Genetically engineered induced pluripotent stem cells and effector cells derived therefrom

Abstract

Provided herein are genetically engineered induced pluripotent stem cells (iPSC's) or derivative cells thereof comprising nucleic acids encoding one or more chimeric antigen binding domains (CARs), and binding domains specific for CD300 and one or more nucleic acids encoding exogeneous promoters operatively linked thereto, and vectors, engineered cells, and compositions comprising the recombinant nucleic acids. Also provided are methods of treating a subject and / or improving a clinical outcome in a subject using the engineered cells.

Description

GENETICALLY ENGINEERED INDUCED PLURIPOTENT STEM CELLS AND EFFECTOR CELLS DERIVED THEREFROMCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U. S. Provisional Patent Application No.63 / 729,469 filed December 9, 2024, U. S. Provisional Patent Application No. 63 / 772,702 filed on March 16, 2025, and U. S. Provisional Patent Application No, 63 / 912,625, filed on November 6, 2025. The contents of each are incorporated herein by reference in their entirety.REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on December 9, 2025, is named “066461-28WO1 Sequence Listing. xml” and is 422,831 bytes in size.FIELD OF THE INVENTION

[0003] The present disclosure provides recombinantly engineered induced pluripotent stem cells expressing one or more chimeric antigen receptors and one or more binding domains specific for CD300a under the control of an exogeneous promoter, and methods of using the cells for the treatment of disease.INCORPORATION BY REFERENCE

[0004] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BACKGROUND

[0005] Adoptive T cell immunotherapy is a rapidly growing field, in particular in cancer treatments. Chimeric antigen receptor (CAR) T cell therapy represents a breakthrough in the field of immunooncology. In general, CAR T cell engagement of, e g, CD19- or CD20-expressing cancer cells, results in T cel l activation, proliferation and secretion of inflammatory cytokines and chemokines resulting in tumor cell lysis. However, autologous CAR T cell therapy presents multiple challenges,14909-7891-2381.1such as time for manufacturing and need for interim therapies in progressing patients, wide variations in terms of quality and quantity of T cells, and difficulties in obtaining enough cells for redosing. “Off-the-shelf’ allogeneic CAR T cells generated by differentiation of pluripotent stem cells (such as induced pluripotent stem cells or human embryonic stem cells) into T cells or derived from third-party donor T cells may provide solutions to these varied problems. For example, off- the-shelf allogenic CAR T cells can be expanded in high numbers prior to treatment and made quickly available to patients. This high-volume manufacturing allows for an opportunity to quickly and easily re-dose patients with a new off-the-shelf cell therapy without delays due to manufacturing or scale of production. Additionally, pretreatment manufacturing protocols allow for multiple edits, manipulations, or T cell receptor selection strategies that otherwise could be difficult to accommodate in the autologous setting.

[0006] However, allogeneic T cells possess foreign immunological identities that can lead to histocompatibility considerations such as graft-versus-host disease (GvHD) and rejection or low- persistence of the allogeneic cells. For example, iPSC-derived T cells that have not undergone engineering to make the cells immuno-compatible or hypoimmunogenic cannot be applied to third-party patients due to the presence of endogenous T cell receptor (TCR) and / or human leukocyte antigen (HLA) mismatch. GvHD is believed to be largely driven by donor- or iPSC-derived T cell recognition of host peptide-HLA complexes through the αβ T cell receptor complex (αβTCR), Rejection is mainly driven by host NK cells, CD8+ T-cells, CD4+ T cells, and, to a lesser extent, by macrophages. In the context of CAR T-cell therapy, the relative contribution of these cell types to allograft rejection may vary depending on their absolute numbers and reconstitution kinetics following preconditioning regimen. Because, in many cases, host NK cells and host CD8+ T cells appear to recover their initial levels more quickly than CD4+ T cells, CD8+ T cells and NK cells may play a larger role in controlling the length of the allogeneic CAR T-cell therapeutic window by being the first and primary contributors to rejection.

[0007] Because of the complexity of the immune system, engineering hypoimmunogenic T cells to enable adoptive cell transfer in an allogeneic setting has been a challenge. Some strategies for preventing or reducing GvHD and increasing therapeutic cell persistence include engineering approaches such as knocking out or disrupting the native alpha beta T cell receptor by gene editing of the T cell receptor alpha constant (TRAC) locus or by insertion of the CAR transgene into the TRAC locus, incorporating a receptor to target activated alloreactive host T cells, or disrupting 24909-7891-2381.1HLA expression on CAR T cells by knocking out beta 2 microglobulin (β2m) and evading NK cell- mediated killing by expressing HLA class I histocompatibility antigen alpha chain E (HLA-E). Recently, the inventors have disclosed an additional method for evading NK-cell cytotoxicity that can be compatible with adoptive cell transfer in an allogeneic setting by inserting a transgene encoding a construct for inhibiting NK cell cytotoxicity comprising a CD300a binding domain (International Publication No. WO2024 / 238934). To date, automated, scalable, and efficient manufacturing of therapeutic immune cells, along with effective mechanisms for protecting these cells against host rejection, remain significant challenges to widespread access to CAR T cells for patients.SUMMARY

[0008] Provided herein is an engineered induced pluripotent stem cell (iPSC), or a derivative cell thereof, comprising:(a) one or more exogenous polynucleotides encoding:i. one or more chimeric antigen receptors (CARs), said one or more CARs comprising a first antigen binding domain, and optionally a second antigen binding domain, ii. a CD300a TASR binding domain comprising:1. a CD300a light chain variable region (CD300a VL); and2. a CD300a heavy chain variable region (CD300a VH);iii. And, optionally, a humoral response-inhibition construct comprising an IgG- degrading enzyme of Streptococcus pyogenes (IdeS), and(b) an exogenous promoter operably linked to one or more of (x) the one or more CARs, (y) the CD300a TASR binding domain, and (z) the IdeS such that expression of the one or more exogenous polynucleotides is at least in part under the control of the exogenous promoter; and(c) a deletion or reduced expression of one or more of Beta 2 Microglobulin (B2M), Class II Major Histocompatibility Complex Transactivator (CIITA), and T Cell Receptor Alpha Constant (TRAC) genes;wherein the one or more exogenous polynucleotides are inserted at a locus of one or more of the B2M gene, the CIITA gene, and the TRAC gene, thereby deleting or reducing the expression of one or more of the B2M gene, the CIITA gene, and / or the TRAC gene.34909-7891-2381.1

[0009] In some embodiments, the exogenous polynucleotide encoding the one or more CARs is inserted at the locus of the TRAC gene, thereby deleting or reducing the expression of the TRAC gene.

[0010] In some embodiments, the exogenous polynucleotide encoding the exogeneous promoter is operably linked to the polynucleotide encoding the one or more CARs.

[0011] In some embodiments, the exogenous polynucleotide encoding the exogeneous promoter is operably linked to the polynucleotide encoding the CD300a TASR binding domain, and, optionally, the IdeS.

[0012] In some embodiments, the exogenous polynucleotide encoding the exogeneous promoter is operably linked to the polynucleotide encoding the CARs, and the CD300a TASR binding domain, and, optionally, the IdeS.

[0013] In some embodiments, the iPSC or the derivative cell comprises a plurality of exogenous polynucleotides, comprising:(a) a first polynucleotide encoding an exogeneous promoter operably linked to the polynucleotide encoding the one or more CARs; and(b) a second exogenous polynucleotides encoding an exogeneous promoter operably linked to the polynucleotide encoding the CD300a TASR binding domain and, optionally, the IdeS.

[0014] In certain embodiments, the one or more first or second exogenous polynucleotides are inserted at the locus of a gene independently selected from the group consisting of the B2M gene and the CIITA gene, thereby deleting or reducing the expression of one or both of the B2M gene and CIITA gene.

[0015] In some embodiments, the exogeneous promoter is independently selected from the group consisting of EFla promoter, a CAG promoter, a PGK promoter, or a CMV promoter, or a fragment of any thereof.

[0016] In certain embodiments, the promoter comprises the EFla promoter or a fragment thereof.

[0017] In some embodiments, the iPSC or derivative cell comprises a deletion or reduced expression of two or more of the B2M gene, the CIITA gene, and the TRAC gene.

[0018] In certain embodiments, the iPSC or derivative cell comprises a deletion or reduced expression of the B2M gene, the CIITA gene, and the TRAC gene.44909-7891-2381.1

[0019] In some embodiments, the first antigen binding domain and the second antigen binding domain of the CARs specifically bind to an antigen independently selected from the group consisting of Cluster of Differentiation (CD) 19 antigen, CD22 antigen, B cell maturation antigen (BCM A), and Nectin-4.

[0020] In some embodiments, the one or more exogenous polynucleotides encode both of the CD300a TASR binding domain and the IdeS, and wherein the CD300a TASR binding domain and the IdeS are operably linked by an autoprotease peptide. In certain embodiments, the autoprotease peptide is a 2A peptide.

[0021] In some embodiments, the polynucleotide encoding the CD300a TASR antibody or a fragment thereof comprises a polynucleotide encoding a single chain variable fragment (scFv) or a VHH. In certain embodiments, the VI II I comprises the VI I domain of a camelid heavy chain antibody.

[0022] In some embodiments, the CD300a binding domain comprises a VHH (CD300a VHH).

[0023] In some embodiments, the CD300a VHH comprises:(a) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, I, or 2 mutations relative to an amino acid sequence comprising AAKPGEDVY (SEQ ID NO: 166);(b) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLSQFAS (SEQ ID NO: 167);(c) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPRSGWGL (SEQ ID NO: 168);(d) a CDRI comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an 54909-7891-2381.1amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKTRYES (SEQ ID NO: 169);(e) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 158), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ IS NO: 163), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising NTRLAHGRDVLGGVAYDI (SEQ ID NO: 170);(f) a CDRI comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 158), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ IS NO: 163), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising NTRRLGRSGDLVQDY (SEQ ID NO: 171);(g) a CDRI comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSRYY (SEQ ID NO: 159), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPDRDY (SEQ ID NO: 172);(h) a CDRI comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLPDVLPLEY (SEQ ID NO: 173),(i) a CDRI comprising 0, I, or 2 mutations relative to an amino acid sequence comprising GFTFSSYW (SEQ ID NO: 160), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ IS NO: 163), and a CDR3 comprising 0, I, or 2 mutations relative to an amino acid sequence comprising ATKVDGSYGIVTEL (SEQ ID NO: 174);(j) a CDRI comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 158), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising INGSGGST (SEQ IS NO: 174), and a CDR3 comprising 0,64909-7891-2381.11, or 2 mutations relative to an amino acid sequence comprising HTRRSGTSMAMDV (SEQ ID NO: 175),(k) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR.3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKLTMVY (SEQ ID NO: 176);(l) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, I, or 2 mutations relative to an amino acid sequence comprising VTKLTNEY (SEQ ID NO: 177);(m) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, I, or 2 mutations relative to an amino acid sequence comprising VTKVRPSYEY (SEQ ID NO: 178);(n) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSPYY (SEQ ID NO: 161), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VAKPGYEY (SEQ ID NO: 179);(o) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGRT (SEQ IS NO: 165), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPGEDVY (SEQ ID NO: 180);(p) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKSNMVY (SEQ ID 74909-7891-2381.1NO: 181);(q) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ IS NO: 163), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising TTKVDGSYGI VTEL (SEQ ID NO: 182); or(r) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYW (SEQ ID NO: 160), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising INGSGGST (SEQ IS NO: 164), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising AAARDRERDY (SEQ ID NO: 183).

[0024] In certain embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ IDNOs: 139-156.

[0025] In certain embodiments, the CD300a VHH comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 139-156.

[0026] In some embodiments, the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 121-138.

[0027] In certain embodiments, the CD300a VHH comprises or consists of the nucleotide sequence of any one of SEQ ID NOs: 121-138.

[0028] In some embodiments, the CD300a TASR binding domain comprises a scFv.

[0029] In some embodiments:(a) the exogenous polynucleotide encoding the CD300a VL is codon optimized to reduce or prevent undesired recombination events; and / or(b) the exogenous polynucleotide encoding the CD300a VH is codon optimized to reduce or prevent undesired recombination events.

[0030] In some embodiments, the iPSC or the derivative cell further encodes a polynucleotide encoding a transmembrane domain.84909-7891-2381.1

[0031] In some embodiments, the transmembrane domain is a human transmembrane domain or a murine transmembrane domain.

[0032] In certain embodiments, the transmembrane domain comprises or consists of a CD8, a CD80, an ITGA, an HLA-B57, a proCAR-4, a CD28, a KIR2DL1, a PDGFRB, or a CD86 transmembrane domain.

[0033] In some embodiments, the transmembrane domain comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 67-76 and 96. In certain embodiments, the transmembrane domain comprises or consists of any one of SEQ ID NOs: 66-75 and 93.

[0034] In some embodiments, a nucleotide sequence encoding the transmembrane domain comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 56-65. In certain embodiments, a nucleotide sequence encoding the transmembrane domain comprises or consists of any one of SEQ ID NOs: 56-65.

[0035] In some embodiments, the iPSC or the derivative cell further encodes a polynucleotide encoding a cytoplasmic domain,

[0036] In some embodiments, the cytoplasmic domain is a human cytoplasmic domain or a murine cytoplasmic domain.

[0037] In some embodiments, the cytoplasmic domain comprises or consists of a CD8v2, a CD8vl, a mCD80, a CD80, a CD86, or an HLA-B57 cytoplasmic domain.

[0038] The application also relates to a vector comprising a polynucleotide of any one of the previous embodiments.

[0039] In some embodiments, the vector is a DNA vector, an RNA vector, a plasmid, a lenti virus vector, an adenoviral vector, an adeno associated viral vector, a Rous sarcoma viral (RSV) vector, or a retrovirus vector.

[0040] In some embodiments, the derivative cell is an early hematopoietic progenitor cell or a CD34+ progenitor cell.

[0041] In certain embodiments, the cell that has been differentiated from a stem cell or the cell that has been differentiated from a progenitor cell is an engineered T cell.94909-7891-2381.1

[0042] In some embodiments, the engineered cell is an engineered T cell.

[0043] In some embodiments, the engineered T cell is a chimeric antigen receptor (CAR) T cell.

[0044] In certain embodiments, the CAR is a CD 19 CAR, a BCMA CAR, a CD22 CAR or a Nectin-4 CAR.

[0045] In some embodiments, the engineered cell is T cell receptor alpha constant (TRAC) deficient.

[0046] In some embodiments, a T cell receptor alpha constant (TRAC) locus is disrupted in the engineered cell.

[0047] In some embodiments, the CAR is inserted into a T cell receptor alpha constant (TRAC) locus in the engineered cell.

[0048] The application also relates to a composition comprising the derivative cell of any one of the previous embodiments, and one or more of a cell culture media and a buffer.

[0049] The application also relates to a pharmaceutical composition comprising the derivative cell of any one of the previous embodiments, and a pharmaceutically acceptable carrier.

[0050] The application also relates to a method of improving a clinical outcome in a subject undergoing a T cell therapy, comprising administering to the subject an effective amount of the derivative cell of any one of the previous embodiments, or the pharmaceutical composition comprising the derivative cell of any one of the previous embodiments.

[0051] The application further relates to a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of the derivative cell of any one of the previous embodiments, or the pharmaceutical composition comprising the derivative cell of any one of the previous embodiments.

[0052] In some embodiments, the disease or disorder is a cancer.

[0053] In some embodiments, the cancer is a hematologic cancer selected from B-cell acute lymphoid leukemia (B-ALL), T cell acute lymphoid leukemia (T-ALL), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL), B cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt’s lymphoma, diffuse large B cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell-follicular lymphoma, large cell-follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, Marginal zone lymphoma, multiple myeloma (MM), myelodysplasia, myelodysplastic syndrome, non-Hodgkin’s 104909-7891-2381.1lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, or preleukemia.

[0054] In some embodiments, the cancer is fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, and other sarcomas, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, lymphoid malignancy, pancreatic cancer, breast cancer, lung cancer, ovarian cancer, prostate cancer, colorectal cancer, hepatocellular carcinoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, pheochromocytomas sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, Wilms' tumor, cervical cancer, testicular tumor, seminoma, bladder carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma or retinoblastoma.

[0055] In some embodiments, the disease or disorder is an autoimmune disease or disorder

[0056] In some embodiments, the autoimmune disease or disorder is myasthenia gravis, neuromyelitis optica spectrum disorder, Sjogren’s syndrome, scleroderma, immune nephritis, systemic lupus erythematosus, arthritis, an autoimmune-induced fibrotic condition, pemphigus vulgaris, multiple sclerosis, colitis, graft-versus-host disease, atherosclerosis, or mucosal-dominant PV.

[0057] In some embodiments, the disease or disorder is selected from the group consisting of a rheumatological disorder, a neurological disorder, a hematological disorder, or a nephrological disorder. In some embodiments, the disease or disorder is a rheumatological disorder selected from the group consisting of rheumatoid arthritis (RA), systemic lupus erythematosus / lupus nephritis (SLE / LN), systemic sclerosis (SSc), idiopathic inflammatory myopathies (IIM), anti-phospholipid syndrome, Sjogren's syndrome, IgG4-related disease (IgG4RD), vasculitis, and autoimmune hepatitis. In some embodiments, the disease or disorder is a neurological disorder selected from the group consisting of neuromyotonia, chronic inflammatory demyelinating polyneuropathy (CIDP), Guillain-Barre syndrome, Lambert-Eaton myasthenic syndrome (LEMS), myasthenia gravis (MG), neuromyelitis optica spectrum disorder (NMOSD), and stiff person syndrome. In some embodiments, the disease or disorder is a hematological disorder selected from the group consisting of acquired anemia, catastrophic antiphospholipid syndrome (CAPS), warm autoimmune hemolytic 114909-7891-2381.1anemia and cold autoimmune hemolytic anemia (wAIHA & cold AIHA), cryoglobulinemia, Felty syndrome, Evans syndrome, chronic graft-versus-host disease (cGVHD), and immune thrombocytopenic purpura (ITP). In some embodiments, the disease or disorder is a nephrological disorder selected from the group consisting of lupus nephritis class III / IV, primary membranous nephropathy, cryoglobulinemia, steroid-dependent focal segmental glomerulosclerosis (FSGS), proliferative glomerulonephritis with monoclonal immunoglobulin deposits (PGMCID), fibrillary glomerulonephritis (GN), and immunotactoid glomerulonephritis.

[0058] In some embodiments, the subject is a human.

[0059] Further aspects, features and advantages of the present invention will be better appreciated upon a reading of the following detailed description of the invention and claims.BRIEF DESCRIPTION OF THE DRAWINGS

[0060] The foregoing and other objects, aspects, features, and advantages of exemplary embodiments will become more apparent and may be better understood by referring to the following description taken in conjunction with the accompanying drawings.

[0061] FIG. 1 illustrates four exemplary', non-limiting recombinant nucleic acid designs described herein, including a promoter (e.. EFla), a CD300a TASR and / or a NKG2A TASR, and a linker (e.g., P2A) in exemplary' designs comprising both a CD300a TASR and a NKG2A TASR.

[0062] FIGS. 2A-2B illustrate MHC class I deficient T cell killing by NK cells. FIG. 2A shows phenotyping of wild-type, CD59 knockout, CIITA knockout, and B2M knockout cells using flow cytometry. FIG. 2B shows the results of aNK cell cytotoxicity assay, wherein % T cell survival was lowest for B2M knockout cells.

[0063] FIGS 3 A-3B illustrate the efficacy of two NKG2A construct designs as compared to the control HLA-E single chain trimer. FIG. 3A shows a schematic of the HLA-E and NKG2A scFv mRNA constructs, and phenotyping of cells electroporated with the constructs, using flow cytometry. FIG. 3B shows that both of the NKG2A scFv constructs provided protection against NK cell killing. The NKG2A construct comprising the CD8 transmembrane domain appeared to perform better than the NKG2A construct comprising IgGlB7.

[0064] FIGS. 4A-4B illustrate that the efficacy of the NKG2A scFv construct comprising the CD8 transmembrane domain (referred to hereafter as the Z199 construct) was reproducible. FIG. 4A shows a schematic of the HLA-E and NKG2A scFv mRNA constructs, and phenotyping of cells 124909-7891-2381.1electroporated with the constructs, using flow cytometry. The negative control shown data displayed in this figure are the same as that shown in FIG, 8A FIG. 4B shows that the Z199 construct provided some protection against NK cell killing. The HLA-E and GFP data shown in this figure are the same as that shown in FIG. 8B.

[0065] FIGS. 5A-5B illustrate that a humanized anti-NKG2A (clone Z270) scFv is stable and functional. FIG. 5A shows phenotyping of cells electroporated with the Z199, humanized Z 199, humanized Z270, and negative control constructs, using flow cytometry. FIG. 5B shows that all of the NKG2A scFv constructs provided protection against NK cell killing.

[0066] FIGS, 6A-6B illustrate that the efficacy of the NKG2A humanized Z270 construct was reproducible. FIG. 6A shows phenotyping of cells electroporated with the Z270 or HLA-E constructs using flow cytometry. The negative control shown data displayed in this figure are the same as that shown in FIG. 10A FIG. 6B shows that the humanized Z270 construct provided protection against NK cell killing.

[0067] FIGS. 7A-7B illustrate the efficacy of two CD300a construct designs as compared to the control HLA-E single chain trimer. FIG. 7A shows a schematic of the HLA-E and CD300a scFv mRNA constructs, and phenotyping of cells electroporated with the constructs, using flow cytometry. FIG. 7B shows that both of the CD300a scFv constructs provided protection against NK cell killing. The CD300a construct comprising the CD8 transmembrane domain appeared to perform better than the CD300a construct comprising IgGlB7.

[0068] FIGS. 8A-8B illustrate that the efficacy of the CD300a scFv construct comprising the CD8 transmembrane domain (referred to hereafter as the TX49 construct) was reproducible. FIG. 8A shows a schematic of the CD300a scFv mRNA construct, and phenotyping of cells electroporated with the constructs, using flow cytometry. The negative control shown data displayed in this figure are the same as that shown in FIG. 4A FIG. 8B shows that the TX49 construct provided some protection against NK cell killing. The HLA-E and GFP data shown in this figure are the same as that shown in FIG. 4B.

[0069] FIGS. 9A-9C illustrate successful humanization of the TX49 construct. FIG. 9 A shows amino acid sequence alignments of the VH (top) and VL (bottom) regions of the mouse anti-human CD300a antibody clone TX49, along with two humanized versions named humTX49 y 1” and “humTX49_v2”. FIG. 9B shows phenotyping of cells electroporated with the constructs, using flow134909-7891-2381.1cytometry. FIG. 9C shows that both humanized TX49 constructs (vl and v2) provided protection against NK cell killing.

[0070] FIGS. 10A-10B illustrate that the efficacy of the humanized TX49 construct vl (hereinafter referred to as humTX49) was reproducible. FIG. 10A shows phenotyping of cells electroporated with the humTX49 and HLA-E constructs, using flow cytometry. The negative control shown data displayed in this figure are the same as that shown in FIG. 6A. FIG. 10B shows that the humTX49 construct provided some protection against NK cell killing.

[0071] FIGS. 11 A-l IB illustrate that humTX49 synergizes with humZ270 to provide NK protection, FIG. 11 A shows phenotyping of cells electroporated with the humTX49, hum270, and / or HLA-E constructs, using flow cytometry. Left panel shows staining of the HLA-E construct electroporated or non-electroporated cells with HLA-E. Middle panel shows staining of the indicated electroporated cells with CD300a. Right panel shows staining of the indicated electroporated cells with NKG2A. FIG. 1 IB shows that cells expressing both the humZ270 and humTX49 constructs were protected against K cell killing comparably to cells expressing the HLA-E construct.

[0072] FIGS. 12A-12C illustrate the efficacy of a bispecific humTX49-humZ270 construct for protecting engineered cells against NK cell cytotoxicity relative to HLA-E. FIG. 12A shows a schematic of two candidate bi specific constructs. “humTX49-humZ270” comprises a hum an GM-CSF signal peptide (‘" GM-CSF SP”), followed by the humTX49 scFV (vL-Linker-vH orientation), a (G4S)5 linker, then the humZ270 scFV (vL-Linker-vH orientation), followed by a CD8 stalk and transmembrane region (“CD8 TAI”), a T2A self-cleaving peptide, and GFP. The “humZ270- humTX49” construct is similar, except the two scFVs are swapped in order. FIG. 12B shows phenotyping of cells electroporated with the constructs, using flow cytometry. FIG. 12C shows that cells expressing either construct were protected against NK cell killing, with the humTX49-humZ270 construct providing protection comparable to that of HLA-E

[0073] FIGS. 13A-13B illustrate that agonism by TX49 (CD300a) and Z270 (NKG2A) was synergistic and more potent than that by HLA-E. FIGS. 13 A and 13B show superior protection against NK cell killing in cells expressing the hu TX49-humZ270 construct as compared to HLA- E. Cells electroporated with equimolar amounts of humTX49 and humZ270 constructs separately were also protected against NK cell killing.144909-7891-2381.1

[0074] FIGS. 14A-14C illustrate that physiological expression of the HLA-E, humZ270, and humZ270-humTX49 constructs at the AAVS1 locus in human primary T cells provide comparable protection against NK cell killing. FIG. 14A shows that cells expressing HLA-E, humZ270, or humZ270-humTX49 constructs, but not cells expressing the humTX49 construct, were equivalently protected against NK cell killing. FIG. 14B shows levels of NKG2A / NKG2C expression in donor NK cells. FIG. 14C shows a diagram of the expression constructs, and phenotyping of cells electroporated with the constructs, using flow cytometry.

[0075] FIGS. 15A-15D illustrate that the mCD80 transmembrane domain impacts 11TX49 mediated NK inhibition, and that transmembrane homodimerization does not appear to impact function. FIG. 15A shows a schematic of the experimental set up. FIG. 15B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 15C shows expression levels as measured for each construct. FIG. 15D shows that cells expressing any of the constructs were protected against NK cell killing.

[0076] FIGS. 16A-16D illustrate that the cytoplasmic tails of CD8a or mCD80 appear to enhance expression and function of hTX49. FIG. 16A shows a schematic of the experimental set up. FIG. 16B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG.16C shows expression levels as measured for each construct. FIG. 16D shows that cells expressing any of the constructs (with the exception of the t8 construct, which did not comprise transmembrane or cytoplasmic domains) were protected against NK cell killing.

[0077] FIGS. 17A-17D illustrate that reduced hinge size increases the functional potency of hTX49. FIG. 17A shows a schematic of the experimental set up. FIG. 17B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 17C shows expression levels as measured for each construct. FIG. 17D shows that cells expressing any of the constructs (with the exception of the t8 construct, which did not comprise transmembrane or cytoplasmic domains) were protected against NK cell killing.

[0078] FIGS. 18A-18D illustrate that an optimized CD300a TASR design (t!2) improves cell persistence over the original construct (vl). FIG. 18 A shows a schematic of the experimental set up. FIG. 18B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 18C shows expression levels as measured for each construct, using flow cytometry. FIG. 18D shows that cells expressing any of the constructs (with the tl 2 construct not comprising a hung region performing best) were protected against NK cell killing.154909-7891-2381.1

[0079] FIGS. 19A-19D illustrate that an optimized CD300a TASR design (t 12) shows protection against NKG2C+ / NKG2A- NK cell donors. FIG, 19 A shows a schematic of the experimental set up. FIG. 19B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 19C shows expression levels as measured for each construct, using flow cytometry. FIG.19D shows that cells expressing both the NKG2A_vl and CD300a_tl2 constructs were protected against NK cell killing

[0080] FIGS. 20A-20D illustrate that mouse CD80 cytoplasmic domain enhanced expression and function of CD300a TASR irrespective of the transmembrane domain. FIG. 20A shows a schematic of the experimental set up. FIG 20B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 20C shows expression levels as measured for each construct, using flow cytometry. FIG. 20D shows that cells expressing the V2, tl 2, 114, or tl 8 constructs comprising a mouse CD80 cytoplasmic domain were protected against NK cell killing.

[0081] FIGS. 21A-21C illustrate that the mCD80 cytoplasmic domain outperformed hCD80 and hCD86 cytoplasmic domains in TASR expression. FIG. 21 A shows a schematic of the experimental set up. FIG. 2 IB shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 21C shows expression levels as measured for each construct, using flow cytometry / .

[0082] FIGS. 22A-22D illustrate that hCD80 and hCD86 cytoplasmic domains do not perform as well as mCD80. FIG, 22A shows a schematic of the experimental set up. FIG. 22B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 22C shows expression levels as measured for each construct, using flow cytometry. FIG. 22D shows that cells expressing t23, t24, and t29 constructs comprising a hCD80 cytoplasmic domain were not as protected against NK cell killing as cells expressing the 112 and v2 constructs comprising a mCD80 cytoplasmic domain.

[0083] FIGS. 23A-23D illustrate that an optimized CD300a TASR with fully human extra-cellular domains (v2) showed enhanced functional potency over the original design (vl). FIG. 23A shows a schematic of the experimental set up. FIG. 23B shows a schematic illustrating the CD300a TASR constructs used to electroporate T cells. FIG. 23C shows expression levels as measured for each construct, using flow cytometry. FIG. 23D shows that cells expressing the v2 construct were more protected against NK cell killing than cells expressing the vl and tl 3 constructs.

[0084] FIGS. 24A-24D illustrate a repeated experiment using additional NK cell donors and showing that optimized CD300a TASR with fully human extra-cellular domains (v2) enhanced 164909-7891-2381.1functional potency over the original design (vl). FIG. 24A shows a schematic of the experimental set up. FIG. 24B shows a schematic illustrating the CD300a TASK constructs used to electroporate T cells. FIG. 24C shows expression levels as measured for each construct, using flow cytometry. FIG. 24D shows that cells expressing the v2 construct were more protected against NK cell killing than cells expressing the vl construct.

[0085] FIGS. 25A-25D illustrate an antibody-dependent cellular cytotoxicity (ADCC) assay using rituximab, showing that NKG2A_vl, CD300a_v2, and HLA-E single chain trimer constructs are capable of reducing ADCC FIG. 25A shows a schematic of the experimental set up. FIG. 25B shows a schematic illustrating the CD300a TASR and the NKG2A TASR used to electroporate T cells. FIG. 25C shows expression levels of CD300a TASR, NKG2A TASR, HLA-E, and RQR8 on T cells electroporated with the indicated niRNA constructs, using flow cytometry. FIG. 25D shows that the humTX49_v2 and humZ270_vl constructs were capable of reducing ADCC.

[0086] FIGS. 26A-26D illustrate that reduced affinity variants of an NKG2A TASR provided enhanced functional protection against NK cell killing. FIG. 26A shows a schematic of the experimental set up. FIG. 26B shows a schematic illustrating the NGK2A TASR constructs (with amino acid mutations noted) used to electroporate T cells. FIG. 26C shows expression levels as measured for each construct, using flow cytometry. FIG. 26D shows that cells expressing the mutated constructs that had reduced affinity for NKG2A were more protected against NK cell killing than cells expressing the non-mutated (wild-type) construct or the vl construct.

[0087] FIGS. 27A-27E illustrate that reduced affinity variants of an NKG2A TASR provided enhanced functional protection and reduced NKG2A downregulation on effector NK cells. FIG.27A shows a schematic of the experimental set up. FIG. 27B shows a schematic illustrating the NGK2A TASR constructs (with amino acid mutations noted) used to electroporate T cells. FIG.27C shows expression of the NKG2A TASR on T cells electroporated with the indicated mRNA constructs. FIG. 27D shows that cells expressing the mutated constructs that had reduced affinity for NKG2A were more protected against NK cell killing than cells expressing the vl or v2 construct. FIG. 27E shows a flow cytometry gating scheme showing that NK cells tend to downregulate NKG2A expression when exposed to T cells expressing NKG2A TASRs toward the end of the NK cytotoxicity assay.

[0088] FIGS. 28A-C show that B2M KO rescues allogeneic T cells from T cell al loreactivity but introduces NK cell alloreactivity. Survival of w'ildtype (WT) and B2M knockout (KO) T cells from 174909-7891-2381.1an HLA-A2+ donor challenged with NK and allo-T cell effectors from HLA-A2- donors was assessed. Allo-T cells contained the AHIII T cell receptor reactive to EMC7 peptide, ALWGFFPVL (SEQ ID NO: 200), presented by HLA-A2. FIG. 28A shows survival curves of WT and B2M KO target T cells challenged with the indicated effectors. N = 1 technical replicate curves per condition. FIG. 28B shows HLA-I expression of WT and B2M KO T cells by flow cytometry, gated live single cell lymphocytes. B2M KO specifically renders T cells susceptible to NK cell rejection. FIG.28C shows NK challenge assay with multiple NK donors and multiple target T cell effectors. The bar graph represents a summary of IC50 values, where each data point represents a WT or B2M KO T cell from one T cell donor against the indicated NK cell donor. Groups were compared using the Mann Whitney U-Test.

[0089] FIGS. 29A-C show discovery and assessment of CD300a TASR. FIG 29A shows screening of strategies to inhibit NK alloreactivity against B2M KO T cells. Ligands are expressed by mRNA electroporation, and knock-out performed by CRISPR / Cas9. Dotted line indicates no protection. N = 1 curve of T cell survival at 6-7 E: T ratios challenged with one NK cell donor (See FIG. 30) FIG. 29B shows an NK challenge assay. The graph legend indicates the cloaking transgene integrated into the B2M locus of human primary T cells, challenged with one of three NK cell donors. Inset shows the NK cell phenotype by flow cytometry, gated CD3-CD56+. N=2 technical replicate curves per condition. FIG, 29C shows a competition assay of pooled T cells from FIG. 29B, along with HLA-I+ control, all derived from one HLA-A2+ donor, against allo-T and NK cells from HLA-A2- donors. The frequency of the indicated T cell member after challenge with indicated NK and / or allo-T cell effector is shown. Allo-T contained a T cell receptor reactive against HA-2 peptide presented on HLA-A2. N = 3 technical replicates per condition.

[0090] FIGS. 30A-C show functional screening and validation of NK cloaking strategies by mRNA electroporation and CRISPR KO of B2M KO T cells, related to FIG. 29A. Cloaking ligands were expressed by mRNA electroporation (EP) and functionally validated. Each graph shown in FIG. 30A represents a single screen member. CRISPR KOs are in addition to B2M KO. The negative control was GFP mRNA for mRNA EP conditions or B2M KO only for the CRISPR KO screen. For each graph, the left inset plot shows the expression profile by flow cytometry, gated on single cell lymphocytes. “NC” indicates the negative control peak. For each graph, the right inset plot shows the NK challenge assay of the given screen member. FIGS. 30C-D show validation of CD300a TASR VI and NKG2A TASR VI by mRNA titration. FIG. 30B shows expression of 184909-7891-2381.1NKG2A and CD300a TASR by flow cytometry on B2M KO T cells 1-day post electroporation (EP) with the given quantity of respective mRNA. FIG. 30C shows survival of B2M KO T cells electroporated CD300a or NKG2A TASR at the indicated mRNA dose as in FIG. 30B, challenged with NK cells at a constant E: T ratio of 4.6. N = 3 technical replicates per condition.

[0091] FIGS. 31 A-D show that Mouse B7-I (also interchangeably referred to herein as “mCD80” and “mB7-l”) domains enhance TASR expression. FIG. 31A shows an experimental overview. TASRs were constructed with various transmembrane and cytoplasmic domains, in-vitro transcribed into mRNA, and then transiently transfected into primary T cells to test for expression. All constructs were bicistronic with GFP to enable normalized comparison between difference constructs. FIG. 3 IB shows various TM+Cyt domains tested. CD8 TM is from the VI scaffold. FIG. 31C shows GFP versus TASR median fluorescence intensity (MFI) of a KIR2D specific TASR with the various TM+Cyt domains from FIG. 3 IB as assessed by flow cytometry, gated on single cell lymphocytes. mB7-l provides higher TASR expression normalized to GFP translation. FIG. 3 I D shows expression of KIR2D, NKG2A, and CD300a TASRs with either CD8TM or mCD80 TM+Cyt after mRNA EP as measured by flow cytometry, gated on single cell lymphocytes.

[0092] FIGS. 32A-G show an inverse correlation of hinge length and functional potency of CD300a TASR but not NKG2A TASR, and further optimization of CD300a TASR (V2). FIGS, 32A-E show the effect of hinge length on function of CD300a TASR and NKG2A TASR. FIG. 32A shows the structures of CD300a TASR variants with different hinge domains with the indicated amino acid lengths and their expression level after mRNA EP into B2M KO T cells. FIG. 32B shows NK challenge assay of B2M KO T ceils expressing the indicated CD300a TASR variants. N = 2 technical replicate curves per condition. FIG. 32C shows correlation of NK protection from FIG. 32B as defined by IC50 value with hinge length. Error bar indicates 95% confidence interval of IC50 value. FIG. 32D shows structures of NKG2A TASR VI and a no hinge variant, along with their expression level by mRNA EP into B2M KO T cells. FIG. 32E shows NK challenge assay of B2M KO T cells expressing the indicated NKG2A TASR variant, N = 1 technical replicate curve per condition. FIGS. 32F-G show' further optimization of CD300a TASR. FIG. 32F shows CD300a TASR VI and two optimized variants tested for expression via mRNA EP of B2M KO T cells. FIG.32G shows NK challenge assay of B2M KO T cells expressing the indicated CD300a TASR variant. N = 1 technical replicate curve per condition.194909-7891-2381.1

[0093] FIGS. 33A-B show an experimental overview and phenotype of non-viral CRISPR-mediated targeted integration of cloaking transgenes into human primary T cells, related to FIG, 29B. FIG. 33 A shows a process for non-viral targeted integration and purification of T cells at the B2M and AAVS1 locus. Stimulated primary T cells were edited with CRISPR Cas9 at the indicated locus, along with linear double stranded DNA (dsDNA) homology repair template. For AAVS1 homology directed repair (HDR), the transgene encodes EFla promoter, cloaking transgene, P2A self-cleavable peptide, RQR8 epitope tag, and BGH poly A sequence. For B2M HDR, the cloaking transgene is integrated at the start codon of the B2M gene under control of endogenous B2M promoter and followed by BGH poly A sequence. Edited T cells were further purified by magnetic bead-based enrichment using antibody to either the cloaking transgene or RQR8 epitope tag if present, and then subjected to 1-2 additional rounds of stimulation and expansion prior to cryopreservation. FIG. 33B shows the phenotype of engineered T cells containing the indicated transgenes in the top label by flow cytometry, gated on single cell lymphocytes. Histograms shown in dotted lines indicate negative control T cells stained with the same markers.

[0094] FIGS. 34A-F show that CD300a TASR outperforms CD47 in B2M KO T cells in additional model systems and with IL-2 activated NK cells. FIGS. 34A-B show a comparison of CD300a TASR and CD47 via mRNA electroporation. FIG. 34A shows flow cytometry phenotypes of T cells electroporated with the indicated mRNA. FIG. 34B shows NK challenge assay of T cells in FIG. 34A against four NK cell donors with either 2Adom or 2Cdom NK phenotype. N = 1 technical replicate curves per condition. FIGS. 34C-D show a comparison of CD300a TASR and CD47 expressed from the AAVS1 loci of B2M KO T cells in both NK and PBMC challenge assays. FIG. 34C shows flow cytometry phenotypes of T cells containing the indicated cloaking transgene or GFP, gated single cell lymphocytes. FIG. 34D shows NK and PBMC challenge assay of T cells from with either NK or PBMC donors as indicated. N = 1 technical replicate curves per condition. FIG. 34E shows the effect of cytokine concentration and culture time on the expression of SIRPa on cultured human NK cells. Cryopreserved NK cells were thawed and cultured at the indicated concentrations of IL-2 or 10 ng / mL IL- 15, used for the standard NK challenge assay, for 3 and 5 days. Monocytes served as positive staining control. NK cells were gated CD3-CD56+, Monocytes were gated CD14+FSChiSSChi. FIG. 34F shows NK challenge assay with B2M KO T cells expressing the indicated cloaking transgene at B2M loci as in FIG. 29C NK cells were cultured for204909-7891-2381.1the indicated duration with the indicated cytokine, during both initial culture as in FIG. 34E and during the 20-hour co-culture. N 2 technical replicate curves per condition.

[0095] FIGS. 35A-B show that CD300a TASR outperforms TIM3 Engager. CD300a TASR with TIM3 engager were expressed in T cells via mRNA electroporation. Both ligands were bicistronic with GFP. FIG. 35 A shows cloaking ligand expression assessed using both ligand-based staining and antibody-based staining by flow cytometry with the indicated mRNA, gated on single cell lymphocytes. FIG. 35B shows NK challenge assay of T cells from FIG. 35A challenged with the four indicated NK cell donors. N = 2 technical replicate curves per condition.

[0096] FIGS, 36A-F show' a flow cytometry gating scheme and additional conditions for Allo-T + NK competition assay, related to FIG. 29D. FIGS. 36A-E show representative gating strategies and readouts for competition assays. All samples were resuspended and acquired at equal volumes. FIG.36A shows pre-gating on CD3+ single-cell lymphocytes. FIG. 36B shows gating for target pool only with no effector cells. Labels designate the pool member. FIG. 36C shows gating with NK effector challenge only. The gate indicated by the dashed lines highlights depleted population relative to FIG 36B. FIG. 36D shows gating with allo-T cell effector challenge only. The gate indicated by the dashed lines and label highlights depleted population relative to FIG. 36B FIG. 36E shows gating wdth allo-T and NK cell challenge, red gate highlights CD300a TASR V2 survival relative to other members and FIG. 36B. FIG. 36F shows additional NK and Allo-T challenge conditions as in FIG. 29. Allo-TEMC7 contains the AHIII T cell receptor reactive to EMC7 peptide, ALWGFFPVL (SEQ ID NO: 200), presented by HLA-A2. NK and allo-T cells effectors are mixed 1:1. N = 3 technical replicates per condition

[0097] FIGS. 37A-E show that CD300a TASR universally protects against NK cells and enhances CAR-T functional potency. FIG. 37A shows the study design and demographic overview of the 45 PBMC donors used. FIG. 37B shows a PBMC challenge assay. Each datapoint represents the IC50 value of a 7 -point curve of T cell survival with the indicated cloaking ligand against PBMCs from one donor. N = 45 donors. Groups were compared using Wilcoxon matched pairs signed rank test. FIG. 37C shows Association of PBMC donor demographics from FIG. 37A with functional data from FIG. 37B Groups were compared using the following statistical analyses: Kruskal-Wallis for Ethnicity, Mann-Whitney for age, gender, and CMV status. Y-axis represents ratio of IC50 between CD300a TASR and HLA-E cloaking ligand, with one indicating equal protection. FIG. 37D shows the relationship between functional potency and adaptive NK cell frequency for CD300a TASR 214909-7891-2381.1(left) and HLA-E (right). The dashed line represents linear fit of log-log transformed data. N = 45 PBMC donors. FIG. 37E (bottom row) shows a B cell lysis assay for engineered CAR-T cell therapy containing the indicated cloaking transgene against the indicated PBMC donors. N = 2 technical replicate curves per condition. FIG. 37E (top row) shows phenotypes of NK cells from the respective PBMC donor.

[0098] FIGS. 38A-D show a PBMC challenge assay and validation, related to FIGS. 37A-D. FIG 38 A shows the experimental overview of the PBMC challenge assay. PBMC phenotyping was performed post-thaw, and cloaked T cell phenotyping performed at time of co-culture. FIG. 38B shows the co-culture plate map and gating scheme for fluorescent barcoding flow cytometry readout in the PBMC challenge assay. FIG. 38C shows inter-assay variation of the PBMC challenge assay of one PBMC donor against one B2M KO T cell source. Experiments were performed on separate days. N = 1 technical curve per experiment. Right panel shows the coefficient of variation (CV) of the IC50 values. FIG. 38D shows phenotypes of the three engineered T cells targets used in FIG. 37, gated on live single lymphocytes Histograms shown in dotted lines indicate negative control. Label indicates cloaking transgene.

[0099] FIGS. 39A-D showNK cell phenotyping of PBMC challenge assay, related to FIGS. 37A-D. FIG. 39A shows a gating scheme for bulk NK cells and adaptive NK cell phenotype. FIG. 39B shows representative expression of NK cell phenotyping markers from one PBMC donor, “NC” indicates CD300a fluorescence-minus-one negative control staining. In panels where “NC” is not indicated due to peak overlap, the negative control is the leftmost peak. FIG. 39C shows the percentage of NK cells expressing the indicated marker with gating from FIG. 39B. N = 45 PBMC donors. FIG. 39D shows adaptive NK cell frequency gated as in FIG. 39A between CMV seropositive (N = 21 donors) and seronegative donors (N = 24 donors), Mann Whitney LTTest.[000100] FIGS. 40A-D show survival curves of 45 donor PBMC challenge assay, related to FIGS.37A-D. Number indicates PBMC donor. Brackets indicate the CMV serostatus of the donor. N = 1 technical replicate curves per condition.[000101] FIGS. 41 A-G show generation of TASR-expressing CAR-T cells by multiplexed non-viral HDR into primary T cells and use in B cell killing assay, related to FIG. 37E. FIG. 41 A shows knock-in efficiency of T cells 4-days post-editing with CD300a TASR atB2M loci and anti-CD19 CAR at TRAC loci. Post-electroporation, T cells were plated into either standard media or media containing the indicated small molecules for 24 hours prior to exchange back into standard media.224909-7891-2381.1(right panel) fold enhancement in editing efficiency of single KI cells after small molecule treatment as in FIG. 41A. N::= 3 editing runs. FIG. 41B shows the process overview for generation, purification, and expansion cloaked CAR-T cells. Stimulated T cells are edited in a single step with B2M and TRAC RNPs and linear dsDNA HDR templates encoding cloaking ligand and anti-CDl 9 CAR. The cloaking HDR template integrates at the start codon of B2M gene while the CAR integrates into TRAC in bicistronic format via 2A cleavable peptide. Both constructs code for BGH polyA tail. Cloak ligand expressing cells are enriched using an appropriate antibody by magnetic enrichment, followed by selection expansion of CAR expressing cells by addition of mitomycin-C treated Raji feeder cells FIG. 41C shows phenotypes of three engineered CAR-T cells by flow cytometry expressed the indicated cloaking transgene at B2M locus, gated live single cells. The same anti-CDl 9 CAR is used for all CAR-T cells FIG. 41 D shows a co-culture plate map and fluorescent barcoding scheme. PBMCs are seeded at 250,000 per well, and then edited CAR-T cells are added to the indicated CAR-T: PBMC ratio. Every column represents one unique PBMC: CAR-T cell pair and cytotoxicity curve. At the end of co-culture, rows are barcoded by staining with unique combinations of fluorescent anti-CD45 antibody along with phenotyping antibodies. The plate is washed and then every' column is pooled into one well and acquired on flow cytometry. FIG. 41E shows a gating scheme for identification of B cells and barcode demultiplexing. FIG. 41F shows B cell counts in the absence of CAR-T cells. PBMCs are seeded at equal density in one column, fluorescently barcoded, and then counted on flow cytometry as in FIGS. 41D-E. FIG. 41G shows a comparison of B cell lysis measurements by conventional and fluorescent barcoding flow cytometry. RQR8-expressing CAR-T cells were co-cultured with PBMCs at the indicated ratios in four identical columns on the plate. Two columns underwent fluorescent barcoding flow cytometry as in FIGS. 41D-E, while two other columns were directly stained with phenotyping antibodies and then acquired on flow cytometry without pooling. N = 2 technical replicates per kill curve.[000102] FIGS. 42A-B show CD300a TASR expressed in a model allogeneic anti-CD19 CAR-T cell exhibit enhanced protection from NK cells with no effect on CAR-mediated killing potency, related to FIG. 37E. FIG. 42A shows cytotoxicity of indicated cloaked anti-CDl 9 CAR-T cells against CD19-expressing Raji cells. Negative control (“No CAR”) contains CD300a TASR integrated into the B2M loci without anti-CDl 9 CAR. N = 3 technical replicate curves per condition. FIG 42B shows NK challenge of cloaked anti-CD19 CAR-T cells containing the234909-7891-2381.1indicated cloaking transgene with a 2Adom and 2Cdom NK donor. N = 3 technical replicates curves per condition.[000103] FIG. 43 shows that CD300a cloaking (using a CD300a TASR) outperforms HLA-E across 45 donor samples in an NK challenge assay. Target DKO edited T cells without inhibitory factor, with HLA-E, or with the CD300a TASR were co-cultured for 72 hours with effector cells (PBMCs, including NK cells) from 45 different donors selected for diverse age, gender, ethnicity, and CMV status. The graph plots the IC50 values of the 3 different edited T cell conditions across 5 bins of NK donors based on the percentage of NKG2A- / NKG2C+ NK ceils from a respective donor. Only T cells comprising the CD300a TASR were protected when challenged with NK cells across all 5 bins of donors.[000104] FIG. 44 shows that the B2M / CIITA KO + CD300a TASR outcompetes other cloaking configurations against T and NK cell alloreactivity. Similar to the experiment described in figure 29 and example 27, a pool of DKO T cells that have been engineered to express the identified transgene or wt cells is challenged with either T effectors, NK effectors, or NK and T effectors. Only target T cells comprising the CD300a TASR were protected when challenged with both T and NK cells.[000105] FIGS. 45A-C show that the CD300a TASR comprising a CD300a VHH can provide equal or better protection in comparison to a CD300a scFv TASR, FIG. 45 A provides an experimental overview of the survival assay and schematics of the VHH and scFv CD300a TASRs that were tested forNK protection. FIG. 45B shows phenotypes of T cells that express the indicated CD300a TASRs 24 hours (DI ) after electroporation with mRNA encoding the identified TASR or GFP (control). The amino acid sequences of the VHHs of CD300a TASRs 20-37 are provided in SEQ ID NOs: 155-172, respectively. FIG. 45C shows survival of engineered T cells containing the indicated CD300a TASRs in an NK cell cytotoxicity assay.[000106] FIGS. 46A-D shows (FIG. 46A) an exemplary differentiation scheme wherein iPSCs were engineered to express a CAR, a NK cell inhibitor, CD300a TASR, and an Immuglobulin degrading protein, IdeS. Additionally, the expression of MHC class I and II molecules and ab T cell receptor (TCRab) w7as eliminated in order to prevent T cell recognition and GvHD; (FIG. 46B) iPSC differentiation through Stage 1 produces CD34+CD43+ progenitors; (FIG. 46C) WT iPSC-derived T cells robustly generate CD4+ / CD8+ immature T cells that express CD3 and TCRab; and (FIG.244909-7891-2381.146D) through a controllable process, WT iPSC-derived T cells mature into CD8SP and CD4SP «PT cells.[000107] FIGS. 47A-C show how iPSC-CAR-T differentiation platforms of the present disclosure offer tunable control over the mix of CD4+ and CD8+ single positive a[3T cells. (FIG. 47A) Schematic of the challenge posed by elimination of the TCR and introduction of the CAR into engineered cells; (FIG. 47B) Engineered iPSC-CAR-T lines include an anti-BCMA or anti-CD19 CAR in addition to a) IDP and CD300a TASR, b) knockout of HLA class I and II, and c) knockout of the TCRab complex; (FIG. 47C) CAR engineered, Stage 2 CD4+CD8+ DP T cells are differentiated to single positive CD4+ or CD8+ T cells in a process that can be purposely shifted by alteration of conditions; (FIG. 47D) Fold change from iPSCs and ending viability.[000108] FIGS. 48A-D show iPSC-CAR-T differentiation traverses biologically recognizable path. (FIG. 48A) UMAP projection of a subset of a public thymocyte development atlas (Park et al.) integrated with internal adult primary T cell data. Colored by development stage; (FIG. 48B) Projection of iPS-CAR-T data into reference UMAP space; (FIG. 48C) Differential expression of progenitor versus mature T cell genes across differentiation stage; and (FIG. 48D) Gene set enrichment analysis of Stage 4 versus Stage 2 cells reveals enrichment of cytokine, interleukin, and interferon gamma signaling.[000109] FIGS. 49A-B shows post-stage 4 iPSC-CAR-T express expected receptor and secreted protein profiles. (FIG. 49A) Surface receptor gene expression is shown across differentiation stages for iPS-CAR-T through Stage 4; and (FIG. 49B) CAR expression is shown by differentiation stage on iPS-CAR-T cells.[000110] FIGS. 50A-E show iPSC-CAR-T cells demonstrate robust T cell function in vitro. (FIG.50A) 24-hour cytotoxicity assay demonstrates short-term cytotoxicity on par with primary CAR-T in a cell engineered with anti-BCMA CAR; (FIG. 50B) IL-2 production detected by ELISA after 48-hour exposure to targets is similar to primary T cell levels; (FIG 50C) Quantification of secreted proteins important for cytotoxicity show similar levels between iPS-CAR-T and primary CAR-T. Data shown for effectors with CD 19 CAR challenged with Nalm6 targets for 48 hours, and (FIG.50D) Antigen-mediated effector expansion evident after 5-day target exposure by cell counts after verification that targets are absent. (FIG. 50E) IPSC-CAR-T cells kill multiple rounds of tumor challenge at an E: T 0.5:1 in an assay without cytokine support254909-7891-2381.1[000111] FIGS. 51A-D show acute tumor clearance in disseminated Nalm6 model, subsequent transient response in tumor rechallenge. (FIG. 51 A) Disseminated Nalm6 tumor model after 3-day engraftment period. Effector dose: 5e6 cells (!’ CAR-T), 30e6 cells (iPS-CAR-T). Tumor challenge and rechallenge occurred at relative day -3 and 36, respectively, and are denoted by dashed red vertical lines; (FIG. 51B) BLI whole body imaging of mice in study; (FIG. 51C) BLI intensity quantified over time show tumor clearance in 1 ’ CAR-T and iPS-CAR-T cohorts. Tumor rechallenge at post-effector infusion day 36 elicits a transient response from anti-CD19 iPS-CAR-T before loss of control. Effector infusion colored orange. Shaded area marks rechallenge timepoints highlighted in next panel; and (FIG. 5 ID) Overlay of all groups shows partial response of iPS-CAR-T to Nalm6 rechallenge.[000112] FIGS. 52A-C show extended persistence of circulating iPS-CAR-T cells in vivo. (FIG. 52A) Disseminated Nalm6 tumor model after 3-day engraftment period. Effector dose: 5e6 cells (1 ’ CAR-T), 30e6 cells (iPS-CAR-T). (FIG. 52B) BLI traces quantified over time show sustained tumor control in iPS-CAR-T cohorts compared to 1’ CAR-T cell cohort. Dotted red line indicates time of tumor challenge. Effector infusion colored orange. (FIG. 52C) Circulating effector cells quantified by flow cytometry at day 7 and 21 post-effector infusion. iPS-CAR-T and 1’ CAR-T cohorts have detectable circulating effectors at both time points.[000113] FIGS. 53A-C show cytolysis of NALM6 (FIG, 53A), Raji (FIG. 53B), and OCI-Lyl (FIG.53C) target cells after 72 hours of co-culture with CNTY-308 cells at E: T ratios of 2: 1, 1:1, and 0.5:1, normalized to target-only controls.[000114] FIGS. 54A-C show supernatants collected from 48-hour co-cultures of CNTY-308 effector cells with 3 different WT and CD19-KO cell lines at a 1:1 E: T ratio and CNTY-308 effector cells alone (No Targets). Concentrations of IFN-y (FIG. 54A), IL -2 (FIG. 54B), and TNFa (FIG. 54C) were quantified by Luminex assay.[000115] FIGS. 5.5A-C show supernatants collected from 48-hour co-cultures of CNTY-308 effector cells with 3 different WT and CD19-KO cell lines at a 1:1 E: T ratio and CNTY-308 effector cells alone (No Targets). Concentrations of granzyme A (FIG. 55A), granzyme B (FIG. 55B), and perforin (FIG. 55C) were quantified by Luminex assay.[000116] FIGS. 56A-B show supernatants collected from 48-hour co-cultures of CNTY-308 effector cells wdth 3 different WT and CD19-KO cell lines at a 1:1 E: T ratio and CNTY-308264909-7891-2381.1effector cells alone (No Targets). Concentrations of IL-ip (FIG. 56A) and IL-6 (FIG. 56B) were quantified by Luminex assay.[000117] FIG. 57 shows CNTY-308 was co-cultured with healthy donor PBMCs at varying T cell to PBMC ratios. B cell killing was measured 72 hours later using flow cytometry.[000118] FIGS. 58A-B show CNTY-308 was co-cultured with 3 SLE (FIG. 58A) and 2 LN (FIG.58 B) donor PBMCs at varying T cell to PBMC ratios. B cell killing was measured 72 hours later using flow cytometry.[000119] FIGS. 59A-F show supernatants collected from 48-hour co-cultures of CNTY-308 effector cells with no PBMC targets or PBMCs from 3 healthy donors at a 1:1 E: T ratio Concentrations of IFN-y (FIG. 59A), IL-2 (FIG. 59B), TNFa (FIG. 59C), granzyme A (FIG. 59D), granzyme B (FIG. 59E), or perforin (FIG. 59F) were quantified by Luminex assay.[000120] FIGS. 60A-F show supernatants were collected from 48-hour co-cultures of CNTY-308 effector cells with no PBMC targets or PBMCs from 3 SLE (FIGS. 60A-B) and 2 LN (FIGS. 60D- F) donors at a 1:1 E: T ratio. Concentrations of IFN-y (FIGS. 60 / X & 60D), IL-2 (FIGS. 60B & 60E), and TNFa (FIGS. 60C & 60F) were quantified by Luminex assay.[000121] FIGS. 61 A-F show7supernatants were collected from 48-hour co-cultures of CNTY-308 effector cells with no PBMC targets or PBMCs from 3 SLE (FIGS. 61 A-B) and 2 LN (FIGS. 61D- F) donors at a 1:1 E: T ratio. Concentrations of granzyme A (FIGS. 61 A & 6 ID), granzyme B (FIGS. 61B & 6IE), and perforin (FIGS. 61C & 6IF) were quantified by Luminex assay.[000122] FIG. 62 shows primary' CAR T and CNTY-308 cells were assessed by flow cytometry for expression of HLA-I and HLA-II.[000123] FIG. 63 shows CNTY-308, K562, and PBMCs from 3 different healthy donors were cultured in the presence or absence of IFN-y. HLA-I expression was assessed 3 days later by flow cytometry.[000124] FIG. 64 shows CNTY-308, K562, and PBMCs from 3 different healthy donors were cultured in the presence or absence of IL-2. HLA-II expression was assessed 3 days later by flow cytometry.[000125] FIG. 65 shows CNTY-308, primary' CAR-T cells, and B2. WK0 primary T cells were stained with CTV dye and cultured with 3 different NK donor cells at various NK to T cell ratios. The number of CT\ / + cells was determined 24 hours later. Percent survival was calculated to assess NK cell toxicity against each T cell subset.274909-7891-2381.1[000126] FIG. 66 shows primary7CD19 CAR-T and CNTY-308 cells were incubated with anti-CD52 antibody. Antibody cleavage was measured by flow cytometry.[000127] FIG. 67 shows CNTY-308 and primary CAR-T cells were stained with CTV dye, incubated in the presence or absence of anti-CD52 antibody, and co-cultured with 3 different NK donor cells at various E: T ratios in triplicate The number of CTV+ cells was determined 24 hours later. Percent survival was calculated to assess NK-mediated ADCC against each T cell subset.[000128] FIG. 68 shows CNTY-308 and primary CAR-T cells were incubated with anti-CD52 antibody for 15 minutes at 37°C. Complement was added and samples were analyzed 2 hours later for viability using CellTiter-Glo®. Percent specific lysis was calculated to assess complement- mediated cytotoxicity against CNTY-308.[000129] FIGS. 69A-C show percent body weight change of tumor-bearing mice treated with CNTY-308. Alice were treated with 10 / 106CNTY-308 cells (FIG. 69 A), 20x]06CNTY-308 cells (FIG. 69B), or 40×106CNTY-308 cells (FIG. 69C). Some groups were treated with a single dose of CNTY-308 (A) or a repeat dose of CNTY-308 (■), while other groups served as vehicle controls (O, O). Arrows represent dosing days.[000130] FIGS. 70A-C show tumor burden by BLI of mice treated with a single dose of CNTY-308. Mice were treated with 10 106CNTY-308 cells (FIG. 70A), 20x 106CNTY-308 cells (FIG.70B), or 40×106CNTY-308 cells (FIG. 70C). Some groups were treated with a single dose of CNTY-308 (A), while other groups served as vehicle controls (O, O). Data are represented as the individual BLI measurements of each mouse over time. Grey zone represents ± 3 standard deviations of the mean baseline total flux (Day -4) prior to NAL 6 tumor cell engraftment. Dashed line marks Day 0, when animals were dosed with CNTY-308.[000131] FIGS. 71 A-C show tumor burden by BLI of mice treated with a repeat dose of CNTY-308. Mice were treated with 10x 10 CNTY-308 cells (FIG. 71 A), 20x 106CNTY-308 cells (FIG.7 IB), or 40x 106CNTY-308 cells (FIG. 71 C) Some groups were treated with a repeat dose of CNTY-308 (■), while other groups served as vehicle controls (O, O). Data are represented as the individual BLI measurements of each mouse over time. Grey zone represents ± 3 standard deviations of the mean baseline total flux (Day -4) prior to NALM6 tumor cell engraftment. Dashed line marks Day 0 and Day 7, when animals were dosed with CNTY-308.[000132] FIG. 72 shows endpoint tumor burden (Day 21) in mice treated with CNTY-308. Alice were treated with increasing doses of CNTY-308, administered either as a single dose or repeated 284909-7891-2381.1dose. Data are represented as the total flux [photons / second] ± SEM. Grey zone represents ± 3 standard deviations of the mean baseline total flux (Day -4) prior to NALM6 tumor cell engraftment.[000133] FIGS. 73A-C show presence of CNTY-308 cells in the blood of tumor-bearing NSG mice. Mice were treated with 10x 106CNTY-308 cells (FIG. 73 A), 20 I ()'' CNTY-308 cells (FIG. 73B), or 40 It)6CNTY-308 cells (FIG. 73C). Some groups were treated with a single dose of CNTY-308 (A) or repeated dose of CNTY-308 (■ ), while other groups served as vehicle controls (O, O).[000134] FIGS. 74A-B show area under the curve of CNTY-308 cells persisting in the blood of tumor-bearing mice. Mice were treated with increasing doses of CNTY-308, administered either as a single dose or repeated dose. FIG. 74A compares the effect of dose on CNTY-308 persistence. FIG. 74B compares the effect of dose schedule on CNTY-308 persistence.[000135] FIG. 75 shows CNTY-308 and Jurkat cells were thawed and plated at a total of 1x107cells in 10 mL of CFP media in triplicate T25 flasks. 50 lU / 'mL of IL-2 was added at the start of the experiment and replenished every 3 to 4 days during the experiment, to each replicate flask of each cell type, in one arm of the experiment. The other arm received no cytokine to each replicate flask of each cell type. The number of viable cells was determined by NC-200 counts every 3 to 4 days. Total viable cells were calculated to assess autonomous cell growth in the presence or absence of IL. -2.[000136] FIG. 76 shows CTV labeling was used to measure T-cell proliferation after 5 days of stimulation with plate-bound aCD3 antibody at 100 ng / mL. The percentage of proliferating cells within the CD2+T cell population for 3 healthy PBMC donors and CNTY-308 were determined. All PBMC donors exhibited significant proliferation by Day 5, whereas CNTY-308 showed no detectable proliferation.[000137] FIG. 77 shows human RBCs were incubated with serial dilutions of CNTY-308 to assess hemolytic activity. The %HR was calculated relative to a complete lysis control (10% Triton X-100) and a negative control (phosphate-b offered saline),[000138] FIGS. 78A-F shows in vivo B cell killing in a primary B cell target model in the context of active GVFID B cell depletion observed in the blood of PBMC-engrafted mice treated with iPS-CAR-T (FIG. 78A). iTs detectable in the blood, generally higher in mice that were not engrafted with PBMCs (FIG. 78B). iT cells in blood are nearly uniformly CART (FIG. 78C). Substantial B cell depletion observed in the spleens of PBMC-engrafted mice treated with iPS-CAR-T (FIG.294909-7891-2381.178D). iTs detectable in the spleen, generally higher in mice engrafted with PBMCs (FIG. 78E). B cell depletion reduced spleen mass by -50% to that of naive spleens (FIG, 78F),[000139] FIGS. 79A-B show histology (FIG. 79A) and quantification (FIG. 79B) of near-complete ablation of B cells in the spleen 7 days after iPS-CAR-T treatment.[000140] FIGS. 80A-B show histology (FIG. 80A) and quantification (FIG. 80B) of near-complete ablation of B cells in bone marrow from femur 7 days after iPS-CAR-T treatment.[000141] Various aspects of the embodiments of the present disclosure now will be described more fully hereinafter. Such aspects may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey its scope to those skilled in the art.DETAILED DESCRIPTION OF CERTAIN EMBODIMENTSI. Definitions[000142] Reference will now be made in detail to certain embodiments of the invention. While the invention will be described in conjunction with such embodiments, it will be understood that they are not intended to limit the invention to those embodiments On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents, which may be included within the invention as defined by the appended claims.[000143] Before describing the present teachings in detail, it is to be understood that the disclosure is not limited to specific compositions or process steps, as such may vary. It should be noted that, as used in this specification and the appended claims, the singular form “a”, “an” and “the” include plural references unless the context clearly dictates otherwise. Thus, for example, reference to “a nucleic acid” includes a plurality of nucleic acids, reference to “a cell” includes a plurality of cells, and the like.[000144] Numeric ranges are inclusive of the numbers defining the range. Measured and measurable values are understood to be approximate, taking into account significant digits and the error associated with the measurement. Also, the use of “comprise”, “comprises”, “comprising”, “contain”, “contains”, “containing”, “include”, “includes”, and “including” are not intended to be limiting. It is to be understood that both the foregoing general description and detailed description are exemplary and explanatory only and are not restrictive of the teachings.304909-7891-2381.1[000145] Unless specifically noted in the above specification, embodiments in the specification that recite “comprising” various components are also contemplated as “consisting of’ or “consisting essentially of’ the recited components; embodiments in the specification that recite “consisting of’ various components are also contemplated as “comprising” or “consisting essentially of” the recited components; and embodiments in the specification that recite “consisting essentially of’ various components are also contemplated as “consisting of’ or “comprising” the recited components (this interchangeability does not apply to the use of these terms in the claims).[000146] The section headings used herein are for organizational purposes and are not to be construed as limiting the disclosed subject matter in any way. In the event that any document or other material incorporated by reference contradicts any explicit content of this specification, including definitions, this specification controls.[000147] The term “or” is used in the inclusive sense, i.e., equivalent to “and / or,” unless the context requires otherwise.[000148] As used herein, “antibody” refers to a protein, or polypeptide sequences derived from an immunoglobulin molecule, which specifically binds to an antigen. Antibodies can be intact immunoglobulins of polyclonal or monoclonal origin, or fragments thereof, and can be from natural or from recombinant sources, or from a library with random or intentionally designed CDR sequences in an antibody construct[000149] As used herein, “CDR” refers to the complementarity determining region amino acid sequences of an antibody (or antibody fragment) which are the hypervariable domains of immunoglobulin heavy and light chains. There are three heavy chain and three light chain CDRs (or CDR regions) in the variable portion of an immunoglobulin. Thus, “CDRs” as used herein may refer to all three heavy chain CDRs, or all three light chain CDRs (or both all heavy and all light chain CDRs, if appropriate). CDRs provide the majority of contact residues for the binding of the antibody to the antigen or epitope. The CDR sequences of antibodies can be determined by the Kabat numbering system (Kabat et al; (Sequences of proteins of Immunological Interest NIH, 1987); alternatively they can be determined using the Chothia numbering system (Al-Lazikani et al., (1997) JMB 273, 927-948), the contact definition method (MacCallum R. M., and Martin A. C R. and Thornton J. M, (1996), Journal of Molecular Biology, 262 (5), 732-745) or any other established method for numbering the residues in an antibody and determining CDRs known to the skilled man in the art. Other numbering conventions for CDR sequences available to a skilled 314909-7891-2381.1person include “AbM” (University of Bath) and “contact” (University College London) methods. The minimum overlapping region using at least two of the Kabat, Chothia, AbM and contact methods can be determined to provide the “minimum binding unit.” The minimum binding unit may be a sub-portion of a CDR.[000150] As used herein, “antigen-binding domain” means the portion of an antibody that is capable of specifically binding to an antigen or epitope. One example of an antigen-binding domain is an antigen-binding domain formed by a VH -VL dimer of an antibody. Another example of an antigen-binding domain is an antigen-binding domain formed by diversification of certain loops from the tenth fibronectin type III domain of an Adnectin.[000151] As used herein, “antibody fragment” or “antibody binding domain” refer to at least one portion of an antibody, or recombinant variants thereof, that, contains the antigen binding domain, i.e., an antigenic determining variable region of an intact antibody, that is sufficient to confer recognition and specific binding of the antibody fragment to a target, such as an antigen and its defined epitope. Examples of antibody fragments include, but are not limited to, Fab, Fab’, F(ab’)2, and Fv fragments, single-chain (sc) variable (“scFv”) antibody fragments, linear antibodies, single domain antibodies (abbreviated “sdAb”) (either VL or VH), diabodies, minibodies, nanobodies (also known as variable domain on a heavy chain (VHH) antibodies, such as camelid VHH antibodies, See, e.g., Bever et al,, Analytical and Bioanalytical Chemistry. 408(22): 5985-6002 (2016)), and multi-specific antibodies formed from antibody fragments. Exemplary binding domains useful in the disclosed embodiments can also include a cytokine, a ligand, or a peptide (such as an adnectin or a designed ankyrin repeat protein (DARPin) (See, e.g., Rafiq et al., Nat Rev Clin Oncol. 2020;17:147-167). In certain embodiments, an NKG2A binding domain or a CD300a binding domain provided herein is an antibody fragment. For a review of certain antibody fragments, see Hudson et al. Nat. Med. 9:129-134 (2003). For a review of scFv fragments, see, e.g., Pluckthün, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., (Springer- Verlag, New York), pp. 269-315 (1994); see also WO 93 / 16185; and U. S. Patent Nos. 5,571,894 and 5,587,458. Diabodies are antibody fragments with two antigen-binding sites that may be bivalent or bispecific. See, for example, EP 404,097; WO 1993 / 01161; Hudson et al., Nat. Med.9:129-134 (2003); and Hollinger et al., Proc. Natl. Acad. Sei. USA 90: 6444-6448 (1993).Triabodies and tetrabodies are also described in Hudson et al., Nat. Med. 9:129-134 (2003). Single¬ domain antibodies are antibody fragments comprising all or a portion of the heavy chain variable 324909-7891-2381.1domain or all or a portion of the light chain variable domain of an antibody. In certain embodiments, a single-domain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see, e.g., U. S. Patent No. 6,248,516 Bl). In certain embodiments, an antibody provided herein is a chimeric antibody Certain chimeric antibodies are described, e.g., in U. S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)). In one example, a chimeric antibody comprises a non-human variable region (e.g., a variable region derived from a mouse, rat, hamster, rabbit, or non-human primate, such as a monkey) and a human constant region. In a further example, a chimeric antibody is a “class switched” antibody in which the class or subclass has been changed from that of the parent antibody. Chimeric antibodies include antigen-binding fragments thereof.[000152] As used herein, “VHH,” “VHH antibody,” or “nanobody” refers to the antigen binding fragment of heavy chain only antibodies (HcAb), which lack VL domains (See, e.g., Bever et al., Analytical and Bioanalytical Chemistry. 408(22): 5985-6002 (2016)). Heavy chain only antibodies are produced in nature by camelids and sharks. A variable domain on a heavy chain (VHH) typically comprise a single polypeptide chain. Compared to mAbs and other antibody fragments, VHHs are typically smaller in size.[000153] As used herein, “scFv” refers to a fusion protein comprising at least one antibody fragment comprising a variable region of a light chain and at least one antibody fragment comprising a variable region of a heavy chain, wherein the light and heavy chain variable regions are contiguously linked via a short flexible polypeptide linker, and capable of being expressed as a single polypeptide chain, and wherein the scFv retains the specificity of the intact antibody from which it is derived.[000154] “Heavy chain variable region” or “VH” (or, in the case of single domain antibodies, e.g., nanobodies, “VHH”) with regard to an antibody refers to the fragment of the heavy chain that contains three CDRs interposed between flanking stretches known as framework regions, these framework regions are generally more highly conserved than the CDRs and form a scaffold to support the CDRs. Unless specified, as used herein an scFv may have the VL and VH variable regions in either order, e.g., with respect to the N-terminal and C-terminal ends of the polypeptide, the scFv may comprise VL-linker-VH or may comprise VH-linker-VL.334909-7891-2381.1[000155] As used herein, ‘’antibody heavy chain,” refers to the larger of the two types of polypeptide chains present in antibody molecules in their naturally occurring conformations, and which normally determines the class to which the antibody belongs.[000156] As used herein, “antibody light chain,” refers to the smaller of the two types of polypeptide chains present in antibody molecules in their naturally occurring conformations. Kappa (“K”) and lambda (“X”) light chains refer to the two major antibody light chain isotypes.[000157] As used herein, “antigen” or “Ag” refers to a molecule that is capable of being bound specifically by an antibody, or otherwise provokes an immune response. This immune response may involve either antibody production, or the activation of specific immunologically competent cells, or both.[000158] “Humanized” forms of non-human antibodies are chimeric antibodies that contain minimal sequence derived from the non-human antibody. A humanized antibody is generally a human antibody (recipient antibody) in which residues from one or more CDRs are replaced by residues from one or more CDRs of a non-human antibody (donor antibody). The donor antibody can be any suitable non-human antibody, such as a mouse, rat, rabbit, chicken, or non-human primate antibody having a desired specificity, affinity, or biological effect. In some instances, selected framework region residues of the recipient antibody are replaced by the corresponding framework region residues from the donor antibody. Humanized antibodies may also comprise residues that are not found in either the recipient antibody or the donor antibody. Such modifications may be made to further refine antibody function. For further details, see Jones et al., Nature, 1986, 321:522-525; Riechmann et al., Nature, 1988, 332:323-329; and Presta, Curr. Op. Struct. Biol., 1992, 2:593-596, each of which is incorporated by reference in its entirety.[000159] A “human antibody” is one which possesses an amino acid sequence corresponding to that of an antibody produced by a human or a human cell, or derived from a non-human source that utilizes a human antibody repertoire or human antibody-encoding sequences (e.g., obtained from human sources or designed de novo). Human antibodies specifically exclude humanized antibodies.[000160] “Affinity” refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., an antibody) and its binding partner (e.g., an antigen or epitope). Unless indicated otherwise, as used herein, “affinity” refers to intrinsic binding affinity, which reflects a 1: 1 interaction between members of a binding pair (e g., antibody and antigen or epitope). The affinity of a molecule X for its partner Y can be represented by the dissociation 344909-7891-2381.1equilibrium constant (KD). The kinetic components that contribute to the dissociation equilibrium constant are described in more detail below. Affinity can be measured by common methods known in the art, including those described herein, such as surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®).[000161] With regard to the binding of an antibody or fragment thereof to a target molecule, the terms “bind,” “specific binding,” “specifically binds to,” “specific for,” “selectively binds,” and “selective for” a particular antigen (e.g., a polypeptide target) or an epitope on a particular antigen mean binding that is measurably different from a non-specific or non-selective interaction (e.g., with a non-target molecule). Specific binding can be measured, for example, by measuring binding to a target molecule and comparing it to binding to a non-target molecule. Specific binding can also be determined by competition with a control molecule that mimics the epitope recognized on the target molecule. In that case, specific binding is indicated if the binding of the antibody to the target molecule is competitively inhibited by the control molecule.[0001 2] The term “autologous” refers to any material derived from the same individual to whom it is later to be re-introduced into the individual.[0001 3] The term “allogeneic” refers to any material derived from a different animal of the same species or different patient as the individual to whom the material is introduced. Two or more individuals are said to be allogeneic to one another when the genes at one or more loci are not identical. In some aspects, allogeneic material from individuals of the same species may be sufficiently unlike genetically to interact antigenically.[000164] The term “treating” (and variations thereof such as “treat” or “treatment”) refers to clinical intervention in an attempt to alter the natural course of a disease or condition in a subject in need thereof. Treatment can be performed both for prophylaxis and during the course of clinical pathology. Desirable effects of treatment include preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis. As used herein, “preventing” refers to the prevention of the disease or condition, e.g., tumor formation, in the patient. For example, if an individual at risk of developing a tumor or other form of cancer is treated with the methods of the present disclosure and does not later develop the tumor or other form of cancer, then the disease has been prevented, at least over a period of time, in that individual.354909-7891-2381.1[000165] As used herein, the term “subject” means a mammalian subject. Exemplary subjects include humans, monkeys, dogs, cats, mice, rats, cows, horses, camels, goats, rabbits, and sheep. In certain embodiments, the subject is a human. A “patient” is a subject suffering from or at risk of developing a disease, disorder or condition or otherwise in need of the compositions and methods provided herein. In some embodiments, the subject has cancer, e.g., a cancer described herein.[000166] The terms “cancer” or “tumor” as used herein refer to a disease characterized by the rapid and uncontrolled growth of aberrant cells. Cancer cells can spread locally or through the bloodstream and lymphatic system to other parts of the body. Examples of various cancers are described herein and include but are not limited to, hematological cancers, including both lymphoid and myeloid malignancies. Myeloid malignancies include, but are not limited to, acute myeloid (or myelocytic or myelogenous or myeloblastic) leukemia (undifferentiated or differentiated), acute promyeloid (or promyelocytic or promyelogenous or promyeloblastic) leukemia, acute myelomonocytic (or myelomonoblastic) leukemia, acute monocytic (or monoblastic) leukemia, erythroleukemia and megakaryocytic (or megakaryoblastic) leukemia. These leukemias may be referred together as acute myeloid (or myelocytic or myelogenous) leukemia (AML). Myeloid malignancies also include myeloproliferative disorders (MPD) which include, but are not. limited to, chronic myelogenous (or myeloid) leukemia (CML), chronic myelomonocytic leukemia (CMML), essential thrombocythemia (or thrombocytosis), and polycythemia vera (PCV). Myeloid malignancies also include myelodysplasia (or myelodysplastic syndrome or MDS), which may be referred to as refractory anemia (RA), refractory anemia with excess blasts (RAEB), and refractory' anemia with excess blasts in transformation (RAEBT), as well as myelofibrosis (MFS) with or without agnogenic myeloid metaplasia.[000167] Hematopoietic cancers also include lymphoid malignancies, which may affect the lymph nodes, spleens, bone marrow, peripheral blood, and / or extranodal sites. Lymphoid cancers include B-cell malignancies, which include, but are not limited to, B-cell non-Hodgkin’s lymphomas (B-NHLs). B-NHLs may be low-grade (or indolent), intermediate-grade (or aggressive) or high-grade (very aggressive). Indolent B-cell lymphomas include follicular lymphoma (FL); small lymphocytic lymphoma (SLL); marginal zone lymphoma (MZL) including nodal MZL, extranodal N1ZL, splenic MZL and splenic MZL with villous lymphocytes; lymphoplasmacytic lymphoma (LPL); and mucosa-associated-lymphoid tissue (MALT or extranodal marginal zone) lymphoma. Intermediategrade B-NHLs include mantle cell lymphoma (MCL) with or without leukemic involvement,364909-7891-2381.1diffuse large cell lymphoma (DLBCL), follicular large cell (or grade 3 or grade 3B) lymphoma, and primary mediastinal lymphoma (PML). High-grade B-NHLs include Burkitt's lymphoma (BL), Burkitt-like lymphoma, small non-cleaved ceil lymphoma (SNCCL) and lymphoblastic lymphoma. Other B-NHLs include immunoblastic lymphoma (or immunocytoma), primary effusion lymphoma, HI V associated (or / AIDS related) lymphomas, and post-transplant lymphoproliferative disorder (PTLD) or lymphoma. B-cell malignancies also include, but are not limited to, chronic lymphocytic leukemia (CLL), prolymphocytic leukemia (PLL), Waldenstrom’s macroglobulinemia (WM), hairy cell leukemia (HCL), large granular lymphocyte (LGL) leukemia, acute lymphoid (or lymphocytic or lymphoblastic) leukemia, and Castleman's disease. NHL may also include T-cell non-Hodgkin’s lymphomas (T-NHLs), which include, but are not limited to T-cell non-Hodgkin’s lymphoma not otherwise specified (NOS), peripheral T-cell lymphoma (PTCL), anaplastic large cell lymphoma ( ALCL), angioimmunoblastic lymphoid disorder (AILD), nasal natural killer (NK) cell / T-cell lymphoma, gamma / delta lymphoma, cutaneous T cell lymphoma, mycosis fungoides, and Sezary' syndrome.[000168] Hematopoietic cancers also include Hodgkin's lymphoma (or disease) including classical Hodgkin's lymphoma, nodular sclerosing Hodgkin's lymphoma, mixed cellularity Hodgkin's lymphoma, lymphocyte predominant (LP) Hodgkin's lymphoma, nodular LP Hodgkin's lymphoma, and lymphocyte depleted Hodgkin's lymphoma. Hematopoietic cancers also include plasma cell diseases or cancers such as multiple myeloma (MM) including smoldering MM, monoclonal gammopathy of undetermined (or unknown or unclear) significance (MGUS), plasmacytoma (bone, extramedullary), lymphoplasmacytic lymphoma (LPL), Waldenstrom's Macroglobulinemia, plasma cell leukemia, and primary amyloidosis (AL). Hematopoietic cancers may also include other cancers of additional hematopoietic cells, including polymorphonuclear leukocytes (or neutrophils), basophils, eosinophils, dendritic cells, platelets, erythrocytes and natural killer cells. Tissues which include hematopoietic cells referred herein to as “hematopoietic cell tissues” include bone marrow; peripheral blood; thymus; and peripheral lymphoid tissues, such as spleen, lymph nodes, lymphoid tissues associated with mucosa (such as the gut-associated lymphoid tissues), tonsils, Peyer's patches and appendix, and lymphoid tissues associated with other mucosa, for example, the bronchial linings.[000169] Exemplary solid cancers that may be treated using a method provided herein include brain cancer, glioma, glioblastoma, neuroblastoma, prostate cancer, colorectal cancer, pancreatic cancer,374909-7891-2381.1Medulloblastoma, melanoma, cervical cancer, gastric cancer, ovarian cancer, lung cancer, cancer of the head, thyroid carcinoma, cholangiocarcinoma, pancreatic adenocarcinoma, skin cutaneous melanoma, colon adenocarcinoma, rectum adenocarcinoma, stomach adenocarcinoma, esophageal carcinoma, head and neck squamous cell carcinoma, breast invasive carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, non-small cell lung carcinoma, mesothelioma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, uterine cancer, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, thyroid cancer, neuroblastom, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms of the endocrine or exocrine pancreas, medullary thyroid cancer, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, or prostate cancer.[000170] The term “pharmaceutical composition” refers to a preparation which is in such form as to permit the biological activity of an active ingredient contained therein to be effective in treating a subject, and which contains no additional components which are unacceptably toxic to the subject in the amounts provided in the pharmaceutical composition.[000171] The terms “increase” and “activate” refer to an increase of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or greater in a recited variable.[000172] The terms “reduce” and “inhibit” refer to a decrease of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 2-fold, 3- fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or greater in a recited variable.[000173] The term “agonize” refers to the activation of receptor signaling to induce a biological response associated with activation of the receptor. An “agonist” is an entity that binds to and agonizes a receptor.[000174] The term “antagonize” refers to the inhibition of receptor signaling to inhibit a biological response associated with activation of the receptor. An “antagonist” is an entity that binds to and antagonizes a receptor.[000175] The term “conservative sequence modifications” refers to amino acid modifications that do not significantly affect or alter the binding characteristics of the antibody or antibody fragment containing the amino acid sequence. Such conservative modifications include amino acid384909-7891-2381.1substitutions, additions and deletions. Modifications can be introduced into an antibody or antibody fragment of the disclosure by standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions are ones in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, one or more amino acid residues within a TFP of the disclosure can be replaced with other amino acid residues from the same side chain family and the altered TFP can be tested using the functional assays described herein.[000176] The term “encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (e.g., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene, cDNA, or RNA, encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, can be referred to as encoding the protein or other product of that gene or cDNA.[000177] Unless otherwise specified, a “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some version contain one or more introns.[000178] The term “endogenous” refers to any material from or produced inside an organism, cell, tissue or system.394909-7891-2381.1[000179] The term “exogenous” refers to any material introduced from or produced outside an organism, cell, tissue or system.[000180] The term “expression” refers to the transcription and / or translation of a particular nucleotide sequence, such as driven by a promoter.[000181] A “vector” and related terms used herein refers to a nucleic acid molecule (e.g., DNA or RNA) which can be operably linked to foreign genetic material (e.g., nucleic acid transgene).Vectors can be used as a vehicle to introduce foreign genetic material into a cell (e.g., host cell). Vectors can include at least one restriction endonuclease recognition sequence for insertion of the transgene into the vector. Vectors can include at least one gene sequence that confers antibiotic resistance or a selectable characteristic to aid in selection of host cells that harbor a vector-transgene construct Vectors can be single-stranded or double-stranded nucleic acid molecules, and can be linear or circular nucleic acid molecules. A donor nucleic acid used for gene editing methods employing zinc finger nuclease, TALEN or CRISPR / Cas can be a type of a vector. One type of vector is a “plasmid,” which refers to a linear or circular double stranded extrachromosomal DNA molecule which can be linked to a transgene, and is capable of replicating in a host cell, and transcribing and / or translating the transgene. A viral vector typically contains viral RNA or DNA backbone sequences which can be linked to the transgene. The viral backbone sequences can be modified to disable infection but retain insertion of the viral backbone and the co-linked transgene into a host cell genome. Examples of viral vectors include retroviral, lentiviral, adenoviral, adeno-associated, baculoviral, papovaviral, vaccinia viral, herpes simplex viral and Epstein Barr viral vectors. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors comprising a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. A vector may be a transfer vector, i e., a composition of matter which comprises an isolated nucleic acid and which can be used to deliver the isolated nucleic acid to the interior of a cell. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “transfer vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to further include non-plasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, a polylysine compound, liposome, and the like.404909-7891-2381.1Examples of viral transfer vectors include, but are not limited to, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like. A vector may be an expression vector, i.e., a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression can be supplied by the host cell or in an in vitro expression system. Further examples of regulatory sequences and other elements for expression are described in, for example, Goeddel, 1990, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, Calif, and Baron et al., 1995, Nucleic Acids Res. 23:3605-3606 Expression vectors include all those known in the art, including cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide.[000182] A transgene is “operably linked” to a vector when there is linkage between the transgene and the vector to permit functioning or expression of the transgene sequences contained in the vector. In one embodiment, a transgene is “operably linked” to a regulatory sequence when the regulatory sequence affects the expression (e.g., the level, timing, or location of expression) of the transgene.[000183] The terms “transfected” or “transformed” or “transduced” or other related terms used herein refer to a process by which exogenous nucleic acid (e.g., transgene) is transferred or introduced into a host cell (e.g., an engineered cell described herein). A “transfected” or “transformed” or “transduced” host cell is one which has been transfected, transformed or transduced with exogenous nucleic acid (transgene). The host cell includes the primary subject cell and its progeny.[000184] The terms “peptide,” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that can comprise a protein’s or peptide’s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for414909-7891-2381.1example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. A polypeptide includes a natural peptide, a recombinant peptide, or a combination thereof.[000185] The term “promoter” refers to a DNA sequence recognized by the transcription machinery of the cell, or introduced synthetic machinery, that can initiate the specific transcription of a polynucleotide sequence. The term “constitutive” promoter refers to a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell under most or all physiological conditions of the cell. The term “inducible” promoter refers to a nucleotide sequence which, when operably linked with a polynucleotide which encodes or specifies a gene product, causes the gene product to be produced in a cell substantially only when an inducer which corresponds to the promoter is present in the cell. In some embodiments, a promoter is an endogenous promoter. In some embodiments, such as certain embodiments wherein a recombinant nucleic acid or vector disclosed herein is inserted into a gene locus (such as a B2M or TRAC locus) in a cell, the promoter is an endogenous B2M promoter or an endogenous TRAC promoter.[000186] The term “linker,” as used herein, refers to a chemical group or a molecule linking two adjacent molecules or moieties. Typically, the linker is positioned between, or flanked by, two groups, molecules, or other moieties and connected to each one via a covalent bond. The term “linker” as used in the context of a scFv can refer to a peptide linker comprising amino acids such as glycine and / or serine residues used alone or in combination, to link variable heavy and variable light chain regions together. In one embodiment, the flexible polypeptide linker is a Gly / Ser linker and comprises the amino acid sequence (Gly-Gly-Gly-Gly-Ser)n, where n is a positive integer equal to or greater than 1. For example, n=l, n=2, n=3, n=4, n=5, n=6, n=7, n=8, n=9 and n=10. In one embodiment, the flexible polypeptide linkers include, but are not limited to, (Gly4Ser)5 or (Gly4Ser)4. In another embodiment, the linkers include multiple repeats of (GlySer), (Gly2Ser), or (Gly3Ser). Also included within the scope of the disclosure are linkers described in WO2012 / 138475 (incorporated herein by reference). In some instances, the linker sequence comprises (G4S)n, wherein n=3 to 6. In some instances, the linker sequence comprises GGGGSGGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 30). In another embodiment, the linker is the Whitlow / 218 linker, GSTSGSGKPGSGEGSTKG (SEQ ID NO: 31). A linker may also be used 424909-7891-2381.1to link one or more binding domains (such as one or more scFvs) together, such as within a bispecific TASR as described herein. Such linkers can comprise the linkers described above and / or a cleavable peptide linker, such as those described elsewhere herein.[000187] As used herein, “NKG2A” refers to a particular member of the NKG2 family that dimerizes with CD94 to form an inhibitory receptor (CD94 / NKG2A). The NKG2 family (also known as CD 159) includes seven members: A, B, C, D, E F, and H. CD94 / NKG2 receptors are C-type lectin receptors which are expressed predominantly on the surface of NK cells and a subset of CD8+ T-lymphocytes. These receptors stimulate or inhibit cytotoxic activity ofNK cells, therefore they are divided into activating and inhibitory receptors according to their function. NKG2A, -B, - C, -E and -H form heterodimers with CD94, linked by disulfide bonds, whereas NKG2D forms homodimers. Inhibitory molecule NKG2A and its splice variant NKG2B contain immunoreceptor tyrosine-based inhibition motifs (ITIMs) in the intracellular part of the molecule. Activating molecules NKG2C and NKG2E and its splice variant NKG2H contain a positively charged residue in their transmembrane regions through which they interact with adaptor molecules containing ITAMs. Inhibitory NKG2 molecules containing ITIMs recruit the Src homology 2 domain containing phosphatases SHP-1 and SHP-2, which leads to the inhibition of cytotoxicity. Ligands of CD94 / NKG2 heterodimeric molecules include nonclassical MHC class I molecules, such as HLA-E in humans.[000188] As used herein, “cluster of differentiation 300a” or “CD300a”, also known as CMRF-35H, inhibitory receptor protein (Irp60) and IgSF12, is a member of the CD300 glycoprotein family of cell surface proteins that regulate a diverse array of immune cell processes. CD300a is found on the surface of leukocytes including neutrophils, basophils, eosinophils, mast cells, monocytes, B lymphocytes, NK cells and dendritic cells, and regulates their proliferation, differentiation, apoptosis and immunity. CD300a is an inhibitory receptor that has three classical and one non- classical ITIM motifs in its cytoplasmic tail. The ITIMs are phosphorylated by Src-family kinases, resulting in recruitment of src homology 2 domain containing protein tyrosine phosphatases (SHPs) or SH2 domain-containing inositol phosphatase (SHIP), CD300a mainly regulates the function of leukocytes by recruiting SHP-1 phosphatase. Signaling through CD300a is complex and involves many downstream signaling components. For example, CD300a engagement by agonist monoclonal antibodies has been shown to inhibit IgE-dependent Ca2+mobilization and mediator release from mast cells, SCF-mediated mast cell activation, differentiation, and survival, and downregulate NK 434909-7891-2381.1cell cytolytic activity. CD300a signaling is involved in allergy response, autoimmune disease and viral infection, and CD300a is considered a therapeutic target in these diseases.[000189] As used herein, a “signal peptide” (sometimes referred to as signal sequence, targeting signal, localization signal, localization sequence, transit peptide, leader sequence, or leader peptide) is a short peptide (such as a peptide 10-50 amino acids in length) present at the N-terminus (or nonclassically at the C-terminus or internally) to direct a synthesized protein of interest to the surface of a cell (such as to the cell membrane). Thus, signal peptides function to prompt a cell to translocate the protein, e.g., to the cellular membrane. This signal sequence is sometimes cleaved off by the cell in the maturation of a polypeptide,[000190] As used herein, an “engineered cell” or a “population of engineered cells” or related terms as used herein refer to a cell (or a population thereof) into which foreign (exogenous or transgene) nucleic acids have been introduced. The foreign nucleic acids can include an expression vector operably linked to a transgene, and the host cell can be used to express the nucleic acid and / or polypeptide encoded by the foreign nucleic acid (transgene). An engineered cell (or a population thereof) can be a cultured cell or can be extracted from a subject. The engineered cell (or a population thereof) includes the primary subject cell and its progeny without regard for the number of passages. Engineered cells encompass progeny cells. In embodiments, an engineered cell describes any cell (including its progeny) that has been modified, transfected, transduced, transformed, and / or manipulated in any way to express a recombinant nucleic acid as disclosed herein. In one example, the engineered cell (or population thereof) can be introduced with an expression vector operably linked to a nucleic acid encoding the desired recombinant nucleic acid described herein. Engineered cells and populations thereof can harbor an expression vector that is stably integrated into the host’s genome or can harbor an extrachromosomal expression vector. In embodiments, engineered cells and populations thereof can harbor an extrachromosomal vector that is present after several cell divisions or is present transiently and is lost after several cell divisions.[000191] As used herein, “hypoimmunogenic” and “hypoimmune” refer to a reduction in immunogenicity. A hypoimmunogenic cell (such as an engineered cell described herein that is hypoimmunogenic, such as a hypoimmunogenic iPSC) gives rise to a reduced immunological rejection response when transferred into an allogeneic host. For example, a hypoimmunogenic pluripotent cell is a pluripotent cell that retains its pluripotent characteristics and yet gives rise to a reduced immunological rejection response when transferred into an allogeneic host. A444909-7891-2381.1hypoimmunogenic cell described herein may induce a reduced immune response (such as compared to cell that is not hypoimmunogenic) or no immune response. Thus, “hypoimmunogenic” or “hypoimmune” refers to any amount of reduced or eliminated immune response when compared to the immune response of a parental (i.e., “wild-type”) cell prior to engineering (e.g., prior to introduction of a recombinant nucleic acid comprising an NKG2A binding domain, a CD300a binding domain, or both, as described herein). For example, relative to a wild-type cell, such a hypoimmunogenic cell may be at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% less prone to immune rejection (e.g., cytolysis) by a subject into which such cells are transplanted. In the context of an engineered cell (such as a pluripotent stem cell, as used herein, such as an iPSC), “wild-type” means a cell that that may comprise some nucleic acid changes, but did not undergo the gene editing procedures of the present disclosure (i.e., introduction of a recombinant nucleic acid comprising an NKG2A binding domain, a CD300a binding domain, or both, as described herein) to achieve hypoimmunogenicity[000192] As used herein, “stem cells” refers to cells capable of going through numerous cycles of cell division, maintaining an undifferentiated state, and having the capacity to differentiate into specialized cell types. Stem cells are further classified into three categories: totipotent, pluripotent, or multipotent somatic. A totipotent cell has the ability to form an entire organism (e.g,, a fertilized egg). As used herein, “pluripotent stem cells” refers to stem cells that lack the ability to form extraembryonic tissue and are therefore unable to generate a fetus, but have the potential to differentiate into any of the three germ layers: endoderm (e.g., the stomach lining, gastrointestinal tract, lungs, etc ), mesoderm (e.g., muscle, bone, blood, urogenital tissue, etc.) or ectoderm (e.g., epidermal tissues and nervous system tissues). Pluripotent stem cells encompass embryonic stem cells (ESCs). Exemplary' human embryonic stem cell (hESC) lines include those made available through the National Institutes of Health Human Embryonic Stem Cell Registry and the Howard Hughes Medical Institute HUES collection (as described in Cowan, C. A. et. al, New England J. Med. 350: 13. (2004), incorporated by reference herein in its entirety). Pluripotent stem cells also encompass “induced pluripotent stem cells” (iPSCs), a type of pluripotent stem cell derived from a non-pluripotent cell, typically an adult somatic cell, by inducing the expression of certain regulatory genes or by the exogenous application of certain proteins (through a process known as somatic cell “reprogramming”). Methods for the derivation of iPSCs by somatic cell reprogramming are known 454909-7891-2381.1in the art and are further described in, e.g., Takahashi and Yamanaka, Cell 126 (4): 663-76 (2006); Yu et al., Science 324(5928)1797-801 (2009), Zhou et al., Stem Cells 27 (11): 2667-74 (2009), Huangfu et al., Nature Biotechnol. 26 (7): 795 (2008); Woltjen et al., Nature 458 (7239): 766-770 (2009); and Zhou et al., Cell Stem Cell 8:381-384 (2009), each of which is incorporated by reference herein in their entirety.[000193] As used herein, the terms ‘‘inhibiting” or “reducing,” such as in the context of inhibiting or reducing NK cell cytotoxicity to an engineered cell as disclosed herein, refer to any amount of reduced or eliminated NK cell cytotoxicity to the engineered cell when compared to levels of NK cell cytotoxicity toward a parental (i.e., “wild-type”) cell prior to the engineering (e.g., prior to introduction of a recombinant nucleic acid comprising an NKG2A binding domain, a CD300a binding domain, or both, as described herein). For example, NK cell cytotoxicity to an engineered cell is “reduced” or “inhibited” when it is decreased, relative to a wild-type cell, by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%.[000194] As used herein, the term “MHC class I” refers to a class of major histocompatibility complex (MHC) molecules that are displayed on the surface of nucleated cells and platelets in vertebrates. MHC class I molecules function as part of the adaptive immune system by binding fragments of cytosolic foreign (non-self) proteins and displaying these antigens on the cell surface for recognition of cells of the immune system, e.g., cytotoxic T cells. In humans, MHC is also referred to as human leukocyte antigen (HLA). Rejection of allogenic therapeutic cells (e.g., in GvHD), such as CAR T cells, is believed to be largely driven by donor- or iPSC-derived T cell recognition of host peptide-HLA complexes through the αβ T cell receptor complex (αβTCR). Rejection is mainly driven by host NK cells, CD8+ T-cells, CD4+ T cells, and, to a lesser extent, by macrophages. HLAs include HLA-A, HLA-B, HLA-C, HLA-E, HLA-F, and HLA-G. HLA class I histocompatibility antigen alpha chain E (HLA-E) is a non-classical HI, A molecule that plays a role in recognition by NK cells. Without being bound by a particular theory, using the heterodimeric receptor CD94 / NKG2A / B / C, NK cells recognize HLA-E molecules bound to antigen at the cell surface. CD94 / NKG2A or CD94 / NKG2B engagement results in an inhibitory effect on the cytotoxic activity of the NK cell, thereby preventing cell lysis, while simultaneously causing NK cell activation.4909-7891-2381.1[000195] As used herein, the term “P2 microglobulin” (B2M) refers to a particular polypeptide component of MHC class I molecules. The B2M protein is encoded by the B2M gene. MHC class I molecules are heterodimers comprised of two polypeptide chains, a and P2-microglobulin, that are noncovalently linked via interaction of B2M and the a3 domain. Without being bound to a particular theory, p2 microglobulin is required for cell surface expression of MHC class I molecules and for stability of the peptide-binding groove. Absence of B2M expression leads to significant reductions in MHC class I molecules detectable on the cell surface.[000196] As used herein, the term “T cell receptor alpha constant” (TRAC) refers to the constant region of the T cell receptor (TCR) alpha chain. The T cell receptor (TCR) is a membrane-anchored heterodimeric protein typically consisting of the highly variable alpha (a) and beta (P) chains (encoded by TRA and TRB, respectively) expressed as part of a complex with invariant CD3 chain molecules. TCRs are found on the surface of T cells, or T lymphocytes, and recognize antigens bound to MHC molecules. Removal of the endogenous TCR by targeting TRAC (e.g., through CAR transgene knock-in) has been used to address histocompatibility barriers associated with cells derived from unrelated donors.[000197] As used herein, the term “deficient” can refer to reduced or eliminated expression and / or functionality of a particular gene product (e.g., as measured by RNA and / or protein detection methods, and / or by functional assays), such as an MHC class I molecule component (e.g., B2M), a TCR component (e.g., TRA, such as via disruption of the TRAC locus), or a molecule that regulates or controls MHC class II expression (such as a Class II major histocompatibility complex transactivator (C1ITA) gene product). In this context, “deficient” refers to any amount of reduced or eliminated expression or functionality when compared to levels of the same gene product in a parental (i.e., “wild-type”) cell prior to the deficiency (e.g., prior to engineered disruption of the gene, such as by any suitable means described herein or known in the art). In some embodiments, in a cell that is deficient in a particular gene product, the gene product is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell. In some embodiments, in a cell that is deficient in a particular gene product, the gene product is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% relative 474909-7891-2381.1to a wild-type cell. “Deficient” can also refer to a reduction or elimination of a detectable molecule in a cell, such as an MHC class I molecule or an MHC class II molecule. For example, a cell is MHC class I deficient when expression of one or more components of an MHC class I molecule (such as B2M) is reduced, relative to a wild-type cell, by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, resulting in reduced or eliminated detectable MHC class I on a cell surface by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild¬ type cell. By way of another example, a cell is MHC class II deficient when expression of one or more components of an MHC class II molecule or of a molecule that regulates or controls expression of an MHC class II molecule (such as CIITA) is reduced, relative to a wild-type cell, by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, resulting in reduced or eliminated detectable MHC class II on a cell surface by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell. By way of another example, a cell is TRAC deficient when expression of the TRAC locus is reduced, relative to a wild-type cell, by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%. In some embodiments, in a cell that is deficient in a particular gene product, the gene product is not detectable, such as by PCR, immunohistochemical staining, ELISA, western blotting, or other suitable nucleic acid or protein detection methods. In some embodiments, a gene locus is “disrupted,” resulting in a cell that is deficient for the gene product encoded by the gene. For example, insertion of a recombinant nucleic acid or vector disclosed herein into a gene locus (e.g., a B2M and / or TRAC locus) can disrupt the locus, thereby reducing or eliminating expression of the gene at the locus. Methods of disrupting a gene locus are known in the art, such as, but not limited to, homologous recombination, CRISPR- Cas9, and TALENs. Further, disruption can be achieved with (e.g, through gene knock out methods) or without (e.g., through insertion of a nucleotide sequence into the gene locus) removal of all or a portion of the targeted gene locus. Thus, methods disclosed herein can include disrupting a gene locus (such as a B2M and / or TRAC locus) using any suitable means known in the art.484909-7891-2381.1II. Recombinant Nucleic AcidsA. Overview[000198] Rejection of allogenic therapeutic cells (e g., in GvHD), such as CAR T cells, is believed to be largely driven by donor- or iPSC-derived T cell recognition of host peptide-HLA complexes through the a£ T cell receptor complex (apTCR). Rejection is mainly driven by host NK cells, CD8+ T-cells, CD4+ T cells, and, to a lesser extent, by macrophages. In the context of CAR T-cell therapy, the relative contribution of these cell types to allograft rejection may vary depending on their absolute numbers and reconstitution kinetics following preconditioning regimen. In three cohorts (acute lymphoblastic leukemia (ALL) and large B-cell malignancies), patients showed significant differences in CD8+ T cell, CD4+ T cell, and NK-cell reconstitution kinetics following autologous CD 19 CAR T-cell treatment and a preconditioning regimen (cyclophosphamide and fludarabine). (Jo et al. Nat Commun. 2022; 13(1):3453; Strati et al. Haematologica. 2021; 106(10); Wang et al. Int J Lab Hematol. 2021;43: 250- 258; Wang et al. Blood.2019;134(Supplement 1): 1301) CD8+ T cells and NK cells recovered to their initial levels within 3-4 weeks, whereas CD4+ T cells showed a significantly slower recovery rate and, in some patients, had not returned to their initial levels even 1 to 2 years after treatment onset. Thus, CD8+ T cells and NK cells may play a larger role in controlling the length of the allogeneic CAR T-cell therapeutic window by being the first and primary contributors to rejection. NK cells are a key component of the innate immune system and influence adaptive immune responses, e.g., via cytokine secretion. The activity of NK cells is thought to be controlled by a balance of inhibitory and activating signals delivered via NK cell-surface receptors. (Crew et al. Molecular Immunology, 2005;42(l): 1205-1214). Eliminating ligands for NK cell activation receptors on allogenic cells or increasing the level of ligands for inhibitory cell receptors could reduce or prevent host NK cell-mediated destruction of allogenic immune cell therapies.[000199] There are two classes of NK cell inhibitory receptors: the immunoglobulin-like KIR and LIR receptors and the C-type lectin-like receptors, CD94 / NKG2 heterodimers. In humans, the ligands for the KIR receptor family members are the classical class I antigens, HLA-A, -B, and -C and the ligand for one LIR (LIR-2) is the nonclassical class I antigen HLA-G. The major ligand for CD94 / NKG2 receptors is the nonclassical class I antigen HLA-E. Among the NK cell inhibitory receptors, CD94 / NKG2A appears to be the most widely expressed on NK cells. (Crew et al.Molecular Immunology, 2005;42(l): 1205-1214)494909-7891-2381.1[000200] Although CD8+ T cells need to go through a clonal expansion to mount an efficient alloresponse, they may be the first subset to reject allogeneic CAR T cells. In the case of CD8+ T cell-mediated rejection, a primary approach has been to eliminate expression of HLA class I molecules on CAR T cells. For example, deletion of the conserved gene [32m completely removes surface expression of HLA class I. (Wang et al. Stem Cells Transl Med. 2015;4(10): 1234-1245). In this way, the inactivation of HLA class I at the surface of CAR T cells could efficiently blunt such rejection and offer an initial therapeutic window to eradicate cancer cells. However, while immunogenic recognition by CD8+ T cells is reduced by this approach, the complete loss of HLA class I molecules increases the risk that host NK cells will recognize and destroy the allogeneic CAR T cells (the so-called “missing self” response). Embedding an NK inhibitor within CAR T cells could further extend their persistence. NK cell mediated destruction of HLA-edited T cells has been proposed to be prevented by expression on CAR T cells of nonpolymorphic HLA molecules such as HLA-E that will bind inhibitor}' receptors on NK cells. (Gornalusse et al. Nat Biotechnol.2017; 35(8):765–772)[000201] HLA-E can inhibit host NK cell cytotoxic activity and thus reduce or prevent immune therapeutic cell lysis. Crew et al. (Molecular Immunology, 2005 42(1):1205-1214) found that expression of an a single chain trimer (SCT) of HLA-E consisting of (from N- to C-terminus) the leader peptide of human [32m, VMAPRTLIL (an HLA-E -binding peptide), a 15 amino acid linker, mature human [32m, a 20 amino acid linker, and mature HLA-E heavy chain reduced NK cell- mediated rejection of porcine xenografts by providing an inhibitory ligand for human NK cells expressing CD94ZNKG2A. SCT HLA-E expression was subsequently found to reduce NK cell cytotoxicity in HLA class I-depleted pluripotent stem cells and in universal CAR T cells.(Gornalusse et al. Nat Biotechnol. 2017;35(8):765-772; Guo et al. Eur J Immunol. 2021;51:2513–2521; Jo et al. Nat Commun. 2022;13(1):3453) SCT HLA-E-expressing CAR T cells are presently in clinical trials for treating cancer patients.[000202] However, SCT HLA-E expression in immune therapy cells presents several drawbacks. While this construct offers some level of protection against NK cells, SCT HLA-E expression makes the therapeutic cells more vulnerable to rejection by HLA-E-restricted CD8+ T cells.Further, SCT HLA-E expression is only effective against NKG2A-expressing NK cells.Additionally, HLA-E also binds the NKG2C activating receptor on NK cells, causing NK cell activation and subsequently increased rejection of therapeutic cells.504909-7891-2381.1[000203] In contrast, the presently disclosed recombinant nucleic acids allow for development of, e.g., immune-evasive universal CAR T-cells that can evade host CD8+- T cell and NK-cell cytotoxicity and can be compatible with adoptive cell transfer in an allogeneic setting. In some embodiments, a disclosed recombinant nucleic acid encodes a construct for inhibiting NK cell cytotoxicity comprising a CD300a binding domain, a NKG2A binding domain, or both a CD300a binding domain and aNKG2A binding domain. In some embodiments, the recombinant nucleic acid comprises a CD300a binding domain and does not encode a NKG2A binding domain. In other embodiments, the recombinant nucleic acid comprises a NKG2A binding domain and does not encode a CD300a binding domain. In a particular embodiment, the CD300a binding domain and the NKG2A binding domain are comprised in the same nucleic acid. In another particular embodiment, the CD300a binding domain and the NKG2A binding domain are comprised in at least two different nucleic acids. A disclosed recombinant nucleic acid comprising both a CD300a binding domain and a NKG2A binding domain may encode the binding domains in any order. Thus, in some embodiments, the nucleotide sequence encoding the NKG2A binding domain is located 5’ of the nucleotide sequence encoding the CD300a binding domain. In other embodiments, the nucleotide sequence encoding the NKG2A binding domain is located 3' of the nucleotide sequence encoding the CD300a binding domain. In embodiments comprising both a NKG2A binding domain and a CD300a binding domain encoded in the same recombinant nucleic acid, the recombinant nucleic acid may further comprise a linker (such as any suitable linker, such as a linker described herein) that links the NKG2A binding domain and the CD300a binding domain.[000204] In some embodiments, the CD300a binding domain comprises an antibody or a fragment thereof, a VHH, a cytokine, a ligand, or a peptide. In some embodiments, the NKG2A binding domain comprises an antibody or a fragment thereof, a VHH, a cytokine, a ligand, or a peptide. In particular embodiments, (a) the antibody or a fragment thereof comprises a single chain variable fragment (scFv) or a VHH, or (b) the peptide is an adnectin or a design ankyrin repeat protein (DARPin). In other particular embodiments, the VHH comprises the VH domain of a camelid heavy chain antibody[000205] In some embodiments, the CD300a binding domain comprises a VHH (CD300a VHH) and / or the NKG2A binding domain comprises a VHH (NKG2A VHH). In particular embodiments, the construct for inhibiting NK cell cytotoxicity comprises or consists of the CD300a binding domain, and the CD300a binding domain comprises a VHH (CD300a VHH). In certain514909-7891-2381.1embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to the amino acid sequence of any one of SEQ ID NOs: 157-161. In certain embodiments, the CD300a VHH comprises a CDR2 comprising 0, 1, or 2 mutations relative to the amino acid sequence of any one of SEQ ID NOs: 162-165. In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to the amino acid sequence of any one of SEQ ID NOs: 166-183.[000206] In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising AAKPGEDVY (SEQ ID NO: 166). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLSQFAS (SEQ ID NO: 167). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPRSGWGL (SEQ ID NO: 168). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKTRYES (SEQ ID NO: 169). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 158), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ IS NO: 163), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising NTRLAHGRDVLGGVAYDI (SEQ ID NO: 170). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 158), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid 524909-7891-2381.1sequence comprising ITGSGGST (SEQ IS NO: 163), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising NTRRLGRSGDLVQDY (SEQ ID NO: 171). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSRYY (SEQ ID NO: 159), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPDRDY (SEQ ID NO: 172). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLPDVLPLEY (SEQ ID NO: 173). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYW (SEQ ID NO: 160), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ IS NO: 163), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKVDGSYGIVTEL (SEQ ID NO: 174). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 158), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising INGSGGST (SEQ IS NO: 164), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising HTRRSGTSMAMDV (SEQ ID NO: 175). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKLTMVY (SEQ ID NO: 176). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLTNEY (SEQ ID NO: 177). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS 534909-7891-2381.1NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKVRPSYEY (SEQ ID NO: 178). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSPYY (SEQ ID NO: 161), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VAKPGYEY (SEQ ID NO: 179). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGRT (SEQ IS NO: 165), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPGEDVY (SEQ ID NO: 180). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ IS NO: 162), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKSNMVY (SEQ ID NO: 181). In some embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 157), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ IS NO. 163), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising TTKVDGSYGIVTEL (SEQ ID NO: 182). In certain embodiments, the CD300a VHH comprises a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYW (SEQ ID NO: 160), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising INGSGGST (SEQ IS NO: 164), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising AAARDRERDY (SEQ ID NO: 183).[000207] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 139-156. In particular embodiments, the CD300a VHH comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 139-156. CD300a TASRs #20-37 disclosed herein (e.g., as shown in FIG. 45) comprise the CD300a VHHs of SEQ ID NOs: 155-172, respectively.544909-7891-2381.1[000208] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 139. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 139.[000209] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%>, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 140. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 140.[000210] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 141. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 141.[000211] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 142. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 142.[000212] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 143. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 143.[000213] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 144. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 144.554909-7891-2381.1[000214] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 145. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 145.[000215] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%>, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 146. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 146.[000216] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 147. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 147.[000217] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 148. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 148.[000218] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 149. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 149.[000219] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 150. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 150.564909-7891-2381.1[000220] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 151. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 151.[000221] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%>, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 152. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 152.[000222] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 153. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 153.[000223] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 154. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 154.[000224] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 155. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 155.[000225] In some embodiments, the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 156. In particular embodiments, the CD300a VHH comprises or consists of an amino acid sequence of SEQ ID NO: 156.574909-7891-2381.1[000226] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 121-138. In particular embodiments, a nucleotide sequence encoding the CD300a VHH comprises or consists of the nucleotide sequence of any one of SEQ ID NOs: 121-138.[000227] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 121. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 121.[000228] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 122. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 122.[000229] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 123. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 123.[000230] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 124. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 124.[000231] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 125. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 125.584909-7891-2381.1[000232] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 126. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 126.[000233] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 127. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 127.[000234] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 128. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 128.[000235] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 129. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 129.[000236] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 130. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 130.[000237] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%>, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 131. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 131.594909-7891-2381.1[000238] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 132. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 132.[000239] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 133. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 133.[000240] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 134. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 134.[000241] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 135. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 135.[000242] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 136. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 136.[000243] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%>, at least 97%, at least 98%, or at least 99% sequence identity' to SEQ ID NO: 137. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 137.604909-7891-2381.1[000244] In some embodiments, a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 138. In particular embodiments, the nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence of SEQ ID NO: 138.[000245] In some embodiments, a recombinant nucleic acid encodes a construct for inhibiting NK cell cytotoxicity comprising (a) a NKG2A binding domain comprising a NKG2A light chain variable region (NKG2A VL) and a NKG2A heavy chain variable region (NKG2A VH); and / or [000246] (b) a CD300a binding domain comprising a CD300a light chain variable region (CD300a VL) and a CD300a heavy chain variable region (CD300a VH). In a particular embodiment, the recombinant nucleic acid encodes a NK. G2A binding domain comprising a NKG2A VL and a NKG2A VH. In another particular embodiment, the recombinant nucleic acid encodes a CD300a binding domain comprising a CD300a VL and CD300a VH.[000247] in certain embodiments, the NKG2A VL comprises a VL CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising RASENIYSYLA (SEQ ID NO: 94), a VL CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising NAKTLAE (SEQ ID NO: 95), and a VL CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising QHHYGTPRT (SEQ ID NO: 96). In certain embodiments, theNKG2A VH comprises a VH CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising SYWMN (SEQ ID NO: 97), a VH CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising RIDPYDSETHYAQKLQG (SEQ ID NO: 98), and a VH CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GGYDFDVGTLYWFFDV (SEQ ID NO: 99). In some embodiments, the CD300a VL comprises a VL CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising RASQDISNYLN (SEQ ID NO: 100), a VL CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising YTSRLHS (SEQ ID NO: 101), and a VL CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising QQGNTLPWT (SEQ ID NO: 102). In some embodiments, the CD300a VH comprises a VH CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising SYWMQ (SEQ ID NO: 103), a VH CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising614909-7891-2381.1EIDPSDSYTNYNQKFKG (SEQ ID NO: 104), and a VH CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising WGMAYGTSSYWYFDV (SEQ ID NO: 105).[000248] In particular embodiments, the NKG2A VL comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 1 or SEQ ID NO: 2. In particular embodiments, the NKG2A VL comprises or consists of an amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 2.[000249] In particular embodiments, a nucleotide sequence encoding the NKG2A VL comprises a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 9 or SEQ ID NO: 10. In particular embodiments, the nucleotide sequence encoding the NKG2A VL comprises or consists of SEQ ID NO: 9 or SEQ ID NO: 10.[000250] In particular embodiments, the NKG2A VH comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 3-8. In particular embodiments, the NKG2A VH comprises or consists of an amino acid sequence of any one of SEQ ID NOs: 3-8.[000251] In particular embodiments, a nucleotide sequence encoding the NKG2A VH comprises a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 11-16. In particular embodiments, the nucleotide sequence encoding the NKG2A VH comprises or consists of any one of SEQ ID NOs: 11-16[000252] In particular embodiments, the CD300a VL comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID 624909-7891-2381.1NO: 17. In particular embodiments, the CD300a VL comprises or consists of the amino acid sequence of SEQ ID NO: 17.[000253] In particular embodiments, a nucleotide sequence encoding the CD300a VL comprises a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 19. In particular embodiments, the nucleotide sequence encoding the CD300a VL comprises or consists of SEQ ID NO: 19.[000254] In particular embodiments, the CD300a VH comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at. least. 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 18. In particular embodiments, the CD300a VH comprises or consists of the amino acid sequence of SEQ ID NO: 18[000255] In particular embodiments, a nucleotide sequence encoding the CD300a VH comprises a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at. least 95%, at least 96%, at least 97%, at. least 98%, or at least 99% sequence identity to SEQ ID NO: 20. In particular embodiments, the nucleotide sequence encoding the CD300a VH comprises or consists of SEQ ID NO: 20.[000256] In some embodiments, all or a portion of a nucleic acid sequence encoding a NKG2A binding domain is codon optimized to reduce or prevent undesired recombination events. In some embodiments, all or a portion of a nucleic acid sequence encoding a CD300a binding domain is codon optimized to reduce or prevent undesired recombination events. In some embodiments, all or a portion of a nucleic acid sequence encoding a NKG2A VL, a NKG2A VH, a CD300a VL, and / or a CD300a VH is codon optimized, such as to reduce or prevent undesired recombination events. In some embodiments, codon optimization encompasses replacing one or more 3-nucleotide sequences encoding for a particular amino acid with a different 3-nucleotide sequence (such as a 3-nucleotide sequence that differs from the initial 3-nucleotide sequence at the first, second, and / or third nucleotide position) encoding for the same amino acid. For example, the 3-nucleotide sequences (codons) GGA, GGG, GGT, GGC(GGU) all encode the amino acid glycine and each may be 634909-7891-2381.1replaced with one of the remaining three in a given nucleic acid sequence in a codon optimization process.[000257] In some embodiments, a codon optimized nucleotide sequence encoding the NKG2A binding domain has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 109. In particular embodiments, the codon optimized nucleotide sequence encoding the NKG2A binding domain comprises or consists of SEQ ID NO: 109.[000258] In some embodiments, a codon optimized nucleotide sequence encoding the NKG2A VL has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 106. In particular embodiments, the codon optimized nucleotide sequence encoding the NKG2A VL comprises or consists of SEQ ID NO: 106.[000259] In some embodiments, a codon optimized nucleotide sequence encoding the NKG2A VH has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 108. In particular embodiments, the codon optimized nucleotide sequence encoding the NKG2A VH comprises or consists of SEQ ID NO: 108.[000260] In some embodiments, the recombinant nucleic acid comprises a first linker (such as a first linker described herein) that links the nucleic acid encoding a NKG2A VL and the nucleic acid encoding a NKG2A VH. In other examples, the recombinant nucleic acid comprises a first linker (such as a first linker described herein) that links the nucleic acid encoding a CD300a VL and the nucleic acid encoding a CD300a VH In some embodiments wherein the recombinant nucleic acid encodes a NKG2A binding domain and a CD300a binding domain, the recombinant nucleic acid comprises a first linker (such as a first linker described herein) that links the nucleic acid encoding a NKG2A VL and the nucleic acid encoding a NKG2A VH, and a second linker (such as a second linker described herein) that links the nucleic acid encoding the CD300a VL and the nucleic acid encoding the CD300a VH.644909-7891-2381.1[000261] An NKG2A binding domain or a CD300a binding domain of the present disclosure can comprise any suitable structure, such as, but not limited to, an antibody or a fragment thereof, or a VHH. Commercially available NKG2A antibodies include, but are not limited to, those produced by R& D Systems (human NKG2A / CD 159a, Catalog #: MAB1059), Creative Biolabs (human anti- NKG2A recombinant antibody, scFv fragment (HPAB-1355-FY-S(P)) (CAT#: HPAB-1355-FY- S(P)), human anti-NKG2A recombinant antibody (HPAB-1355-FY) (CAT#: HPAB-1355-FY); human anti -NKG2A recombinant antibody; Fab fragment (HP AB-1355-FY-F(E)) (CAT#. HPAB- 1355-FY-F(E)); recombinant anti-human KLRC1 antibody scFv fragment (CAT#: MOB-604- S(P))), and Miltenyl Biotec (CD159a (NKG2A) Antibody, anti-human, REAfinity™, Catalog # 130-122-329). Commercially available CD300a antibodies include, but are not limited to, those produced by R& D Systems (human CD300a / LMIR1 antibody, Catalog #s: MAB2640, MAB26401) and ThermoFisher Scientific (CD300a monoclonal antibody (MEM-260), PE, Catalog # A15778; CD300a monoclonal antibody (7H8E4), Catalog # MA5-38479; CD300a monoclonal antibody (2F9C5), Catalog # 67242-1-IG). Exemplary binding domains useful in the disclosed embodiments can also include a cytokine, a ligand, or a peptide (such as an adnectin or a designed ankyrin repeat protein (DARPin) (See, e.g., Rafiq et al., Nat Rev Clin Oncol. 2020;17:147-167).[000262] In some embodiments wherein the NKG2A binding domain is an scFv, the NKG2A scFv comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 113. In particular embodiments, the NKG2A scFv comprises or consists of the amino acid sequence of SEQ ID NO: 113.[000263] In some embodiments wherein the NKG2A binding domain is an scFv, a nucleotide sequence encoding the NKG2A scFv comprises a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 109 or 115. In particular embodiments, the nucleotide sequence encoding the NKG2A scFv comprises or consists of any one of SEQ ID NOs: 109 or 115.[000264] In some embodiments wherein the CD300a binding domain is an scFv, the CD300a scFv comprises an amino acid sequence having at least 80%, at least 81%, at least 82%, at least 83%, at 654909-7891-2381.1least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 114. In particular embodiments, the CD300a scFv comprises or consists of the amino acid sequence of SEQ ID NO: 114.[000265] In some embodiments wherein the CD300a binding domain is an scFv, a nucleotide sequence encoding the CD300a scFv comprises a nucleotide sequence having at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 116. In particular embodiments, the nucleotide sequence encoding the CD300a scFv comprises or consists of SEQ ID NO: 116.[000266] A disclosed recombinant nucleic acid comprising a CD300a scFv and / or a NKG2A scFv may encode the NKG2A VL and VH and the CD300a VL and VH in any suitable order. Thus, in some embodiments, the nucleotide sequence encoding the NKG2A VL is located 5’ of the nucleotide sequence encoding the NKG2A VH. In other embodiments, the nucleotide sequence encoding the NKG2A VL is located 3’ of the nucleotide sequence encoding the NKG2A VH, Similarly, in some embodiments, the nucleotide sequence encoding the CD300a VL is located 5’ of the nucleotide sequence encoding the CD300a VH, In other embodiments, the nucleotide sequence encoding the CD300a VL is located 3’ of the nucleotide sequence encoding the CD300a VH. In some embodiments comprising both a CD300a scFv and a NKG2A scFv, the nucleotide sequence encoding the NKG2A scFv is located 3’ of the nucleotide sequence encoding the CD300a scFv. In other embodiments comprising both a CD300a scFv and a NKG2A scFv, the nucleotide sequence encoding the NKG2A scFv is located 5’ of the nucleotide sequence encoding the CD300a scFv. In some embodiments comprising both a CD300a scFv and a NKG2A scFv, the recombinant nucleic acid encodes a third linker that links the NKG2A scFv and the CD300a scFv.B. Linkers[000267] Embodiments of a recombinant nucleic acid disclosed herein may include one or more linkers, such as 1, 2, 3, 4, 5, 6, 7, or more linkers, or no linker. As described herein, a disclosed recombinant nucleic acid (such as a recombinant nucleic acid that comprises a NKG2A binding domain and does not comprise a CD300a binding domain) can comprise a first linker that links a 664909-7891-2381.1nucleic acid encoding a NKG2A VL and a nucleic acid encoding a NKG2A VH. In other embodiments, a disclosed recombinant nucleic acid (such as a recombinant nucleic acid that comprises a CD300a binding domain and does not comprise a NKG2A binding domain) can comprise a first linker that links the nucleic acid encoding a CD300a VL and the nucleic acid encoding a CD300a VH. In embodiments wherein the recombinant nucleic acid encodes a NKG2A binding domain and a CD300a binding domain, the recombinant nucleic acid can comprise a first linker that links the nucleic acid encoding a NKG2A VL and the nucleic acid encoding a NKG2A VH, and a second linker that links the nucleic acid encoding the CD300a VL and the nucleic acid encoding the CD300a VH. Such a “first linker"’ may be 5’ or 3’ of such a “second linker” in the recombinant nucleic acid, such that the NKG2A scFv is encoded 5’ or 3’ of the CD300a scFv in the recombinant nucleic acid. Further, embodiments of a disclosed recombinant nucleic acid that encodes a NKG2A binding domain (such as a NKG2A scFv) and a CD300a binding domain (such as a CD300a scFv) can comprise a third linker that links the NKG2A binding domain and the CD300a binding domain. In particular embodiments, a first linker, a second linker, and / or a third linker comprise a peptide linker (such as a cleavable peptide linker), a glycine-serine linker (e.g., GGGGSGGGGSGGGGSGGGGSGGGGS, SEQ ID NO: 30), or a Whitlow / 218 linker (GSTSGSGKPGSGEGSTKG, SEQ ID NO: 31). The first, second, and third linkers may be the same linker type, different linker types, or any combination thereof.[000268] A first linker, a second linker, and / or a third linker may be any suitable linker. In certain embodiments, the linker includes at least a linear group comprising groups selected from alkyl, amide, disulfide, polyethylene glycol, ether, thioether (-S-) and hydroxylamino (-O-N(H)-) groups. In certain embodiments, the linear group comprises groups selected from alkyl, amide, and ether groups. In certain embodiments, the linear group comprises one or more alkyl groups. In some embodiments, the one or more linkers comprise one or more amino acids (e.g., a peptide linker). Such linkers can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40 or more amino acids in length. An illustrative peptide linker has a length from 3 amino acids (AA) to 90 AA, from 3 AA to 80 AA, from 3 AA to 70 AA, from 3 AA to 60 AA, from 3 AA to 50 AA, from 3 AA to 40 AA, from 3 AA to 30 AA, from 3 AA to 20 AA, from 3 AA to 10 AA. For example, the linker can have a length from 3 AA to 5 AA, from 5 AA to 10 AA, from 10 AA to 15 AA, from 15 AA to 20 AA, from 20 AA to 25 AA, from 25 AA to 30 AA, from 30 AA to 35 AA, from 35 AA to 40 AA, from 40 AA to 50 AA, from 50 AA to 60 AA, from 60 AA to 70674909-7891-2381.1AA, from 70 AA to 80 AA, from 80 AA to 90 AA, or from 90 AA to 100 AA. In some embodiments, the peptide linker is at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, or at least 15 amino acids in length.[000269] A peptide linker can comprise any of a variety of sequences. For example, a peptide linker can comprise one or more serine (S), glycine (G), and / or threonine (T) residues. Glycine may impart flexibility in a given linker, whereas serine and / or threonine may improve solubility. In some embodiments, the linker is a glycine-serine linker. In particular embodiments, the glycine-serine linker comprises (Glym-Ser)n, where m is 1 to 10 (such as 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 9, 2 to 8, 2 to 7, 2 to 6, 2 to 5, 2 to 4, 2 to 3, 3 to 7, 3 to 6, 3 to 5, or 3 to 4) and n is 1 to 10 (such as 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 7, 2 to 6, 2 to 5, 2 to 4, or 2 to 3). In specific, non-limiting embodiments, the first linker, the second linker, and / or the third linker is a glycine-serine linker that comprises (Glym-Ser)n, wherein m is 3 to 6 and n is 1 to 10. In other specific, non-liming embodiments, m=4 and n=5.[000270] In some embodiments wherein the first, second, and / or third linker is a cleavable peptide, the first, second, and / or third linker is a self-cleaving peptide. In particular embodiments, the selfcleaving peptide is a T2A peptide (e.g., GSGEGRGSLLTCGDVEENPGP, SEQ ID NO: 32), a P2A peptide (GSGATNFSLLKQAGDVEENPGP, SEQ ID NO: 33), an E2A peptide (GSGQCTNYALLKLAGDVESNPGP, SEQ ID NO: 117), or an F2A peptide (VKQTLNFDLLKLAGDVESNPGP, SEQ ID NO: 29). In some embodiments, the self-cleaving peptide optionally comprises the amino acids glycine-serine-glycine (GSG) at an N-terminus.[000271] In particular embodiments, the first linker, the second linker, and / or the third linker comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 29-33 and 117. In other particular embodiments, the first linker, the second linker, and / or the third linker comprises any one of SEQ ID NOs: 29-33 and 117. In other particular embodiments, the first linker, the second linker, and / or the third linker consists of any one of SEQ ID NOs: 29-33 and 117,[000272] In some embodiments, a nucleotide sequence encoding the first linker, the second linker, and / or the third linker comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 34-37, 107, 111, or 112.684909-7891-2381.1In particular embodiments, a nucleotide sequence encoding the first linker, the second linker, and / or the third linker comprises any one of SEQ ID NOs: 34-37, 107, 111, or 112. In particular embodiments, a nucleotide sequence encoding the first linker, the second linker, and / or the third linker consists of any one of SEQ ID NOs: 34-37, 107, 111, or 112.[000273] SEQ ID NO: 34 (encoding the glycine- serine linker of SEQ ID NO: 30):GGTGGAGGAGGTTCTGGTGGTGGAGGATCAGGAGGCGGTGGAAGCGGAGGTGGAGGA1 C l GGT GGAGGTGGA I’C AC. Signal Peptides[000274] In some embodiments, a disclosed recombinant nucleic acid comprises a signal peptide, wherein the signal peptide is a cell surface expression signal peptide that directs the protein product of the recombinant nucleic acid to the surface of a cell. Any suitable signal peptide that directs the protein product of the recombinant nucleic acid to the surface of a cell can be of use in the disclosed embodiments. In some embodiments, the signal peptide is a GMCSF signal peptide. In some such embodiments, the signal peptide comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence MVLLVTSLLLCELPHPAFLLIP (SEQ ID NO: 38). In particular embodiments, the signal peptide consists of the amino acid sequence of SEQ ID NO: 38. In some embodiments of a disclosed recombinant nucleic acid, a nucleotide sequence encoding the signal peptide comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 39-41. In particular embodiments, a nucleotide sequence encoding the signal peptide comprises or consists of any one of SEQ ID NOs: 39-41. In other particular embodiments, a nucleotide sequence encoding the signal peptide comprises or consists of any one of SEQ ID NOs: 39-41.[000275] In some embodiments, the signal peptide is a CDS signal peptide. In some such embodiments, the signal peptide comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to the amino acid sequence MALPVTALLLPLALLLHAARP (SEQ ID NO: 92). In particular embodiments, the signal peptide 694909-7891-2381.1consists of the amino acid sequence of SEQ ID NO: 92. In some embodiments of a disclosed recombinant nucleic acid, a nucleotide sequence encoding the signal peptide comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 110. In particular embodiments, a nucleotide sequence encoding the signal peptide comprises SEQ ID NO: 110. In other particular embodiments, a nucleotide sequence encoding the signal peptide consists of SEQ ID NO: 110.[000276] In some embodiments, a nucleotide sequence encoding the signal peptide is located 5’ of a nucleotide sequence encoding a NKG2A binding. In some embodiments, a nucleotide sequence encoding the signal peptide is located 5’ of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, a nucleotide sequence encoding the signal peptide is located 5’ of both a nucleotide sequence encoding a NKG2A binding domain and of a nucleotide sequence encoding a CD300a binding domain.D. Hinge Regions[000277] Embodiments of the disclosed recombinant nucleic acids may include one or more extracellular hinge regions, which may or may not share sequence homology with immunoglobulin hinge regions and which provide flexibility to protein products of the recombinant nucleic acids. In some embodiments, a hinge region separates a binding domain (such as a CD300a binding domain (such as a disclosed CD300a VHH or CD300a scFv) or a NKG2A binding domain (such as a disclosed NKG2A scFv)) from a transmembrane domain. In some embodiments, the disclosed recombinant nucleic acids do not encode a hinge region. In some embodiments, a hinge region comprises a CD8 hinge, an hlgGl hinge, an hlgG2 hinge, an hlgG3 hinge, a FACD hinge, or any combination thereof. In some embodiments, the hinge region comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 43-48. In particular embodiments, the hinge region comprises any one of SEQ ID NOs: 43-48. In particular embodiments, the hinge region consists of any one of SEQ ID NOs: 43-48.[000278] In some embodiments of a disclosed recombinant nucleic acid, a nucleotide sequence encoding the hinge region comprises a nucleotide sequence having at least 80%, at least 85%, at 704909-7891-2381.1least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 50-55. In particular embodiments, a nucleotide sequence encoding the hinge region comprises any one of SEQ ID NOs: 50-55. In particular embodiments, a nucleotide sequence encoding the hinge region consists of any one of SEQ ID NOs: 50-55.[000279] In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the hinge region is located 3’ of the nucleotide sequence encoding the signal peptide. In other embodiments, the nucleotide sequence encoding the hinge region is located 3’ of a nucleotide sequence encoding a NKG2A binding domain In other embodiments, the nucleotide sequence encoding the hinge region is located 3’ of a nucleotide sequence encoding a CD300a binding domain. In other embodiments, the nucleotide sequence encoding the hinge region is located 3’ of a nucleotide sequence encoding a NKG2A binding domain and of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, the nucleotide sequence encoding the hinge region is located 3’ of the nucleotide sequence encoding the signal peptide, and 3’ of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, the nucleotide sequence encoding the hinge region is located 3’ of the nucleotide sequence encoding the signal peptide, and 3’ of a nucleotide sequence encoding a NKG2A binding domain. In some embodiments, the nucleotide sequence encoding the hinge region is located 3’ of the nucleotide sequence encoding the signal peptide, 3’ of a nucleotide sequence encoding aNKG2A binding domain, and 3’ of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, the nucleotide sequence encoding the hinge region is located (a) 3’ of a nucleotide sequence encoding a signal peptide, (b) 3’ of a nucleotide sequence encoding a NKG2A binding domain and of a nucleotide sequence encoding a CD300a binding domain; or (c) both (a) and (b).E. Transmembrane Domains[000280] Some embodiments of a disclosed recombinant nucleic acid comprise one or more transmembrane domains. Any suitable transmembrane domain can be of use in the present disclosure. In some embodiments, a transmembrane domain is a human transmembrane domain or a murine transmembrane domain. In particular embodiments, the transmembrane domain comprises or consists of a CD8, a CD80, an mCD80, an ITGA, an HLA-B57, a proCAR-4, a CD28, a KIR2DL1, a PDGFRB, or a CD86 transmembrane domain.714909-7891-2381.1[000281] In some embodiments, the transmembrane domain comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 66-75 or 93. In particular embodiments, the transmembrane domain comprises any one of SEQ ID NOs: 66-75 or 93. In other particular embodiments, the transmembrane domain consists of any one of SEQ ID NOs: 66-75 or 93.[000282] In some embodiments of a disclosed recombinant nucleic acid, a nucleotide sequence encoding the transmembrane domain comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 56-65. In particular embodiments, the nucleotide sequence encoding the transmembrane domain comprises any one of SEQ ID NOs: 56-65. In other particular embodiments, the nucleotide sequence encoding the transmembrane domain consists of any one of SEQ ID NOs: 56-65.[000283] in some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding a signal peptide. In some embodiments, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding a NKG2A binding domain. In some embodiments, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding aNKG2A binding domain and of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding a spacer. In some embodiments, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding a hinge region. In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the transmembrane domain is located 3?of a nucleotide sequence encoding a signal peptide, and 3’ of a nucleotide sequence encoding a CD300a binding domain. In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding a signal peptide, 3’ of a nucleotide sequence encoding a CD300a binding domain, and 3’ of a nucleotide sequence encoding a spacer. In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the transmembrane domain is located 3’ of a724909-7891-2381.1nucleotide sequence encoding a signal peptide, and 3’ of a nucleotide sequence encoding a NKG2A binding domain. In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the transmembrane domain is located 3’ of a nucleotide sequence encoding a signal peptide, 3’ of a nucleotide sequence encoding a NKG2A binding domain, and 3’ of a nucleotide sequence encoding a spacer. In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the transmembrane domain is located (a) 3’ of a nucleotide sequence encoding a signal peptide, (b) 3’ of a nucleotide sequence encoding a NKG2A binding domain and of a nucleotide sequence encoding a CD300a binding domain, (c) 3’ of a nucleotide sequence encoding a hinge region, or (d) any combination of (a)-(c).F. Cytoplasmic Domains[000284] Embodiments of a disclosed recombinant nucleic acid can comprise one or more cytoplasmic domains, wherein the cytoplasmic domain is a human cytoplasmic domain or a murine cytoplasmic domain Any suitable cytoplasmic domain can be of use in the disclosed embodiments. In some embodiments, the cytoplasmic domain comprises or consists of a CD8v2, a CD8vl, a mCD80, a CD80, a CD86, or an HLA-B57 cytoplasmic domain. In some embodiments, the cytoplasmic domain comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 82-87. In particular embodiments, the cytoplasmic domain comprises any one of SEQ ID NOs: 82-87. In other particular embodiments, the cytoplasmic domain consists of any one of SEQ ID NOs: 82-87.[000285] In some embodiments of a disclosed recombinant nucleic acid, a nucleotide sequence encoding the cytoplasmic domain comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 76-81. In particular embodiments, a nucleotide sequence encoding the cytoplasmic domain comprises any one of SEQ ID NOs: 76-81. In other particular embodiments, a nucleotide sequence encoding the cytoplasmic domain consists of any one of SEQ ID NOs: 76-81.[000286] In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the cytoplasmic domain is located 3’ of a nucleotide sequence encoding a signal peptide. In some embodiments, the nucleotide sequence encoding the cytoplasmic domain is located 3’ of a 734909-7891-2381.1nucleotide sequence encoding a NKG2A binding domain. In some embodiments, the nucleotide sequence encoding the cytoplasmic domain is located 3’ of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, the nucleotide sequence encoding the cytoplasmic domain is located 3’ of a nucleotide sequence encoding a NKG2A binding domain and of a nucleotide sequence encoding a CD300a binding domain. In some embodiments, the nucleotide sequence encoding the cytoplasmic domain is located 3’ of a nucleotide sequence encoding a hinge region. In some embodiments, the nucleotide sequence encoding the cytoplasmic domain is located 3’ of a nucleotide sequence encoding a transmembrane domain. In some embodiments of a disclosed recombinant nucleic acid, the nucleotide sequence encoding the cytoplasmic domain is located (a) 3' of a nucleotide sequence encoding a signal peptide, (b) 3’ of a nucleotide sequence encoding a NKG2A binding domain and of a nucleotide sequence encoding a CD300a binding domain, (c) 3’ of a nucleotide sequence encoding a hinge region, (d) 3’ of a nucleotide sequence encoding a transmembrane domain, or (e) any combination of (a)-(d).G. Exemplary Trans Antigen Signaling Receptors (TASRs)[000287] The present disclosure discloses a novel class of genetic constructs, “TASRs” (Trans Antigen Signaling Receptors) that, when expressed on the surface of a mammalian cell, can enhance the persistence of said cell by reducing or preventing its rejection by, e.g., autologous NK cells. TASRs are modular constructs comprising the following domains or functional parts: 1) a signal peptide to present the molecule to the cell surface, 2) a binding domain that binds to an inhibitory receptor expressed on NK cells, for example a VHH (such as a CD300a VHH disclosed herein), or a single chain variable fragment (scFv) that comprises a variable light / heavy chain linked to a variable heavy / light chain by a linker, 3) an optional extracellular hinge region, 4) a transmembrane region, and / or 5) an optional cytoplasmic region. TASRs disclosed herein can bind inhibitory' receptors on an NK cell, e.g., NKG2A or CD300a. The present disclosure and Examples provided herein demonstrate certain TASR designs having surprising efficacy in reducing or preventing immune therapy cell destruction by, e.g., NK cells, although the disclosure is not limited to the particular designs examined within the Examples section.[000288] In some embodiments, a recombinant nucleic acid disclosed herein encodes one or more components of such a TASR, In some embodiments, the TASR comprises a NKG2A binding domain (an “NKG2A TASR”), such as any suitable NKG2A binding domain, such as an NKG2A 744909-7891-2381.1binding domain described herein. In some embodiments, the TASR comprises a CD300a binding domain (a “CD300a TASR”), such as any suitable CD300a binding domain, such as a CD300a binding domain described herein. In some embodiments, the TASR is defined by the formula:[000289] SP-VL-Linker-VH-Hinge-TM-Cyt or SP-VH-Linker-VL-Hinge-TM-Cyt; wherein:[000290] “SP” represents an optional signal peptide,[000291] “VL” represents a light chain variable region (such as a NKG2A VL or a CD300a VL)[000292] “ Linker” represents a linker;[000293] “VH” represent a heavy chain variable region (such as a NKG2A VH or a CD300a VH);[000294] “Spacer” represents a spacer;[000295] “Hinge” represents an optional hinge region;[000296] “TM” represents a transmembrane domain; and[000297] “ Cyt” represents an optional cytoplasmic domain.[000298] In some embodiments, the TASR is defined by the formula:[000299] SP-VL-Linker-VH-Spacer-TM-Cyt or SP-VH-Linker-VL-Spacer-TM-Cyt.[000300] In some embodiments, the TASR is defined by the formula:[000301] SP-VHH-Spacer-TM-Cyt (such as SP-CD300a VHH-Spacer-TM-Cyt).[000302] In some embodiments, the TASR is defined by the formula:[000303] SP-VHH-Hinge-TM-Cyt (such as SP-CD300a VHH-Hinge-TM-Cyt).[000304] In some embodiments, the TASR is defined by the formula:[000305] SP-VHH-TM-Cyt (such as SP-CD300a VHH-TM-Cyt).[000306] In some embodiments of a disclosed nucleic acid, the recombinant nucleic acid encodes a spacer. In some embodiments, a spacer of a TASR disclosed herein comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 42 (AAATTTP). In particular embodiments, the spacer comprises or consists of SEQ ID NO: 42. In some embodiments, a recombinant nucleic acid encoding a spacer (of a TASR disclosed herein) comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 49 (GCAGCCGCAACAACGACACCT). In some embodiments, the nucleotide sequence encoding the spacer comprises or consists of SEQ ID NO: 49. In some embodiments of a disclosed recombinant 754909-7891-2381.1nucleic acid, the nucleotide sequence encoding the spacer is located (a) 3’ of the nucleotide sequence encoding the signal peptide, (b) 3' of the nucleotide sequence encoding the CD300a binding domain, 3’ of the nucleotide sequence encoding the NKG2A binding domain, or 3’ of the nucleotide sequence encoding the NKG2A binding domain and of the nucleotide sequence encoding the CD300a binding domain; or (c) both (a) and (b). In some embodiments wherein the TASR comprises a spacer, the TASR does not further comprise a hinge.[000307] FIGS. 1, 31, 32, and 45 provide diagrams of exemplary formulas for TASRs of use herein. A TASR described herein (such as a TASR of FIGS. 1, 31, 32, or 45) can comprise any suitable signal peptide, linker, spacer, hinge, transmembrane domain, or cytoplasmic domain, or combination thereof, such as any of the signal peptides, linkers, spacers, hinges, transmembrane domains, or cytoplasmic domains described in detail herein. In particular embodiments, the linker is a Whitlow / 218 linker as described herein (such as the linker of SEQ ID NO: 31 ).[000308] In some embodiments, a CD300a TASR comprises a CD300a VHH, such as a CD300a VHH having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs. 139-156. In some embodiments, such a CD300a TASR further comprises a signal peptide that directs the TASR (such as a protein product of a disclosed recombinant nucleic acid encoding a TASR comprising a CD300a VHH) to the surface of a cell, such as any of the signal peptides disclosed herein. In some embodiments, the signal peptide is a CD8 signal peptide. In some embodiments, such a CD300a TASR further comprises a transmembrane domain, such as any suitable transmembrane domain, such as any transmembrane domain disclosed herein. In particular embodiments, the transmembrane domain is a CD8 transmembrane domain or a mCD80 transmembrane domain. In particular embodiments, the transmembrane domain is a CD8 transmembrane domain. In some embodiments, such a CD300a TASR further comprises a cytoplasmic domain, such as any suitable cytoplasmic domain, such as any cytoplasmic domain disclosed herein. In particular embodiments, the transmembrane domain is a mCD80 cytoplasmic domain. In some embodiments, such a CD300a TASR further comprises a spacer, such as a spacer of SEQ ID NO: 42. In such embodiments, the spacer is located between the CD300a VHH and a transmembrane domain in the TASR. In particular embodiments, the hinge region is a CDS hinge region. In some embodiments, such a CD300a TASR further comprises a hinge region, such as any of the hinge regions disclosed herein. In such embodiments, the hinge region is located between the 764909-7891-2381.1CD300a VHH and a transmembrane domain in the TASR. In particular embodiments, the hinge region is a CD8 hinge region. In some embodiments, such a CD300a TASR does not comprise a hinge region.[000309] In particular embodiments, a CD300a TASR (e.g., a CD300a TASR encoded by a disclosed recombinant nucleic acid) comprises a signal peptide, a CD300a VHH (such as a CD300a VHH having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs 139-156), a spacer, a transmembrane domain, and a cytoplasmic domain, optionally further comprising a hinge region separating the CD300a VHH or the spacer and the transmembrane domain. In particular embodiments, the CD300a TASR (e.g., as encoded by a disclosed recombinant nucleic acid) comprises a CD8 signal peptide (such as a CD8 signal peptide having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 92), a CD300a VHH (such as a CD300a VHH having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs 139-156), a spacer having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 42, a CD8 transmembrane domain (such as a transmembrane domain having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 66), and a mCD80 cytoplasmic domain (such as a cytoplasmic domain having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 84). In a particular embodiment, the CD300a TASR (e.g., as encoded by a disclosed recombinant nucleic acid) comprises the CD8 signal peptide of SEQ ID NO: 92, a CD300a VHH of any one of SEQ ID NOs 139-156, the spacer of SEQ ID NO: 42, the CD8 transmembrane domain of SEQ ID NO: 66, and the mCD80 cytoplasmic domain of SEQ ID NO: 84.[000310] In particular embodiments, a recombinant nucleic acid encoding a CD300a TASR encodes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at 774909-7891-2381.1least 99% sequence identity to the amino acid sequence of any one of SEQ ID NOs: 204-221. In particular embodiments, the recombinant nucleic acid encoding the CD300a TASR encodes an amino acid sequence comprising or consisting of the amino acid sequence of any one of SEQ ID NOs: 204-221.[000311] In particular embodiments, a recombinant nucleic acid encoding a CD300a TASR has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity' to the nucleic acid sequence of any one of SEQ ID NOs: 186-203. In particular embodiments, the recombinant nucleic acid encoding the CD300a TASR comprises or consists of the nucleic acid sequence of any one of SEQ ID NOs: 186-203.[000312] In some embodiments, an NK. G2A TASR comprises a NKG2A VL and a NKG2A VH, such as any NKG2A VL and any NKG2A VH disclosed herein (such as comprised in a NKG2A scFv disclosed herein). In particular embodiments, a recombinant nucleic acid encoding a NKG2A TASR encodes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 21. In particular embodiments, the recombinant nucleic acid encoding the NKG2A TASR encodes an amino acid sequence comprising or consisting of SEQ ID NO: 21.[000313] In particular embodiments, the recombinant nucleic acid encoding a NKG2A TASR comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 23. In other particular embodiments, the recombinant nucleic acid encoding the NKG2A TASR comprises or consists of SEQ ID NO: 23.[000314] In some embodiments, a CD300a TASR comprises a CD300a VL and a CD300a VH, such as any CD300a VL and any CD300a VH disclosed herein (such as comprised in a CD300a scFv disclosed herein). In particular embodiments, the recombinant nucleic acid encoding a CD300a TASR encodes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 22. In particular embodiments the recombinant nucleic acid encoding a CD300a TASR encodes an amino acid sequence comprising or consisting of SEQ ID NO: 22.784909-7891-2381.1[000315] In particular embodiments, the recombinant nucleic acid encoding a CD300a TASR comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 24. In other particular embodiments, the recombinant nucleic acid encoding the CD300a TASR comprises or consists of SEQ ID NO: 24.H. Exemplary Bispecific TASRs[000316] In some embodiments, a TASR disclosed herein is a bispecific TASR, wherein the bispecific TASR comprises two or more binding domains, and accordingly, may bind two or more inhibitory' receptors on an NK cell, e g., both NKG2A and CD300a. Such bispecific TASRs can comprise two scFvs, such as a first scFv that specifically binds a first inhibitory' receptor expressed on NK cells (such as CD300a), and a second scFv that specifically binds a second inhibitory receptor expressed on NK cells (such as NKG2A). In some embodiments, a bispecific TASR is defined by the formula:[000317] SP-VL 1 -Linker! -VH 1 -Linker3-VL2-Linker2-VH2-Hinge-TM-Cyt; wherein:[000318] “SP” represents an optional signal peptide,[000319] “VL1” represents a first light chain variable region (such as a NKG2A VL or a CD300a VL)[000320] “Linkerl” represents a first linker;[000321] “VH1” represent a first heavy chain variable region (such as a NKG2A VH or a CD300a VH);[000322] “Linker3 ” represents a third linker;[000323] “VL2” represents a second light chain variable region (such as a NKG2A VL or a CD300a VL)[000324] “Linker2” represents a second linker;[000325] “VH2” represent a second heavy chain variable region (such as a NKG2A VH or a CD300a VH);[000326] “Hinge” represents an optional hinge region;[000327] “TM” represents a transmembrane domain; and[000328] “Cyt” represents an optional cytoplasmic domain.794909-7891-2381.1[000329] A bispecific TASR disclosed herein may alternatively be defined by any one of the following formulas: SP-VH1-Linker1-VL1-Linker3-VL2-Linker2-VH2-Hinge-TM-Cyt; SP-VL1-Linker1-VH1-Linker3-VH2-Linker2-VL2-Hinge-TM-Cyt; or SP-VH1-Linker1-VL1-Linker3-VH2-Linker2-VL2-Hinge-TM-Cyt FIG. 1 provides a diagram of exemplary formulas for bispecific TASRs of use herein. A bispecific TASR described herein (such as a bispecific TASR of FIG. 1) can comprise any suitable signal peptide, linker, hinge, transmembrane domain, or cytoplasmic domain, such as an of the signal peptides, linkers, hinges, transmembrane domains, or cytoplasmic domains described in detail herein. In particular embodiments, the third linker comprises a cleavable peptide linker, such as a T2A linker described herein (such as the T2A linker of SEQ ID NO: 32), or a P2A linker described herein (such as the P2A linker of SEQ ID NO: 33).[000330] In some embodiments of the disclosed bispecific TASRs, VL1-Linker1-VH1 (or VH1-L1-VL1) is a CD300a scFv (such as comprising a CD300a VL, a linker, and a CD300a VH disclosed herein) or is a NKG2A scFv (such as comprising a NKG2A VL., a linker, and a NKG2A VH disclosed herein). Similarly, in some embodiments, VL2-Linker2-VH2 (or VH2-Linker-VL2) is a CD300a scFv (such as a CD300a scFv disclosed herein, such as a CD300a scFv comprising a CD300a VL, a linker, and a CD300a VH disclosed herein) or is a NKG2A scFv (such as a NKG2A scFv disclosed herein, such as a NKG2A scFv comprising a NKG2A VL, a linker, and a NKG2A VH disclosed herein). In some embodiments, if VLl-Linkerl -VH1 (or VH1-L1-VLI) is a CD300a scFv, then VL2-Linker2-VH2 (or VH2 -Linker- VL2) is a NKG2A scFv. In other embodiments, if VL1 -Linker 1-VH1 (or VH1-L1-VL1) is a NKG2A scFv, then VL2-Linker2-VH2 (or VH2-Linker-VL2) is a CD300a scFv.[000331] In some embodiments, the nucleotide sequence of the NKG2A VL, the NKG2A VH, the CD300a VL, and / or the CD300a VH of a disclosed recombinant nucleic acid encoding a bispecific TASR is codon optimized, such as to reduce or prevent undesired recombination events. In some embodiments, a codon optimized nucleotide sequence encoding an NKG2A scFv of the bispecific TASR has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 109. In particular embodiments, the codon optimized nucleotide sequence encoding the NKG2A scFv comprises or consists of SEQ ID NO: 109.4909-7891-2381.1[000332] In some embodiments, a codon optimized nucleotide sequence encoding the NKG2A VL of the bispecific TASR has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 106. In particular embodiments, the codon optimized nucleotide sequence encoding the NKG2A VL comprises or consists of SEQ ID NO: 106.[000333] In some embodiments, a codon optimized nucleotide sequence encoding the NKG2A VH of the bispecific TASR has at least 80%, at least 81%, at least 82%, at least 83%, at least 84%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 108. In particular embodiments, the codon optimized nucleotide sequence encoding the NKG2A VH comprises or consists of SEQ ID NO: 108.[000334] In particular embodiments, a recombinant nucleic acid encoding a bispecific TASR encodes an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 25 or SEQ ID NO: 26. In particular embodiments, the recombinant nucleic acid encoding theNKG2A TASR encodes an amino acid sequence comprising or consisting of SEQ ID NO: 25 or SEQ ID NO: 26.[000335] In particular embodiments, the recombinant nucleic acid encoding the bispecific TASR comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 27 or SEQ ID NO: 28. In other particular embodiments, the recombinant nucleic acid encoding the bispecific TASR comprises or consists of SEQ ID NO: 27 or SEQ ID NO: 28.I. Humoral Response-Inhibition Constructs[000336] Provided herein is an engineered iPSC, or derivative cell thereof, that expresses one or more molecules that protects the engineered cell from an immune system. In some embodiments, the engineered cell comprises one or more modifications that increases expression of the one or more tolerogenic factors. The one or more molecules can generally comprise a nucleic acid, a protein, an enzyme, or both. In some embodiments, the nucleic acid comprises a recombinant 814909-7891-2381.1nucleic acid encoding one or more constructs, such as a humoral response-inhibition construct. In some embodiments, the humoral response-inhibition construct encodes an enzyme. In some embodiments, the enzyme can degrade an antibody. In some instances, the antibody comprises IgG antibody. In some instances, enzyme comprises an IgG-degrading enzyme.[000337] In some embodiments, an engineered iPSC, or a derivative cell thereof, further comprises one or more exogenous polynucleotides encoding a humoral response-inhibition construct. In some embodiments, the humoral response-inhibition construct comprises an IgG-degrading enzyme or a fragment thereof. In some embodiments, the IgG-degrading enzyme or the fragment thereof comprises IgG-degrading enzyme of S. pyogenes (IdeS) or a fragment thereof, IgG-degrading enzyme of S. equi subsp. zooepidemicus (IdeZ) or a fragment thereof, IgG-degrading enzyme of S. equi subsp equi. (IdeE) or a fragment thereof an endoglycosidase from Streptococcus pyogenes (EndoS) or a fragment thereof, or streptococcal cysteine proteinase from Streptococcus pyogenes (SpeB) or a fragment thereof. In some embodiments, IdeS or the fragment thereof comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs. 292 and 293. In some embodiments, the IdeS or the fragment thereof comprises or consists of an amino acid sequence set forth in any one of SEQ ID NOs: 292 and 293. In some embodiments, the IdeS or the fragment thereof is operably linked to a transmembrane domain. In some embodiments, the transmembrane domain is a CD8 transmembrane domain.[000338] In some embodiments, the IgG-degrading enzyme or the fragment thereof comprises IgG- degrading enzyme of S. pyogenes (IdeS) or a fragment thereof. The IdeS or the fragment thereof can comprise an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 292. The IdeS or the fragment thereof can comprise or consist of an amino acid sequence set forth in SEQ ID NO: 292. The IdeS or the fragment thereof can comprise an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 293. The IdeS or the fragment thereof can comprise or consist of an amino acid sequence set forth in SEQ ID NO: 293.824909-7891-2381.1[000339] The IdeS of the fragment thereof can be operably linked to a transmembrane domain. The IdeS or the fragment thereof can be attached to the engineered cell via the transmembrane domain. The transmembrane domain can be a CD8 transmembrane domain.[000340] An exogenous polynucleotide described herein can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to one or more of SEQ ID NOs: 17-18, 31, 38, 42, 92, 93, 143, 146, 2873-297. The exogenous polynucleotide can encode a polypeptide comprising or consisting of any amino acid sequence of any one of SEQ ID NOs: 17-18, 31, 38, 42, 92, 93, 143, 146, 287-297. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%>, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 38. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 31. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 42. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 286. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 287. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 93. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%,834909-7891-2381.1at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 288. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 289. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 290. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 92. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 291. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 292. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 293. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 17, The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 18. The exogenous polynucleotide can encode a polypeptide comprising an 844909-7891-2381.1amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 143. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 146. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 294. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 295. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 296. The exogenous polynucleotide can encode a polypeptide comprising an amino acid sequence at having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to an amino acid sequence set forth in SEQ ID NO: 297.[000341] The one or more exogenous polynucleotides described herein can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence of any one of SEQ ID NOs: 39, 298-303. The one or more exogenous polynucleotides can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence set forth in SEQ ID NO: 39. The one or more exogenous polynucleotides can comprise or consist of a nucleic acid sequence set forth in SEQ ID NO: 39. The one or more exogenous polynucleotides can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 854909-7891-2381.193%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence set forth in SEQ ID NO: 298. The one or more exogenous polynucleotides can comprise or consist of a nucleic acid sequence set forth in SEQ ID NO: 298. The one or more exogenous polynucleotides can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence set forth in SEQ ID NO: 299. The one or more exogenous polynucleotides can comprise or consist of a nucleic acid sequence set forth in SEQ ID NO: 299. The one or more exogenous polynucleotides can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence set forth in SEQ ID NO: 300. The one or more exogenous polynucleotides can comprise or consist of a nucleic acid sequence set forth in SEQ ID NO: 300. The one or more exogenous polynucleotides can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence set forth in SEQ ID NO: 301. The one or more exogenous polynucleotides can comprise or consist of a nucleic acid sequence set forth in SEQ ID NO: 301. The one or more exogenous polynucleotides can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence set forth in SEQ ID NO: 302. The one or more exogenous polynucleotides can comprise or consist of a nucleic acid sequence set forth in SEQ ID NO: 302. The one or more exogenous polynucleotides can comprise a nucleic acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a nucleic acid sequence set forth in SEQ ID NO: 303. The one or more exogenous polynucleotides can comprise or consist of a nucleic acid sequence set forth in SEQ ID NO: 303.[000342] Table 1. Sequence TableConstruct SEQ Amino Acid or Nucleic Acid SequenceID NO:GMCSFR 38 MVLLVTSLLLCELPHPAFLLIP864909-7891-2381.1signalpeptidescFv 31 GSTSGSGKPGSGEG STKGlinkersequenceCDS hinge 42 AAATTTPCD 8 tm 286 IYIWAPLAGTCGVLLLSLVITCD8 hinge 287 AAATTTP1YIWAPLAGTCGVLLLSLVIT+ tmCD8 tm 93 Pm’PAPRPPTPAPTIASQPLSLRl’EACRPAAGGAVHTRGLDFACDIYlWAP LAGTCGVLLLSLVITLYCN*CD8 tm 288 SDSNASIGMKKYFVGVNSAGKVA1SAKE1KEDNIGAQVLGLFTLSTGQDS WNQTNPTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACD1YIWAPLAGTCGVLLLSLVITLYCN*mCD80 289 LYCNKCFCKHRSCFRRNEASRETNNSLTFGPEEALAEQ TVFLP2A 290 ATNFSLLKQAGDVEENPGPCD8 92 MALPVTALLLPLALLLHAARPsignalpeptidepCART12 291 DSFSANQEIRYSEVTPYHVTSVWTKGVTPPANFTQGEDVFHAPYVANQG 4 WYDITKTFNGKDDLLCGAATAGNMLHWWFDQNKDQIKRYLEEHPEKQ KINFNGEQMFDVKEAIDTKNHQLDSKLFEYFKEKAFPYLSTKHLGVFPDH VIDMFINGYRLSLTNHGPTPVKEGSKDPRGGIFDAVFTRGDQSKLLTSRHD FKEKNLKEISDLIKKELTEGKALGLSHTYANVRINHVINLWGADFDSNGN LKAIYVTDIdeS 292 MALPVTALLLPLALLLHAARPDSFSANQEIRYSEVTPYHVTSVWTKGVTP PANFTQGEDVFHAPYVANQGWYDITKTFNGKDDLLCGAATAGNMLHW WFDQNKDQIKRYLEEHPEKQKINFNGEQMFDVKEAIDTKNHQLDSKLFE YFKEKAFPYLSTKHLGVFPDHVIDMFINGYRLSLTNHGPTPVKEGSKDPR GGIFDAVFTRGDQSKLLTSRHDFKEKNLKEISDLIKKELTEGKALGLSHTY ANVRINHVTNLWGADFDSNGNLKAIYVTDSDSNASIGMKKYFVGVNSAG KVAISAKEIKEDNIGAQVLGLFTLSTGQDSWNQTNIdeS 293 MALPVTALLLPLALLLHAARPDSFSANQEIRYSEVTPYHVTSVWTKGVTP PANFTQGEDVFHAPYVANQGWYDITKTFNGKDDLLCGAATAGNMLHW WFDQNKDQIKRYLEEHPEKQKINFNGEQMFDVKEAIDTKNHQLDSKLFE YFKEKAFPYLSTKHLGVFPDHVIDMFINGYRLSLTNHGPTPVKEGSKDPR GGIFDAVFTRGDQSKLLTSRHDFKEKNLKEISDLIKKELTEGKALGLSHTY ANVRINHVTNLWGADFDSNGNLKAIYVTDSDSNASIGMKKYFVGVNSAG KVAISAKEIKEDNIGAQVLGLFTLSTGQDSWNQTNPTTTPAPRPPTPAPTIA SQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITL YCN*CD300a 17 DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGKAPKLLIYYT TASR VL SRLHSGVPSRFSGSGSGTDYTLTISNLQPEDFATYFCQQGNTLPWTFGQGT KVEIK CD300a 18 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYWMQWVRQAPGQGLEW TASR VH IGEIDPSDSYTNYNQKFKGRATLTVDTSTSTTYMELSSLTSEDTAVYYCAR WGMAYGTSSYWYFDVWGRGTLVTVSS CD300a 143 EVQLVESGGGLVQPGGSLRLSCAASGFTFSDYAMSWVRQAPGKQREWVTASR SAITGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCNT 874909-7891-2381.1VHH RLAHGRDVLGGVAYDTWGQGTLVTVSSCD300a 146 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYYMSWVRQAPGKQREWVS TASR AISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVTK VHH EPDVLPLEYWGQGTEVTVSS VHH 294 MVLLVTSLLLCELPHPAFLLIPEVQLVESGGGLVQPGGSLRLSCAASGFTF SDYAMSWVRQAPGKQREWVSAITGSGGSTYYADSVKGRFTISRDNSKNT LYLQMNSLRAEDTAVYYCNTRLAHGRDVLGGVAYDIWGQGTLVTVSSA AATTTPIYIWAPLAGTCGVLLLSLVITLYCNKCFCKHRSCFRRNEASRETN NSLTFGPEEALAEQTVFLGSGATNFSLLKQAGDVEENPGPMALPVTALLL PLALLLHAARPDSFSANQEIRYSEVTPYHVTSVWTKGVTPPANFTQGEDV FHAPYVANQGWYDITKTFNGKDDLLCGAATAGNMLHWWFDQNKDQIK RYLEEHPEKQKINFNGEQMFDVKEAIDTKNHQLDSKLFEYFKEKAFPYLS TKHLGVFPDHVIDMFINGYRLSLTNHGPTPVKEGSKDPRGGIFDAVFTRG DQSKLLTSRHDFKEKNLKEISDLIKKELTEGKALGLSHTYANVRINHVINL WGADFDSNGNLKAIYVTDSDSNASIGMKKYFVGVNSAGKVAISAKEIKE DNIGAQVLGLFTLSTGQDSWNQTNPTTTPAPRPPTPAPTIASQPLSLRPEAC RPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCN* CD300a 295 DIQMTQSPSSLSASVGDRVTITCRASQDISNYLNWYQQKPGKAPKLLIYYT TASR SRLHSGVPSRFSGSGSGTDYTLTISNLQPEDFATYFCQQGNTLPWTFGQGTscFv KVEIKGSTSGSGKPGSGEGSTKGQVQLVQSGAEVKKPGASVKVSCKASG YTFTSYWMQWVRQAPGQGLEWIGEIDPSDSYTNYNQKFKGRATLTVDTS TSTIYMELSSLTSEDTAVYYCARWGMAYGTSSYWYFDVWGRGTLVTVS SAAATTFPIYIWAPLAGTCGVLLLSLVITLYCNKCFCKHRSCFRRNEASRE TNNSLTFGPEEALAEQTVTLIdeS-CD8 296 MALPVTALLLPLALLLHAARPDSFSANQEIRYSEVTPYHVTSVWTKGVTP tm PANFTQGEDVFHAPYVANQGWYDITKTFNGKDDLLCGAATAGNMLHW WFDQNKDQIKRYLEEHPEKQKINFNGEQMFDVKEAIDTKNHQLDSKLFE YFKEKAFPYLSTKHLGVFPDHVIDMFINGYRLSLTNHGPTPVKEGSKDPR GGIFDAVFTRGDQSKLLTSRHDFKEKNLKEISDLIKKELTEGKALGLSHTY ANVRINHVTNLWGADFDSNGNLKAIYVTDSDSNASIGMKKYFVGVNSAG KVAISAKEIKEDNIGAQVLGLFTLSTGQDSWNQTNPTTTPAPRPPTPAPTIA SQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITL YCN*CD300a 297 MVLLVTSLLLCELPHPAFLLIPEVQLVESGGGLVQPGGSLRLSCAASGFTF TASR- SSYYMSWVRQAPGKQREWVSAISGSGGSTYYADSVKGRFT1SRDNSKNT IdeS LYLQMNSLRAEDTAVYYCVTKLPDVLPLE^GQGTLVTVSSAAATTTPI YIWAPLAGTCGVLLLSLVITLYCNKCFCKHRSCFRRNEASRETNNSLTFGP EEALAEQTXTLELGSGATNFSLLKQAGDVEENPGPMAEPVTALELPLALL LHAARI’DSFSANQEIRYSEVTPYHVTSXAVrKGVTPPANFrQGEDVFHAPY VAN QGWYDITKTFNGKDDLLCGAATAGN MLHWWFDQNKDQIKRYLEE HPEKQKINFNGEQMFDVKEAIDTKNHQLDSKLFEYFKEKAFPYLSTKHLG VFPDIWIDMFINGYRLSLTNHGPTPVKEGSKDPRGGIFDAVFTRGDQSKLL TSRHDFKEKNI. XFJSDIJKKEI> TEGKALGLSHTYANVRINHV'INLWGADF DSNGNLKAIYVTDSDSNASIGMKKYFVGVNSAGKVAISAKEIKEDN1GAQ VLGLFTLSTGQDSVVNQTNPTTrPAPRI’PTPAPTIASQPLSLRl’EACRI’AAG GAVHTRGLDFACDIY1WAPLAGTCGVLLLSLVITLYCN*GMCSFR 39 ATGGTGCTCTTGGTGACTTCCCTTTTGCTGTGCGAGCTGCCTCATCCTG signal CGTTCCTGCTGATTCCCpeptideCD8 tm 298 CCCACCACCACCCCCGCGCCACGCCCTCCAACTCCTGCCCCCACAATA 884909-7891-2381.1GCATCACAACCGCTTAGTCTTCGACCCGAGGCTTGTCGGCCTGCTGCT GGAGGGGCGGTACA TACCCGCGGTCTCGATTTCGCGTGCGACATTTA T A TTIGGGCACCCTTGGCTGGCACCTGCGGCGTATTGTrGCTTAGCC TTG TGATCACGTTGTACTGCAACTGAIdeS 299 ATGGCACTCCCAGTCACCGCGCTTTGCTGCCCCTCGCTCTGCTTCTCC ATGCAGCAAGGCCAGATTCCTTTTCTGCGAATCAGGAGATTCGCTATA GTGAAGTGACACCCTATCACGTAACCTCAGTATGGACAAAGGGAGTT ACTCCTCCGGCCAACTTCACCCAAGGGGAAGACGTATTCCATGCTCCG TACGTGGCAAATCAGGGATGGTACGACATTACTAAGACCTTTAATGGC AAGGATGATCTGTTGTGTGGAGCGGCAACAGCAGGAAATATGCTCCA CTGGTGGTTCGACCAAAATAAGGACCAGATAAAACGCTATTTGGAGG AGCACCCCGAGAAGCAGAAGATCAATTTTAATGGAGAACAAATGTTC GACGTGAAGGAAGCTATCGACACTAAAAATCATCAACTTGATAGTAA GCTTTTCGAGTACTTCAAGGAAAAAGCCTTTCCATATTTGTCCACAAA GCATCTCGGAGTTTTCCCAGACCACGTAATTGATATGTTTATTAACGG CTATCGACTCAGTCTCACTAATCACGGCCCAACGCCGGTTAAAGAAGG GTCTAAGGACCCAAGGGGCGGTATCTTTGACGCCGTGTTTACCAGGGG GGACCAGTCAAAACTTCTCACGAGCAGGCACGACTTTAAAGAGAAGA ACTTGAAGGAGATTTCCGATCTGATTAAAAAGGAACTGACAGAAGGG AAAGCTCTGGGGCTCAGTCACACTTACGCCAACGTGCGAATAAACCAC GTTATAAATCTCTGGGGTGCAGACTTCGACTCCAACGGGAACCTTAAA GCTATATATGTCACTGATTCCGACTCTAACGCCTCCATCGGGATGAAA AAGTACTTCGTCGGGGTCAATAGTGCAGGTAAGGTAGCAATCAGCGC AAAGGAGATAAAGGAGGATAATATAGGGGCACAGGTCTTGGGACTGT TTACACTCTCTACAGGCCAAGACAGCTGGAATCAAACGAAT CD300a 300 GACATCCAGATGACCCAGAGCCCGTCCTCTCTGTCCGCCTCCGTGGGG TASR GACAGGGTCACCATCACCTGCCGTGCCTCACAGGACATCTCGAACTACscFv CTCAATTGGTACCAGCAGAAGCCCGGCAAAGCCCCCAAGCTGCTCATC TATTACACCTCCCGCCTTCACTCTGGTGTGCCCTCTCGCTTCTCGGGTA GTGGC TCCGGAACAGACTACACTCTGACCA ITAGCAACCTGCAGCCAG AGGACTTrGCAACTTACTTCTGTCAACAGGGCAACACGC TACCGTGGA CCTTCGGCCAGGGCACCAAGGTGGAGATCAAGGGATCGACCTCAGGC TCTGGTAAACCTGGTAGTGGGGAGGGCTCCACCAAGGGACAGGTCCA ACTGGTGCAGAGCGGCGCGGAGGTGAAGAAGCCCGGGGCATCCGTAA AGGTGTCATGCAAGGCGTCTGGCTACACGTTCACCAGCTATTGGATGC AGTGGGTCCGCCAGGCCCCTGGCCAGGGCCTGGAGTGGATCGGTGAA ATTGACCCGTCCGACAGCTACACCAACTACAACCAGAAATTTAAAGGC CGCGCTACACTGACCGTGGATACTTCGACCTCCACGACCTACATGGAG CTGTCTTCTCTCACTTCCGAGGACACCGCCGTGTACTACTGTGCTCGGT GGGGAATGGCCTACGGCACCAGCTCGTATTGGTACTTCGACGTGTGGG GCCGTGGTACTTTGGTTACGGTTTCTTCTGCAGCCGCAACAACGACAC CTATC 1ACATC IGGGCTCC’rTrGGCAGGCACCTGCGGGGTGC'IGCTGC TGTCCCTGGTGATCACCCTGTACTGTAACAAGTGTTTCTGCAAGCACA GAAGCTGCTTCCGGCGGAACGAGGCCAGCAGAGAGACAAACAACAGC CTGACATTCGGCCCCGAAGAGGCCCTGGCTGAGCAGACAGTTTTTCTGIdeS-CD 301 ATGGCACTCCCAGTCACCGCGCTTTGCTGCCCCTCGCTCTGCTTCTCC tm8 ATGCAGCAAGGCCAGATTCCTTTTCTGCGAATCAGGAGATTCGCTATA GTGAAGTGACACCCTATCACGTAACCTCAGTATGGACAAAGGGAGTT ACTCCTCCGGCCAACTTCACCCAAGGGGAAGACGTATTCCATGCTCCGTACGTGGCAAATCAGGGATGGTACGACATTACTAAGACCTTTAATGGC 894909-7891-2381.1AAGGATGATCTGTTGTGTGGAGCGGCAACAGCAGGAA ATATGCTCCA CTGGTGGTTCGACCAAAATAAGGACCAGATAAAACGCTATTTGGAGG AGCACCCCGAGAAGCAGAAGATCAATFTTAATGGAGAACAAATGT TC GACGTGAAGGAAGCTATCGACACTAAAAATCATCAACTTGATAGTAA GCTTTTCGAGTACTTCAAGGAAAAAGCCTTTCCATATTTGTCCACAAA GCATCTCGGAGTTTTCCCAGACCACGTAATTGATATGTTTATTAACGG CTATCGACTCAGTCTCACTAATCACGGCCCAACGCCGGTTAAAGAAGG GTCTAAGGACCCAAGGGGCGGTATCTTTGACGCCGTGTTTACCAGGGG GGACCAGTCAAAACTTCTCACGAGCAGGCACGACTTTAAAGAGAAGA ACTTGAAGGAGATTTCCGATCTGATTAAAAAGGAACTGACAGAAGGG AAAGCTCTGGGGCTCAGTCACACTTACGCCAACGTGCGAATAAACCAC GTTATAAATCTCTGGGGTGCAGACTTCGACTCCAACGGGAACCTTAAA GCTATATATGTCACTGATTCCGACTCTAACGCCTCCATCGGGATGAAA AAGI AC TI CG TCGGGGTC AATAGTGC AGGTAAGGTAGCAA TCAGCGC AAAGGAGATAAAGGAGGATAATATAGGGGCACAGGTCTTGGGACTGT TTACACTCTCTACAGGCCAAGACAGCTGGAATCAAACGAATCCCACCA CCACCCCCGCGCCACGCCCTCCAACTCCTGCCCCCACAATAGCATCAC AACCGCTTAGTCTTCGACCCGAGGCTTGTCGGCCTGCTGCTGGAGGGG CGGTACAIACCCGCGGTCTCGATTTCGCGTGCGACATITAIATTTGGG CACCCTTGGCTGGCACCTGCGGCGTA ITG I'TGC TTAGCC TTGTGA TCAC GTTGTACTGCAACTGA CD300a 302 ATGGTGCTGCTGGTCACATCTCIGCTGCIGTGCGAGCTGCCCCATCCTG TASR- CCTTTCTGCTGATTCCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGAC IdeS TTGTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGCCGCCAGCGGCTT CACCTTCAGCAGCTACTACATGAGCTGGGTCCGACAGGCCCCTGGCAA GCAGAGAGAATGGGTTTCCGCCATCTCTGGCAGCGGCGGCAGCACAT ATTACGCCGATTCTGTGAAGGGCAGATTCACCATCAGCCGGGACAACA GCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGAC ACCGCCGTGTACTACTGTGTGACCAAGCTGCCTGACGTGCTGCCCCTG GAATATTGGGGACAGGGCACACTGGTCACCGTGTCTAGTGCCGCAGCC ACCACCACACCTATCTACATTTGGGCCCCTCTGGCCGGCACATGTGGC GTTTTGCTGCTGAGCCTGGTCATCACACTGTACTGCAACAAGTGCTTCT GCAAGCACAGAAGCTGCTTCCGGCGGAACGAGGCCTCCAGAGAGACA AACAACTCCCTGACATI CGGCCCCGAAGAGGCCC TGGCTGAGCAGAC AGTGTTTCTGGAACTTGGCTCCGGCGCCACCAACTTCAGCCTGCTTAA ACAGGCAGGCGACGTGGAAGAGAACCCCGGACCTATGGCTCTGCCTG TGACAGCTCTGCTTCTGCCTCTGGCACTGCTGCTGCATGCCGCCAGAC CTGATAGCTTCAGCGCCAATCAAGAGATCCGGTACAGCGAAGTGACC CCITACCACGIGACCAGCGTGTGGACAAAGGGCGTGACACCrCCTGCC AACTTCACCCAGGGCGAAGATGTGTTTCACGCCCCTTACGTGGCAAAT CAAGGATGGTACGACATCACCAAGACCTTCAACGGCAAGGACGACCT GCTGTGTGGCGCTGCCACAGCCGGAAATATGCTGCACTGGTGGTTCGA CCAGAACAAGGACCAGATCAAGCGCTACCTCGAGGAACACCCCGAGA AGCAGAAGATCAATTTCAACGGCGAGCAGATGTTCGACGTGAAAGAG GCCATCGACACCAAGAATCACCAGCIGGACAGCAAGCTGTICGAGTA CTTCAAAGAGAAGGCATTCCCCTACCTGAGCACCAAGCACCTGGGCGT GTTCCCAGACCACGTGATCGACATGTTCATCAACGGCTACAGACTGAG CCTGACCAATCACGGCCCCACACCTGTGAAAGAAGGCAGCAAGGATC CCAGAGGCGGCATCTTCGATGCCGTGTTCACAAGAGGCGACCAGAGC AAGCTGCTGACCAGCCGGCACGA1TTCAAAGAAAAGAACCTGAAAGAGATCAGCGACCTGATCAAGAAAGAGC TGACCGAGGGCAAAGCCC TGG 904909-7891-2381.1GCCTGTCTCACACATATGCCAACGTGCGGATCAACCATGTGATCAACC TGTGGGGAGCCGACTTCGACAGCAACGGCAATCTGAAGGCCATCTAC GTGACCGACAGCGACTCCAATGCCAGCATCGGCATGAAGAAATACTTC GTGGGCGTGAACAGCGCCGGCAAGGTGGCCATTTCTGCCAAAGAAA TCAAAGAGGACAACATCGGCGCTCAGGTGCTGGGACTGTTCACACTGT CTACCGGCCAGGACAGCTGGAACCAGACAAACCCCACAACAACCCCT GCTCCTCGGCCTCCTACACCAGCTCCTACAATTGCCAGCCAGCCTCTGT CTCTGAGGCCCGAAGCTTGTAGACCTGCTGCTGGCGGAGCCGTGCATA CAAGAGGACTGGATTTCGCCTGCGACATCTATATCTGGGCACCCCTGG CTGGAACCTGCGGAGTTCTGCTGCTCTCTCTCGTGATCACCCTGTATTG CAATTGA VHH 303 ATGGTGCTGCTGGTCACATCTCTGCTGCTGTGCGAGCTGCCCCATCCTG CCTTTCTGCTGATTCCTGAGGTGCAGCTGGTGGAATCTGGCGGAGGAC TTGTTCAGCCTGGCGGCTCTCTGAGACTGTCTTGTGCCGCCAGCGGCTT CACCTTCAGCGATTACGCCATGAGCTGGGTCCGACAGGCCCCTGGAAA ACAGAGAGAAI GGGTTTCCGCCAI CACAGGCAGCGGCGGCAGCACAT ATrACGCCGAlTCTGTGAAGGGCAGAlTCACCATCAGCCGGGACAACA GCAAGAACACCCTGTACCTGCAGATGAACAGCCTGAGAGCCGAGGAC ACCGCCGTGTACTACTGCAATACCAGACTGGCCCACGGCAGGGATGTG CTTGGCGGAGTGGCCTATGATATCTGGGGCCAGGGAACCCTGGTCACC GTTAGTTCTGCCGCTGCCACCACCACACCTATCTACATTTGGGCCCCTC TGGCCGGCACATGTGGCGTmGCTGCTGAGCCTGGTCATCACACTGT ACTGCAACAAGTGCTTCTGCAAGCACAGAAGCTGCTTCCGGCGGAAC GAGGCCAGCAGAGAGACAAACAACTCCCTGACATTCGGCCCCGAAGA GGCCCTGGCTGAGCAGACAGTGTTTCTGGGCTCCGGCGCCACCAACTT CAGCCTGCTTAAACAGGCAGGCGACGTGGAAGAGAACCCCGGACCTA TGGCTCTGCCAGTGACAGCTCTGCTTCTGCCTCTGGCACTGCTGCTGCA TGCCGCCAGACCTGATAGCTTCAGCGCCAATCAAGAGATCCGGTACAG CGAAGTGACCCCTTACCACGTGACCAGCGTGTGGACAAAGGGCGTGA CACCTCCTGCCAACTTCACCCAGGGCGAAGATGTGTTTCACGCCCCTT ACGTGGCAAATCAAGGATGGTACGACATCACCAAGACCTTCAACGGC AAGGACGACCTGCTGTGTGGCGCTGCCACAGCCGGAAATATGCTGCA CTGGTGG IT CGACCAGAACAAGGACCAGA TCAAGCGCTACCTCGAGG AACACCCCGAGAAGCAGAAGATCAAITICAACGGCGAGCAGA IGTTC GACGTGAAAGAGGCCATCGACACCAAGAATCACCAGCTGGACAGCAA GCTGTTCGAGTACITCAAAGAGAAGGCATTCCCCTACCTGAGCACCAA GCACCTGGGCGTGTTCCCAGACCACGTGATCGACATGTTCATCAACGG CTACAGACTGAGCCTGACCAATCACGGCCCCACACCTGTGAAAGAAG GCAGCAAGGATCCCAGAGGCGGCATCTTCGATGCCGTGTTCACAAGA GGCGACCAGAGCAAGCTGCTGACCAGCCGGCACGATTTCAAAGAAAA GAACCTGAAAGAGATCAGCGACCTGATCAAGAAAGAGCTGACCGAGG GCAAAGCCCTGGGCCTGTCTCACACATATGCCAACGTGCGGATCAACC ATGTGATCAACCTGTGGGGAGCCGACTTCGACAGCAACGGCAATCTG AAGGCCATCTACGTGACCGACAGCGACTCCAATGCCAGCATCGGCAT GAAGAAATAClTCGTGGGCGTGAACAGCGCCGGCAAGGTGGCCA'nT CTGCCAAAGAAATCAAAGAGGACAACATCGGCGCTCAGGTGCTGGGA CTGTTCACACTGTCTACCGGCCAGGACAGCTGGAACCAGACAAACCCC ACAACAACCCCTGCTCCTCGGCCTCCTACACCAGCTCCTACAATTGCC AGCCAGCCTCTGTCTCTGAGGCCCGAAGCTTGTAGACCTGCTGCTGGC GGAGCCGTGCATACAAGAGGACTGGAnTCGCCTGCGACATCTATAl’CTGGGCACCCCTGGCTGGAACCTGCGGAGTTCTGCTGCTCTCTCTCGTG 914909-7891-2381.1ATCACCCTGTATTGCAATTGAIdeS- 304 CPPCPAPELLGGPSVFmediatedIgGcleavagesiteIII. Vectors[000343] The present disclosure provides vectors comprising a disclosed recombinant nucleic acid, such as vectors comprising one or more NKG2A and / or CD300a binding domains, such as a CD300a VHH or one or more NKG2A and / or CD300a scFvs as disclosed herein. A vector of the present disclosure can comprise, e.g., an NKG2A binding domain (such as an NKG2A scFv) and / or a CD300a binding domain (such as a CD300a VHH or a CD300a scFv), or aNKG2A TASR and / or a CD300a TASR (such as a CD300a TASR comprising a CD300a VHH, or a bispecific TASR comprising both a NKG2A scFv and a CD300a scFv as described herein).[000344] A vector useful in the present embodiments can be, for example, a DNA vector, an RNA vector, a plasmid, a lentivirus vector, an adenoviral vector, an adeno associated viral vector, a Rous sarcoma viral (RSV) vector, or a retrovirus vector. In particular embodiments of a vector, the recombinant nucleic acid is operably linked to a promoter. In specific, non-limiting embodiments, the promoter is an EFl a promoter, a CAG promoter, a PGK promoter, or a CMV promoter. In some embodiments, the promoter is an EFla promoter and comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 91. In particular embodiments, the promoter is an EFla promoter and comprises a nucleotide sequence of SEQ ID NO: 91.[000345] A recombinant nucleic acid herein can also include other regulatory elements, i.e., transcriptional and translational control sequences, such as enhancers, polyadenylation signals, terminators, protein degradation signals, and the like, that provide for and / or regulate transcription of a coding sequence and / or regulate translation of an encoded polypeptide (such as a TASR or a bispecific TASR described herein). Exemplary regulatory sequences are well known in the art and are described, for example, in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA (1990).924909-7891-2381.1[000346] In some embodiments, a nucleic acid sequence in the vector further comprises a poly(A) sequence. In particular embodiments, the poly(A) sequence comprises a bGH poly(A) signal, such as a bGH poly(A) signal encoded by a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 88.[000347] In some embodiments, a nucleic acid sequence in the vector further comprises a 3’UTR.[000348] In particular embodiments, a vector comprising a disclosed recombinant nucleic acid integrates into a genome at an adeno-associated virus integration site 1 (AAVS1) of the genome. Originally described as a major hotspot for adeno-associated virus (AAV) integration, intron 1 of the protein phosphatase 1, regulatory subunit 12C (PPP1R12C) gene on human chromosome 19 is referred to the AAVS1 locus (Oceguera-Yanez et al., Methods 2016;101:43-55) This locus allows stable, long-term transgene expression in many cell types, including embryonic stem cells. Thus, in some embodiments, a vector disclosed herein comprises an AAVS1 right homology arm and an AAVS1 left homology arm In some embodiments, a nucleic acid encoding the AAVS1 left homology arm comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 89.[000349] In some embodiments, a nucleic acid encoding the AAVS1 right homology arm comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to SEQ ID NO: 90.IV. Engineered Cells[000350] Some embodiments of the present disclosure comprise an engineered (e.g., genetically modified) cell, such as an engineered human cell. In some embodiments, the cell is an allogeneic cell.?\n engineered cell refers to a cell (or progeny of a cell) comprising an engineered genetic modification, e.g., a cell that has been contacted with a gene editing system and genetically modified by the gene editing system. The terms “engineered cell” and “genetically modified cell” are used interchangeably throughout. The engineered human cell may be any of the exemplary cell types disclosed herein.934909-7891-2381.1[000351] In some embodiments, the cell is a stem cell. In some embodiments, the cell is a pluripotent stem cell such as an induced pluripotent stem cell (iPSC) or a human embryonic stem cell (hESC). In some embodiments, the engineered cell is selected from a stem cell, a progenitor cell, or a cell that has been differentiated from a stem cell or a progenitor cell. In some embodiments, the engineered cell is a pluripotent stem cell that is subsequently differentiated into an immune cell. In some embodiments, the cell is an immune cell. As used herein, “immune cell” refers to a cell of the immune system, including e.g., a lymphocyte (e.g, T cell, B cell, natural killer cell (“NK cell”, and NKT cell, or iNKT cell)), monocyte, macrophage, mast cell, dendritic cell, or granulocyte (e.g., neutrophil, eosinophil, and basophil) In some embodiments, the cell is a primary immune cell. In some embodiments, the cell is a pluripotent stem cell-derived immune cell. In some embodiments, the immune system cell may be selected from CD3+, CD4+and CD8+T cells, regulatory T cells (Tregs), B cells, NK cells, and dendritic cells (DC). In some embodiments, the immune cell is allogeneic.[000352] In some embodiments, the cell is a lymphocyte. In some embodiments, the cell is an adaptive immune cell. In some embodiments, the cell is a T cell. In some embodiments, the cell is a B cell. In some embodiments, the cell is a NK cell. In some embodiments, the cell is a macrophage. In some embodiments, the lymphocyte is allogeneic.[000353] As used herein, a T cell can be defined as a cell that expresses a T cell receptor (“TCR” or “aP TCR” or “y§ TCR”), however in some embodiments, the TCR of a T cell may be genetically modified to reduce or delete its expression (e.g., by genetic modification to the TRAC or TRBC genes), therefore expression of the protein CD3 may be used as a marker to identify a T cell by standard flow cytometry methods. CD3 is a multi-subunit signaling complex that associates with the TCR. Thus, a T cell may be referred to as CD3+. In some embodiments, a T cell is a cell that expresses a CD3+ marker and either a CD4+ or CD8+ marker. In some embodiments, the T cell is allogeneic.[000354] In some embodiments, the T cell expresses the glycoprotein CD8 and therefore is CD81 by standard flow cytometry methods and may be referred to as a “cytotoxic” T cell. In some embodiments, the T cell expresses the glycoprotein CD4 and therefore is CD4+ by standard flow cytometry methods and may be referred to as a “helper” T cell. CD4+ T cells can differentiate into subsets and may be referred to as a Thl cell, Th2 cell, Th9 cell, Thl7 cell, Th22 cell, T regulatory (“Treg”) cell, or T follicular helper cells (“Tfh”). Each CD4 - subset releases specific cytokines that 944909-7891-2381.1can have either proinflammatory or anti-inflammatory functions, survival or protective functions. A T cell may be isolated from a subject by CD4+ or CD8+ selection methods.[000355] In some embodiments, the T cell is a memory T cell. In the body, a memory T cell has encountered antigen. A memory T cell can be located in the secondary lymphoid organs (central memory T cells) or in recently infected tissue (effector memory T cells). A memory T cell may be a CD8+ T cell. A memory T cell may be a CD4+ T cell.[000356] As used herein, a “central memory T cell” can be defined as an antigen-experienced T cell, and for example, may express CD62L and CD45RO. A central memory T cell may be detected as CD62L+ and CD45RCH, e g. by flow cytometry. A central memory T cell also expresses CCR7, therefore may be detected as CCR7+ by standard flow cytometry methods.[000357] As used herein, an “early stem-cell memory T cell” (or “Tscm”) can be defined as a T cell that expresses CD27 and CD45RA, and therefore is CD27+ and CD45RA+ by standard flow cytometry methods. A Tscm does not express the CD45 isoform CD45RO, therefore a Tscm will further be CD45RO- if stained for this isoform by standard flow cytometry methods. A CD45RO- CD27+ cell is therefore also an early stem-cell memory T cell. Tscm cells further express CD62L and CCR7, therefore may be detected as CD62L+ and CCR7+ by standard flow cytometry methods. Early stem-cell memory’ T cells have been shown to correlate with increased persistence and therapeutic efficacy of cell therapy products.[000358] In some embodiments, the cell is a B cell. As used herein, a “B cell” can be defined as a cell that expresses CD19 or CD20, or B cell mature antigen (“BCMA”), and therefore a B cell is CD19+, or CD20+, or BCMA+ by standard flow cytometry methods. A B cell is further negative for CD3 and CD56 by standard flow cytometry methods. The B cell may be a plasma cell. The B cell may be a memory B cell. The B cell may be a naive B cell. The B cell may be IgM+ or may have a class-switched B cell receptor (e.g., IgG+, or IgA+). In some embodiments, the B cell is allogeneic.[000359] In some embodiments, the cell is a mononuclear cell, such as from bone marrow or peripheral blood In some embodiments, the cell is a peripheral blood mononuclear cell (“PBMC”). In some embodiments, the cell is a PBMC, e.g. a lymphocyte or monocyte. In some embodiments, the cell is a peripheral blood lymphocyte (“PBL”). In some embodiments, the mononuclear cell is allogeneic.954909-7891-2381.1[000360] In some embodiments, the engineered cell is a stem cell, a progenitor cell, or a primary. Stem cells may be broadly defined as cells capable of going through numerous cycles of cell division, maintaining an undifferentiated state and having the capacity to differentiate into specialized cell types (Tesche et al., Stem Cells Int. 2010;2010:824876). They may go through asymmetric division wherein the stem cell creates a copy of itself and a daughter cell that is capable of differentiation. Stem cells are further classified into three categories: totipotent, pluripotent, or multipotent somatic. A totipotent cell has the ability to form an entire organism (e.g., a fertilized egg). Pluripotent stem cells lack the ability to form extraembryonic tissue and are therefore unable to generate a fetus, but can give rise to any cell type from the three germ cell layers (i.e., endoderm, mesoderm, or ectoderm). Examples of pluripotent stem cells include human embryonic stem cells (hESC) and induced pluripotent stem cells (iPSC). Adult stem cells or mesenchymal stem cells (MSCs) are examples of multipotent stem cells that are isolated from mature tissues and differentiate into other tissue types. Multipotent somatic stem cells are capable of differentiating into a variety of closely related cells within a tissue, but lack the ability to differentiate into other tissues. A much-studied area of multipotent somatic stem cells is the hematopoietic stem cell (HSC) which generates daughter cells that in turn differentiate into all subpopulations of hematopoietic cells (e.g., red blood cells, platelets, etc.).[000361] Accordingly, stem cells useful herein can include pluripotent stem cells (PSCs), induced pluripotent stem cells (iPSCs); embryonic stem cells (ESCs); mesenchymal stem cells (MSCs, e.g., isolated from bone marrow (BM), peripheral blood (PB), placenta, umbilical cord (UC) or adipose); hematopoietic stem cells (HSCs; e.g. isolated from BM or UC); neural stem cells (NSC’s); tissue specific progenitor stem cells (TSPSCs); and limbal stem cells (LSCs). Progenitor and primary cells include mononuclear cells (MNCs, e.g., isolated from BM or PB); endothelial progenitor cells (EPCs, e.g. isolated from BM, PB, and UC); neural progenitor cells (NPCs); and tissue-specific primary cells or cells derived therefrom (TSCs) including chondrocytes, myocytes, and keratinocytes. Cells for organ or tissue transplantations such as cardiomyocytes, thyroid cells, thymocytes, neuronal cells, skin cells, and retinal cells are also included.[000362] In some embodiments, the human cell is isolated from a human subject. In some embodiments, the cell is isolated from human donor PBMCs or leukopaks.[000363] In some embodiments, the cell is from a cell line. In some embodiments, the cell line is derived from a human subject. In some embodiments, the cell is from a cell bank. In some964909-7891-2381.1embodiments, the cell is genetically modified and then transferred into a cell bank. In some embodiments the cell is removed from a subject, genetically modified ex vivo, and transferred into a cell bank. In some embodiments, a genetically modified population of cells is transferred into a cell bank. In some embodiments, a genetically modified population of immune cells is transferred into a cell bank. In some embodiments, a genetically modified population of immune cells comprising a first and second subpopulations, wherein the first and second sub-populations have at least one common genetic modification and at least one different genetic modification are transferred into a cell bank.[000364] A variety of methods for introducing nucleic acids (such as a recombinant nucleic acid disclosed herein) into cells are known in the art, including, but not limited to, electroporation; transfection employing calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, or other substances; viral transfection; non-viral transfection; microprojectile bombardment; lipofection; and infection (e.g, where the vector is an infectious agent).[000365] In some embodiments, a recombinant nucleic acid is introduced into a cell (e.g, to produce an engineered cell) using a CRISPR / Cas system; zinc finger nuclease (ZFN) system; or a transcription activator-like effector nuclease (TALEN) system. Generally, the gene editing systems involve the use of engineered cleavage systems to induce a double strand break (DSB) or a nick (e.g, a single strand break, ori SSB) in a target DNA sequence. Cleavage or nicking can occur through the use of specific nucleases such as engineered ZFN, TALENs, or using the CRISPR / Cas system with an engineered guide RNA to guide specific cleavage or nicking of a target DNA sequence. Further, targeted nucleases are being developed based on the Argonaute system (e.g, from T. thermophilus, known as ‘TtAgo’, see Swarts et al (2014) Nature 507(7491): 258-261), which also may have the potential for uses in gene editing and gene therapy.[000366] In some embodiments, the gene editing system is a TALEN system. Transcription activator-like effector nucleases (TALEN) are restriction enzymes that can be engineered to cut specific sequences of DNA. They are made by fusing a TAL effector DNA-binding domain to a DNA cleavage domain (a nuclease which cuts DNA strands). Transcription activator-like effectors (TALEs) can be engineered to bind to a desired DNA sequence, to promote DNA cleavage at specific locations. The restriction enzymes can be introduced into cells, for use in gene editing or for gene editing in situ, a technique known as gene editing with engineered nucleases. Such974909-7891-2381.1methods and compositions for use therein are known in the art. See, e.g., WO2019147805, W02014040370, WO2018073393, the contents of which are hereby incorporated in their entireties.[000367] In some embodiments, the gene editing system is a zinc-finger system. Zinc-finger nucleases (ZFNs) are artificial restriction enzymes generated by fusing a zinc finger DNA-binding domain to a DNA-cleavage domain. Zinc finger domains can be engineered to target specific desired DNA sequences to enables zinc-finger nucleases to target unique sequences within complex genomes. The non-specific cleavage domain from the type Ils restriction endonuclease FokI is typically used as the cleavage domain in ZFNs. Cleavage is repaired by endogenous DNA repair machinery, allowing ZFN to precisely alter the genomes of higher organisms. Such methods and compositions for use therein are known in the art. See, e.g., WO2011091324, the contents of which are hereby incorporated in their entireties.[000368] In some embodiments, the gene editing system is a CRISPR / Cas system, including e.g., a CRISPR guide RNA comprising a guide sequence and RNA-guided DNA binding agent.[000369] In some embodiments, a disclosed engineered cell comprises a chimeric antigen receptor (e g., the disclosed cell is engineered to express the CAR). Exemplary CAR constructs are described, for example, in Fresnak AD, et al. Nat Rev Cancer. 2016; 16(9):566-81, which is incorporated by reference in its entirety for the teaching of these CAR models. For example, the CAR can be a TRUCK, Universal CAR, Self-driving CAR, Armored CAR, Self-destruct CAR, Conditional CAR, Marked CAR, TenCAR, Dual CAR, or sCAR. TRUCKS (T cells redirected for universal cytokine killing) co-express a chimeric antigen receptor (CAR) and an antitumor cytokine. Cytokine expression may be constitutive or induced by T cell activation. Targeted by CAR specificity, localized production of pro-inflammatory cytokines recruits endogenous immune cells to tumor sites and may potentiate an antitumor response. Universal, allogeneic CAR T cells can be engineered to no longer express endogenous T cell receptor (TCR) and / or major histocompatibility complex (MHC) molecules, thereby reducing or preventing graft-versus-host disease (GVHD) or rejection. Self-driving CARs co-express a CAR and a chemokine receptor, which binds to a tumor ligand, thereby enhancing tumor homing. CAR T cells engineered to be resistant to immunosuppression (Armored CARs) may be genetically modified to no longer express various immune checkpoint molecules (for example, cytotoxic T lymphocyte-associated antigen 4 (CTLA4) or programmed cell death protein 1 ( PD1)), with an immune checkpoint switch receptor, or may be administered with a monoclonal antibody that blocks immune checkpoint signaling. A self-destruct 984909-7891-2381.1CAR may be designed using RNA delivered by electroporation to encode the CAR. Alternatively, inducible apoptosis of the T cell may be achieved based on ganciclovir binding to thymidine kinase in gene-modified lymphocytes or the more recently described system of activation of human caspase 9 by a small- molecule dimerizer. A conditional CAR T cell is by default unresponsive, or switched Off, until the addition of a small molecule to complete the circuit, enabling full transduction of both signal 1 and signal 2, thereby activating the CAR T cell. Alternatively, T cells may be engineered to express an adaptor-specific receptor with affinity for subsequently administered secondary antibodies directed at target antigen. Marked CAR T cells express a CAR plus a tumor epitope to which an existing monoclonal antibody agent binds. In the setting of intolerable adverse effects, administration of the monoclonal antibody clears the CAR T cells and alleviates symptoms with no additional off-tumor effects A tandem CAR (TanCAR) T cell expresses a single CAR consisting of two linked single-chain variable fragments (scFvs) that have different affinities fused to intracellular co-stimulatory domain(s) and a CD3ζ domain. TanCAR T cell activation is achieved only when target cells co-express both targets. A dual CAR T cell expresses two separate CARs with different ligand binding targets; one CAR includes only the CD3ζ domain and the other CAR includes only the co-stimulatory domain(s). Dual CAR T cell activation requires co-expression of both targets on the tumor. A safety CAR (sCAR) comprises extracellular scFv fused to an intracellular inhibitory domain sCAR T cells co-expressing a standard CAR become activated only when encountering target cells that possess the standard CAR target but lack the sCAR target.[000370] The antigen recognition domain of the disclosed CAR may be an scFv or antibody fragment. There are, however, many alternatives. An antigen recognition domain from native T-cell receptor (TCR) alpha and beta single chains have been described, as have simple ectodomains (e.g., CD4 ectodomain to recognize HIV infected cells) and more exotic recognition components such as a linked cytokine (which leads to recognition of cells bearing the cytokine receptor). In fact, almost anything that binds a given target with high affinity can be used as an antigen recognition region. The endodomain is the business end of the CAR that after antigen recognition transmits a signal to the immune effector cell, activating at least one of the normal effector functions of the immune effector cell. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines. Therefore, the endodomain may comprise the “intracellular signaling domain” of a T cell receptor (TCR) and optional co-receptors. While usually the entire 994909-7891-2381.1intracellular signaling domain can be employed, in many cases it is not necessary to use the entire chain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal[000371] Cytoplasmic signaling sequences that regulate primary activation of the TCR complex that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyrosine-based activation motifs (ITAMs). Examples of ITAM containing cytoplasmic signaling sequences include those derived from CD8, CD3ζ, CD35, CD3γ, CD3ε, CD32 (Fc gamma Rlla), DAP10, DAP12, CD79a, CD79b, FcyRly, FceRip (FCERIB), and FceRly (FCERIG).[000372] In some embodiments, the intracellular signaling domain is derived from CD3 zeta (CD3ζ (TCR zeta, GenBank Accession No. BAG36664.1). T-cell surface glycoprotein CD3 zeta (CD3ζ chain, also known as T-cell receptor T3 zeta chain or CD247 (Cluster of Differentiation 247), is a protein that in humans is encoded by the CD247 gene.[000373] First-generation CARs typically had the intracellular domain from the CD3ζ chain, which is the primary transmitter of signals from endogenous TCRs. Second -generation CARs add intracellular signaling domains from various costimulatory protein receptors (e.g., CD27, CD28, 4-1 BB, ICOS) to the endodomain of the CAR to provide additional signals to the T cell. Preclinical studies have indicated that the second generation of CAR designs improves the antitumor activity of T cells. More recent, third-generation CARs combine multiple signaling domains to further augment potency. T cells grafted with these CARs have demonstrated improved expansion, activation, persistence, and tumor-eradicating efficiency independent of costimulatory receptor / ligand interaction. (Imai C, et al. Leukemia 2004 18:676-84; Maher J, et al. Nat Biotechnol 2002 20:70-5)[000374] For example, the endodomain of the CAR can be designed to comprise the CD3ζ signaling domain by itself or combined with any other desired cytoplasmic domain(s) useful in the context of the CAR. For example, the cytoplasmic domain of the CAR can comprise a CD3ζ chain portion and a costimulatory signaling region. The costimulatory signaling region refers to a portion of the CAR comprising the intracellular domain of a costimul atoiy molecule. A costimulatory molecule is a cell surface molecule other than an antigen receptor or their ligands that is required for an efficient response of lymphocytes to an antigen. Examples of such molecules include CD27, CD28, 4-1 BB (CD137), 0X40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA- 1), CD2,1004909-7891-2381.1CD7, LIGHT, NKG2C, B7-H3, and a ligand that specifically binds with CD83, CD8, CD4, b2c, CD80, CD86, DAP10, DAP12, MyD88, BTNL3, and NKG2D. Thus, while the CAR is exemplified primarily with CD28 as the co-stimulatory signaling element, other costimulatory elements can be used alone or in combination with other co-stimulatory signaling elements.[000375] In some embodiments, an engineered cell of the present disclosure comprises a vector comprising a recombinant nucleic acid encoding a construct for inhibiting NK cell cytotoxicity as disclosed herein. In particular embodiments, the construct comprises a NKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both a NKG2A binding domain and a CD300a binding domain.[000376] In some embodiments, a disclosed engineered cell expresses a recombinant nucleic acid (such as a recombinant nucleic acid disclosed herein) encoding a construct for inhibiting NK cell cytotoxicity, wherein the construct comprises a NKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both a NKG2A binding domain and a CD300a binding domain.[000377] In a particular embodiment, an engineered cell comprises a first vector and a second vector, wherein (a) the first vector comprises a first recombinant nucleic acid encoding a first construct for inhibiting NK cell cytotoxicity comprising a NKG2A binding domain comprising (i) a NKG2A light chain variable region (NKG2A VL) and (ii) a NKG2A heavy chain variable region (NKG2A VH): and (b) the second vector comprises a second recombinant nucleic acid encoding a second construct for inhibiting NK cell cytotoxicity comprising a CD300a binding domain comprising (i) a CD300a light chain variable region (CD300a VL) and (ii) a CD300a heavy chain variable region (CD300a VH).[000378] In some embodiments of an engineered cell, a vector comprising a recombinant nucleic acid as disclosed herein is inserted into a safe harbor locus of at least one allele of the engineered cell. In particular embodiments, the safe harbor locus is an AAVS1 locus.[000379] In any of the embodiments of an engineered cell disclosed herein, the engineered cell may be MHC class I, TRAC, and / or MHC class II deficient. In some embodiments, the engineered cell is MHC class I deficient. In some embodiments, the engineered cell is MHC class II deficient. In some embodiments, the engineered cell is TRAC deficient. In some embodiments, the engineered cell is MHC class 1 and MHC class II deficient. In some embodiments, the engineered cell is MHC class I and TRAC deficient. In some embodiments, the engineered cell is MHC class II and TRAC1014909-7891-2381.1deficient. In some embodiments, the engineered cell is MHC class I, MHC class II, and TRAC deficient,[000380] In some embodiments, the engineered cell is MHC class I deficient. In some embodiments, in a cell that is MHC class I deficient, a MHC class I molecule functionality (such as an amount of the MHC class I molecule detectable on a cell surface) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that is not MHC class I deficient. In some embodiments, in a cell that is MHC class I deficient, a MHC class I molecule functionality (such as an amount of the MHC class I molecule detectable on a cell surface) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90- 100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100%, relative to a wild-type cell that is not MHC class I deficient.[000381] In some embodiments of the disclosed engineered cells, a β2 microglobulin (B2M) gene locus of the engineered cell is disrupted. In particular embodiments, a recombinant nucleic acid or vector disclosed herein is inserted into the B2M gene locus of the engineered cell. In some embodiments, in a cell that is MHC class I deficient due to a B2M gene locus disruption, B2M expression and / or functionality (such as functionality of a B2M gene product) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that does not comprise a B2M gene locus disruption. In some embodiments, in a cell that is MHC class I deficient due to a B2M gene locus disruption, B2M expression and / or functionality (such as functionality of a B2M gene product) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% relative to a wild-type cell that does not comprise a B2M gene locus disruption.[000382] In some embodiments wherein a disclosed recombinant nucleic acid or vector is inserted into the B2M locus of a cell, expression of the recombinant nucleic acid or vector is driven by an endogenous B2M promoter of the cell.1024909-7891-2381.1[000383] In some embodiments, the engineered cell is MHC class II deficient. In some embodiments, in a cell that is MHC class II deficient, a MHC class II molecule functionality (such as an amount of the MHC class II molecule detectable on a cell surface) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that is not MHC class II deficient. In some embodiments, in a cell that is MHC class II deficient, a MHC class II molecule functionality (such as an amount of the MHC class II molecule detectable on a cell surface) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80- 100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99- 100%, or 100%, relative to a wild-type cell that is not MHC class II deficient.[000384] In some embodiments of the disclosed engineered cells, a CIITA gene locus of the engineered cell is disrupted. In some embodiments, in a cell that is MHC class II deficient due to a CIITA gene locus disruption, CIITA expression and / or functionality (such as functionality of a CIITA gene product) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that does not comprise a CIITA gene locus disruption. In some embodiments, in a cell that is MHC class II deficient due to a CIITA gene locus disruption, CIITA expression and / or functionality (such as functionality of a CIITA gene product) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82- 100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% relative to a wild-type cell that does not comprise a CIITA gene locus disruption.[000385] In particular embodiments, the engineered cell is a stem cell, a progenitor cell, a cell that has been differentiated from a stem cell, or a cell that has been differenti ted from a progenitor cell For example, the stem cell can be a pluripotent stem cell, such as an induced pluripotent stem cell (iPSC) or a human embryonic stem cell (hESC). In particular embodiments, the cell is a progenitor cell, such as an early hematopoietic progenitor cell or a CD34+ progenitor cell. In some embodiments, the engineered cell is a cell that has been differentiated from a stem cell. In other embodiments the engineered cell is a cell that has been differentiated from a progenitor cell In some embodiments, the engineered cell is an engineered T cell. In particular embodiments, the 1034909-7891-2381.1engineered cell that has been differentiated from a progenitor cell is an engineered T cell. In other particular embodiments, the engineered cell is an induced pluripotent stem cell that is subsequently differentiated into a T cell.[000386] As described above, an engineered T cell can be a chimeric antigen receptor (CAR) T cell. In specific, non-limiting embodiments, the CAR is a BCMA CAR, a CD19 CAR, and / or a CD20 CAR[000387] In some embodiments, the engineered cell is T cell receptor alpha constant (TRAC) deficient. In some embodiments, the TRAC locus is disrupted in the engineered cell. In particular embodiments, the CAR is inserted into the TRAC locus in the engineered cell. In some embodiments, in a cell that is TRAC deficient (such as due to a TRAC gene locus disruption), TRAC expression and / or functionality (such as functionality of a TRAC gene product) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that is not TRAC deficient. In some embodiments, in a cell that is TRAC deficient (such as due to a TRAC gene locus disruption), TRAC expression and / or functionality (such as functionality of a TRAC gene product) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88- 100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% relative to a wild-type cell that is not TRAC deficient. In some embodiments wherein a CAR is inserted into the TRAC locus of a cell, expression of the CAR is driven by an endogenous TRAC promoter of the cell.[000388] In particular embodiments, the engineered T cell is derived from a pluripotent stem cell or a CD34+ progenitor cell. In some such embodiments, the pluripotent stem cell is an induced pluripotent stem cell (iPSC) or a human embryonic stem cell (hESC).[000389] In some embodiments, the engineered cell is an autologous cell. In other embodiments, the engineered cell is an allogenic cell. In particular embodiments, the engineered cell is from a donor or is derived from a stem cell of a donor, wherein the donor is not the subject.[000390] In particular embodiments, the engineered cell is a human cell. In some embodiments, the engineered cell is hypoimmunogenic. Hypoimmunogenicity can result from introduction of a recombinant nucleic acid disclosed herein (i.e., a recombinant nucleic acid comprising an NKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both, as 1044909-7891-2381.1described herein) into the engineered cell. For example, relative to a wild-type cell, such a hypoimmunogenic cell may be at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% or more less prone to immune rejection by a subject into which such cells are transplanted. In particular embodiments, the hypoimmunogenic engineered cell (such as an engineered cell expressing a recombinant nucleic acid comprising an NKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both, as described herein) is resistant to NK cell mediated cellular cytotoxicity.V. Compositions[000391] Disclosed herein are compositions comprising a disclosed recombinant nucleic acid, a vector comprising a disclosed recombinant nucleic acid, and / or an engineered cell comprising a disclosed recombinant nucleic acid. Such compositions comprise one or more of a cell culture media and a buffer.[000392] A pharmaceutical composition comprising the engineered cell of any one of claims further comprises a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers of use herein include chemical components with which an engineered cell disclosed herein may be combined and which, following the combination, can be used to administer the engineered cell to a subject in need thereof. Non-limiting examples of pharmaceutically acceptable carriers include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose, salt solutions (such as Ringer’s solution), and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as stabilizers, emulsifiers, salts for influencing osmotic pressure, and / or buffers and the like that do not deleteriously react with the engineered cells of the disclosure One of ordinary skill in the art will recognize that other pharmaceutical carriers are useful in the present disclosure.VI. MethodsA. Overview[000393] The presently disclosed methods of using the recombinant nucleic acids, vectors, and engineered cells disclosed herein allow for development of, e.g., hypoimmunogenic iPSCs, and / or hypoimmunogenic T-cells (such as CAR T-cells) that can resist host CD8+ T cell and NK-cell cytotoxicity and can be compatible with adoptive cell transfer in an allogeneic setting.1054909-7891-2381.1B. Methods of Producing an Engineered Cell[000394] The present disclosure provides methods of producing engineered cells, such as engineered iPSCs, such as engineered iPSCs that are differentiated into different cell types for subsequent transplantation into subjects. As will be appreciated by those in the art, the methods for differentiation depend on the desired cell type using known techniques. For example, engineered cells may be differentiated in suspension and then put into a gel matrix form, such as Matrigel, gelatin, or fibrin / thrombin forms to facilitate cell survival. Exemplary methods for differentiating pluripotent stem cells into CD34+ progenitor cells and further to T-cells are described, for example, in U. S. Provisional Patent Application No. 63 / 483,814, filing date February 08, 2023, the contents of which are incorporated herein by reference in their entirety. Differentiation is assayed as is known in the art, generally by evaluating the presence of cell-specific markers.[000395] General techniques for recombinant nucleic acid manipulations are described for example in Sambrook et al., in Molecular Cloning: A Laboratory Manual, Vols. 1-3, Cold Spring Harbor Laboratory Press, 2 ed., 1989, or F. Ausubel et al., in Current Protocols in MolecularBiology (Green Publishing and Wiley-Interscience: New York, 1987) and periodic updates, herein incorporated by reference in their entireties. In some embodiments, a nucleic acid (e.g., DNA) comprising a disclosed recombinant nucleic acid is operably linked to an expression vector carrying one or more suitable transcriptional or translational regulatory elements derived from mammalian, viral, or insect genes. Such regulatory elements include a transcriptional promoter, an optional operator sequence to control transcription, a sequence encoding suitable mRNA ribosomal binding sites, and sequences that control the termination of transcription and translation. The expression vector can include an origin or replication that confers replication capabilities in the host cell. The expression vector can include a gene that confers selection to facilitate recognition of transgenic host cells (e.g., transformants). The expression vector construct can be introduced into a cell using a method appropriate for the cell. A variety of methods for introducing nucleic acids into cells are known in the art, including, but not limited to, electroporation; transfection employing calcium chloride, rubidium chloride, calcium phosphate, DEAE-dextran, or other substances; viral transfection; non-viral transfection; microprojectile bombardment; lipofection; and infection (e.g., where the vector is an infectious agent).1064909-7891-2381.1[000396] In some embodiments, a method of producing an engineered T cell comprises differentiating an engineered cell comprising a recombinant nucleic acid disclosed herein into a T cell. In some embodiments, a method of producing an engineered hypoimmunogenic induced pluripotent stem cell (iPSC) comprises expressing in an iPSC a recombinant nucleic acid encoding a NKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both a NKG2A binding domain and a CD300a binding domain, thereby producing the engineered hypoimmunogenic iPSC. In other embodiments, a method of producing an engineered, hypoimmunogenic induced pluripotent stem cell (iPSC) comprises expressing in an iPSC a first recombinant nucleic acid comprising aNKG2A binding domain and a second recombinant nucleic acid comprising a CD300a binding domain, thereby producing the engineered hypoimmunogenic iPSC.[000397] In particular embodiments, a method of producing an engineered iPSC comprises contacting an iPSC with a first vector and a second vector, wherein the first vector comprises a first recombinant nucleic acid encoding a first construct for inhibiting NK cell cytotoxicity comprising a NKG2A binding domain, and the second vector comprises a second recombinant nucleic acid encoding a second construct for inhibiting NK cell cytotoxicity comprising a CD300a binding domain, and wherein the contacting occurs under conditions whereby the first recombinant nucleic acid and the second recombinant nucleic acid are expressed in the iPSC, thereby producing the engineered iPSC.[000398] Some embodiments of a method of producing an engineered hypoimmunogenic T cell comprise (a) expressing in an iPSC a recombinant nucleic acid encoding a CD300a binding domain (such as a CD300a VHH disclosed herein), a NKG2A binding domain, or both a NKG2A binding domain and a CD300a binding domain, thereby producing an engineered hypoimmunogenic iPSC; and (b) differentiating the engineered hypoimmunogenic iPSC into an engineered T cell, thereby producing the engineered T cell. In some embodiments, a method of producing an engineered hypoimmunogenic T cell comprises expressing in a T cell a recombinant nucleic acid encoding a CD300a binding domain (such as a CD300a VHH disclosed herein), a NKG2A binding domain, or both a NKG2A binding domain and a CD300a binding domain, thereby producing the engineered hypoimmunogenic T cell. In other embodiments, the method comprises expressing in a T cell a first recombinant nucleic acid comprising a NKG2A binding domain and a second recombinant nucleic1074909-7891-2381.1acid comprising a CD300a binding domain, thereby producing the engineered hypoimmunogenic T cell.[000399] Some embodiments of a method of producing an engineered T cell comprise contacting a T cell with a first vector and a second vector, wherein the first vector comprises a first recombinant nucleic acid encoding a first construct for inhibiting NK cell cytotoxicity comprising a NKG2A binding domain, and the second vector comprises a second recombinant nucleic acid encoding a second construct for inhibiting NK cell cytotoxicity comprising a CD300a binding domain, and wherein the contacting occurs under conditions whereby the first recombinant nucleic acid and the second recombinant nucleic acid are expressed in the T cell, thereby producing the engineered T cell.[000400] In particular embodiments, the methods comprise expressing in the engineered iPSC, the iPSC, the engineered cell, or the T cell a chimeric antigen receptor (CAR). In particular embodiments, the CAR is a BCMA CAR, a CD 19 CAR, and / or a CD20 CAR.[000401] in some embodiments of the disclosed methods, the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell is T cell receptor alpha constant (TRAC) deficient. In some such embodiments, the TRAC locus is disrupted in the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell. In particular embodiments, a CAR (such as a CD 19 CAR, a CD20 CAR, or a BCMA CAR) is inserted into the TRAC locus in the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell. In some embodiments, in the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell that is TRAC deficient (such as due to a TRAC gene locus disruption), TRAC expression and / or functionality (such as functionality of a TRAC gene product) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that is not TRAC deficient. In some embodiments, in the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell that is TRAC deficient (such as due to a TRAC gene locus disruption), TRAC expression and / or functionality (such as functionality of a TRAC gene product)1084909-7891-2381.1is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90- 100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% relative to a wild-type cell that is not TRAC deficient.[000402] In some embodiments wherein a CAR is inserted into the TRAC locus of the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell, expression of the CAR is driven by an endogenous TRAC promoter of the ceil.[000403] In some embodiments, the NKG2A binding domain and / or the CD300a binding domain comprises a single chain variable fragment (scFv), a VHH (also known herein as a nanobody), a cytokine, a ligand, or a peptide. In particular embodiments, the VHH comprises the VH domain of a camelid heavy chain antibody. In other particular embodiments, the peptide is an adnectin or a DARPin.[000404] in particular embodiments, the NKG2A binding domain comprises a scFv comprising a NKG2A light chain variable region (NKG2A VL) and a NKG2A heavy chain variable region (NKG2A VH), such as any of the NK. G2A VL s or NKG2A VHs disclosed herein. In other particular embodiments, the CD300a binding domain comprises a CD300a VHH disclosed herein. In other particular embodiments, the CD300a binding domain comprises a scFv comprising a CD300a light chain variable region (CD300a VL) and a CD300a heavy chain variable region (CD300a VH), such as any of the CD300a VLs or CD300a VHs disclosed herein.[000405] In some embodiments, a vector used in producing an engineered cell comprises a recombinant nucleic acid comprising a CD300a binding domain (such as a CD300a VHH disclosed herein), a NKG2A binding domain, or both a CD300a binding domain and a NKG2A binding domain. In some embodiments, the recombinant nucleic acid is expressed in the iPSC.[000406] In particular embodiments, contacting a cell as described herein comprises introducing a disclosed recombinant nucleic acid into the cell (such as an engineered cell, iPSC, or T cell) using transfection, electroporation, transduction, or knock-in. In particular embodiments, the transfection comprises contacting the engineered cell, the iPSC, or the T cell with a cationic polymer and the recombinant nucleic acid. In other particular embodiments, the transduction comprises contacting the engineered cell, the iPSC, or the T cell with a lentivirus comprising the recombinant nucleic1094909-7891-2381.1acid. In yet other particular embodiments, the knock-in comprises contacting the engineered cell, the iPSC, or the T cell with an adeno-associated virus comprising the recombinant nucleic acid.[000407] In any of the embodiments of the methods disclosed herein, an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell may be MHC class I, TRAC, and / or MHC class II deficient. In some embodiments, the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell is MHC class I deficient. In some embodiments, the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell is MHC class II deficient. In some embodiments, the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell is TRAC deficient. In some embodiments, the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell is MHC class I and MHC class II deficient. In some embodiments, the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell is MHC class I and TRAC deficient. In some embodiments, the engineered cell is MHC class II and TRAC deficient. In some embodiments, the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell is MHC class I, MHC class II, and TRAC deficient.[000408] In some embodiments of the disclosed methods, a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) is MHC class I deficient. In some such embodiments, a β2 microglobulin (B2M) gene locus of the cell (such as the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell) is disrupted. In some embodiments, a recombinant nucleic acid or vector disclosed herein is inserted into the B2M gene locus of cell (such as the engineered cell, the iPSC, the engineered iPSC, the engineered hypoimmunogenic iPSC, the T cell, the engineered T cell, or the hypoimmunogenic engineered T cell).1104909-7891-2381.1[000409] In some embodiments of the disclosed methods, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class I deficient, a MHC class I molecule functionality (such as an amount of the MHC class I molecule detectable on a cell surface) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that is not MHC class I deficient. In some embodiments, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class I deficient, a MHC class I molecule functionality (such as an amount of the MHC class I molecule detectable on a cell surface) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97- 100%, 98-100%, 99-100%, or 100%, relative to a wild-type cell that is not MHC class I deficient. In some embodiments, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class I deficient due to a B2M gene locus disruption, B2M expression and / or functionality (such as functionality of a B2M gene product) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild¬ type cell that does not comprise a B2M gene locus disruption. In some embodiments, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class I deficient due to a B2M gene locus disruption, B2M expression and / or functionality (such as functionality of a B2M gene product) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95- 100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% relative to a wild-type cell that does not comprise a B2M gene locus disruption.[000410] In some embodiments wherein a disclosed recombinant nucleic acid or vector is inserted into the B2M locus of a cell (such as an engineered cell, an iPSC, an engineered iPSC, an1114909-7891-2381.1engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein), expression of the recombinant nucleic acid or vector is driven by an endogenous B2M promoter of the cell.[000411] In some embodiments of the disclosed methods, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class II deficient, a MHC class II molecule functionality (such as an amount of the MHC class II molecule detectable on a cell surface) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that is not MHC class II deficient. In some embodiments, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class II deficient, a MHC class II molecule functionality (such as an amount of the MHC class II molecule detectable on a cell surface) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%, 91-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96- 100%, 97-100%, 98-100%, 99-100%, or 100%, relative to a wild-type cell that is not MHC class II deficient.[000412] In some embodiments of the disclosed methods, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class II deficient due to a CIITA gene locus disruption, CIITA expression and / or functionality (such as functionality of a CIITA gene product) is reduced by at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99%, relative to a wild-type cell that does not comprise a CIITA gene locus disruption. In some embodiments, in a cell (such as an engineered cell, an iPSC, an engineered iPSC, an engineered hypoimmunogenic iPSC, a T cell, an engineered T cell, or a hypoimmunogenic engineered T cell as disclosed herein) that is MHC class II deficient due to a CIITA gene locus disruption, CIITA expression and / or functionality (such as functionality of a CIITA gene product) is reduced by 10-100%, 20-100%, 30-100%, 40-100%, 50-100%, 60-100%, 70-100%, 80-100%, 81-100%, 82-100%, 83-100%, 84-100%, 85-100%, 86-100%, 87-100%, 88-100%, 89-100%, 90-100%,1124909-7891-2381.191-100%, 92-100%, 93-100%, 94-100%, 95-100%, 96-100%, 97-100%, 98-100%, 99-100%, or 100% relative to a wild-type cell that does not comprise a CIITA gene locus disruption.C. Methods of Inhibiting or Reducing NK Cell Cytotoxicity to an Engineered Cell [000413] As described herein, CD8+ T cells and NK cells may play a large role in controlling the length of the allogeneic T-cell therapeutic window by being the first and primary contributors to rejection. Thus, embodiments of the present disclosure provide methods of inhibiting or reducing NK cell cytotoxicity’ and / or CD8+ T cell cytotoxicity to an engineered therapeutic cell disclosed herein. In some embodiments, methods of inhibiting or reducing NK cell cytotoxicity to an engineered cell comprise expressing in the engineered cell a recombinant nucleic acid encoding a NKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both a NKG2A binding domain and a CD300a binding domain, thereby inhibiting or reducing NK cell cytotoxicity to the engineered cell. In some embodiments, the method comprises expressing in the engineered cell a first recombinant nucleic acid comprising a NKG2A binding domain and a second recombinant nucleic acid comprising a CD300a binding domain thereby inhibiting or reducing NK cell cytotoxicity to the engineered cell.[000414] In some embodiments, a method of inhibiting or reducing NK cell cytotoxicity to a T cell comprises (a) expressing in an iPSC a recombinant nucleic acid encoding aNKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both a NKG2A binding domain and a CD300a binding domain, thereby producing an engineered iPSC; and (b) differentiating the engineered iPSC into a T cell, thereby inhibiting or reducing NK cell cytotoxicity to the T cell.[000415] In other embodiments, the method comprises expressing in a T cell a recombinant nucleic acid encoding a NKG2A binding domain, a CD300a binding domain (such as a CD300a VHH disclosed herein), or both a NKG2A binding domain and a CD300a binding domain, thereby inhibiting or reducing NK cell cytotoxicity to the T cell. In yet other embodiments, the method comprises expressing in a T cell a first recombinant nucleic acid comprising a NKG2A binding domain and a second recombinant nucleic acid comprising a CD300a binding domain, thereby producing the engineered iPSC.[000416] In some embodiments, a method of inhibiting or reducing NK cell cytotoxicity to a T cell further comprises disrupting a B2M, TRAC, and / or CIITA locus (such as using any suitable means 1134909-7891-2381.1disclosed herein or known in the art) in the engineered cell or the engineered iPSC In some embodiments, the method further comprises inserting a recombinant nucleic acid or vector disclosed herein into a B2M locus of the engineered cell or the engineered iPSC, thereby disrupting the B2M locus. In some embodiments wherein a disclosed recombinant nucleic acid or vector is inserted into the B2M locus of the engineered cell or the engineered iPSC, expression of the recombinant nucleic acid or vector is driven by an endogenous B2M promoter of the engineered cell or the engineered iPSC. In some embodiments, the method further comprises inserting a CAR into a TRAC locus of the engineer...

Claims

1. WHAT IS CLAIMED IS:

1. An engineered induced pluripotent stem cell (iPSC), or a derivative cell thereof, comprising:3.(a) one or more exogenous polynucleotides encoding:4.i. one or more chimeric antigen receptors (CARs), said one or more CARs comprising a first antigen binding domain, and optionally a second antigen binding domain;5.ii. a CD300a TASR binding domain comprising:

1. a CD300a light chain variable region (CD300a VL); and2. a CD300a heavy chain variable region (CD300a VH);8.iii. And, optionally, a humoral response-inhibition construct comprising an IgG-degrading enzyme of Streptococcus pyogenes (IdeS); and (b) an exogenous promoter operably linked to one or more of (x) the one or more CARs, (y) the CD300a TASR binding domain, and (z) the IdeS such that expression of the one or more exogenous polynucleotides is at least in part under the control of the exogenous promoter; and9.(c) a deletion or reduced expression of one or more of Beta 2 Microglobulin (B2M), Class II Major Histocompatibility Complex Transactivator (CIITA), and T Cell Receptor Alpha Constant (TRAC) genes;10.wherein the one or more exogenous polynucleotides are inserted at a locus of one or more of the B2M gene, the CIITA gene, and the TRAC gene, thereby deleting or reducing the expression of one or more of the B2M gene, the CIITA gene, and / or the TRAC gene.

2. The iPSC or the derivative cell of claim 1, wherein the exogenous polynucleotide encoding the one or more CARs is inserted at the locus of the TRAC gene, thereby deleting or reducing the expression of the TRAC gene.

3. The iPSC or the derivative cell of claim 1, wherein polynucleotide encoding the exogeneous promoter is operably linked to the polynucleotide encoding the one or more CARs.12.25513.4909-7891-2381.1 4. The iPSC or the derivative cell of claim 1, wherein polynucleotide encoding the exogeneous promoter is operably linked to the polynucleotide encoding the CD300a TASR binding domain, and, optionally, the IdeS.

5. The iPSC or the derivative cell of claim 1, wherein polynucleotide encoding the exogeneous promoter is operably linked to the polynucleotide encoding the CARs, and the CD300a TASR binding domain, and, optionally, the IdeS.

6. The iPSC or the derivative cell of claim 1 comprising a plurality of exogenous polynucleotides, comprising:16.(a) a first polynucleotide encoding an exogeneous promoter operably linked to the polynucleotide encoding the one or more CARs17.(b) a second exogenous polynucleotides encoding an exogeneous promoter operably linked to the polynucleotide encoding the CD300a TASR binding domain and, optionally, the IdeS.

7. The iPSC or the derivative cell of claim 1 or 6, wherein the one or more first or second exogenous polynucleotides are inserted at the locus of a gene independently selected from the group consisting of the B2M gene and the CIITA gene, thereby deleting or reducing the expression of one or both of the B2M gene and CIITA gene.

8. The iPSC or the derivative cell of any one of claims 1-7, wherein the exogeneous promoter is independently selected from the group consisting of EFla, a CAG promoter, a PGK promoter, or a CMV promoter, or a fragment of any thereof.

9. The iPSC or the derivative cell of claim 8, wherein the promoter comprises the EFla promoter or a fragment thereof.

10. The iPSC or the derivative cell of any one of claims 1-9, comprising a deletion or reduced expression of two or more of the B2M gene, the CIITA gene, and the TRAC gene.

11. The iPSC or the derivative cell of claim 10, comprising a deletion or reduced expression of the B2M gene, the CIITA gene, and the TRAC gene.

12. The iPSC or the derivative cell of any one of claims 1-11, wherein the first antigen binding domain and the second antigen binding domain of the CARs specifically bind to an antigen independently selected from the group consisting of Cluster of Differentiation (CD) 19 antigen, CD22 antigen, B cell maturation antigen (BCMA), and Nectin-4.24.25625.4909-7891-2381.1 13. The iPSC or the derivative cell of any one of claims 1-12, wherein the one or more exogenous polynucleotides encode both of the CD300a TASR binding domain and the IdeS, and wherein the CD300a TASR binding domain and the IdeS are operably linked by an autoprotease peptide.

14. The iPSC or the derivative cell according to claim 13, wherein the autoprotease peptide is a 2A peptide15. The iPSC or the derivative cell of any one of claims 1-14 wherein the polynucleotide encoding the CD300a TASR antibody or a fragment thereof comprises a polynucleotide encoding a single chain variable fragment (scFv) or a VHH16. The iPSC or the derivative cell of claim 15, wherein the VHH comprises the VH domain of a camelid heavy chain antibody17. The iPSC or the derivative cell of any one of claims 1-16, wherein the CD300a binding domain comprises a VHH (CD300a VHH).

18. The iPSC or the derivative cell of any one of claims 1-17, wherein the CD300a VHH comprises:31.(a) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising AAKPGEDVY (SEQ ID NO: 182);32.(b) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLSQFAS (SEQ ID NO: 183);33.(c) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPRSGWGL (SEQ ID NO: 184);34.(d) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence35.25736.4909-7891-2381.1 comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKTRYES (SEQ ID NO: 185);37.(e) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 174), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ ID NO: 179), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising NTRLAHGRDVLGGVAYDI (SEQ ID NO: 186);38.(f) a CDRl comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 174), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ ID NO: 179), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising NTRRLGRSGDLVQDY (SEQ ID NO: 187);39.(g) a CDRl comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSRYY (SEQ ID NO: 175), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPDRDY (SEQ ID NO: 188);40.(h) a CDRl comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLPDVLPLEY (SEQ ID NO: 189);41.(i) a CDRl comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYW (SEQ ID NO: 176), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ ID NO: 179), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKVDGSYGIVTEL (SEQ ID NO: 190);42.(j) a CDRl comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSDYA (SEQ ID NO: 174), a CDR2 comprising 0, 1, or 2 mutations 25843.4909-7891-2381.1 relative to an amino acid sequence comprising INGSGGST (SEQ ID NO: 180), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising HTRRSGTSMAMDV (SEQ ID NO: 191);44.(k) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ATKLTMVY (SEQ ID NO: 192);45.(l) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKLTNEY (SEQ ID NO: 193);46.(m) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKVRPSYEY (SEQ ID NO: 194);47.(n) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSPYY (SEQ ID NO: 177), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VAKPGYEY (SEQ ID NO: 195);48.(o) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGRT (SEQ ID NO: 181), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKPGEDVY (SEQ ID NO: 196);49.(p) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ISGSGGST (SEQ ID NO: 178), and a 25950.4909-7891-2381.1 CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising VTKSNMVY (SEQ ID NO: 197),51.(q) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYY (SEQ ID NO: 173), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising ITGSGGST (SEQ ID NO: 179), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising TTKVDGSYGIVTEL (SEQ ID NO: 198); or52.(r) a CDR1 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising GFTFSSYW (SEQ ID NO: 176), a CDR2 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising INGSGGST (SEQ ID NO: 180), and a CDR3 comprising 0, 1, or 2 mutations relative to an amino acid sequence comprising AAARDRERDY (SEQ ID NO: 199).

19. The iPSC or the derivative cell of claim 17, wherein the CD300a VHH comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 155-172.

20. The iPSC or the derivative cell of any one of claims 17 or 18, wherein the CD300a VHH comprises or consists of the amino acid sequence of any one of SEQ ID NOs: 155-172.

21. The iPSC or the derivative cellclaim 17, wherein a nucleotide sequence encoding the CD300a VHH comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 137-154.

22. The iPSC or the derivative cell of claim 17, wherein a nucleotide sequence encoding the CD300a VHH comprises or consists of the nucleotide sequence of any one of SEQ ID NOs: 137-154.

23. The iPSC or the derivative cell of claim 1, wherein the CD300a TASR binding domain comprises a scFv.

24. The iPSC or the derivative cell of claim 1, wherein,59.(a) the exogenous polynucleotide encoding the CD300a VL is codon optimized to reduce or prevent undesired recombination events; and / or60.26061.4909-7891-2381.1 (b) the exogenous polynucleotide encoding the CD300a VH is codon optimized to reduce or prevent undesired recombination events.

25. The iPSC or the derivative cell of any one of claims 1-24, further encoding a transmembrane domain.

26. The iPSC or the derivative cell of claim 25, wherein the transmembrane domain is a human transmembrane domain or a murine transmembrane domain.

27. The iPSC or the derivative cell of claim 25 or 26, wherein the transmembrane domain comprises or consists of a CD8, a CD80, an ITGA, an HLA-B57, a proCAR-4, a CD28, a KIR2DL1, a PDGFRB, or a CD86 transmembrane domain.

28. The iPSC or the derivative cell of claims 25-27, wherein66.(a) the transmembrane domain comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 67-76 and 96; or67.(b) the transmembrane domain comprises or consists of any one of SEQ ID NOs: 67-76 and 96.

29. The recombinant nucleic acid of any one of claims 25-28, wherein69.(a) a nucleotide sequence encoding the transmembrane domain comprises a nucleotide sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to any one of SEQ ID NOs: 56-65, or70.(b) a nucleotide sequence encoding the transmembrane domain comprises or consists of any one of SEQ ID NOs: 56-65.

30. The iPSC or the derivative cell of any one of claims 1-29, further encoding a polynucleotide encoding a cytoplasmic domain.

31. The iPSC or the derivative cell of claim 30, wherein the cytoplasmic domain is a human cytoplasmic domain or a murine cytoplasmic domain.

32. The iPSC or the derivative cell of claim 30 or 31, wherein the cytoplasmic domain comprises or consists of a CD8v2, a CD8vl, a mCD80, a CD80, a CD86, or an HLA-B57 cytoplasmic domain.

33. A vector comprising a polynucleotide of any one of claims 1-32.75.26176.4909-7891-2381.1 34. The vector of claim 33, wherein the vector is a DNA vector, an RNA vector, a plasmid, a lentivirus vector, an adenoviral vector, an adeno associated viral vector, a Rous sarcoma viral (RSV) vector, or a retrovirus vector.

35. The derivative cell of claim 1, wherein the derivative cell is an early hematopoietic progenitor cell or a CD34+ progenitor cell.

36. The derivative cell of claim 35, wherein the cell that has been differentiated from a stem cell or the cell that has been differentiated from a progenitor cell is an engineered T cell.

37. The derivative cell of any one of claims 35-36, wherein the engineered cell is an engineered T cell,38. The derivative cell of any one of claims 35-37 wherein the engineered T cell is a chimeric antigen receptor (CAR) T cell.

39. The derivative cell of claim 38, wherein the CAR is a CD19 CAR, a BCMA CAR, a CD22 CAR or a Nectin-4 CAR.

40. The derivative cell of any one of claims 35-39, wherein the engineered cell is T cell receptor alpha constant (TRAC) deficient.

41. The derivative cell of any one of claims 35-40, wherein a T cell receptor alpha constant (TRAC) locus is disrupted in the engineered cell.

42. The derivative cell of any one of claims 35-41, wherein the C AR is inserted into a T cell receptor alpha constant (TRAC) locus in the engineered cell.

43. A composition comprising the derivative cell of any one of claims 35-42, and one or more of a cell culture media and a buffer.

44. A pharmaceutical composition comprising the derivative cell of any one of claims 35-42, and a pharmaceutically acceptable carrier.

45. A method of improving a clinical outcome in a subject undergoing a T cell therapy, comprising administering to the subject an effective amount of the derivative cell of any one of claims 35-42, or the pharmaceutical composition of claim 44.

46. A method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of the derivative cell of any one of claims 35-42 or the pharmaceutical composition of claim 44.

47. The method of claim 46, wherein the disease or disorder is a cancer.

48. The method of one of claim 47, wherein the cancer is a hematologic cancer selected from 26291.4909-7891-2381.1 B-cell acute lymphoid leukemia (B-ALL), T cell acute lymphoid leukemia (T-ALL), acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL), B cell prolymphocytic leukemia, blastic plasmacytoid dendritic cell neoplasm, Burkitt’s lymphoma, diffuse large B cell lymphoma, follicular lymphoma, hairy cell leukemia, small cell-follicular lymphoma, large cell-follicular lymphoma, malignant lymphoproliferative conditions, MALT lymphoma, mantle cell lymphoma, Marginal zone lymphoma, multiple myeloma (MM), myelodysplasia, myelodysplastic syndrome, non-Hodgkin’s lymphoma, plasmablastic lymphoma, plasmacytoid dendritic cell neoplasm, Waldenstrom macroglobulinemia, or preleukemia.

49. The method of claim 47, wherein the cancer is fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, and other sarcomas, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, lymphoid malignancy, pancreatic cancer, breast cancer, lung cancer, ovarian cancer, prostate cancer, colorectal cancer, hepatocellular carcinoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, pheochromocytomas sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, Wilms' tumor, cervical cancer, testicular tumor, seminoma, bladder carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, melanoma, neuroblastoma or retinoblastoma.

50. The method of claim 46, wherein the disease or disorder is an autoimmune disease or disorder.

51. The method of the claim 50, * wherein the autoimmune disease or disorder is myasthenia gravis, neuromyelitis optica spectrum disorder, Sjogren’s syndrome, scleroderma, immune nephritis, systemic lupus erythematosus, arthritis, an autoimmune-induced fibrotic condition, pemphigus vulgaris, multiple sclerosis, colitis, graft-versus-host disease, atherosclerosis, or mucosal-dominant PV.

52. The method of any one of claims 45-51, wherein the subject is a human.96.26397.4909-7891-2381.1

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