rhinitis A virus (E2A), Thosea asigna virus (T2A), and porcine tescho virus-1 (P2A).

[0149] As used herein, the term “promoter” is a nucleotide sequence that permits binding of RNA polymerase and directs the transcription of a gene. Typically, a promoter is located in the 5’ non-coding region of a gene, proximal to the transcriptional start site of the gene. Sequence elements within promoters that function in the initiation of transcription are often characterized by consensus nucleotide sequences. Examples of promoters include, but are not limited to, promoters from bacteria, yeast, plants, viruses, and mammals (including humans). A promoter can be inducible, repressible, and / or constitutive. Inducible promoters initiate increased levels of transcription from DNA under their control in response to some change in culture conditions, such as a change in temperature.

[0150] As used herein, the term “enhancer” refers to a ty pe of regulatory element that can increase the efficiency of transcription, regardless of the distance or orientation of the enhancer relative to the start site of transcription.

[0151] As used herein, the term “variant” refers to a polynucleotide (or polypeptide) having a sequence substantially similar to a reference polynucleotide (or polypeptide). In the case of a polynucleotide, a variant can have deletions, substitutions, additions of one or more nucleotides at the 5' end, 3 ' end, and / or one or more internal sites in comparison to the reference polynucleotide. Similarities and / or differences in sequences between a variant and the reference polynucleotide can be detected using conventional techniques known in the art, for example polymerase chain reaction (PCR) and hybridization techniques. Variant polynucleotides alsoinclude synthetically derived polynucleotides, such as those generated, for example, by using site- directed mutagenesis. Generally, a variant of a polynucleotide, including, but not limited to, a DNA, can have at least about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%. about 92%, about 93%, about 94%. about 95%, about 96%, about 97%, about 98%. about 99% or more sequence identity to the reference polynucleotide as determined by sequence alignment programs known by skilled artisans. In the case of a polypeptide, a variant can have deletions, substitutions, additions of one or more amino acids in comparison to the reference polypeptide. Similarities and / or differences in sequences between a variant and the reference polypeptide can be detected using conventional techniques known in the art, for example Western blot. Generally, a variant of a polypeptide, can have at least about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99% or more sequence identity to the reference polypeptide as determined by sequence alignment programs known by skilled artisans.

[0152] As used herein, the term “effective amount” refers to an amount sufficient to effect beneficial or desirable biological and / or clinical results.

[0153] As used herein, a “subject” refers to an animal that is the object of treatment, observation or experiment. “Animal” includes cold- and warm-blooded vertebrates and invertebrates such as fish, shellfish, reptiles, and in particular, mammals. “Mammal,” as used herein, refers to an individual belonging to the class Mammalia and includes, but not limited to, humans, domestic and farm animals, zoo animals, sports and pet animals. Non-limiting examples of mammals include mice; rats; rabbits; guinea pigs; dogs; cats; sheep; goats; cows; horses; primates, such as monkeys, chimpanzees and apes, and, in particular, humans. In some embodiments, the mammal is a human. However, in some embodiments, the mammal is not a human.

[0154] As used herein, the term “treatment” refers to an intervention made in response to a disease, disorder or physiological condition manifested by a patient. The aim of treatment may include, but is not limited to, one or more of the alleviation or prevention of symptoms, slowing or stopping the progression or w orsening of a disease, disorder, or condition and the remission of the disease, disorder or condition. The term “treat” and “treatment” includes, for example, therapeutic treatments, prophylactic treatments, and applications in which one reduces the risk that a subject will develop a disorder or other risk factor. Treatment does not require the complete curing of a disorder and encompasses embodiments in which one reduces symptoms or underlying risk factors. In some embodiments, “treatment” refers to both therapeutic treatment and prophylactic or preventative measures. Those in need of treatment include those alreadyaffected by a disease or disorder or undesired physiological condition as well as those in which the disease or disorder or undesired physiological condition is to be prevented. For example, in some embodiments treatment may reduce the level of RAS signaling in the subject, thereby to reduce, alleviate, or eradicate the symptom(s) of the disease(s). As used herein, the term "‘prevention” refers to any activity that reduces the burden of the individual later expressing those RAS-related disease symptoms. This can take place at primary, secondary and / or tertiary prevention levels, wherein: a) primary prevention avoids the development of symptoms / disorder / condition; b) secondary prevention activities are aimed at early stages of the condition / disorder / symptom treatment, thereby increasing opportunities for interventions to prevent progression of the condition / disorder / symptom and emergence of symptoms; and c) tertiary prevention reduces the negative impact of an already established condition / disorder / symptom by, for example, restoring function and / or reducing any condition / disorder / symptom or related complications. The term “prevent” does not require the 100% elimination of the possibility of an event. Rather, it denotes that the likelihood of the occurrence of the event has been reduced in the presence of the compound or method.

[0155] “Pharmaceutically acceptable” carriers are ones which are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. “Pharmaceutically acceptable” carriers can be. but not limited to. organic or inorganic, solid or liquid excipients which is suitable for the selected mode of application such as oral application or injection, and administered in the form of a conventional pharmaceutical preparation, such as solid such as tablets, granules, powders, capsules, and liquid such as solution, emulsion, suspension and the like. Often the physiologically acceptable carrier is an aqueous pH buffered solution such as phosphate buffer or citrate buffer. The physiologically acceptable carrier may also comprise one or more of the following: antioxidants including ascorbic acid, low molecular weight (less than about 10 residues) polypeptides, proteins, such as serum albumin, gelatin, immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone, ammo acids, carbohydrates including glucose, mannose, or dextrins, chelating agents such as EDTA, sugar alcohols such as mannitol or sorbitol, salt-forming counterions such as sodium, and nonionic surfactants such as Tween, polyethylene glycol (PEG), and Pluronics. Auxiliary, stabilizer, emulsifier, lubricant, binder, pH adjuster controller, isotonic agent and other conventional additives may also be added to the carriers.

[0156] Described herein include synthetic protein-level cell death circuits, which can be collectively termed “synpoptosis” circuits. To engineer these circuits, inspiration was taken from natural cell death pathways that use regulated proteolysis along with protein-level caging and degradation mechanisms. Synpoptosis circuits and methods provide for rationally designed,programmable control of mammalian cell death. This disclosure presents a set of naturally inspired synthetic protein-level circuits, collectively termed “synpoptosis” circuits, that programmably control user-selectable death programs in target mammalian cells.

[0157] The ability to kill the right cells in the right way can address a variety of therapeutic challenges, such as cancer, fibrosis, senescence, and infection. Synpoptosis circuits provide a foundation for rationally designed, programmable control of mammalian cell death, and should enable new and broad therapeutic str...

Claims

WHAT IS CLAIMED IS:

1. A synthetic protein circuit comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, wherein two first apoptosis polypeptides are capable of associating with each other to constitute a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell, and optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

2. A synthetic protein circuit comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a first partner domain; and a second apoptosis poly peptide comprising a small subunit of an apoptotic effector protein, a second partner domain capable of binding the first partner domain, a first heterologous protease cleavage site, and a first degron, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second apoptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptosis polypeptide from a second apoptosis polypeptide destabilized state to a second apoptosis polypeptide stabilized state, wherein the first apoptosis polypeptide and the second apoptosis polypeptide in the second apoptosis polypeptide stabilized state are capable of associating via binding of the first partner domain and the second partner domain to form a subunit, and wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state,wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell, and optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

3. A synthetic protein circuit comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, a first partner domain, a first degron, and a first heterologous protease cleavage site; and a second apoptosis polypeptide comprising a small subunit of an apoptotic effector protein and a second partner domain capable of binding the first partner domain, wherein the first apoptosis polypeptide and the second apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a subunit, wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state, and optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

4. The synthetic protein circuit of any one of claims 1-3, wherein: the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase- 4. caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, caspase-14 or any variant, portion, or derivative thereof; the first heterologous protease comprises or is derived from one or more of a prokary otic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first heterologous protease is engineered, optionally wherein the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, optionally the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2;the first heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site, further optionally the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7; the first apoptotic protein complex in the first apoptotic protein complex active state is capable of being inhibited by a small molecule inhibitor of apoptosis, optionally, the small molecule inhibitor comprises Quinoline-Val-Asp- Difluorophenoxymethylketone (Q-VD-OPh), carbobenzoxy-valyl-alanyl-aspartyl-[O- methyl]-fluoromethylketone (Z-VAD-FMK), and / or emricasan; and / or the first apoptosis polypeptide and / or the second apoptosis polypeptide comprise one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization.

5. A synthetic protein circuit comprising: a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domain separated by a first heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, and optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

6. The synthetic protein circuit of claim 5, wherein the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, optionally a bulky domain, further optionally the first inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein.

7. A synthetic protein circuit comprising: a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first degron separated by a first heterologous protease cleavage site, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from a first pyroptosis polypeptide destabilized state to a first pyroptosis polypeptide stabilized state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide stabilized state is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

8. A synthetic protein circuit comprising: a first pyroptosis polypeptide comprising a pyroptosis effector domain, a first degron, and a first heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, and wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state, and optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

9. The synthetic protein circuit of any one of claims 5-8, wherein: the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein;the GSDM protein is from the gasdermin (GSDM) family, including GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59; the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first heterologous protease is engineered, optionally wherein the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, optionally the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; the first heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild ty pe first heterologous protease cleavage site, further optionally the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7; the first pyroptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to the first pyroptosis polypeptide active state in the absence of the first heterologous protease in the first heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization; and / or inducing pyroptosis in the cell causes the cell to induce key signatures of pyroptosis: chromatin condensation and DNA fragmentation, pore formation, cell swelling, and osmotic lysis, followed by release of one or more inflammatory cytokines, optionally the one or more inflammatory cytokines comprise IL-18, IL-ip, IL-6, IL-8, interferon gamma (IFN-y), and / or tumor necrosis factor-alpha (TNF-a).

10. A synthetic protein circuit comprising: one or more apoptosis polypeptides; and / or one or more pyroptosis polypeptides; and / or one or more input polypeptides.

11. A synthetic protein circuit comprising: (i) one or more apoptosis polypeptides orone or more pyroptosis polypeptides and (ii) one or more input polypeptides, configured to form one or more logic gates selected from the group comprising an OR logic gate, AND logic gate, NOR logic gate, NAND logic gate, IMPLY logic gate, NIMPLY logic gate, XOR logic gate, and an XNOR logic gate.

12. The synthetic protein circuit of claim 11. comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, a second heterologous protease cleavage site, and a first degron, wherein the first apoptosis polypeptide is capable of being in a first apoptosis polypeptide destabilized state;(ii) a first input polypeptide comprising a first heterologous protease; and / or(iii) a second input polypeptide comprising a second heterologous protease, wherein the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide, wherein the second heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptosis polypeptide from the first apoptosis polypeptide destabilized state to a first apoptosis polypeptide stabilized state; wherein two of the first apoptosis polypeptides in the first apoptosis polypeptide stabilized state are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, and wherein the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, and wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell.

13. The synthetic protein circuit of claim 11, comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site and a second heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state;(ii) a first input polypeptide comprising a first heterologous protease; and / or(iii) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the second heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state; or(III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state, and / or the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the second heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell.

14. The synthetic apoptosis protein circuit of claim 11, comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, a first partner domain, a first degron, and a first heterologous protease cleavage site;(ii) a second apoptosis polypeptide comprising a small subunit of an apoptotic effector protein, a second partner domain capable of binding the first partner domain, a second heterologous protease cleavage site, and a second degron,(iii) a first input polypeptide comprising a first heterologous protease: and / or(iv) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosispolypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide and thereby releasing the second degron, and wherein the second heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptosis polypeptide from a second apoptosis polypeptide destabilized state to a second apoptosis polypeptide stabilized state, wherein the first apoptosis polypeptide and the second apoptosis polypeptide in the second apoptosis polypeptide stabilized state are capable of associating via binding of the first partner domain and the second partner domain to form a subunit, and wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; or(III) wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from the first apoptosis polypeptide stabilized state to the first apoptosis polypeptide destabilized state, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide and thereby releasing the second degron, and wherein the second heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptosis polypeptide from the second apoptosis polypeptide destabilized state to the second apoptosis polypeptide stabilized state.

15. The synthetic protein circuit of claim 11, comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, a second heterologous protease cleavage site, and a first degron;(ii) a second apoptosis polypeptide comprising the large subunit of the apoptotic effector protein and the small subunit of the apoptotic effector protein separated by the second heterologous protease cleavage site, the first heterologous protease cleavage site, and a seconddegron. wherein two of the first apoptosis polypeptides are capable forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, two of the second apoptosis polypeptides are capable forming a second apoptotic protein complex in a second apoptotic protein complex inactive state, and / or one of the first apoptosis polypeptide and one of the second apoptosis polypeptide are capable forming a third apoptotic protein complex in a third apoptotic protein complex inactive state;(iii) a first input polypeptide comprising a first heterologous protease: and / or(iv) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, and the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second apoptosis polypeptide to expose the second degron, and wherein the second degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from a second apoptosis polypeptide stabilized state to a second apoptosis polypeptide destabilized state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell:(II) wherein the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide, and wherein the second heterologous protease cleavage site of the second apoptosis polypeptide being cut changes the second apoptotic protein complex from the second apoptotic protein complex inactive state to a second apoptotic protein complex active state, and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state, wherein the second apoptotic protein complex in the second apoptotic protein complex active state is capable of inducing apoptosis in the cell; or(III) wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second apoptosis polypeptide to expose the first degron, and wherein the first degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from the second apoptosis polypeptide stabilized state to the second apoptosis polypeptide destabilized state; and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from the first apoptosis polypeptide stabilized state to the first apoptosis polypeptide destabilized state.

16. The synthetic protein circuit of claim 11, comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, and a first partner domain and a first degron separated by a first heterologous protease cleavage site, and a second heterologous protease cleavage site;(ii) a second apoptosis polypeptide comprising a small subunit of an apoptotic effector protein and a second partner domain capable of binding the first partner domain, wherein the first apoptosis polypeptide and the second apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a subunit; wherein two subunits are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell;(iii) a first input polypeptide comprising a first heterologous protease; and / or(iv) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, thereby releasing the first degron;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state.

17. The synthetic protein circuit of claim 11. comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein, a first partner domain, a first degron, and a first heterologous protease cleavage site;(ii) a second apoptosis polypeptide comprising the large subunit of the apoptotic effector protein, the first partner domain, a second degron, and a second heterologous protease cleavage site;(iii) a third apoptosis polypeptide comprising a small subunit of an apoptotic effector protein and a second partner domain capable of binding the first partner domain, wherein the first apoptosis polypeptide and the third apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a first subunit, and / or the second apoptosis polypeptide and the third apoptosis polypeptide are capable of associating via binding of the first partner domain and the second partner domain to form a second subunit, wherein two first subunits, two second subunits, and / or one first subunit and one second subunit are capable of associating with each other to constitute a first apoptotic protein complex capable of being in a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell;(iv) a first input polypeptide comprising a first heterologous protease; and / or(v) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from a first apoptosis polypeptide stabilized state to a first apoptosis polypeptide destabilized state;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide to expose the second degron. and wherein the second degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from a second apoptosis polypeptide stabilized state to a second apoptosis polypeptide destabilized state; or(III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide to expose the first degron, and wherein the first degron of the first apoptosis polypeptide being exposed changes the first apoptosis polypeptide from the first apoptosis polypeptide stabilized state to the first apoptosis polypeptide destabilized state, andthe second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second apoptosis polypeptide to expose the second degron, and wherein the second degron of the second apoptosis polypeptide being exposed changes the second apoptosis polypeptide from the second apoptosis polypeptide stabilized state to the second apoptosis polypeptide destabilized state.

18. The synthetic protein circuit of any one of claims 1-17, wherein: the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase- 4. caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, caspase-14 or any variant, portion, or derivative thereof; the first and / or second heterologous protease comprises or is derived from one or more of a prokary otic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first and / or second heterologous protease is engineered, optionally wherein the first and / or second heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, optionally the first and / or second heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; the first and / or second heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the first and / or the second heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first and / or second heterologous protease cleavage site, further optionally the first and / or second heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7; the first and second heterologous protease are different from each other; the first, second, and / or third apoptotic protein complex in the first, second, and / or third apoptotic protein complex active state is capable of being inhibited by a small molecule inhibitor of apoptosis, optionally, the small molecule inhibitor comprises Quinoline-V al-Asp-Difluorophenoxymethylketone (Q-VD-OPh), carbobenzoxy-valyl- alanyl-aspartyl-[O-methyl]-fluoromethylketone (Z-VAD-FMK), and / or emricasan; and / or the first apoptosis polypeptide, the second apoptosis polypeptide and / or the third apoptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the thirdpartner domain and the fourth partner domain is capable of inhibiting change of the first, second, and / or third apoptotic protein complex in the first, second, and / or third apoptotic protein complex inactive state to the first, second, and / or third apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state and / or the second heterologous protease in the second heterologous protease in the second heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization.

19. The synthetic protein circuit of claim 11. comprising:(i) a first pyroptosis polypeptide comprising a pyroptosis effector domain, a first inhibitory domain, and a second inhibitory domain, wherein the pyroptosis effector domain and the first inhibitory domain are separated by a first heterologous protease cleavage site, and the pyroptosis effector domain and the second inhibitory7domain are separated by a second heterologous cleavage site, wherein the first and / or second inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state;(ii) a first input polypeptide comprising a first heterologous protease; and / or(hi) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut releases the first inhibitory domain from the first pyroptosis polypeptide, thereby the second inhibitory domain is capable of inhibiting the activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in the first pyroptosis polypeptide inactive state;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut releases the second inhibitory domain from the first pyroptosis polypeptide, thereby the first inhibitory domain is capable of inhibiting the activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in the first pyroptosis polypeptide inactive state; or(III) wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of thefirst pyroptosis polypeptide being cut releases the first inhibitory domain from the first pyroptosis polypeptide, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut releases the second inhibitory domain from the first pyroptosis polypeptide, wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut and the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, and wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell.

20. The synthetic protein circuit of claim 19, wherein the first and / or second inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, optionally a bulky7domain, further optionally the first and / or second inhibitory7domain comprises an auto-inhibitory7domain of a gasdermin family of pore-forming proteins or maltose-binding protein.

21. The synthetic protein circuit of claim 11 , comprising:(i) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory7domain separated by a first heterologous protease cleavage site and a second heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state;(ii) a first input polypeptide comprising a first heterologous protease; and / or(iii) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosispolypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell; or(III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, and / or the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in the cell.

22. The synthetic protein circuit of claim 21, wherein the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, optionally a bulky domain, further optionally the first inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein.

23. The synthetic protein circuit of claim 1 1, comprising:(i) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first degron separated by a first heterologous protease cleavage site, a second degron, and a second heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide destabilized state;(ii) a first input polypeptide comprising a first heterologous protease; and / or(iii) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide destabilized state to a first pyroptosis polypeptide stabilized state,wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide stabilized state is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell; or(II) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide destabilized state to the first pyroptosis polypeptide stabilized state, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide in the first pyroptosis polypeptide destabilized state to expose the second degron, and wherein the second degron of the first pyroptosis polypeptide in the first pyroptosis polypeptide stabilized state being exposed changes the first pyroptosis polypeptide from the first pyroptosis polypeptide stabilized state to the first pyroptosis polypeptide destabilized state.

24. The synthetic protein circuit of claim 11. comprising:(i) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domain separated by a first heterologous protease cleavage site, a second heterologous cleavage site, and a first degron, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain of the first pyroptosis polypeptide, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state;(ii) a second pyroptosis polypeptide comprising a pyroptosis effector domain and a second inhibitory domain separated by the second heterologous protease cleavage site, the first heterologous cleavage site, and a second degron, wherein the second inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain of the second pyroptosis polypeptide, thereby the second pyroptosis polypeptide is in a second pyroptosis polypeptide inactive state;(iii) a first input polypeptide comprising a first heterologous protease; and / or(iv) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state,wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, and wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron. and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from a second pyroptosis polypeptide stabilized state to a second pyroptosis polypeptide destabilized state;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second pyroptosis polypeptide, and wherein the second heterologous protease cleavage site of the second pyroptosis polypeptide being cut changes the second pyroptosis polypeptide from the second pyroptosis polypeptide inactive state to a second pyroptosis polypeptide active state, wherein the second pyroptosis polypeptide in the second pyroptosis polypeptide active state is capable of inducing pyroptosis in the cell, and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state; or(III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron, and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from the second pyroptosis polypeptide stabilized state to the second pyroptosis polypeptide destabilized state, and wherein the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from the first pyroptosis polypeptide stabilized state to the first pyroptosis polypeptide destabilized state.

25. The synthetic protein circuit of claim 24, wherein the first and / or second inhibitory’domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, optionally a bulky domain, further optionally the first and / or second inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein.

26. The synthetic protein circuit of claim 11. comprising:(i) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first degron separated by a first heterologous protease cleavage site, and a second heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell;(ii) a first input polypeptide comprising a first heterologous protease; and / or(iii) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide and thereby releasing the first degron from the first pyroptosis polypeptide; or(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron. and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state.

27. The synthetic protein circuit of claim 11, comprising:(i) a first pyroptosis polypeptide comprising a pyroptosis effector domain, a first degron, and a first heterologous protease cleavage site, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell;(ii) a second pyroptosis polypeptide comprising a pyroptosis effector domain, a second degron, and a second heterologous protease cleavage site, wherein the second pyroptosis polypeptide is capable of being in a second pyroptosis polypeptide active state, wherein the second pyroptosis polypeptide in the second pyroptosis polypeptide active state is capable of inducing pyroptosis in the cell;(iii) a first input polypeptide comprising a first heterologous protease: and / or(iv) a second input polypeptide comprising a second heterologous protease, wherein:(I) the first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron. and wherein the first degron of the first pyroptosispolypeptide being exposed changes the first pyroptosis polypeptide from a first pyroptosis polypeptide stabilized state to a first pyroptosis polypeptide destabilized state;(II) the second heterologous protease in a second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron. and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from a second pyroptosis polypeptide stabilized state to a second pyroptosis polypeptide destabilized state; or(III) the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide to expose the first degron, and wherein the first degron of the first pyroptosis polypeptide being exposed changes the first pyroptosis polypeptide from the first pyroptosis polypeptide stabilized state to the first pyroptosis polypeptide destabilized state, and the second heterologous protease in the second heterologous protease active state is capable of cutting the second heterologous protease cleavage site of the second pyroptosis polypeptide to expose the second degron, and wherein the second degron of the second pyroptosis polypeptide being exposed changes the second pyroptosis polypeptide from the second pyroptosis polypeptide stabilized state to the second pyroptosis polypeptide destabilized state.

28. The synthetic protein circuit of any one of claims 1-27, wherein: the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein; the GSDM protein is from the gasdermin (GSDM) family, including GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59; the first and / or second heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first and / or second heterologous protease is engineered, optionally wherein the first and / or second heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP). derivatives thereof, or any combination thereof, optionally the first and / or second heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; the first and / or second heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally acaspase cleavage sequence, optionally wherein the first and / or second heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first and / or second heterologous protease cleavage site, further optionally the first and / or second heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7; the first pyroptosis polypeptide and / or the second pyroptosis polypeptide comprise one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of, inhibiting change of the first and / or second pyroptosis polypeptide from the first and / or second pyroptosis polypeptide inactive state to the first and / or second pyroptosis polypeptide active state, in the absence of the first and / or second heterologous protease in the first and / or second heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization; and / or inducing pyroptosis in the cell causes the cell to release one or more inflammatory cytokines, optionally the one or more inflammatory cytokines comprise IL-18, IL-ip, IL- 6. IL-8, interferon gamma (IFN-y), and / or tumor necrosis factor-alpha (TNF-a).

29. The synthetic protein circuit of any one of claims 1-28, wherein:(i) the first, second, and / or third apoptosis polypeptide;(ii) the first and / or second pyroptosis polypeptide; and / or(iii) the first and / or second input polypeptide, are configured to be in a first localized state, optionally the first localized state comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof, optionally the first, second, and / or third apoptosis polypeptide, the first and / or second pyroptosis polypeptide, and / or the first and / or second input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state.

30. The synthetic protein circuit of claim 29. wherein:(i) the first second, and / or third apoptosis polypeptide;(ii) the first and / or second pyroptosis polypeptide; and / or(iii) the first and / or second input polypeptide, are configured to be in second localized state(s), wherein the first localized state and the second localized state(s) are different, optionally the second localized state(s) comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof, optionally the first, second, and / or third apoptosis polypeptide, the first and / or second pyroptosis polypeptide, and / or the first and / or second input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state.

31. The synthetic protein circuit of any one of claims 29-30. wherein:(i) the first, second, and / or third apoptosis polypeptide;(ii) the first and / or second pyroptosis polypeptide; and / or(iii) the first and / or second input polypeptide comprise a first localization signal, optionally the first localization signal is adjacent to a third degron and / or a third heterologous cleavage site.

32. The synthetic protein circuit of any one of claims 29-31. wherein:(i) the first, second, and / or third apoptosis polypeptide;(ii) the first and / or second pyroptosis polypeptide; and / or(iii) the first and / or second input polypeptide comprise second localization signal(s), optionally the second localization signal is adjacent to a third degron and / or a third heterologous protease cleavage site.

33. The synthetic protein circuit of any one of claims 29-32, wherein the presence of the third degron and / or wherein the third heterologous cleavage site being cut changes:(i) the first, second, and / or third apoptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or to a non-localized state;(ii) the first and / or second pyroptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or to a non-localized state; and / or(iii) the first and / or second input polypeptide from a non-localized state or the first localized state to the second localized state(s) or to a non-localized state.

34. The synthetic protein circuit of any one of claims 29-33, wherein: the synthetic protein circuit further comprises a third input polypeptide comprising a third heterologous protease capable of cutting the third heterologous protease cleavage site, optionally the third heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the third heterologous protease is engineered, optionally wherein the third heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, further optionally the third heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; and / or the third heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the third heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type third heterologous protease cleavage site, further optionally the third heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7.

35. The synthetic protein circuit of any one of claims 29-34, wherein:(a) the first, second, and / or third apoptosis polypeptide in the first localized state;(b) the first, second, and / or third apoptosis polypeptide in the second localized state(s);(c) the first and / or second pyroptosis in the first localized state;(d) the first and / or second pyroptosis polypeptide in the second localized state(s);(e) the first and / or second input polypeptide in the first localized state; and / or(f) the first and / or second input polypeptide in the second localized state(s). is capable of modulating an activation threshold and / or sensitivity of the synthetic protein circuit.

36. The synthetic protein circuit of any one of claims 1-35, wherein the synthetic protein circuit is present in a cell, wherein the cell is: a cell of a subject, optionally a subject suffering from a disease or disorder, optionally the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrantangiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof; a cell derived from a donor; and / or an in vivo cell, an ex vivo cell, an in vitro cell or an in situ cell.

37. A synthetic protein circuit comprising: one or more first, second, or third apoptosis polypeptides; one or more first or second pyroptosis polypeptides; one or more input polypeptides; one or more pyroptosis effector proteins; and / or one or more mutant pyroptosis effector proteins, wherein the synthetic protein circuit is capable of inducing in a cell: apoptosis via a first apoptotic protein complex in first apoptotic protein complex active state; and / or pyroptosis via a first and / or second pyroptosis polypeptide in a first and / or second pyroptosis polypeptide active state or a pyroptosis effector protein in a pyroptosis effector protein active state.

38. A synthetic protein circuit comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, and wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; and(ii) a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory’ domain separated by the first heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide inactive state, wherein the first heterologous protease in the first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first of thefirst pyroptosis polypeptide, wherein the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, and wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of:(a) inducing pyroptosis in the cell; and(b) inhibiting the induction of apoptosis in the cell by the first apoptotic protein complex in the first apoptotic protein complex active state, optionally said inhibition is at least 10%, optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

39. The synthetic protein circuit of claim 38, wherein: the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase- 4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, caspase- 14 or any variant, portion, or derivative thereof; the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein; the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD. GSDME, or DFNB59; the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukary otic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first heterologous protease is engineered, optionally wherein the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, optionally the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; the first heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site, further optionally the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7;the first apoptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization; the first pyroptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to the first pyroptosis polypeptide active state in the absence of the first heterologous protease in the first heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization; and / or the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, optionally a bulky' domain, further optionally the first inhibitory domain comprises an auto-inhibitory domain of a gasdermin family of pore-forming proteins or maltose-binding protein.

40. A synthetic protein circuit comprising: a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein, separated by a first heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptosis protease complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of:(a) inducing apoptosis in a cell expressing a pyroptosis effector protein; and / or(b) cutting the pyroptosis effector protein, wherein the pyroptosis effector protein being cut changes the pyroptosis effector protein from a pyroptosiseffector protein inactive state to a pyroptosis effector protein active state, wherein the pyroptosis effector protein in the pyroptosis effector protein active state is capable of:(c) inducing pyroptosis in the cell; and(d) inhibiting the induction of apoptosis in the cell by the first apoptotic protein complex in the first apoptotic protein complex active state, and optionally the synthetic protein circuit further comprises a first input polypeptide comprising the first heterologous protease.

41. A synthetic protein circuit comprising:(i) a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein, separated by a first heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic protein complex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of:(a) inducing apoptosis in a cell expressing a pyroptosis effector protein; and(b) cutting the pyroptosis effector protein, wherein the pyroptosis effector protein being cut changes the pyroptosis effector protein from a pyroptosis effector protein inactive state to a pyroptosis effector protein active state, wherein the pyroptosis effector protein in the pyroptosis effector protein active state is capable of:(c) inducing pyroptosis in the cell; and(d) inhibiting the induction of apoptosis in the cell by the apoptotic protein complex in the first apoptotic protein complex active state; and(ii) a first pyroptosis polypeptide comprising a mutant pyroptosis effector domain comprising a mutation, capable of inhibiting the pyroptosis effector protein in the pyroptosis effector protein active state, thereby changing the pyroptosis effector protein from the pyroptosis effector protein active state to a pyroptosis effector protein inactive state; thereby the first apoptotic protein complex in the first apoptotic protein complexactive state is capable of inducing apoptosis in the cell, optionally the synthetic protein circuit comprises a first input polypeptide comprising the first heterologous protease.

42. The synthetic protein circuit of any one of claims 40-41. wherein: the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase- 4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, caspase-14 or any variant, portion, or derivative thereof; the first apoptosis polypeptide comprises one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state, in the absence of the first heterologous protease in the first heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization; the pyroptosis effector protein comprises a gasdermin (GSDM) protein; the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59; the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first heterologous protease is engineered, optionally wherein the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, optionally the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; the first heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site, further optionally the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7; the mutant pyroptosis effector domain is derived from an N-terminal domain of a gasdermin (GSDM) protein, optionally the GSDM protein is GSDMA, GSDMB,GSDMC, GSDMD, GSDME, or DFNB59; the one or more mutations comprises a V99N, L101N, L103N, V193E, A195E,G199E, and / or I217N mutation in GSDME; and / or the mutation is an I217N mutation in GSDME.

43. The synthetic protein circuit of any one of claims 38-42, wherein inducing pyroptosis in the cell causes the cell to release one or more inflammatory cytokines, optionally the one or more inflammatory cytokines comprise IL- 18, IL-10, IL-6, IL-8, interferon gamma (IFN- y), and / or tumor necrosis factor-alpha (TNF-a).

44. The synthetic protein circuit of any one of claims 38-43, wherein inhibition of the pyroptosis effector protein by the first pyroptosis polypeptide is dose-dependent, thereby the induction of apoptosis in the cell is dose-dependent.

45. The synthetic protein circuit of claim 44, wherein a concentration of the first pyroptosis peptide is at least two-folder higher than a concentration of the pyroptosis effector protein in the cell, thereby the inhibition of the pyroptosis effector protein by the first pyroptosis polypeptide is increased relative to a cell wherein the concentration of the first pyroptosis polypeptide is not at least two-fold higher than the concentration of the pyroptosis effector protein.

46. The synthetic protein circuit of any one of claims 38-45. wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide, are configured to be in a first localized state, optionally the first localized state comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament. intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof, optionally the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is: (i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state.

47. The synthetic protein circuit of claim 46, wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide, are configured to be in second localized state(s), wherein the first localized state and the second localized state(s) are different, optionally the second localized state(s) comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof, optionally the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is:(i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state.

48. The synthetic protein circuit of any one of claims 46-47, wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide comprise a first localization signal, optionally the first localization signal is adjacent to a third degron and / or a third heterologous cleavage site.

49. The synthetic protein circuit of any one of claims 46-48, wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide comprise second localization signal(s), optionally the second localization signal is adjacent to a third degron and / or a third heterologous protease cleavage site.

50. The synthetic protein circuit of any one of claims 46-49, wherein the presence of the third degron and / or wherein the third heterologous cleavage site being cut changes:(i) the first apoptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state;(ii) the first pyroptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state; and / or(iii) the first input polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state.

51. The synthetic protein circuit of any one of claims 46-50, wherein the synthetic protein circuit further comprises a third input polypeptide comprising a third heterologousprotease capable of cutting the third heterologous protease cleavage site, optionally wherein the third heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the third heterologous protease is engineered, optionally wherein the third heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, further optionally the third heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2.

52. The synthetic protein circuit of claim 51, wherein the third heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the third heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type third heterologous protease cleavage site, further optionally the third heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7.

53. The synthetic protein circuit of any one of claims 46-52. wherein:(a) the first apoptosis polypeptide in the first localized state;(b) the first apoptosis polypeptide in the second localized state(s);(c) the first pyroptosis in the first localized state;(d) the first pyroptosis polypeptide in the second localized state(s);(e) the first input polypeptide in the first localized state; and / or(f) the first second input polypeptide in the second localized state(s), is capable of modulating an activation threshold and / or sensitivity of the synthetic protein circuit.

54. The synthetic protein circuit of any one of claims 38-53, wherein the synthetic protein circuit is present in a cell, wherein the cell is: a cell of a subject, optionally a subject suffering from a disease or disorder, optionally the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof; a cell derived from a donor; and / or an in vivo cell, an ex vivo cell, an in vitro cell or an in situ cell.

55. A synthetic protein circuit comprising: a first polypeptide comprising a first signal transducer binding domain and a first part of a first cell death executioner, wherein the first signal transducer binding domain is capable of binding a first signal transducer to form a first signal transducer-bound polypeptide; a second polypeptide comprising a second signal transducer binding domain and a second part of the first cell death executioner, wherein the second signal transducer binding domain is capable of binding a second signal transducer to form a second signal transducer-bound polypeptide, and wherein the first part of the first cell death executioner and the second part of the first cell death executioner are capable of associating with each other to constitute a first cell death executioner capable of being in a first cell death executioner active state when the first signal transducer and the second signal transducer are in close proximity’ at an association location; and wherein the first cell death executioner in the first cell death executioner active state is capable of inducing apoptosis or pyroptosis in a cell.

56. The synthetic protein circuit of claim 55, wherein: the first signal transducer binding domain of the first polypeptide and the second signal transducer binding domain of the second polypeptide are identical; the first transducer and the second transducer are identical and / or are the same protein; the first cell death executioner comprises an apoptosis effector protein or a pyroptosis effector protein; the first part of the first cell death executioner comprises a large subunit of the apoptotic effector protein and / or wherein the second part of second first cell death executioner comprises a small subunit of the apoptotic effector protein; the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase- 4. caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, caspase- 14 or any variant, portion, or derivative thereof; the pyroptosis effector protein comprises a gasdermin (GSDM) protein; and / or the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59.

57. The synthetic protein circuit of any one of claims 55-56, wherein: the first signal transducer, the second signal transducer, or both, are capable of being localized at the association location; the first signal transducer when in a first signal transducer active state, the secondsignal transducer when in a second signal transducer active state, or both, are capable of being localized at the association location; the first signal transducer when in a first inactive state, the second signal transducer when in a second inactive state, or both, are capable of being localized at the association location; the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer, or both; the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first signal transducer active state, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer in a second signal transducer active state, or both; the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first inactive state, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer in a second inactive state, or both; the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, or both; the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at a first cellular location other than the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at a second cellular location other than the association location, or both; the first cellular location, the second cellular location, or both comprise one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory- vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof; and / orthe association location comprises one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof.

58. The synthetic protein circuit of any one of claims 55-57. wherein: a first concentration of the first signal transducer-bound polypeptide is at least twofold higher at the association location as compared to a first cellular location other than the association location when the first signal transducer is in a first signal transducer active state, and / or wherein a second concentration of the second signal transducer-bound polypeptide is at least two-fold higher at the association location as compared to a second cellular location other than the association location when the second signal transducer is in a second signal transducer active state; a first concentration of the first cell death executioner in the first cell death executioner active state is at least two-fold higher at the association location as compared to a cellular location other than the association location when the first signal transducer is in a first signal transducer active state and / or when the second signal transducer is in a second signal transducer active state; the first part of the first cell death executioner and the second part of the first cell death executioner have the weak association affinity when the first signal transducer is in a first signal transducer inactive state and / or the second signal transducer is in a second signal transducer inactive state; the first part of the first cell death executioner and the second part of the first cell death executioner are incapable of associating to form the first cell death executioner in the first cell death executioner active state when the first signal transducer is in a first signal transducer inactive state and / or the second signal transducer is in a second signal transducer inactive state; a first concentration of the first signal transducer-bound polypeptide and a second concentration of the second signal transducer-bound polypeptide at the association location are insufficient for the first part of the first cell death executioner and the second part of the first cell death executioner to form an active first cell death executioner when the first signal transducer is in a first signal transducer inactive state and / or the secondsignal transducer is in a second signal transducer inactive state; a first concentration of the first signal transducer-bound polypeptide at the association location is comparable to a first cellular location other than the association location when the first signal transducer is in a first signal transducer inactive state, and / or wherein a second concentration of the second signal transducer-bound polypeptide at the association location is comparable to a second cellular location other than the association location when the second signal transducer is in a second signal transducer inactive state; the first part of the first cell death executioner and the second part of the first cell death executioner are capable of associating with each other to form the first cell death executioner in the first cell death executioner active state at a threshold first polypeptide concentration and a threshold second polypeptide concentration at the association location; and / or the threshold first polypeptide concentration and the threshold second polypeptide concentration at the association location are reached at a threshold signal transducer activation level of the signal transducer.

59. The synthetic protein circuit of any one of claims 55-58, wherein: the first signal transducer binding domain and / or the second signal transducer binding domain are identical; the first signal transducer binding domain and / or the second signal transducer binding domain are different; the first signal transducer binding domain and / or the second signal transducer binding domain each is capable of binding molecules of the first signal transducer and / or the second signal transducer; the first signal transducer and / or the second signal transducer belong to a signal transduction pathway; the first signal transducer binding domain and / or the second signal transducer binding domain comprise a RAS binding domain (RBD) and / or RAS association domain (RAD); optionally the RAS binding domain comprises or is derived from a RAS interacting protein, optionally selected from the group comprising AG02, APBB1IP, APPL1, ARAF, ARL1, ARL2, ARRB1, ARRB2, BAIAP2, BCL2, BCL2L1, BRAF, BRAP, BSG. CALM1. CALM3, CALML3. CALML4. CALML5, CALML6, CNKSR1, CNKSR2, CSK, DAB2IP, EGFR, ERBIN, FGA, FGB, FGG, FN1, GRB2, HK1, IFNGR1, IL6, IQGAP1, ITGA2B, ITGB3, KSR1, KSR2, LGALS3, LYN, LZTR1, MAP2K1, MAP2K2, MAPK1, MAPK14, MAPK3, MAPKAP1, MARK2, MARK3, MBP, MSI2, MTOR, NCBP2AS2, NF1, NIBAN2, PDE4DIP, PDE6D, PDPK1, PEBP1, PIK3CA,-21 flPIK3CB, PIK3CD, PIK3RL PIK3R2, PIP5K1A, PLCE1, PPIA, PRKCZ, PTGS2, RAFI, RALB, RALGDS, RAP1A, RAP1B, RAP1GDS1, RASA1, RASA2, RASA3, RASA4, RASAL1, RASAL2, RASAL3, RASSF1, RASSF2, RASSF5, RGL1, RGL3, RIN1, SH0C2, S0S1, SOS2, SPRED1, SPRED2, SPRED3. SRC, SYNGAP1, TIAM1, TLN1, VCL. VWF, YWHAB. or any combination thereof; the first signal transducer binding domain and / or the second signal transducer binding domain comprises a lipid binding domain; the lipid binding domain comprises a Pleckstrin homology (PH) domain; and / or the first signal transducer binding domain and / or the second signal transducer binding domain comprises an antibody, an antibody fragment, a binding domain derived from a natural protein, an scFv, a Fv, a Fab, a (Fab')2, a single domain antibody (SDAB), a VH or VL domain, a camelid VHH domain, a Fab, a Fab1, a F(ab')2, a Fv, a scFv, a dsFv, a diabody, a triabody, a tetrabody, a multispecific antibody formed from antibody fragments, a single-domain antibody (sdAb), a single chain comprising cantiomplementary scFvs (tandem scFvs) or bispecific tandem scFvs, an Fv construct, a disulfide-linked Fv, a dual variable domain immunoglobulin (DVD-Ig) binding protein or a nanobody, an aptamer, an affibody, an affilin, an affitin, an affimer, an alphabody, an anticalin. an avimer, a DARPin, a Fynomer, a Kunitz domain peptide, a monobody, or any combination thereof.

60. The synthetic protein circuit of any one of claims 55-59, wherein: the first signal transducer is capable of binding the first signal transducer binding domain and / or the second signal transducer is capable of binding the second signal transducer binding domain following a modification selected from the group comprising phosphorylation, dephosphorylation, acetylation, methylation, acylation, glycosylation, glycosylphosphatidylinositol (GPI) anchoring, sulfation, disulfide bond formation, deamidation, ubiquitination, sumoylation, nitration of tyrosine, hydrolysis of ATP or GTP, binding of ATP or GTP, cleavage, or any combination thereof; the first signal transducer, the second signal transducer, or both are endogenous proteins; the first signal transducer, the second signal transducer, or both comprise AKT, PI3K, MAPK, p44 / 42 MAP kinase, TYK2. p38 MAP kinase, PKC, PKA. SAPK. ELK, JNK, eJun, RAS, Raf, MEK 1 / 2, MEK 3 / 6, MEK 4 / 7, ZAP-70, LAT, SRC, LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX, PLCy, PLCy, NF-kB, FAK, CREB, aIHP3, FcsRI, BAD, p70S6K, STAT1, STAT2, STAT3, STAT5, STAT6, or any combination thereof;the first signal transducer and / or the second signal transducer are capable of regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology', cell differentiation, apoptosis, or any combination thereof; the first signal transducer, the second signal transducer, or both comprise a RAS protein: the RAS protein is KRAS, NRAS, HRAS, or any combination thereof; the first signal transducer, the second signal transducer, or both are exogenous proteins; the synthetic protein circuit comprises the first signal transducer, the second signal transducer, or both; the first signal transducer, the second signal transducer, or both comprise a lipid; the lipid comprises a phospholipid; and / or the phospholipid is phosphatidylinositol 3 -phosphate.

61. The synthetic protein circuit of any one of claims 55-60, wherein: the synthetic protein circuit is capable of detecting an activity of the first signal transducer and an activity' of the second signal transducer; an activity' of the first cell death executioner correlates with an activity of the first signal transducer and / or an activity’ of the second signal transducer; the synthetic protein circuit is capable of detecting activities of the first signal transducer and activities of the second signal transducer over a period of time; activities of the first cell death executioner correlate with activities of the first signal transducer and activities of the second signal transducer over a period of time; the synthetic protein circuit is capable of detecting an aberrant signaling; aberrant signaling involves an active signal transducer; the aberrant signaling involves an overactive signal transducer; the aberrant signaling involves a constitutively active signal transducer over a period of time; the synthetic protein circuit is capable of detecting an activity' of a signal transducer activator and / or an activity' of a signal transducer repressor; the aberrant signaling involves an active signal transducer repressor and an active signal transducer; the aberrant signaling involves an inactive signal transducer activator and an active signal transducer; the aberrant signaling involves an inactive signal transducer; the aberrant signaling involves an underactive signal transducer;the aberrant signaling involves a constitutively inactive signal transducer over a period of time; the aberrant signaling involves an inactive signal transducer repressor and an inactive signal transducer; the aberrant signaling involves an active signal transducer activator and an inactive signal transducer; the aberrant signaling involves an active signal transducer, and wherein the aberrant signaling comprises an aberrant signal of at least one signal transduction pathway regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof; the synthetic protein circuit is capable of directly or indirectly inducing cell death in the presence of the aberrant signaling; the first cell death executioner is capable of directly or indirectly inducing cell death in the presence of aberrant signaling; the synthetic protein circuit is capable of directly or indirectly inducing cell death when a first level of activation of the first signal transducer is above a first signal transducer activation threshold and / or a second level of activation of the second signal transducer is below a second signal transducer activation threshold; the effector protein is capable of directly or indirectly inducing cell death when a first level of activation of the first signal transducer is above a first signal transducer activation threshold and / or a second level of activation of the second signal transducer is below a second signal transducer activation threshold; the synthetic protein circuit is present in a cell; and / or the cell is: a cell of a subject, optionally a subject suffering from a disease or disorder, optionally the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof; a cell derived from a donor; and / or an in vivo cell, an ex vivo cell, an in vitro cell or an in situ cell.

62. A nucleic acid composition, comprising: one or more polynucleotides encoding the synthetic protein circuit of any one of claims 1-61, optionally the one or more polynucleotides comprise:one or more first polynucleotides encoding a first apoptosis polypeptide, a first pyroptosis polypeptide, or a first polypeptide; one or more second polynucleotides encoding a second apoptosis polypeptide, a second pyroptosis polypeptide, or a second polypeptide; and / or one or more third polynucleotides encoding a third apoptosis polypeptide. optionally, the nucleic acid composition comprises one or more polynucleotides encoding a first and / or second input polypeptide.

63. The nucleic acid composition of claim 62, wherein: at least two of the one or more polynucleotides are operably linked to a tandem gene expression element; the one or more polynucleotides comprise: a 5’UTR and / or a 3’UTR; a tandem gene expression element selected from the group an internal nbosomal entry site (IRES), foot-and-mouth disease virus 2A peptide (F2A), equine rhinitis A virus 2A peptide (E2A), porcine teschovirus 2A peptide (P2A) or Thosea asigna virus 2A peptide (T2A), or any combination thereof; and / or a transcript stabilization element, optionally the transcript stabilization element comprises woodchuck hepatitis post-translational regulatory element (WPRE), bovine growth hormone polyadenylation (bGH-polyA) signal sequence, human growth hormone polyadenylation (hGH-polyA) signal sequence, or any combination thereof; the one or more polynucleotides are operably connected to a promoter selected from the group comprising: a minimal promoter, optionally TATA, miniCMV, and / or miniPromo; a tissue-specific promoter and / or a lineage-specific promoter; and / or a ubiquitous promoter, optionally a cytomegalovirus (CMV) immediate early promoter, a CMV promoter, a viral simian virus 40 (SV40) (e.g., early or late), a Moloney murine leukemia virus (MoMLV) LTR promoter, a Rous sarcoma virus (RSV) LTR, an RSV promoter, a herpes simplex virus (HSV) (thymidine kinase) promoter. H5, P7.5, and Pl 1 promoters from vaccinia virus, an elongation factor 1 -alpha (EF 1 a) promoter, early growth response 1 (EGR1 ), ferritin H (F erH), ferritin L (FerL), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), eukaryotic translation initiation factor 4A1 (EIF4A1), heat shock 70 kDa protein 5 (HSPA5), heat shock protein 90 kDa beta, member 1 (HSP90B1), heat shock protein 70 kDa (HSP70). p-kinesin ( -KIN), the human ROSA 26 locus, aUbi quitin C promoter (UBC), a phosphoglycerate kinase- 1 (PGK) promoter, 3- phosphoglycerate kinase promoter, a cytomegalovirus enhancer, human f>-actin (HBA) promoter, chicken P-actin (CBA) promoter, a CAG promoter, a CASI promoter, a CBH promoter, or any combination thereof; the nucleic acid composition is configured to enhance stability, durability, and / or expression level, optionally a 5' untranslated region (UTR), a 3' UTR, and / or a 5’ cap; optionally one or more modified nucleotides, further optionally selected from the group comprising pseudouridine. N-l-methyl-pseudouridine, 2- aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3 -methyl adenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2- aminoadenosine, C5 -bromouridine, C5-fluorouridine, C5-iodouridine, C5- propynyl-uridine. C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)- methylguanine, and 2-thiocytidine; and / or optionally a modified nucleotide in place of one or more uridines, optionally the modified nucleoside is selected from pseudouridine (\| / ), N 1-methyl- pseudouridine (m IT), and 5-methyl-uridine (m5U); the nucleic acid composition is complexed or associated with one or more lipids or lipid-based carriers, thereby forming liposomes, lipid nanoparticles (LNPs), lipoplexes, and / or nanoliposomes, optionally encapsulating the nucleic acid composition; the nucleic acid composition is, comprises, or further comprises, one or more vectors, optionally at least one of the one or more vectors is a viral vector, a plasmid, a transposable element, a naked DNA vector, a lipid nanoparticle (LNP), a bacterial cell, a bacteriophage, or any combination thereof, optionally the viral vector is an AAV vector, a lentivirus vector, a retrovirus vector, an adenovirus vector, a herpesvirus vector, a herpes simplex virus vector, a cytomegalovirus vector, a vaccinia virus vector, a MV A vector, a baculovirus vector, a vesicular stomatitis virus vector, a human papillomavirus vector, an avipox virus vector, a Sindbis virus vector, a VEE vector, a Measles virus vector, an influenza virus vector, a hepatitis B virus vector, an integration-deficient lentivirus (IDLV) vector, or any combination thereof, and optionally the transposable element is piggybac transposon or sleeping beauty transposon;the one or more polynucleotides are comprised in the one or more vectors, optionally the one or more polynucleotides are comprised in the same vector and / or different vectors, optionally the one or more polynucleotides are situated on the same nucleic acid and / or different nucleic acids; the nucleic acid composition comprises circular mRNA, circular DNA, selfamplifying RNA, self-amplifying RNA, and / or mRNA; the nucleic acid composition is configured to achieve relative levels of the first apoptotic polypeptide, the second apoptotic polypeptide, the third apoptotic polypeptide, the first input polypeptide, the second input polypeptide, the first polypeptide and / or the second polypeptide desired by a user; the nucleic acid composition is configured to achieve relative levels of the first pyroptotic polypeptide, the second pyroptotic polypeptide, the first input polypeptide, the second input polypeptide, the first polypeptide and / or the second polypeptide desired by a user; the expression of one or more of the first apoptotic polypeptide, the second apoptotic polypeptide, the third apoptotic polypeptide, first pyroptotic polypeptide, the second pyroptotic polypeptide, the first input polypeptide, and / or the second polypeptide is configured to be dosage invariant and / or robust to tissue tropism and stochastic expression; and / or the induction of apoptosis or the induction of pyroptosis can be tuned by adjusting the relative levels of the first apoptotic polypeptide, the second apoptotic polypeptide, the third apoptotic polypeptide, first pyroptotic polypeptide, the second pyroptotic polypeptide, the first input polypeptide, and / or the second polypeptide.

64. A composition, comprising: a first population of sender cells comprising:(i) one or more first polynucleotide(s) encoding a first apoptosis polypeptide comprising a large subunit of an apoptotic effector protein and a small subunit of an apoptotic effector protein separated by a first heterologous protease cleavage site, wherein two of the first apoptosis polypeptides are capable of forming a first apoptotic protein complex in a first apoptotic protein complex inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first apoptosis polypeptide, and wherein the first heterologous protease cleavage site of the first apoptosis polypeptide being cut changes the first apoptotic proteincomplex from the first apoptotic protein complex inactive state to a first apoptotic protein complex active state, and wherein the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in a cell; and(ii) a first vector; and a second population of sender cells comprising:(i) one or more second polynucleotide(s) encoding a first input polypeptide; and(ii) a second vector.

65. The composition of claim 64, wherein the first vector of the first population of sender cells and the second vector of the second population of sender cells are capable of delivering the one or more first polynucleotide(s) and the one or more second polynucleotide(s) to receiver cells.

66. The composition of claim 65, wherein the first apoptosis polypeptide and the first input polypeptide are expressed in the receiver cells, thereby the first apoptotic protein complex in the first apoptotic protein complex active state is capable of inducing apoptosis in the receiver cells.

67. The composition of any one of claims 64-66, wherein: the apoptotic effector protein comprises caspase-1, caspase-2, caspase-3, caspase- 4, caspase-5, caspase-6, caspase-7, caspase-8, caspase-9, caspase-10, caspase-14 or any variant, portion, or derivative thereof; the first heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first heterologous protease is engineered, optionally wherein the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, optionally the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; the first heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site, further optionally the first heterologous protease cleavage site comprises the sequence of any one of SEQ IDNOs: 3-7; and / or the first apoptosis polypeptide, the first input polypeptide, or both comprise a membrane-localization domain, optionally the membrane-localization domain comprises a CAAX domain.

68. A composition, comprising: a first population of sender cells comprising:(i) one or more first polynucleotide(s) encoding a first pyroptosis polypeptide comprising a pyroptosis effector domain and a first inhibitory domain separated by a first heterologous protease cleavage site, wherein the first inhibitory domain is capable of inhibiting activity of the pyroptosis effector domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state, wherein a first heterologous protease in a first heterologous protease active state is capable of cutting the first heterologous protease cleavage site of the first pyroptosis polypeptide, and wherein the first heterologous protease cleavage site of the first pyroptosis polypeptide being cut changes the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in a cell, and(ii) a first vector; and a second population of sender cells comprising:(i) one or more second polynucleotide(s) encoding a first input polypeptide comprising the first heterologous protease; and(ii) a second vector.

69. The composition of claim 68, wherein: the first vector of the first population of sender cells and the second vector of the second population of sender cells are capable of delivering the one or more first polynucleotide(s) and the one or more second polynucleotide(s) to receiver cells; the first pyroptosis polypeptide and the first input polypeptide are expressed in the receiver cells, thereby the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in the receiver cells; the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein; the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59; the first heterologous protease comprises or is derived from one or more of aprokaryotic protease, bacterial protease, archaeal protease, eukaryotic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the first heterologous protease is engineered, optionally wherein the first heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, optionally the first heterologous protease cleavage site comprises the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; the first heterologous protease cleavage site is natural or engineered, wherein the first heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wild type first heterologous protease cleavage site, further optionally the first heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7; and / or the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, optionally a bulky domain, further optionally the first inhibitory' domain comprises an auto-inhibitory' domain of a gasdermin family of pore-forming proteins or maltose-binding protein.

70. The composition of any one of claims 68-69, wherein the first apoptosis polypeptide or the first pyroptosis polypeptide comprise one or more third partner domains and one or more fourth partner domains capable of binding the third partner domain, wherein binding of the third partner domain and the fourth partner domain of the first apoptosis polypeptide is capable of inhibiting change of the first apoptotic protein complex from the first apoptotic protein complex inactive state to the first apoptotic protein complex active state in the absence of the first heterologous protease in the first heterologous protease active state, or wherein binding of the third partner domain and the fourth partner domain of the first pyroptosis polypeptide is capable of inhibiting change of the first pyroptosis polypeptide from the first pyroptosis polypeptide inactive state to the first pyroptosis polypeptide active state in the absence of the first heterologous protease in the first heterologous protease active state, optionally wherein said third partner domains and fourth partner domains are capable of multimerization.

71. A composition, comprising: a first population of sender cells comprising:(i) one or more first polynucleotide(s) encoding a first pyroptosispolypeptide comprising a pyroptosis effector domain and a first partner domain; and(ii) a first vector, wherein the first population of sender cells express a silencer polypeptide comprising a first inhibitory domain and a second partner domain capable of binding the first partner domain, and wherein the inhibitor domain of the silencer polypeptide is capable of inhibiting the first pyroptosis polypeptide when the first pyroptosis polypeptide associates with the silencer polypeptide via binding of the first partner domain and the second partner domain, thereby the first pyroptosis polypeptide is in a first pyroptosis polypeptide inactive state.

72. The composition of claim 71, wherein: the first vector is capable of delivering the one or more first polynucleotide(s) to receiver cells; the first pyroptosis polypeptide is expressed in the receiver cells, wherein the first pyroptosis polypeptide is capable of being in a first pyroptosis polypeptide active state, wherein the first pyroptosis polypeptide in the first pyroptosis polypeptide active state is capable of inducing pyroptosis in the receiver cells; the receiver cells do not express the silencer polypeptide; the first inhibitory domain comprises a protein domain configured to reduce or abrogate the activity of the pyroptosis effector domain, optionally a bulky7domain, further optionally the first inhibitory domain comprises an auto-inhibitory7domain of a gasdermin family of pore-forming proteins or maltose-binding protein; the pyroptosis effector domain comprises an N-terminal domain of a gasdermin (GSDM) protein; and / or the GSDM protein is GSDMA, GSDMB, GSDMC, GSDMD, GSDME, or DFNB59.

73. The composition of any one of claims 68-72, wherein inducing pyroptosis in a receiver cell causes the receiver cell to release one or more inflammatory cytokines, optionally the one or more inflammatory7cytokines comprise IL-18, IL-ip, IL-6, IL-8, interferon gamma (IFN-y), and / or tumor necrosis factor-alpha (TNF-a).

74. The composition of any one of claims 68-73, wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide, are configured to be in a first localized state, optionallythe first localized state comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof, optionally the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is: (i) tethered to an intracellular organelle and / or membrane: or (ii) fused to a polypeptide that recruits to said localized state.

75. The composition of any one of claims 64-74, wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide, are configured to be in second localized state(s), wherein the first localized state and the second localized state(s) are different, optionally: the second localized state(s) comprises a state created by phase separation, a state defined by the proximity to a given protein, and / or localization to one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, Golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof, optionally the first apoptosis polypeptide, the first pyroptosis polypeptide, and / or the first input polypeptide is:(i) tethered to an intracellular organelle and / or membrane; or (ii) fused to a polypeptide that recruits to said localized state.

76. The synthetic protein circuit of any one of claims 64-75, wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide comprise a first localization signal, optionally the first localization signal is adjacent to a third degron and / or a third heterologous cleavage site.

77. The composition of any one of claims 64-76, wherein:(i) the first apoptosis polypeptide;(ii) the first pyroptosis polypeptide, and / or(iii) the first input polypeptide comprise second localization signal(s), optionally the second localization signal is adjacent to a third degron and / or a third heterologous protease cleavage site.

78. The composition of any one of claims 64-77, wherein the presence of the third degron and / or wherein the third heterologous cleavage site being cut changes:(i) the first apoptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state;(ii) the first pyroptosis polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state; and / or(iii) the first input polypeptide from a non-localized state or the first localized state to the second localized state(s) or a non-localized state.

79. The composition of any one of claims 64-78, wherein: the synthetic protein circuit further comprises a third input polypeptide comprising a third heterologous protease capable of cutting the third heterologous protease cleavage site, optionally wherein the third heterologous protease comprises or is derived from one or more of a prokaryotic protease, bacterial protease, archaeal protease, eukary otic protease, fungal protease, plant protease, mammalian protease, and human protease, optionally the third heterologous protease is engineered, optionally wherein the third heterologous protease comprises tobacco etch virus (TEV) protease, tobacco vein mottling virus (TVMV) protease, hepatitis C virus protease (HCVP), derivatives thereof, or any combination thereof, further optionally the third heterologous protease cleavage site compnses the sequence of SEQ ID NO: 1 or SEQ ID NO: 2; and / or wherein the third heterologous protease cleavage site is natural or engineered, optionally the cleavage sequence of a human apoptotic effector protease, optionally a caspase cleavage sequence, optionally wherein the third heterologous protease cleavage site is configured to modulate an activation threshold of the synthetic protein circuit, optionally wherein said configuring comprises introducing one or more amino acid insertions, substitutions, or deletions relative to a wi Id type third heterologous protease cleavage site, further optionally the third heterologous protease cleavage site comprises the sequence of any one of SEQ ID NOs: 3-7.

80. The composition of any one of claims 64-79, wherein:(a) the first apoptosis polypeptide in the first localized state;(b) the first apoptosis polypeptide in the second localized state(s);(c) the first pyroptosis in the first localized state;(d) the first pyroptosis polypeptide in the second localized state(s);(e) the first input polypeptide in the first localized state; and / or(f) the first second input polypeptide in the second localized state(s), is capable of modulating an activation threshold and / or sensitivity of the synthetic protein circuit.

81. The composition of any one of claims 64-80, wherein the first and / or second vector is a viral vector, a plasmid, a naked DNA vector, a naked RNA vector, a lipid nanoparticle, or any combination thereof, optionally the viral vector is an AAV vector, a lentivirus vector, a retrovirus vector, an integration-deficient lentivirus (IDLV) vector, further optionally the AAV vector comprises single-stranded AAV (ssAAV) vector or a self-complementary AAV (scAAV) vector.

82. The composition of any one of claims 64-81 , wherein the receiver cell or the sender cell is: a cell of a subject, optionally a subject suffering from a disease or disorder, optionally the disease or disorder is a blood disease, an immune disease, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof: a cell derived from a donor; and / or an in vivo cell, an ex vivo cell, or an in situ cell.

83. The composition of any one of claims 64-82, wherein the first and / or the second vector is capable of delivering the one or more first polynucleotides and / or the one or more second polynucleotides to one or more tissues of a subject, optionally: the receiver cells are situated within one or more tissues of a subject; and / or the one or more tissues comprise adrenal gland tissue, appendix tissue, bladder tissue, bone, bowel tissue, brain tissue, breast tissue, bronchi, coronal tissue, ear tissue, esophagus tissue, eye tissue, gall bladder tissue, genital tissue, heart tissue, hypothalamus tissue, kidney tissue, large intestine tissue, intestinal tissue, larynx tissue, liver tissue, lung tissue, lymph nodes, mouth tissue, nose tissue, pancreatic tissue, parathyroid gland tissue, pituitary' gland tissue, prostate tissue, rectal tissue, salivary gland tissue, skeletal muscle tissue, skin tissue, small intestine tissue, spinal cord, spleen tissue, stomach tissue, thymus gland tissue, trachea tissue, thyroid tissue, ureter tissue, urethra tissue, soft and connective tissue, peritoneal tissue, blood vessel tissue, fat tissue, or any combination thereof.

84. The composition of any one of claims 64-83, wherein the receiver cell comprises a unique cell type and / or a unique cell state.

85. The composition of claim 84, wherein the unique cell type and / or the unique cell state comprises a unique gene expression pattern, optionally the unique cell ty pe and / or unique cell state comprises (i) one or more mutations of a protein, (ii) structural variants and / or copy -number alternations of one or more protein-coding genes, (iii) epigenetic signature(s), and / or (iv) a unique anatomic location, further optionally the unique cell type and / or the unique cell state comprises anatomically locally unique gene expression; wherein the unique cell type and / or the unique cell state is caused by hereditable, environmental, and / or idiopathic factors; wherein the unique cell type and / or the cell in the unique cell state (i) causes and / or aggravates a disease or disorder and / or (ii) is associated with the pathology of a disease or disorder; and / or wherein the unique cell state comprises a senescent cell state induced by a tumor microenvironment, optionally the senescent cell state induced by a tumor microenvironment comprises expression of CD57, K.R.LG1, TIGIT, p21, p53, phospho- p53, DECI , PPP1 A, yII2AX, 53BPI, Rad 17, ATR, ATM, MDC1, TIF, IL-6, IL-8, CXCR2, IGF2, IGFBP3, IGFBP5, IGFBP7, STC 1 , GDF15, SERPIN, ICAM-1, DEP1, B2MG, N0TCH3, DcR2, or any combination thereof.

86. The composition of any one of claims 84-85, wherein the unique cell state and / or unique cell type is characterized by one or more of: aberrant signaling of one or more signal transducer(s); cell proliferation, stress pathways, oxidative stress, stress kinase activation, DNA damage, lipid metabolism, carbohydrate regulation, metabolic activation including Phase I and Phase II reactions, Cytochrome P-450 induction or inhibition, ammonia detoxification, mitochondrial function, peroxisome proliferation, organelle function, cell cycle state, morphology, apoptosis, DNA damage, metabolism, signal transduction, cell differentiation, cell-cell interaction and cell to non-cellular compartment; acute phase stress, cell adhesion, AH-response, anti-apoptosis and apoptosis, antimetabolism, anti- proliferation, arachidonic acid release, ATP depletion, cell cycle disruption, cell matrix disruption, cell migration, cell proliferation, cell regeneration, cellcell communication, cholestasis, differentiation, DNA damage, DNA replication, early response genes, endoplasmic reticulum stress, estogenicity, fatty liver, fibrosis, general cell stress, glucose deprivation, growth arrest, heat shock, hepatotoxicity, hypercholesterolemia, hypoxia, immunotox, inflammation, invasion, ion transport, liver regeneration, cell migration, mitochondrial function, mitogenesis, multidrug resistance,nephrotoxicity, oxidative stress, peroxisome damage, recombination, ribotoxic stress, sclerosis, steatosis, teratogenesis, transformation, disrupted translation, transport, and tumor suppression; and nutrient deprivation, hypoxia, oxidative stress, hyperproliferative signals, oncogenic stress. DNA damage, ribonucleotide depletion, replicative stress, and telomere attrition, promotion of cell cycle arrest, promotion of DNA-repair, promotion of apoptosis, promotion of genomic stability , promotion of senescence, and promotion of autophagy7, regulation of cell metabolic reprogramming, regulation of tumor microenvironment signaling, inhibition of cell sternness, survival, and invasion.

87. The composition of any one of claims 84-86, wherein the unique cell state comprises: a physiological state, optionally a cell cycle state, a differentiation state, a development state a metabolic state, or a combination thereof; and / or a pathological state, optionally a disease state, a human disease state, a diabetic state, an immune disorder state, a neurodegenerative disorder state, an oncogenic state, or a combination thereof.

88. The composition of any one of claims 64-87. wherein the receiver cell is characterized by aberrant signaling of one or more signal transducers, and wherein the aberrant signaling involves: an overactive signal transducer; a constitutively active signal transducer over a period of time; an active signal transducer repressor and an active signal transducer; an inactive signal transducer activator and an active signal transducer; an inactive signal transducer; an underactive signal transducer; a constitutively inactive signal transducer over a period of time; an inactive signal transducer repressor and an inactive signal transducer; and / or an active signal transducer activator and an inactive signal transducer.

89. The composition of claim 88, wherein the aberrant signaling comprises an aberrant signal of at least one signal transduction pathway regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof, optionally the signal transduscer(s) is AKT, PI3K, MAPK, p44 / 42 MAP kinase, TYK2, p38 MAP kinase, PKC, PKA, SAPK, ELK, JNK, eJun, RAS, Raf, MEK 1 / 2, MEK 3 / 6, MEK 4 / 7, ZAP-70, LAT, SRC, LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX. PLCy, PLCy, NF-kB, FAK, CREB, allip3, FcsRI, BAD, p70S6K, STAT1,STAT2, STAT3, STAT5, STAT6, or any combination thereof.

90. A method of selectively killing a target cell comprising: expressing the synthetic protein circuit of any one of claims 1-61 or the nucleic acid composition of any one of claims 62-63 in the target cell, wherein the synthetic protein circuit is configured to be responsive to a unique cell type and / or unique cell state of the target cell, optionally wherein the first and / or second heterologous protease is configured to be in the first and / or second heterologous protease active state in response to the unique cell ty pe and / or unique cell state of the target cell.

91. The method of claim 90, wherein: the unique cell type and / or the unique cell state comprises a unique gene expression pattern, optionally the unique cell type and / or unique cell state comprises: (i) one or more mutations of a protein, (ii) structural variants and / or copy -number alternations of one or more protein-coding genes, (iii) epigenetic signature(s), and / or (iv) a unique anatomic location, further optionally the unique cell ty pe and / or the unique cell state comprises anatomically locally unique gene expression; the unique cell type and / or the unique cell state is caused by hereditable, environmental, and / or idiopathic factors; the unique cell type and / or the cell in the unique cell state (i) causes and / or aggravates a disease or disorder and / or (ii) is associated with the pathology of a disease or disorder; the unique cell state comprises a senescent cell state induced by a tumor microenvironment, optionally the senescent cell state induced by a tumor microenvironment comprises expression of CD57, KR.LG1, TIGIT, p21, p53. phospho- p53, DECI, PPP1 A. yH2AX, 53BPI, Radi 7, ATR, ATM, MDC1, TIF, IL-6, IL-8, CXCR2, IGF2, IGFBP3, IGFBP5, IGFBP7, STC1, GDF15, SERPIN. ICAM-1, DEP1, B2MG, NOTCH3, DcR2, or any combination thereof: the unique cell state and / or unique cell type is characterized by aberrant signaling of one or more signal transducer(s); the unique cell state comprises: a physiological state, optionally a cell cycle state, a differentiation state, a development state a metabolic state, or a combination thereof; and / or a pathological state, optionally a disease state, a human disease state, a diabetic state, an immune disorder state, a neurodegenerative disorder state, an oncogenic state, or a combination thereof; the unique cell state and / or unique cell type is characterized by one or more of cell proliferation, stress pathways, oxidative stress, stress kinase activation, DNA damage,lipid metabolism, carbohydrate regulation, metabolic activation including Phase I and Phase II reactions, Cytochrome P-450 induction or inhibition, ammonia detoxification, mitochondrial function, peroxisome proliferation, organelle function, cell cycle state, morphology’, apoptosis, DNA damage, metabolism, signal transduction, cell differentiation, cell-cell interaction and cell to non-cellular compartment; the unique cell state and / or unique cell type is characterized by one or more of acute phase stress, cell adhesion, AH-response, anti-apoptosis and apoptosis, antimetabolism, anti- proliferation, arachidonic acid release, ATP depletion, cell cycle disruption, cell matrix disruption, cell migration, cell proliferation, cell regeneration, cellcell communication, cholestasis, differentiation, DNA damage, DNA replication, early response genes, endoplasmic reticulum stress, estogenicity, fatty' liver, fibrosis, general cell stress, glucose deprivation, growth arrest, heat shock, hepatotoxicity, hypercholesterolemia, hypoxia, immunotox, inflammation, invasion, ion transport, liver regeneration, cell migration, mitochondrial function, mitogenesis, multidrug resistance, nephrotoxicity, oxidative stress, peroxisome damage, recombination, ribotoxic stress, sclerosis, steatosis, teratogenesis, transformation, disrupted translation, transport, and tumor suppression; the unique cell state and / or unique cell type is characterized by one or more of nutrient deprivation, hypoxia, oxidative stress, hyperproliferative signals, oncogenic stress, DNA damage, ribonucleotide depletion, replicative stress, and telomere attrition, promotion of cell cycle arrest, promotion of DNA-repair, promotion of apoptosis, promotion of genomic stability, promotion of senescence, and promotion of autophagy, regulation of cell metabolic reprogramming, regulation of tumor microenvironment signaling, inhibition of cell sternness, survival, and invasion; the aberrant signaling involves: an overactive signal transducer; a constitutively active signal transducer over a period of time; an active signal transducer repressor and an active signal transducer; an inactive signal transducer activator and an active signal transducer; an inactive signal transducer; an underactive signal transducer; a constitutively inactive signal transducer over a period of time; an inactive signal transducer repressor and an inactive signal transducer; and / or an active signal transducer activator and an inactive signal transducer; and / or wherein the aberrant signaling comprises an aberrant signal of at least one signal transduction pathway regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof, optionally the signal transducer(s) is AKT, PI3K, MAPK, p44 / 42MAP kinase, TYK2, p38 MAP kinase, PKC, PKA, SAPK, ELK, JNK, eJun, RAS, Raf, MEK 1 / 2, MEK 3 / 6, MEK 4 / 7, ZAP-70, LAT, SRC, LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX, PLCy, PLCy, NF-kB, FAK, CREB, aIII 3, FceRI, BAD, p70S6K, STAT1, STAT2, STAT3, STAT5, STAT6.

92. The method of any one of claims 90-91, wherein configuring the first and / or the second heterologous protease to be in the first and / or the second heterologous protease active state in response to the unique cell type and / or the unique cell state of the target cell comprises: expressing a second synthetic protein circuit in the target cell, wherein the second synthetic protein circuit comprises: a first polypeptide comprising a first signal transducer binding domain and a first part of a first protease domain of the first or second heterologous protease, wherein the first signal transducer binding domain is capable of binding a first signal transducer to form a first signal transducer-bound polypeptide; a second polypeptide comprising a second signal transducer binding domain and a second part of the first protease domain of the first or second heterologous protease, wherein the second signal transducer binding domain is capable of binding a second signal transducer to form a second signal transducer-bound polypeptide, wherein the first part of the first protease domain and the second part of the first protease domain have weak association affinity, and wherein the first part of the first protease domain and the second part of the first protease domain are capable of associating with each other to constitute the first or second heterologous protease capable in a first or second heterologous protease active state capable of cutting:(i) the first, second, or third apoptosis polypeptide at the first or second heterologous protease cleavage site when the first signal transducer and the second signal transducer are in close proximity at an association location; or(ii) the first or second pyroptosis polypeptide at the first or second heterologous protease cleavage site when the first signal transducer and the second signal transducer are in close proximity at an association location.

93. The method of any one of claims 90-92, wherein: the first signal transducer binding domain of the first polypeptide and the second signal transducer binding domain of the second polypeptide are identical; the first signal transducer and the second signal transducer are identical and / or are the same protein; the first signal transducer, the second signal transducer, or both, are capable of being localized at the association location;the first signal transducer when in a first signal transducer active state, the second signal transducer when in a second signal transducer active state, or both, are capable of being localized at the association location; the first signal transducer when in a first inactive state, the second signal transducer when in a second inactive state, or both, are capable of being localized at the association location; the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer, or both; the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first signal transducer active state, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer in a second signal transducer active state, or both; the first signal transducer binding domain of the first polypeptide is capable of binding to the first signal transducer in a first inactive state, wherein the second signal transducer binding domain of the second polypeptide is capable of binding to the second signal transducer in a second inactive state, or both; the first signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at the association location, or both; the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at a first cellular location other than the association location, wherein the signal transducer binding domain of the first polypeptide is capable of binding the first signal transducer to form the first signal transducer-bound polypeptide at a second cellular location other than the association location, or both; the first cellular location, the second cellular location, or both comprise one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclearmembrane, chloroplast, cell wall, or any combination thereof; the association location comprises one or more of a cell membrane, lipid raft, mitochondrion, peroxisome, cytosol, vesicle, lysosome, plasma membrane, nucleus, nucleolus, inner mitochondrial matrix, inner mitochondrial membrane, intermembrane space, outer mitochondrial membrane, secretory vesicle, endoplasmic reticulum, golgi body, phagosome, endosome, exosome, microtubule, microfilament, intermediate filament, filopodium, ruffle, lamellipodium, sarcomere, focal contact, podosome, ribosome, microsome, plasma membrane, nuclear membrane, chloroplast, cell wall, or any combination thereof; the first signal transducer binding domain and the second signal transducer binding domain are different; the first signal transducer binding domain and / or the second signal transducer binding domain each is capable of binding molecules of the first signal transducer and / or the second signal transducer; the first signal transducer and / or the second signal transducer belong to a signal transduction pathway; the first signal transducer binding domain and / or the second signal transducer binding domain comprise a RAS binding domain (RBD) and / or RAS association domain (RAD), optionally the RAS binding domain comprises or is derived from a RAS interacting protein, optionally selected from the group comprising AGO2, APBB1IP, APPL1, ARAF, ARL1, ARL2, ARRB1, ARRB2, BAIAP2, BCL2, BCL2L1, BRAF, BRAP, BSG. CALM1. CALM3, CALML3. CALML4, CALML5, CALML6, CNKSR1, CNKSR2, CSK, DAB2IP, EGFR, ERBIN, FGA, FGB, FGG, FN1, GRB2, HK1, IFNGR1, IL6, IQGAP1, ITGA2B, ITGB3, KSR1, KSR2, LGALS3, LYN, LZTR1, MAP2K1, MAP2K2, MAPK1, MAPK14, MAPK3, MAPKAP1, MARK2, MARK3, MBP, MSI2, MTOR, NCBP2AS2, NF1, NIBAN2, PDE4DIP, PDE6D, PDPK1, PEBP1, PIK3CA, PIK3CB. PIK3CD. PIK3R1. PIK3R2, PIP5K1A, PLCE1. PPIA, PRKCZ. PTGS2, RAFI, RALB, RALGDS, RAP1A, RAP1B, RAP1GDS1, RASA1, RASA2, RASA3, RASA4, RASAL1, RASAL2, RASAL3, RASSF1, RASSF2, RASSF5, RGLL RGL3, RIN1, SHOC2, S0S1, S0S2, SPRED1, SPRED2, SPRED3. SRC, SYNGAP1, TIAM1, TLN1, VCL. VWF, YWHAB. or any combination thereof; the first signal transducer binding domain and / or the second signal transducer binding domain comprises a lipid binding domain; the lipid binding domain comprises a Pleckstrin homology (PH) domain; the first signal transducer binding domain and / or the second signal transducerbinding domain comprises an antibody, an antibody fragment, a binding domain derived from a natural protein, an scFv, a Fv, a Fab, a (Fab')2, a single domain antibody (SDAB), a VH or VL domain, a camelid VHH domain, a Fab, a Fab1, a F(ab')2, a Fv, a scFv, a dsFv, a diabody, a triabody, a tetrabody, a multispecific antibody formed from antibody fragments, a single-domain antibody (sdAb). a single chain comprising cantiomplementary scFvs (tandem scFvs) or bispecific tandem scFvs, an Fv construct, a disulfide-linked Fv, a dual variable domain immunoglobulin (DVD-Ig) binding protein or a nanobody, an aptamer, an affibody, an affilin, an affitin, an affimer, an alphabody, an anticalin, an avimer, a DARPin, a Fynomer, a Kunitz domain peptide, a monobody, or any combination thereof; the first signal transducer is capable of binding the first signal transducer binding domain and / or the second signal transducer is capable of binding the second signal transducer binding domain following a modification selected from the group comprising phosphorylation, dephosphorylation, acetylation, methylation, acylation, glycosylation, glycosylphosphatidylinositol (GPI) anchoring, sulfation, disulfide bond formation, deamidation, ubiquitination, sumoylation, nitration of tyrosine, hydrolysis of ATP or GTP, binding of ATP or GTP, cleavage, or any combination thereof; the first signal transducer, the second signal transducer, or both are endogenous proteins; the first signal transducer, the second signal transducer, or both comprise AKT, PI3K, MAPK, p44 / 42 MAP kinase, TYK2, p38 MAP kinase, PKC, PKA, SAPK, ELK, JNK, eJun, RAS, Raf, MEK 1 / 2, MEK 3 / 6. MEK 4 / 7, ZAP-70, LAT, SRC. LCK, ERK 1 / 2, Rsk 1, PYK2, SYK, PDK1, GSK3, FKHR, AFX, PLCy, PLCy, NF-kB, FAK, CREB, cdlip3, FCERI. BAD, p70S6K, STAT1, STAT2, STAT3, STAT5, STAT6, or any combination thereof; the first signal transducer and / or the second signal transducer are capable of regulating cell survival, cell growth, cell proliferation, cell adhesion, cell migration, cell metabolism, cell morphology, cell differentiation, apoptosis, or any combination thereof; the first signal transducer, the second signal transducer, or both comprise a RAS protein, a CTNNB1 protein, or a TP53 protein; the RAS protein is KRAS, NHAS. HRAS. or any combination thereof, optionally the RAS protein comprises a G12 mutation, G13 mutation, a Q61 mutation, and / or an A146 mutation, optionally the G12 mutation is selected from the group comprising G12A, G12C, G12D, G12R, G12S, G12V, and any combination thereof, optionally the G13 mutation is selected from the group comprising G13C, G13D, G13dup, G13R, G13S,G13V, and any combination thereof, optionally the Q61 mutation is selected from the group comprising Q61H, Q61K, Q61L, Q61R, Q61E, Q61P, Q61*. and any combination thereof, optionally the A146 mutation is selected from the group comprising A146P, A146T. A146V, and any combination thereof; the signature detected by the input polypeptide(s) is correlated with any other cellular signature, protein state, cell type, and / or cell state capable of being read out as a biomarker; the signal transducer is CTNNB1, and wherein the cell state is defined by CTNNB1 mutation(s) and / or localization and / or concentration and / or protein turnover and / or multimerization and / or PTM(s); the signal transducer is TP53, and wherein the cell state is defined by TP53 mutation(s) and / or elevated TP53 concentration and / or altered TP53 oligomerization / multimerization state and / or TP53 localization pattern and / or PTM(s) and / or turnover; the signal transducer(s) are associated with disease, optionally cancer; the first signal transducer, the second signal transducer, or both are exogenous proteins; the second synthetic protein circuit comprises the first signal transducer, the second signal transducer, or both; and / or the first signal transducer, the second signal transducer, or both comprise a lipid, optionally the lipid comprises a phospholipid, further optionally the phospholipid is phosphatidylinositol 3-phosphate.

94. A method of treating or preventing a disease or disorder in a subject in need thereof, comprising: expressing the synthetic protein circuit of any one of claims 1-61 or the first synthetic protein circuit and / or second synthetic protein circuit of any one of claims 90- 93, in a cell of the subject.

95. A method of treating or preventing a disease or disorder in a subject in need thereof, comprising: administering to the subject an effective amount of the nucleic acid composition of any one of claims 62-63 or the composition of any one of claims 64-89, thereby treating or preventing the disease or disorder in the subject.

96. The method of claim 95, wherein administering comprises:(i) isolating one or more cells from the subject;(ii) contacting said one or more cells with the nucleic acid composition of any oneof claims 62-63, thereby generating engineered cells, optionally the contacting comprises transfection; and(iii) administering the one or more engineered cells into a subject after the contacting step.

97. The method of any one of claims 90-96, wherein the disease or disorder is a blood disease, an immune disease, a neurological disease or disorder, a cancer, an infectious disease, a genetic disease, a disorder caused by aberrant mtDNA, a metabolic disease, a disorder caused by aberrant cell cycle, a disorder caused by aberrant angiogenesis, a disorder cause by aberrant DNA damage repair, or any combination thereof, optionally a solid tumor.

98. The synthetic protein circuit, nucleic acid composition, or method of any one of claims 1-97, further comprising a supplementary protein circuit comprising: a first polypeptide comprising an optional first supplementary' domain and a first part of a first protease domain of a supplementary heterologous protease; a second polypeptide comprising a second supplementary domain and a second part of the first protease domain of the supplementary protease, wherein the first part of the first protease domain and the second part of the first protease domain have weak association affinity, and wherein the first part of the first protease domain and the second part of the first protease domain are capable of associating with each other to constitute the supplementary heterologous protease, optionally the first and / or second supplementary domain is a signal transducer binding domain, optionally the supplementary heterologous protease in a supplementary heterologous protease active state is capable of cutting (i) the first, second, or third apoptosis polypeptide at the first or second heterologous protease cleavage site, and / or (ii) the first or second pyroptosis polypeptide at the first or second heterologous protease cleavage site, further optionally when a first signal transducer and a second signal transducer are in close proximity at an association location, optionally the supplementary heterologous protease is the first or second heterologous protease.

99. The synthetic protein circuit, nucleic acid composition, or method of any one of claims 1-98, wherein the induction of apoptosis and / or pyroptosis is dependent on the dose of one or more synthetic protein circuit components, optionally cell death is triggered when a threshold amount of one or more of the following is reached: (i) the first, second, and / or third apoptoticprotein complex in the first, second, and / or third apoptotic protein complex active state; (ii) the first and / or second pyroptosis polypeptide in a first and / or second pyroptosis polypeptide active state; and / or (iii) a pyroptosis effector protein in a pyroptosis effector protein active state.

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