Circular RNA compositions and methods of use thereof for targeted delivery

Abstract

The present disclosure provides compositions, methods of making, and use of a circular ribonucleic acid (circRNA) therapeutic for targeted delivery to immune cells, including T cells, for effector functions such as B cell depletion, providing, among other things, methods of treating B cell-mediated diseases such as autoimmune disease and cancer, meeting the medical need for efficacious and safe therapies that overcome the disadvantages of ex vivo CAR T therapy. Provided by the present disclosure is a circRNA therapeutic comprising: (i) a circRNA comprising a ribosome recruiting element, and a coding sequence encoding a CAR comprising, for example, an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) a targeting moiety (e.g, anti-CD5) that binds to an immune cell (e.g., a T cell), wherein the circRNA is in a delivery vehicle, and a targeting moiety that binds to a T cell is conjugated to the delivery vehicle (e.g, a lipid nanoparticle).

Description

ATTORNEY DOCKET NO. ORB-019WO1CIRCULAR RNA COMPOSITIONS AND METHODS OF USE THEREOF FOR TARGETED DELIVERYCROSS REFERENCE TO RELATED APPLICATIONS[1] The present application claims priority to, and the benefit of U. S. Provisional Application No: 63 / 763,317, filed on February 26, 2025, U. S. Provisional Application No: 63 / 846,699, filed on July 18, 2025, and U. S. Provisional Application No: 63 / 929,029, filed on December 2, 2025, the contents of which are incorporated herein by reference in their entireties.INCORPORATION BY REFERENCE OF SEQUENCE LISTING[2] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. The sequence listing file, created on February 20, 2026, is named ORB-019WOl.xml and is 630,784 bytes in size.BACKGROUND[3] In various diseases such as autoimmune disease and cancer, B cells contribute to disease pathogenesis via local production of inflammatory cytokines or by acting as antigen presenting cells (Sanz, 2010, Sarvaria et al., 2017). Autoantibodies produced by B cells are pathogenic in certain disease types (Ramirez -Valle, 2024). Most patients with autoimmune diseases fail to eliminate autoreactive B cells and autoantibodies due to impaired B cell tolerance (Pillai, 2011). Some clinical studies have shown that depletion of B cells has been associated with positive patient outcomes (Arnold, 2024; English, 2024; Blache, 2023).[4] Ex vivo chimeric antigen receptor (CAR) T cell therapy has several disadvantages, such as manufacturing processes that are complex, laborious, and expensive. Ex vivo CAR therapy requires patients’ or donors’ T cells to be isolated from peripheral blood via apheresis, activated, and subsequently transduced by a retrovirus (e.g., lentivirus or gammaretrovirus) comprising a chimeric antigen receptor (CAR) construct to create an engineered CAR T cell. CAR T cells are further expanded before infusing the engineered T cells into a patient’s bloodstream to attack target cells. Further, ex vivo CAR therapy requires lymphodepletion before administration, and is also associated with serious side effects, including cytokine release syndrome (CRS), which can cause multi-organ failure and evenATTORNEY DOCKET NO. ORB-019WO1death, and immune effector cell-associated neurotoxicity syndrome (ICANS), which is a neuropsychiatric syndrome that can cause confusion and seizures (Bui, 2024; Mullard, 2024).[5] There is an unmet need for efficacious and safe therapies to treat diseases such as autoimmune disease and cancer that overcome the disadvantages of ex vivo CAR therapy.SUMMARY OF THE INVENTION[6] The present disclosure provides, among other things, an improved in vivo CAR therapy for treating B cell-mediated diseases based on circular ribonucleic acids (circRNA). The present disclosure provides, among other things, targeted in vivo delivery of therapeutic genes to select cell types (e.g., immune cells, or, more specifically, T cells) for treating B cell-mediated diseases (e.g., autoimmune disease or cancer). The present disclosure provides, in part, a circular ribonucleic acid for targeted delivery comprising: (i) a circRNA comprising a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti -CD 19 antibody or antigen-binding fragment thereof, (ii) a targeting moiety, and (iii) a delivery vehicle, wherein the targeting moiety is conjugated to the delivery vehicle (e.g., lipid nanoparticle, viral particle, among others).[7] CircRNA therapeutics provide multiple advantages compared to linear RNA. For example, circRNA therapeutics provides enhanced stability. Without wishing to be bound by any mechanism, since circRNA is a closed molecule with no free ends, it confers resistance to exonucleases, which leads to RNA durability. In addition, the absence of open 5’ and 3’ ends leads to potential reduction of an innate immune response by the patient. Moreover, circular RNA allows for efficient translation compared to linear RNA; IRES-mediated translation (cap-independent) leads to high synthesis of the protein encoded by the circular RNA.[8] The present disclosure provides, in part, a circular ribonucleic acid for targeted delivery comprising: (i) a circRNA comprising a ribosome recruiting element (e.g., an Internal Ribosome Entry Site (IRES), IRES transacting factor (ITAF) recruiting element, polypyrimidine tract binding protein (PTB), branched cap or qRNA, etc.), and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (ii) a targeting moiety, wherein the targeting moiety binds to a T cell, wherein the circRNA is in a delivery vehicle, and a targeting moiety that binds to a T cell is conjugated to the delivery vehicle (e.g., a nanoparticle, including for example, lipid nanoparticle, viral particle, among others).ATTORNEY DOCKET NO. ORB-019WO1[9] Exemplary targeting moieties of the present disclosure for targeting immune cells, more specifically, T cells, include, without limitation, anti-CD2, anti-CD3, anti-CD5, anti-CD7, or anti-CD8 antibody or antigen-binding fragment thereof (e.g., more specifically, anti-CD5 antibody or antigen-binding fragment thereof). The present disclosure thus provides for specific targeting of a circRNA that comprises a chimeric antigen receptor comprising an anti-CD19 antibody or antigen-binding domain to T cells, thus localizing the anti-CD19 CAR to a specific cell type required for its action and minimizing off-target effects, thereby increasing safety and efficacy. In some embodiments, safety is further enhanced by the inclusion of microRNA binding sites in the untranslated regions (UTR) of the circRNA to minimize toxicity due to off-target effects.

[0010] In some embodiments, a single microRNA binding site (e.g., miR-122) is included in the 3’ UTR. In some embodiments, the microRNA binding site is flanked by an upstream spacer and a downstream spacer. Without wishing to be bound by any particular theory, the upstream and the downstream spacer facilitate circularization of the precursor RNA into circular RNA. In some embodiments, the upstream spacer and the downstream spacer are designed to have poly A sequences that are interrupted by insertion of a cytosine. In some embodiments, the upstream spacer and the downstream spacer are designed to have a hydropathy index that favors limiting interaction of the encapsulated circRNA with the lipid nanoparticle, facilitating release into a target cell.

[0011] In some embodiments, the circRNAs and circRNA therapeutics of the present disclosure were precisely engineered for in vivo CAR therapy. In the present disclosure, the circRNAs and circRNA therapeutics stably and durably express CAR to high levels, in part, due to the increased stability of circRNA relative to a linear RNA; they are codon-optimized for high expression of a CAR construct, for example, anti-CD19 CAR construct; include variant 5' and / or 3' untranslated regions; and ribosome recruiting elements, for example, cell type-optimized Internal Ribosome Entry Site (IRES) elements that drive unexpectedly high expression of the CAR construct, including anti-CD19 CAR coding sequence, in select cell types, for example, T cells.

[0012] Expression of anti-CD19 CAR by T cells causes activation of T cells and specific binding to target B cells to carry out B cell depletion. Such a B cell compartment immune reset depletes the existing population of B cells, allowing a new generation of naive B cell development from the bone marrow, thereby resetting the B cell immune population by, forATTORNEY DOCKET NO. ORB-019WO1example, depleting autoreactive B cells in autoimmune diseases and malignant B cells in B cell-mediated cancers, thereby treating the B cell-mediated disease.

[0013] In some embodiments of the present disclosure, a targeting moiety is conjugated to a delivery vehicle (e.g., lipid nanoparticles or viral particles) by various exemplary conjugations, including as non-limiting examples, chemical conjugations (thiol-maleimide, azide-dibenzocyclooctyne (DBCO), transcyclooctene (TCO)-tetrazine), and enzymatic conjugations (such as transglutaminase-catalyzed glutamine-amine conjugation, and sortase-catalyzed conjugation (such as a sortase-catalyzed LPXT / (G)n-(G)n conjugation, wherein, for e.g., n=l-5 and X is any amino acid residue), among other conjugation strategies. Exemplary conjugation strategies are described in FIG. 13 A and exemplary conjugation handles are provided in Table 12.

[0014] LNP delivery, among other things, increases stability, bioavailability, and distribution of circRNA to target tissues. The LNP formulations disclosed herein ensure that the circRNA encapsulated by the LNP are protected from degradation during delivery, and allow for efficient and targeted delivery to target cells, e.g., T cells. Provided herein are efficient methods of making stable lipid nanoparticles with sufficient LNP properties, such as size, poly dispersity and efficacy in encapsulating circRNA (as measured by percent encapsulation) for therapeutic applications. The conjugation handles and conjugation methods disclosed herein for conjugating the lipid nanoparticle to a targeting moiety minimizes toxicity from the delivery and accumulation of circRNA at unintended, off-target cells, and facilitates specific targeting.

[0015] In some embodiments, lipid nanoparticles of the present disclosure contain multiple polyethylene glycol (PEG)-lipids. In some embodiments, lipid nanoparticles of the present disclosure contain two or more PEG-lipids. In some embodiments, lipid nanoparticles of the present disclosure contain two PEG-lipids, wherein one PEG is used for conjugation and the other PEG is structural. Although PEG was used for conjugation, increasing the total PEG used for conjugation did not increase targeting. Counterintuitively, a lower total PEG content (e.g., 0.1% PEG) was associated with higher CAR expression.

[0016] Further, in some embodiments, the conjugating PEG comprises a long chain lipid anchor component between about C14 and C22 (e.g., C18 PEG). As described in more detail in the Examples, in some embodiments, a higher mole percent of total C18 PEG (conjugating PEG and structural PEG) was found to decrease LNP distribution to the liver (off-target organ) and increase LNP distribution to the spleen (target organ).ATTORNEY DOCKET NO. ORB-019WO1

[0017] In further embodiments, ionizable lipids and the ratios of different lipid components were optimized to achieve specific targeting to T cells. In some embodiments, increasing phospholipid mole percentage (mol%), for example, increasing distearoylphosphatidylcholine (DSPC) mole percentage (mol%), increased potency and decreased off-target expression in liver.

[0018] In summary, the present disclosure provides, in part, a highly stable, durable and targeted in vivo CAR therapy encoded by circRNA that is achieved through an unexpected and surprising synergy of a combination of precisely engineered features in the circRNA, delivery vehicle (e.g., lipid nanoparticle), and targeting moiety conjugated to the delivery vehicle. In some embodiments, the targeted in vivo CAR therapy in the present disclosure provides for in vivo expression of the CAR therapy in specific cells, and use of circRNA improves the duration and magnitude of CAR expression on T cells. As described herein, the targeted in vivo CAR therapy reaches the specific cell types and tissues, e.g., T cells, and allows for enhanced in vivo CAR T generation and improves potency for B cell depletion. Among other things, the targeted in vivo CAR T therapy of the present disclosure requires no apheresis or lymphodepletion and is associated with greater ease of manufacturing and lower costs. The patient’s own body serves as a manufacturer of CAR-T cells, and this in vivo CAR therapy may be less prone to cytokine release syndrome and toxicity effects.

[0019] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: (i) a circRNA comprising a ribosome recruiting element, and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) a targeting moiety, wherein the circRNA is encapsulated in a delivery vehicle, wherein the targeting moiety is conjugated to the delivery vehicle, and wherein the targeting moiety comprises an anti-CD5 antibody heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89.

[0020] In some embodiments, the CDR1 sequence comprises SEQ ID NO: 64, the CDR2 sequence comprises SEQ ID NO: 65, and the CDR3 sequence comprises SEQ ID NO: 66.

[0021] In some aspects, provided herein is a circular ribonucleic acid (circRNA) for targeted delivery comprising: (i) a circRNA comprising: a ribosome recruiting element, and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) a targeting moiety, wherein the targeting moiety binds to a T cell, wherein the circRNA is in a delivery vehicle, wherein theATTORNEY DOCKET NO. ORB-019WO1targeting moiety that binds to a T cell is conjugated to the delivery vehicle, and wherein the targeting moiety is an anti-CD5 antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising complementarity determining region (CDR) sequencesGFTFSMYS (SEQ ID NO: 64), ISTGARDT (SEQ ID NO: 65) and GDLRYGPDGYDY (SEQ ID NO: 66).

[0022] In some embodiments, the delivery vehicle is a lipid nanoparticle (LNP).

[0023] In some embodiments, the anti-CD5 antibody or antigen-binding fragment comprises a sequence having at least 95% identity to SEQ ID NO: 89.

[0024] In some embodiments, the anti-CD5 antibody or antigen-binding fragment comprises SEQ ID NO: 89.

[0025] In some embodiments, the anti-CD5 antibody or antigen-binding fragment is a VHH.

[0026] In some embodiments, the anti-CD5 antibody or antigen-binding fragment is conjugated to a conjugating PEG-lipid in the delivery vehicle.

[0027] In some embodiments, the anti-CD5 antibody or antigen-binding fragment is covalently attached to a conjugation handle.

[0028] In some embodiments, the conjugation handle is covalently attached to the C-terminus of the heavy chain of the anti-CD5 antibody or antigen-binding fragment.

[0029] In some embodiments, the conjugation handle is 5-10 amino acids in length.

[0030] In some embodiments, the conjugation handle comprises SEQ ID NO: 120.

[0031] In some embodiments, the conjugation handle consists of SEQ ID NO: 120.

[0032] In some embodiments, the anti-CD5 antibody or antigen-binding fragment is covalently attached to the conjugation handle, and the sequence of the anti-CD5 antibody or antigen-binding fragment with attached conjugation handle comprises SEQ ID NO: 122.

[0033] In some embodiments, the anti-CD5 antibody or antigen-binding fragment is conjugated to the conjugating PEG-lipid in the delivery vehicle via a thiol-maleimide conjugation.

[0034] In some embodiments, the anti-CD5 antibody or antigen-binding fragment is conjugated to the conjugating PEG-lipid in the delivery vehicle via a thiol-maleimide conjugation between the conjugating PEG-lipid and the conjugation handle.

[0035] In some embodiments, the delivery vehicle is an LNP comprising two PEG-lipids, one of which is the conjugating PEG-lipid.ATTORNEY DOCKET NO. ORB-019WO1

[0036] In some embodiments, the conjugating PEG-lipid comprises a long chain fatty acid.

[0037] In some embodiments, the conjugating PEG-lipid is DSPE-PEG 2000-maleimide.

[0038] In some embodiments, the LNP further comprises one or more ionizable lipids, one or more phospholipids, and one or more sterol lipids.

[0039] In some embodiments, the LNP comprises the one or more ionizable lipids at 30 to 49 mol% (e.g., at 30 to 45 mol%), the one or more phospholipids at 5 to 40 mol% (e.g., at 15 to 35 mol%), the one or more sterol lipids at 15 to 50 mol% (e.g., at 30 to 50 mol%), the PEG lipid (e.g., the DMG-PEG 2000) at 0 to 10 mol% (e.g., at 0 to 5 mol%), and the conjugating PEG-lipid (e.g., the DSPE-PEG 2000-maleimide) at 0.01-1 mol%.

[0040] In some embodiments, the LNP comprises the one or more ionizable lipids at 30 to 49 mol%, the one or more phospholipids at 5 to 40 mol%, the one or more sterol lipids at 15 to 50 mol%, the DMG-PEG 2000 lipid at 0 to 10 mol%, and the DSPE-PEG 2000-maleimide lipid at 0.01-1 mol%.

[0041] In some embodiments, the LNP comprises the one or more ionizable lipids at 30 to 45 mol%, the one or more phospholipids at 15 to 35 mol%, the one or more sterol lipids at 15 to 50 mol%, the DMG-PEG 2000 lipid at 0 to 5 mol%, and the DSPE-PEG 2000-maleimide lipid at 0.01-1 mol%.

[0042] In some embodiments, the LNP comprises the one or more ionizable lipids, the one or more phospholipids, the one or more sterol lipids, the DMG-PEG 2000 lipid, and the DSPE-PEG 2000-maleimide lipid at a ratio of 39: 20: 39: 1.9: 0.1 mol%.

[0043] In some embodiments, the LNP comprises the one or more ionizable lipids, the one or more phospholipids, the one or more sterol lipids, the DMG-PEG 2000 lipid, and the DSPE-PEG 2000-maleimide lipid combined at a molar ratio of 39: 20: 39: 1.9: 0.1.

[0044] In some embodiments, the one or more ionizable lipids is(CompoundATTORNEY DOCKET NO. ORB-019WO1

[0045] In some embodiments, the one or more phospholipids is 1,2-distearoyl-snglycero-3-phosphocholine (DSPC).

[0046] In some embodiments, the sterol lipid is cholesterol.

[0047] In some embodiments, the ribosome recruiting element is an Internal Ribosome Entry Site (IRES), wherein the IRES comprises a sequence selected from Table 1.

[0048] In some embodiments, the IRES comprises a sequence selected from SEQ ID NO: 1-11 of Table 1.

[0049] In some embodiments, the IRES comprises SEQ ID NO: 1.

[0050] In some embodiments, the coding sequence further encodes a CD8 signal peptide, andthe CAR comprises a CD8 hinge, a CD8 transmembrane domain, a 4-1BB costimulatory domain, and a CD3 zeta signaling domain.

[0051] In some embodiments, the anti-CD19 antibody or antigen-binding fragment thereof comprises a VL sequence of SEQ ID NO: 54 and a VH sequence of SEQ ID NO: 55.

[0052] In some embodiments, the coding sequence encodes an amino acid sequence of SEQ ID NO: 58.

[0053] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: a circular RNA comprising a ribosome recruiting element and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA is encapsulated in an LNP, wherein a targeting moiety is conjugated to the LNP, wherein the targeting moiety is an anti-CD5 VHH, the anti-CD5 VHH comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89, wherein the LNP comprises one or more ionizable lipids at 30 to 49 mol% (e.g, at 30 to 45 mol%), one or more phospholipids at 5 to 40 mol% (e.g., at 15 to 35 mol%), one or more sterol lipids at 15 to 50 mol% (e.g., at 30 to 50 mol%), DMG-PEG 2000 at 0 to 10 mol% (e.g., at 0 to 5 mol%), and DSPE-PEG 2000-maleimide at 0.01 to 1 mol%, and wherein the VHH is conjugated to the LNP.

[0054] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: a circular RNA comprising a ribosome recruiting element and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA is encapsulated in an LNP, wherein a targeting moiety is conjugated to the LNP, wherein the targeting moiety is an anti-CD5 VHH, the anti-CD5 VHH comprising CDR1, CDR2, andATTORNEY DOCKET NO. ORB-019WO1CDR3 sequences from SEQ ID NO: 89, wherein the LNP comprises one or more ionizable lipids at 30 to 49 mol%, one or more phospholipids at 5 to 40 mol%, one or more sterol lipids at 15 to 50 mol%, a DMG-PEG 2000 lipid at 0 to 10 mol%, and a DSPE-PEG 2000-maleimide lipid at 0.01 to 1 mol%, and wherein the VHH is conjugated to the LNP.

[0055] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: a circular RNA comprising a ribosome recruiting element and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA is encapsulated in an LNP, wherein a targeting moiety is conjugated to the LNP, wherein the targeting moiety is an anti-CD5 VHH, the anti-CD5 VHH comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89, wherein the LNP comprises one or more ionizable lipids at 30 to 45 mol%, one or more phospholipids at 15 to 35 mol%, one or more sterol lipids at 30 to 50 mol%, a DMG-PEG 2000 lipid at 0 to 5 mol%, and a DSPE-PEG 2000-mal eimide lipid at 0.01 to 1 mol%, and wherein the VHH is conjugated to the LNP.

[0056] In some embodiments, the VHH has a length of 110 to 130 (e.g., 115 to 125) amino acids. In some embodiments, the VHH has a length of 110 to 130 amino acids. In some embodiments, the VHH has a length of 115 to 125 amino acids. In some embodiments, the VHH has a length of about 121 amino acids.

[0057] In some embodiments, the VHH is conjugated to the LNP via a thiol-maleimide conjugation, and the thiol-maleimide conjugation is between the DSPE-PEG 2000-maleimide and a conjugation handle that is covalently attached to the VHH.

[0058] In some embodiments, the N-terminus of the conjugation handle is covalently attached to the C-terminus of the VHH.

[0059] In some embodiments, the VHH with the attached conjugation handle has a length of 120 to 140 amino acids (e.g., 125 to 135 amino acids). In some embodiments, the VHH with the attached conjugation handle has a length of 120 to 140 amino acids. In some embodiments, the VHH with the attached conjugation handle has a length of 125 to 135 amino acids.

[0060] In some embodiments, the VHH with the attached conjugation handle has a length of about 129 amino acids.

[0061] In some embodiments, the VHH comprises SEQ ID NO: 89.

[0062] In some embodiments, the conjugation handle comprises SEQ ID NO: 120.ATTORNEY DOCKET NO. ORB-019WO1

[0063] In some embodiments, the sequence of the VHH with the attached conjugation handle comprises SEQ ID NO: 122.

[0064] In some embodiments, the sequence of the VHH with the attached conjugation handle is SEQ ID NO: 122.

[0065] In some embodiments, the LNP comprises the one or more ionizable lipids, the one or more phospholipids, the one or more sterol lipids, the DMG-PEG 2000 lipid, and the DSPE-PEG 2000-maleimide lipid at a ratio of 39: 20: 39: 1.9: 0.1 mol%.

[0066] In some embodiments, the one or more ionizable lipids is Compound 7.

[0067] In some embodiments, the one or more phospholipids is 1,2-distearoyl-snglycero-3-phosphocholine (DSPC).

[0068] In some embodiments, the one or more sterol lipids is cholesterol.

[0069] In some embodiments, provided herein is a circRNA therapeutic of the present disclosure comprising a circRNA sequence having at least 85% identity to SEQ ID NO: 264.

[0070] In some embodiments, provided herein is a circRNA therapeutic of the present disclosure comprising a circRNA sequence according to SEQ ID NO: 264.

[0071] In some aspects, provided herein is a circRNA therapeutic comprising(i) a circular RNA (circRNA) comprising a coding sequence encoding a CD 19 CAR, wherein the circRNA is encapsulated in an LNP,

[0072] (ii) the LNP, wherein the LNP comprises Compound 7 at 39 mol%, DSPC at 20 mol%, Cholesterol at 39 mol%, DMG-PEG 2000 lipid at 1.9 mol%, and DSPE-PEG 2000-maleimide lipid at 0.1 mol%, and(iii) a VHH with a conjugation handle comprising the sequence of SEQ ID NO: 122, wherein the VHH with the conjugation handle is attached to the LNP via thiol-maleimide conjugation of the cysteine in the conjugation handle with the DSPE-PEG 2000-maleimide lipid.

[0073] In some embodiments, the circRNA comprises a sequence according to SEQ ID NO: 264.

[0074] In some aspects, provided herein is a circRNA therapeutic, comprising: (i) a circRNA comprising in order: (a) a 5’ exon fragment and a 5' UTR, (b) an IRES having at least 85% identity to SEQ ID NO: 1, (c) a coding sequence encoding a chimeric antigen receptor (CAR) that targets CD 19, (d) a 3' UTR and a 3’ exon fragment, and (ii) an anti-CD5 T cell targeting moiety comprising the complementarity determining region (CDR) sequences CDR1, CDR2, and CDR3, wherein the CDR1, the CDR2, and the CDR3 amino acid sequences are GFTFSMYS (SEQ ID NO: 64), ISTGARDT (SEQ ID NO: 65), andATTORNEY DOCKET NO. ORB-019WO1GDLRYGPDGYDY (SEQ ID NO: 66), respectively, wherein the circRNA is encapsulated in a lipid nanoparticle, and wherein an anti-CD5 T cell targeting moiety is conjugated to the lipid nanoparticle.

[0075] In some embodiments, the coding sequence encoding a CAR comprises SEQ ID NO: 125.

[0076] In some embodiments, the CAR comprises an anti-CD19 scFv,a CD8a hinge, a CD8a transmembrane domain, a 4-1BB costimulatory domain, and a CD3 zeta signaling domain.

[0077] In some embodiments, the CAR comprises an amino acid sequence of SEQ ID NO: 58.

[0078] In some embodiments, the anti-CD19 scFv comprises a VL sequence of DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGV PSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT (SEQ ID NO: 54), linked by a linker having the amino acid sequence of GGGGSGGGGSGGGGS (SEQ ID NO: 56), and a VH sequence of EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETT YYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQ GTSVTVSS (SEQ ID NO: 55).

[0079] In some aspects, provided herein isa circular ribonucleic acid (circRNA) comprising in order: (a) a 5’ exon fragment and 5’ untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES) comprising a sequence at least 85% identical to any one of SEQ ID NO: 1-11, (c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (d) a 3’ UTR and a 3’ exon fragment, and (e) a single microRNA binding site within the 5’ UTR or the 3’ UTR.

[0080] In some embodiments, the 3’ UTR comprises the single microRNA binding site.

[0081] In some embodiments, the microRNA binding site comprises SEQ ID NO: 40.

[0082] In some embodiments, the single microRNA binding site is flanked by an upstream spacer and a downstream spacer, and wherein the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3.

[0083] In some embodiments, the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.4.

[0084] In some embodiments, the sequence of each of the upstream spacer has a hydropathy index of -0.6 to -0.5.ATTORNEY DOCKET NO. ORB-019WO1

[0085] In some embodiments, each of the upstream spacer and downstream spacer comprise consecutive adenines but do not comprise more than five (5) consecutive adenines (As).

[0086] In some embodiments, each of the upstream spacer and downstream spacer comprise consecutive adenines but do not comprise more than four (4) consecutive adenines (As).

[0087] In some embodiments, each of the upstream spacer and downstream spacer comprise consecutive adenines but do not comprise more than three (3) consecutive adenines (As).

[0088] In some aspects, provided herein is a circular ribonucleic acid (circRNA) comprising, in order: (a) a 5’ exon fragment and a 5’ untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES), (c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (d) a 3’ UTR comprising an upstream spacer, a microRNA binding site, and a downstream spacer, and a 3’ exon fragment, wherein each of the upstream spacer and downstream spacer comprise multiple instances of consecutive adenines (As) but do not comprise more than five (5) consecutive adenines (As).

[0089] In some embodiments, each of the upstream spacer and the downstream spacer do not comprise more than four (4) consecutive adenines (As).

[0090] In some embodiments, each of the upstream spacer and the downstream spacer do not comprise more than three (3) consecutive adenines (As).

[0091] In some embodiments, the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3.

[0092] In some embodiments, the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.4.

[0093] In some embodiments, each of the upstream spacer and downstream spacer comprise one or more instances of a cytosine (C) interrupting consecutive As.

[0094] In some embodiments, each of the upstream spacer and downstream spacer comprise no uracil.

[0095] In some embodiments, the upstream spacer sequence and the downstream spacer sequence consist only of As and Cs.

[0096] In some embodiments, each of the upstream spacer and downstream spacer have a length of not more than 18 nucleotides.ATTORNEY DOCKET NO. ORB-019WO1

[0097] In some embodiments, each of the upstream spacer and downstream spacer have a length of 12-18 nucleotides.

[0098] In some embodiments, each of the upstream spacer and downstream spacer have a length of 12-15 nucleotides.

[0099] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 4 nucleotides.

[0100] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 3 nucleotides.

[0101] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 2 nucleotides.

[0102] In some embodiments, the microRNA binding site is a microRNA-122 (miR-122) binding site.

[0103] In some embodiments, the 3’ UTR comprises TAATAGaacaaacacaaacCAAACACCATTGTCACACTCCAaacaacaaacaaacaACCACACA AATGGTCGCCGA (SEQ ID NO: 349).

[0104] In some embodiments, the 5’ UTR comprises TCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCAT ATGACCTCTGCGTGTGCCTGGTAATTGACTAAGATATC (SEQ ID NO: 348).

[0105] In some embodiments, the 5’ UTR comprises SEQ ID NO: 348 and the 3’ UTR comprises SEQ ID NO: 349.

[0106] In some embodiments, the 5’ exon fragment comprises SEQ ID NO: 382 and a 3’ exon fragment comprises SEQ ID NO: 389.

[0107] In some embodiments, the IRES comprises a sequence having at least 90% identity to any one of SEQ ID NO: 1-6.

[0108] In some embodiments, the IRES comprises the sequence of SEQ ID NO: 1.

[0109] In some embodiments of the circRNA, wherein: the coding sequence further encodes a CD8 signal peptide, and the CAR further comprises a CD8 hinge, a CD8 transmembrane domain, a 4-1BB costimulatory domain, and a CD3 zeta signaling domain.

[0110] In some embodiments, the anti-CD19 antibody or antigen-binding fragment thereof comprises a VL sequence of SEQ ID NO: 54, and a VH sequence of SEQ ID NO: 55.

[0111] In some embodiments, the VL and the VH are joined by a linker.

[0112] In some embodiments, the linker comprises SEQ ID NO: 56.ATTORNEY DOCKET NO. ORB-019WO1

[0113] In some embodiments, the coding sequence encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 58.

[0114] In some embodiments, the coding sequence encodes an amino acid sequence having at least 95% identity to SEQ ID NO: 58.

[0115] In some embodiments, the coding sequence encodes an amino acid sequence of SEQ ID NO: 58.

[0116] In some aspects, provided herein is a circRNA comprising a sequence having at least 85% identity to SEQ ID NO: 264.

[0117] In some embodiments, provided herein is a circRNA comprising a sequence having at least 90% identity to SEQ ID NO: 264.

[0118] In some embodiments, provided herein is a circRNA comprising a sequence having at least 95% identity to SEQ ID NO: 264.

[0119] In some embodiments, provided herein is a circRNA comprising a sequence having at least 98% identity to SEQ ID NO: 264.

[0120] In some embodiments, provided herein is a circRNA comprising a sequence having at least 99% identity to SEQ ID NO: 264.

[0121] In some embodiments, the circRNA comprises SEQ ID NO: 264.

[0122] In some embodiments, provided herein is a circRNA therapeutic, comprising circRNA of the present disclosure.

[0123] In some embodiments, the circRNA is as provided by the present disclosure.

[0124] In some embodiments, the circRNA therapeutic comprises the circRNA of the present disclosure encapsulated in an LNP.

[0125] In some embodiments, the LNP is conjugated to a targeting moiety, wherein the targeting moiety binds to T cells.

[0126] In some embodiments, the targeting moiety comprises an anti-CD5 antibody or antigen-binding fragment thereof.

[0127] In some aspects, provided herein is a precursor RNA comprising, from 5' to 3' end: (i) a 5' intron fragment and a 5’ exon fragment, (ii) a 5' untranslated region (UTR), (iii) an IRES, (iv) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (v) a 3' untranslated region (UTR), and (vi) a 3' exon fragment and a 3’ intron fragment, wherein the 5' and 3' intron fragments are derived from a Twort-ORF142 Group I intron, and wherein the 3’ UTR comprises a single microRNA binding site.ATTORNEY DOCKET NO. ORB-019WO1

[0128] In some embodiments, the single microRNA binding site is flanked by an upstream spacer and a downstream spacer.

[0129] In some embodiments, each of the upstream spacer and the downstream spacer comprise no more than five (5) consecutive adenines (As).

[0130] In some embodiments, each of the upstream spacer and the downstream spacer do not comprise more than four (4) consecutive adenines (As).

[0131] In some embodiments, each of the upstream spacer and the downstream spacer do not comprise more than three (3) consecutive adenines (As).

[0132] In some embodiments, the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3.

[0133] In some embodiments, the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.3.

[0134] In some embodiments, each of the upstream spacer and downstream spacer comprise one or more instances of a cytosine (C) interrupting consecutive As.

[0135] In some embodiments, each of the upstream spacer and downstream spacer comprise no uracil.

[0136] In some embodiments, the upstream spacer sequence and the downstream spacer sequence consist only of As and Cs.

[0137] In some embodiments, each of the upstream spacer and downstream spacer have a length of not more than 18 nucleotides.

[0138] In some embodiments, each of the upstream spacer and downstream spacer have a length of 12-18 nucleotides.

[0139] In some embodiments, each of the upstream spacer and downstream spacer have a length of 12-15 nucleotides.

[0140] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 4 nucleotides.

[0141] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 3 nucleotides.

[0142] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 2 nucleotides.

[0143] In some embodiments, the microRNA binding site is a microRNA-122 (miR-122) binding site.ATTORNEY DOCKET NO. ORB-019WO1

[0144] In some aspects, provided herein is a precursor RNA comprising, from 5' to 3' end: (i) a 5' intron fragment and a 5’ exon fragment, (ii) a 5' untranslated region (UTR), (iii) an IRES, (iv) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (v) a 3' untranslated region (UTR), and (vi) a 3' exon fragment and 3’ intron fragment, wherein the 5' and the 3' intron fragments are derived from a Twort-ORF142 Group I intron, wherein the 3’ UTR comprises a microRNA binding site flanked by an upstream spacer and a downstream spacer, and wherein each of the upstream spacer and the downstream spacer comprise no more than five (5) consecutive adenines (As).

[0145] In some embodiments, each of the upstream spacer and the downstream spacer comprise no more than four (4) consecutive adenines (As).

[0146] In some embodiments, each of the upstream spacer and the downstream spacer comprise no more than three (3) consecutive adenines (As).

[0147] In some embodiments, each of the upstream spacer and the downstream spacer comprise one or more instances of a cytosine (C) interrupting consecutive As.

[0148] In some embodiments, each of the upstream spacer and the downstream spacer comprise no uracil.

[0149] In some embodiments, each of the upstream spacer and the downstream spacer comprise 12-18 nucleotides.

[0150] In some embodiments, each of the upstream spacer and downstream spacer have a length of 12-15 nucleotides.

[0151] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 4 nucleotides.

[0152] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 3 nucleotides.

[0153] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 2 nucleotides.

[0154] In some embodiments, the microRNA binding site is a microRNA-122 (miR-122) binding site.

[0155] In some embodiments, the 5’ intron fragment comprises SEQ ID NO: 381.

[0156] In some embodiments, the 5’ exon fragment comprises SEQ ID NO: 382.

[0157] In some embodiments, the 5’ UTR comprises SEQ ID NO: 348.ATTORNEY DOCKET NO. ORB-019WO1

[0158] In some embodiments, the IRES comprises a sequence having at least 90% identity to any one of SEQ ID NO: 1-6.

[0159] In some embodiments, the IRES comprises the sequence of SEQ ID NO: 1.

[0160] In some embodiments, the coding sequence encoding the CAR comprising the anti-CD19 antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 355.

[0161] In some embodiments, the 3’ UTR comprises SEQ ID NO: 349 or 350.

[0162] In some embodiments, the 3’ exon fragment comprises SEQ ID NO: 389.

[0163] In some embodiments, the 3’ intron fragment comprises SEQ ID NO: 390.

[0164] In some embodiments, the 5’ UTR comprises SEQ ID NO: 348 and the 3’ UTR comprises SEQ ID NO: 349.

[0165] In some embodiments, the 5’ intron fragment and the 5’ exon fragment together comprises SEQ ID NO: 276 and the 3’ exon fragment and the 3’ intron fragment together comprises SEQ ID NO: 277.

[0166] In some aspects, provided herein is a precursor RNA comprising, from 5' to 3' end: (i) a 5' intron fragment and a 5’ exon fragment together comprising SEQ ID NO: 276, (ii) a 5' untranslated region (UTR), (ii) an IRES, (iii) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (iv) a 3' untranslated region (UTR), and (v) a 3’ exon fragment and a 3' intron fragment together comprising SEQ ID NO: 277.

[0167] In some embodiments, provided herein is a circular RNA circularized from the precursor RNA of the present disclosure.

[0168] In some embodiments, provided herein is a circular RNA (circRNA) therapeutic comprising the circular RNA of the present disclosure.

[0169] In some embodiments, provided herein is a method of depleting B cells comprising administering to a subject in need thereof the circRNA therapeutic of the present disclosure.

[0170] In some embodiments, provided herein is a method of treating a B cell-mediated disease comprising administering to a subject in need thereof the circRNA therapeutic of the present disclosure.

[0171] In some embodiments, the B cell-mediated disease is an autoimmune disease. In some embodiments, the autoimmune disease is one or more of lupus (e.g., systemic lupus erythematosus), an inflammatory myopathy, systemic sclerosis, rheumatoid arthritis, Sjogrens disease, multiple sclerosis (e.g., PPMS, RMS), myasthenia gravis, autoimmuneATTORNEY DOCKET NO. ORB-019WO1hemolytic anemia, immune thrombocytopenia, and inflammatory bowel disease (e.g., Crohn’s disease, ulcerative colitis).

[0172] In some embodiments, the B cell-mediated disease is cancer.BRIEF DESCRIPTION OF THE DRAWINGS

[0173] The drawings are for illustration purposes only and are not meant to be limiting.

[0174] FIG. 1A is a schematic showing a template DNA (e.g., plasmid), which is transcribed to synthesize an exemplary linear precursor RNA (from which the circular RNA of the present disclosure is derived). In some embodiments, as shown in FIG. IB, autocatalytic self-splicing of two halves of a Permuted Group I Intron-Exon or PIE construct (e.g., a 5’ intron fragment and a 3’ intron fragment) leads to the formation of the singlestranded circular RNA (circRNA) drug substance of the present disclosure that encodes an anti-CD19 chimeric antigen receptor (CAR) for treatment of B-cell-driven diseases (e.g., autoimmune disease and cancer). The circRNA sequence comprises, for example, a 5’ exon fragment and 5’ untranslated region (UTR), a ribosome recruiting element (e.g., an internal ribosome entry site (IRES)), a coding sequence (CDS), and a 3’ UTR and 3’ exon fragment, with the 5’ and / or 3’ UTRs optionally containing one or more spacers, and the 5' exon fragment and 3' exon fragment are joined at a splice junction. As shown in more detail in FIG. 1C, after autocatalytic self-splicing of two halves of a PIE construct, the 5’ exon fragment and 3’ exon fragment are ligated, and the intron is excised from the circular RNA. The splice junction shows the boundary after self-splicing and circularization, leaving the 5' exon fragment and 3' exon fragment adjacent to each other at the splice junction. The nucleobase composition of the drug substance comprises adenine (represented by A’s), cytosine (represented by C’s), guanine (represented by G’s), and uracil (represented by T’s or U’s; in the context of RNA, it is understood that T’s and U’s both represent uracil). In some embodiments, the circRNA is 2000 to 2700 nucleotides in length. In some embodiments, the circRNA is 2000 to 2500 nucleotides in length. In some embodiments, the circRNA is 2200 to 2500 nucleotides in length. In some embodiments, the circRNA is 2300 to 2400 nucleotides in length. In some embodiments, the circRNA is 2300 to 2350 nucleotides in length. In some embodiments, the circRNA is about 2000 nucleotides long. In some embodiments, the circRNA is about 2100 nucleotides long. In some embodiments, the circRNA is about 2200 nucleotides long. In some embodiments, the circRNA is about 2300 nucleotides long. In some embodiments, the circRNA is about 2330 nucleotides long. In some embodiments, the circRNA is 2332 nucleotides long. In some embodiments, theATTORNEY DOCKET NO. ORB-019WO1circRNA is about 2400 nucleotides long. In some embodiments, the circRNA is about 2500 nucleotides long.

[0175] FIG. ID shows a schematic of a circRNA drug product of the present disclosure comprising a circRNA encoding for, for example, an anti-CD19 CAR, encapsulated by a lipid nanoparticle (LNP) containing an exemplary ionizable lipid (e.g., Compound 7) conjugated to a targeting moiety, for example, an anti-CD5 VHH (nanobody) for robust targeted delivery to T cells, including CD4+and CD8+T cells.

[0176] FIG. IE is a schematic of the circRNA that shows an exemplary mechanism of action of the circRNA drug product without wishing to be bound by any particular theory. Although not bound by any mechanism, the targeting moiety, e.g., anti-CD5 nanobody, allows specific targeting of the circRNA encapsulated in the LNP to the targeting receptor, e.g., T cells that express CD5. Thus, the circRNA encoding the anti-CD19 CAR is delivered to the T cell, which uses the T cell’s own machinery to translate the anti-CD19 CAR and express the CAR on the surface of the T cell. The CAR T cell then binds to its target, e.g., B cells expressing CD 19, via the antigen-binding domain of the CAR. Antigen binding to the CAR triggers intracellular signaling domains, leading to T cell activation that induces secretion of cytokines and cytotoxic molecules. These molecules destroy and eliminate target B cells, resulting in B cell depletion, which is the intended on-target effect for the therapeutic treatment of B cell-mediated diseases.

[0177] FIG. 2A is a plot of area under the curve showing reporter gene expression from circRNA constructs comprising multiple exemplary IRESs. The performance of various IRESs was evaluated in vitro in comparison to a reference IRES (IRES090).

[0178] FIG. 2B is a plot of area under the curve (average mean fluorescence per cell) for circRNA constructs comprising various exemplary IRESs.

[0179] FIG. 2C is a graph of percentage of CAR-positive activated and resting T cells (of total live CD3-positive cells), two days post-transfection of circRNA constructs comprising various exemplary IRESs.

[0180] FIG. 2D is a graph of total CAR molecules per CAR T cell in resting and activated T cells showing CAR expression per cell, two days post-transfection of circRNA constructs comprising various exemplary IRESs.

[0181] FIG. 2E is a graph of gene expression (gMFI) of circRNA comprising exemplary IRESs on day 1 post-transfection of the circRNA comprising exemplary IRESs relative to a reference IRES (IRES090). The different data points represent different T cell donors.ATTORNEY DOCKET NO. ORB-019WO1

[0182] FIG. 2F is a graph of in vivo anti-CD19 CAR expression in CD3+T cells from spleens of hPBMC (human peripheral blood mononuclear cells)-engrafted NSGDKO mice as a percentage of total CD3+T cells at 48 hours after administration of circRNA (with various exemplary IRESs) formulated in LNP.

[0183] FIG. 2G is a graph of in vivo anti-CD19 CAR expression in CD3+T cells from bone marrow of hPBMC-engrafted NSG DKO mice as a percentage of total CD3+T cells at 48 hours after administration of circRNA (with various exemplary IRESs) formulated in LNP.

[0184] FIG. 2H is a graph of in vivo anti-CD19 CAR expression in CD3+T cells from peripheral blood of hPBMC-engrafted NSG DKO mice as a percentage of total CD3+T cells at 48 hours after administration of circRNA (with various exemplary IRESs) formulated in LNP.

[0185] FIG. 21 is a graph of proportion of B cells to mean B cell count of vehicle control in spleens of hPBMC-engrafted NSG DKO mice at 48 hours after administration of circRNA (with various exemplary IRESs) formulated in LNP.

[0186] FIG. 2J is a graph of proportion of B cells to mean B cell count of vehicle control in bone marrow of hPBMC-engrafted NSG DKO mice at 48 hours after administration of circRNA (with various exemplary IRESs) formulated in LNP.

[0187] FIG. 2K is a graph of proportion of B cells to mean B cell count of vehicle control in peripheral blood of hPBMC-engrafted NSG DKO mice at 48 hours after administration of circRNA (with various exemplary IRESs) formulated in LNP.

[0188] FIG. 3A is a schematic of circRNA showing, amongst the other elements described in FIG. 1A, spacers within the 5’ UTR and the 3’ UTR.

[0189] FIG. 3B is a graph of area under the curve of eGFP expression in HEK293T cells upon insertion of exemplary 3’ spacers in the circRNA construct.

[0190] FIG. 3C is a graph of CAR expression in human T cells from a circRNA comprising CAA40 spacer in the 3’ UTR. Three replicates are shown. A “no RNA” control and a scrambled IRES (negative control) were also tested.

[0191] FIG. 4A is a graph of percent CAR expression in T cells from three donors transfected with a circRNA encapsulated in a LNP comprising Compound 7, with an anti-CD5 VHH319 targeting moiety, wherein the circRNA had no miR-122 binding site, one miR-122 binding site, or three miR-122 binding sites in the 3’ UTR.ATTORNEY DOCKET NO. ORB-019WO1

[0192] FIG. 4B is a graph of CAR expression (measured as geometric Mean Fluorescence Intensity, gMFI) in T cells from three donors transfected with a circRNA encapsulated in a LNP comprising Compound 7, with an anti-CD5 VHH319 targeting moiety, wherein the circRNA had no miR-122 binding site, one miR-122 binding site, or three miR-122 binding sites in the 3 ’ UTR.

[0193] FIG. 4C is a graph of percent CAR expression in Huh7 immortalized hepatocytes (off-target liver cells) transfected with a circRNA encapsulated in a LNP comprising Compound 7, with an anti-CD5 VHH319 targeting moiety, wherein the circRNA had no miR-122 binding site, one miR-122 binding site, or three miR-122 binding sites in the 3’ UTR.

[0194] FIG. 4D is a graph of CAR expression (gMFI) in Huh7 immortalized hepatocytes (off-target liver cells) transfected with a circRNA encapsulated in a LNP comprising Compound 7, with an anti-CD5 VHH319 targeting moiety, wherein the circRNA had no miR-122 binding site, one miR-122 binding site or three miR-122 binding sites in the 3’ UTR.

[0195] FIG. 4E is a graph of luciferase expression in spleens of CD34+HSC-engrafted humanized NSG mice at 24 hours after administration of various circular RNA constructs comprising one or three miR-122 binding sites, relative to circRNA constructs comprising no miR-122 binding site in the 3’ UTR.

[0196] FIG. 4F is a graph of luciferase expression in livers of CD34+HSC-engrafted humanized NSG mice at 24 hours after administration of various circular RNA constructs comprising one or three miR-122 binding sites, relative to circRNA constructs comprising no miR-122 binding site in the 3' UTR.

[0197] FIG. 4G shows a schematic of the miR-122 binding site in the UTR of the circRNA constructs with re-designed flanking sequences (miR-122_l spacer and miR-122_2 spacer).

[0198] FIG. 4H is a graph of percent circularization of in vitro transcribed RNA. As shown in FIG. 4H, inclusion in the 3’ UTR of either miR-122_l spacer (within SEQ ID NO: 228) or miR-122_2 spacer (within SEQ ID NO: 264) showed comparable circularization, with an improvement seen with the inclusion of miR-122_2 spacer as compared to the miR-122 1 spacer. Overall, these results showed that interrupting poly A tracts (for example, by replacing A with cytosine) resulted in some improvement in circularization of RNA.ATTORNEY DOCKET NO. ORB-019WO1

[0199] FIG. 41 is a graph of hepatic knockdown by circRNA comprising UTR with a miR-122_2 spacer relative to miR-122_l spacer. In Huh7 hepatocytes as measured by flow cytometry, 24 hours after transfection, percent CAR expression was comparably decreased by inclusion of either miR-122_l spacer or miR-122_2 spacer relative to no miR-122 binding site in the 3 ’ UTR of the circRNA. “Full control” refers to a no treatment control (with no buffer or reagent addition, no RNA, and no electroporation). “EP Only” refers to an empty electroporation control where cells were electroporated in the absence of RNA).

[0200] FIG. 4J is a graph of hepatic knockdown by circRNA comprising UTR with a miR-122_2 spacer relative to miR-122_l spacer. In Huh7 hepatocytes as measured by flow cytometry, 24 hours after transfection, CAR gMFI levels were comparably decreased by inclusion of either miR-122_l spacer or miR-122_2 spacer relative to no miR-122 binding site. “Full control” refers to a no treatment control (with no buffer or reagent addition, no RNA and no electroporation). “EP Only” refers to an empty electroporation control where cells were electroporated in the absence of RNA.

[0201] FIG. 4K is a graph of anti-CD19 CAR positive cells as a percentage of CD3+T cells from spleens of CD34+HSC-engrafted humanized NSG mice at 24 hours after administration of circRNA constructs formulated in LNP. This data compares an exemplary circRNA construct without a miRNA binding site (SEQ ID NO: 175) and a circRNA construct with one miRNA-122 binding site in the 3’ UTR of the circRNA (SEQ ID NO: 264).

[0202] FIG. 4L is a graph of anti-CD19 CAR expression in blood as a percentage of total CD3+ T cells following administration of circRNA including no miRNA binding site or miR-122_2 spacer in the 3' UTR; the circRNA is conjugated to an anti-CD5 targeting moiety (VHH319) and encapsulated in two exemplary LNPs, (LNP-11 and LNP-15, as described in Table 16).

[0203] FIG. 4M is a graph of proportion of B cells relative to vehicle control in blood following administration of circRNA including no miRNA binding site (SEQ ID NO: 175) or one miR-122_2 spacer in the 3' UTR (SEQ ID NO: 264). The circRNAs were encapsulated in LNPs conjugated to an anti-CD5 targeting moiety (VHH319) and were formulated in two exemplary formulations LNP-11 and LNP-15 (as described in Table 16).

[0204] FIG. 4N is a graph of anti-CD19 CAR expression in spleen as a percentage of total CD3+ T cells following administration of circRNA including no miRNA binding site (SEQ ID NO: 175) or one miR-122_2 spacer in the 3' UTR (SEQ ID NO: 264). The circRNAsATTORNEY DOCKET NO. ORB-019WO1were encapsulated in LNPs conjugated to an anti-CD5 targeting moiety (VHH319) and were formulated in two exemplary formulations LNP-11 and LNP-15 (as described in Table 16).

[0205] FIG. 40 is a graph of proportion of B cells relative to vehicle control in the spleen following administration of circRNA including no miRNA binding site (SEQ ID NO: 175) or one miR-122_2 spacer in the 3' UTR (SEQ ID NO: 264). The circRNAs were encapsulated in LNPs conjugated to an anti-CD5 targeting moiety (VHH319) and were formulated in two exemplary formulations LNP-11 and LNP-15 (as described in Table 16).

[0206] FIG. 5 is a graph of CAR expression measured by flow cytometry showing that various exemplary circRNA comprising codon optimized coding sequences demonstrated CAR expression in activated T cells in vitro.

[0207] FIG. 6A is a schematic of exemplary anti-CD19 CAR architectures comprising an FMC63 scFv, and various exemplary transmembrane domains, costimulatory domains, and signal domains; and exemplary signaling peptides.

[0208] FIG. 6B is a graph of anti-CD19 CAR expression in CD3+T cells from spleens of CD34+HSC-engrafted NSG humanized mice at 24 hours after administration of circRNA (encoding various exemplary CAR architectures) encapsulated in targeted LNP.

[0209] FIG. 6C is a graph of anti-CD19 CAR expression in circulating CD3+T cells of CD34+HSC-engrafted NSG humanized mice at 24 hours after administration of circRNA (encoding various exemplary CAR architectures) encapsulated in targeted LNP.

[0210] FIG. 6D is a graph of the number of anti-CD19 CAR positive T cells per 5 x 105splenocytes from spleens of CD34+HSC-engrafted NSG humanized mice at 24 hours after administration of circRNA (encoding with exemplary CAR architectures) encapsulated in targeted LNP.

[0211] FIG. 6E is a graph of proportion of B cells to mean B cell count of vehicle control present in spleens of CD34+HSC-engrafted NSG humanized mice at 24 hours after administration of circRNA (encoding exemplary CAR architectures) encapsulated in targeted LNP.

[0212] FIG. 6F is a graph of proportion of B cells to mean B cell count of vehicle control in peripheral blood of CD34+HSC-engrafted NSG humanized mice at 24 hours after administration of circRNA (encoding exemplary CAR architectures) encapsulated in targeted LNP.

[0213] FIG. 7A is a graph of total mGL+ cells relative to percentage of CD3+ T cells 24 hours after administration of an exemplary circRNA encapsulated in an LNP (“RNA-LNP”)ATTORNEY DOCKET NO. ORB-019WO1at 0.5 mg / kg, wherein the LNP comprises 0.1% C18 PEG (LNP-19), 0.5% C18 PEG (LNP-20) or 1% C18 PEG (LNP-21).

[0214] FIG. 7B is a graph showing mGL expression in liver (off-target) relative to spleen 24 hours after administration of an exemplary RNA-LNP at 0.5 mg / kg, wherein the LNP comprises 0.1% C18 PEG (LNP-19), 0.5% C18 PEG (LNP-20) or 1% C18 PEG (LNP-21), as described in Table 16.

[0215] FIG. 7C is a graph showing total luciferase flux in the liver of mice administered circRNA encoding a reporter protein (luciferase) encapsulated in non-targeted LNP-1, LNP-18 and LNP-22, as described in Table 16.

[0216] FIG. 7D is a graph showing total luciferase flux in the spleen of mice administered circRNA encoding a reporter protein (luciferase) encapsulated in non-targeted LNP-1, LNP-18 and LNP-22, as described in Table 16.

[0217] FIG. 7E is a graph showing circRNA levels in the liver of the mice, normalized to the mean value for circRNA encapsulated in LNP-1, as described in Table 16.

[0218] FIG. 7F is a graph showing circRNA levels in the spleen of the mice, normalized to the mean value for circRNA encapsulated in LNP-1, as described in Table 16.

[0219] FIG. 8A is a graph showing mGreenLantem (mGL) expression in CD45.2+ leukocytes in various tissues, including bone marrow, lungs, lymph nodes, peripheral blood, and spleen.

[0220] FIG. 8B shows cellular distribution as percentage of mGL+ cells in tissues after administration of RNA-LNP, wherein the LNP comprises Compound 2.

[0221] FIG. 8C shows cellular distribution as percentage of mGL+ cells in tissues after administration of RNA-LNP comprising mGreenLantem (mGL) and firefly luciferase (ffluc) reporter RNAs, wherein the LNP comprised Compound 7.

[0222] FIG. 8D is a graph that shows anti-CD19 CAR expression in spleen as a percentage of CD3+ T cells. RNA-LNPs (SEQ ID NO: 175), wherein the LNPs comprised both Compound 2 and Compound 7 showed targeting to spleen with the inclusion of an anti-CD5 targeting moiety (VHH319) relative to the corresponding base LNP alone (base LNP refers to LNP with no targeting moiety).

[0223] FIG. 8E is a graph that shows corresponding B cell depletion in spleen, after administration of exemplary RNA-LNPs (wherein LNPs comprised both Compound 2 and Compound 7 comprising SEQ ID NO: 175), was comparable.ATTORNEY DOCKET NO. ORB-019WO1

[0224] FIG. 8F is a graph that shows anti-CD19 CAR positive cells relative to percentage of total CD3+ T cells in multiple studies carried out with RNA encapsulated in a base LNP, which refers to a non-targeted LNP, compared to a RNA-targeted LNP comprising SEQ ID NO: 175 and Compound 2. Also included was a PBS-treated vehicle control.

[0225] FIG. 8G is a graph that shows anti-CD19 CAR MFI relative to total CD3+ T cells in multiple studies carried out with RNA-LNP comprising SEQ ID NO: 175 and Compound 2.

[0226] FIG. 8H is a graph that shows B cell depletion in multiple studies carried out with RNA-LNP comprising SEQ ID NO: 175 and Compound 2.

[0227] FIG. 8I-FIG. 8K are flow cytometry data showing the increased targeting efficiency ofcircRNA formulated in a CD5-targeted LNP targeting to CD4+ and CD8+ T cells in the spleen of mice. The mice were administered via intravenous administration 0.5 mg / kg of circRNA (encoding mGL) formulated in an anti-CD5 targeted LNP or a vehicle control, and the level of mGL expression was measured.

[0228] FIG. 8L-FIG. 8N are flow cytometry data showing the increased targeting efficiencyof circRNA formulated in a CD5-targeted LNP targeting to CD4+ and CD8+ T cells in the lymph node of mice. The mice were administered via intravenous administration 0.5 mg / kg of circRNA (encoding mGL) or a vehicle control, and the level of mGL expression was measured.

[0229] FIG. 9A is a graph of mGreenLantem gMFI expression in Jurkat T cells using circRNA (encoding an mGreenLantem reporter) encapsulated by LNP with various exemplary LNP formulations described in Table 16 and shows that increasing the DSPC from the standard amount of about 10% DSPC to about 15-25% DSPC increased mGL expression and increasing to greater than 25% DSPC further increased the mGL expression in Jurkat T cells.

[0230] FIG. 9B is a graph of % mGL positive (among live cells) in Jurkat T cells by circRNA encoding mGreenLantem reporter encapsulated by LNP, with various exemplary LNP formulations described in Table 16. A reference 10% DSPC showed about 75% mGL positive cells. At 15-25% DSPC, most formulations tested showed increased reporter expression compared to the reference 10% DSPC. At greater than 25% DSPC, all LNP formulations tested showed greater than 90% mGL positive cells.ATTORNEY DOCKET NO. ORB-019WO1

[0231] FIG. 9C is a graph that shows anti-CD19 CAR positive cells relative to percentage of mCD45+ cells.

[0232] FIG. 9D is a graph that shows B cell depletion in multiple studies carried out using various exemplary LNP formulations, including formulations comprising Compound 7.

[0233] FIG. 10A is a schematic that shows exemplary targeting moiety formats, e.g., single-chain variable fragment (scFv), VHH and full-length antibodies.

[0234] FIG. 10B is a graph of percent B cells remaining after 24 hours upon administration of RNA-LNPs comprising various exemplary targeting moieties (anti-CD2, anti-CD5, anti-CD7 and anti-CD8) relative to vehicle control. Anti-CD5 VHH performed as well as full-length anti-CD2, anti-CD7 and anti-CD8 antibodies.

[0235] FIG. 10C is a graph showing proportion of B cells remaining 24 hours after administration of RNA-LNP comprising two exemplary targeting moieties, anti-CD5 and anti-CD8 VHHs, with a “G3S-Cys-AAA” (SEQ ID NO: 120) conjugation handle or “G4S-Cys-AAA” (SEQ ID NO: 343) conjugation handle.

[0236] FIG. 10D is a graph showing anti-CD19 CAR MFI relative to CD4+ T cells by administration of RNA-LNP comprising RNA of SEQ ID NO: 175, exemplary anti-CD5 VHH319 comprising a “G3S-Cys-AAA” (SEQ ID NO: 120) conjugation handle and a reference conjugation handle.

[0237] FIG. 10E is a graph showing anti-CD19 CAR MFI relative to CD8+ T cells by administration of RNA-LNP comprising exemplary anti-CD5 VHH319 comprising a “G3S-Cys-AAA” (SEQ ID NO: 120) conjugation handle and a reference conjugation handle.

[0238] FIG. 10F is a graph showing CD69 MFI as a marker of T cell activation relative to CD4+ T cells by administration of RNA-LNP comprising exemplary anti-CD5 VHH319 comprising a “G3S-Cys-AAA” (SEQ ID NO: 120) conjugation handle and a reference conjugation handle.

[0239] FIG. 10G is a graph showing CD69 MFI as a marker of T cell activation relative to CD8+ T cells by administration of RNA-LNP comprising exemplary anti-CD5 VHH319 comprising a “G3S-Cys-AAA” (SEQ ID NO: 120) conjugation handle and a reference conjugation handle.

[0240] FIG. 11A is a graph showing the percentage of anti-CD19 CAR+ cells relative to total human CD3+ T cells 24 hours after RNA-LNP administration for LNPs with various exemplary anti-CD5 VHH nanobodies as targeting moieties: VHH319, VHH322, VHH325, VHH329, VHH330 and VHH331 described in Table 10.ATTORNEY DOCKET NO. ORB-019WO1

[0241] FIG. 11B is a graph showing the proportion of B cells remaining after 24 hours of RNA-LNP administration relative to vehicle control for LNPs with various exemplary anti-CD5 VHH nanobodies as targeting moieties: VHH319, VHH322, VHH325, VHH329, VHH330 and VHH331 described in Table 10.

[0242] FIG. 12A is a schematic of various conjugation handles used to conjugate the targeting moiety to the delivery vehicle, e.g., LNPs (sequences shown in Table 12).

[0243] FIG. 12B is a graph showing percentage of anti -CD 19 CAR+ cells relative to total CD3+ T cells in blood.

[0244] FIG. 12C is a graph showing percentage of anti-CD19 CAR CD3+ T cells (MFI) in blood.

[0245] FIG. 12D is a graph showing percentage of anti-CD19 CAR+ cells relative to total CD3+ T cells in spleen.

[0246] FIG. 12E is a graph showing percentage of anti-CD19 CAR CD3+ T cells (MFI) in spleen.

[0247] FIG. 12F is a graph showing number of anti-CD19 CAR+ T cells per 5 x 105splenocytes.

[0248] FIG. 12G is a graph showing proportion of B cells remaining in spleen relative to vehicle control when administered an anti-CD19 RNA-LNP (SEQ ID NO: 175), with exemplary modified anti-CD5 VHH319 targeting moieties - an anti-CD5 VHH319 comprising a reference conjugation handle, for example, at the C-terminal, and an anti-CD5 VHH319 comprising a “G3S-CYS-AAA” (SEQ ID NO: 120) conjugation handle were compared.

[0249] FIG. 12H is a graph showing proportion of B cells remaining in spleen relative to vehicle control when administered an anti-CD19 RNA-LNP with exemplary modified anti-CD5 VHH319 targeting moieties - an anti-CD5 VHH319 comprising S7C, S70C and S82bC, and a control anti-CD5 VHH319 comprising an exemplary C-terminal reference conjugation handle.

[0250] FIG. 121 is a graph showing proportion of B cells remaining in blood relative to vehicle control when administered an LNP-anti-CD19 RNA comprising exemplary modified anti-CD5 VHH319 targeting moieties - VHH319 comprising S7C, S70C and S82bC and a control VHH319 comprising a reference conjugation handle.

[0251] FIG. 12J is a graph showing anti-CD19 CAR expression in spleen as a percentage of CD3+ T cells when administered anti-CD19 circRNA conjugated to an LNP using variousATTORNEY DOCKET NO. ORB-019WO1exemplary conjugation chemistries, namely, 0.1% PEG maleimide C-terminal conjugation, with and without a histidine tag, 0.1% PEG-maleimide C-terminal “G3S-Cys-AAA” (SEQ ID NO: 120) or G4SG4S-Cys-AS (SEQ ID NO: 345) conjugation handle, 0.1% PEG maleimide S82bC conjugation with and without histidine tag and a 0.5% PEG maleimide S82bC conjugation without histidine tag relative to a vehicle control and base LNP control.

[0252] FIG. 12K is a graph showing proportion of B cells remaining in spleen relative to vehicle control when administered anti-CD19 circRNA conjugated to an LNP comprising exemplary conjugation chemistries, namely, 0.1% PEG maleimide C-terminal conjugation, with and without a histidine tag, 0.1% PEG-maleimide C-terminal “G3S-Cys-AAA” (SEQ ID NO: 120) conjugation handle, 0.1% PEG maleimide S82bC conjugation with and without histidine tag and a 0.5% PEG maleimide S82bC conjugation without histidine tag relative to a vehicle control and base LNP control.

[0253] FIG. 13A shows a schematic of exemplary conjugation chemistries in conjugating a targeting moiety to the delivery vehicle, e.g., LNP.

[0254] FIG. 13B shows flow cytometry data showing eGFP positive cells in conjugates using thiol-maleimide and azide-DBCO conjugations relative to an unconjugated (base) LNP control.

[0255] FIG. 13C shows flow cytometry data showing mGreenLantern positive cells in conjugates using sortase-catalyzed LPXT / G-glycine conjugation, wherein X is any amino acid residue.

[0256] FIG. 14A is a graph of mGL positive cells as a percentage of total CD45.2+ leukocytes.

[0257] FIG. 14B is a graph of cellular biodistribution of gene expression in cells administered linear RNA in an LNP formulation comprising Compound 2.

[0258] FIG. 14C is a graph of cellular biodistribution of gene expression in cells administered circular RNA in an LNP formulation comprising Compound 2.

[0259] FIG. 14D is a graph of anti-CD19 CAR positive T cells derived from flow cytometry data shown in FIG. 14F on day 1, day 4, and day 7 comparing the number of CAR positive cells from linear RNA (SEQ ID NO: 275) and circRNA (SEQ ID NO: 230).

[0260] FIG. 14E is flow cytometry data from day 1, day 4, and day 7 comparing anti-CD19 CAR positive cells from linear RNA (SEQ ID NO: 275) and circRNA (SEQ ID NO: 230).ATTORNEY DOCKET NO. ORB-019WO1

[0261] FIG. 14F is a graph of anti-CD19 CAR expression in spleen on day 1, day 2 and day 3 comparing expression from linear mRNA and circRNA.

[0262] FIG. 14G is a plot showing area under the curve (AUC) for mGreenLantern (mGL) reporter protein expression by circRNAs comprising an exemplary IRES and circRNA comprising a reference IRES sequence (“circular RNA control”), compared to a linear RNA encoding mGL reporter protein in activated primary human T cells (from one donor).

[0263] FIG. 14H is a graph that showed a time course of anti-CD19 CAR+ T cells measured over 7 days in mice administered 1 mg / kg of circRNA encoding anti-CD19 CAR formulated in a targeted LNP (comprising Compound 7), and demonstrated that anti-CD19 CAR+ T cell populations were sustained in vivo.

[0264] FIG. 15A is a graph of the percentage of anti-CD19 CAR positive cells relative to total CD3 positive T cells in the blood of humanized mice, 24 hours after administration of each of three doses of RNA-LNP relative to vehicle control.

[0265] FIG. 15B is a graph of the percentage of anti-CD19 CAR positive cells relative to total CD3 positive T cells in the spleen of humanized mice, 24 hours after administration of each of three doses of RNA-LNP relative to vehicle control.

[0266] FIG. 15C is a graph of the percentage of B cells (relative to vehicle control) in blood after administration of three doses each of 0.5 mg / kg of RNA-LNP in mice, as measured 24 hours after administration of each dose, on day 1, 8 and 15.

[0267] FIG. 15D is a graph of the percentage of B cells (relative to vehicle control) in spleen after administration of three doses each of 0.5 mg / kg of RNA-LNP in mice, as measured 24 hours after administration of each dose, on day 1, 8 and 15.

[0268] FIG. 15E is a graph of the percentage of CD69-positive CD4+ T cells in blood 24 hours after administration of each of three doses of RNA-LNP.

[0269] FIG. 15F is a graph of the percentage of CD69-positive CD8+ T cells in blood 24 hours after administration of each of three doses of RNA-LNP.

[0270] FIG. 15G is a graph of the percentage of CD69-positive CD4+ T cells in spleen 24 hours after administration of each of three doses of RNA-LNP.

[0271] FIG. 15H is a graph of the percentage of CD69-positive CD8+ T cells in spleen 24 hours after administration of each of three doses of RNA-LNP. The results in FIG. 15E-FIG.15H showed T cell activation in blood and spleen after RNA-LNP administration.ATTORNEY DOCKET NO. ORB-019WO1

[0272] FIG. 16A is a graph of the percentage of anti-CD19 CAR positive cells as a percentage of total CD3 positive T cells in blood. The left panel shows anti-CD19 CAR positive cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of circRNA encoding anti-CD19 CAR relative to vehicle control. The right panel shows anti-CD19 CAR positive cells, 24 hours after administration of 0.5 mg / kg dose of circRNA encoding anti-CD19 CAR relative to vehicle control on days 1, 2 and 7 after administration.

[0273] FIG. 16B is a graph of the percentage of anti-CD19 CAR positive cells as a percentage of total CD3 positive T cells in spleen. The left panel shows anti-CD19 CAR positive cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of circRNA (encoding anti-CD19 CAR) relative to vehicle control. The right panel shows anti-CD19 CAR positive cells, 24 hours after administration of 0.5 mg / kg dose of RNA-LNP relative to vehicle control on days 1, 2 and 7 after administration.

[0274] FIG. 16C is a graph of the percentage of remaining B cells (relative to vehicle control) in blood. The left panel shows percent remaining B cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of RNA-LNP relative to vehicle control. The right panel shows percent remaining B cells, 24 hours after administration of 0.5 mg / kg dose of RNA-LNP relative to vehicle control on days 1, 2 and 7 after administration.

[0275] FIG. 16D is a graph of the percentage of remaining B cells (relative to vehicle control) in spleen. The left panel shows percent remaining B cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of RNA-LNP relative to vehicle control. The right panel shows percent remaining B cells, 24 hours after administration of 0.5 mg / kg dose of RNA-LNP relative to vehicle control on days 1, 2 and 7 after administration.

[0276] FIG. 17A is a graph of the percentage of anti-CD19 CAR positive cells as a percentage of total CD3 positive T cells in blood. The left panel shows anti-CD19 CAR positive cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of RNA-LNP relative to vehicle control. The right panel shows anti-CD19 CAR positive cells, 24 hours after administration of 0.5 mg / kg dose of RNA-LNP relative to vehicle control on days 1, 2 and 7 after administration.

[0277] FIG. 17B is a graph of the percentage of anti-CD19 CAR positive cells as a percentage of total CD3 positive T cells in spleen. The left panel shows anti-CD19 CAR positive cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of RNA-LNP relative to vehicle control. The right panel shows anti-CD19 CAR positive cells,ATTORNEY DOCKET NO. ORB-019WO124 hours after administration of 0.5 mg / kg dose of circRNA (encoding anti-CD19 CAR) relative to vehicle control on days 1, 2 and 7 after administration.

[0278] FIG. 17C is a graph of the percentage of remaining B cells (relative to vehicle control) in blood. The left panel shows percent remaining B cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of circRNA (encoding antiCD 19 CAR) relative to vehicle control. The right panel shows percent remaining B cells, 24 hours after administration of 0.5 mg / kg dose of circRNA (encoding anti-CD19 CAR) relative to vehicle control on days 1, 2 and 7 after administration.

[0279] FIG. 17D is a graph of the percentage of B cells (relative to vehicle control) in spleen. The left panel shows percent remaining B cells, 24 hours after administration of 0.1 mg / kg, 0.5 mg / kg and 1.0 mg / kg doses of RNA-LNP relative to vehicle control. The right panel shows percent remaining B cells, 24 hours after administration of 0.5 mg / kg dose of RNA-LNP relative to vehicle control on days 1, 2 and 7 after administration.

[0280] FIG. 17E is a bar graph shown as an alternate representation of the right panel of FIG. 17B. FIG. 17E shows anti-CD19 CAR positive cells (as a percentage of CD3+ T cells), 24 hours after administration of 0.5 mg / kg dose of circRNA (encoding anti-CD19 CAR) formulated in targeted LNP relative to vehicle control on days 1, 2 and 7 after administration.

[0281] FIG. 17F is a bar graph shown as an alternate representation of FIG. 17D. FIG. 17F shows percent remaining B cells (relative to vehicle) after B cell count was normalized to total live splenocytes, 24 hours after administration of 0.5 mg / kg dose of circRNA (encoding anti-CD19 CAR) formulated in targeted LNP relative to vehicle control on days 1, 2 and 7 after administration.

[0282] FIG. 18 depicts immunohistochemistry staining in the left panel that shows targeted circRNA delivery to the white pulp in the spleen, where T cells are located. The right panel shows a schematic of the anatomical sites within the spleen, such as white pulp, marginal zone, red pulp, and B cell follicle. An expanded view in the right panel shows T cells localized within the white pulp of the spleen.

[0283] FIG. 19A is a graph of the percentage of B cells remaining (compared to vehicle), after administration of three doses at 1 mg / kg circRNA (encoding anti-CD19 CAR) in CD34+NSG mice.

[0284] FIG. 19B is an image that showed immunohistochemistry staining of B cells using a CD20 marker in the spleen of CD34+NSG mice administered a vehicle control depicting high density of B cells.ATTORNEY DOCKET NO. ORB-019WO1

[0285] FIG. 19C is an image that showed immunohistochemistry staining of B cells using a CD20 marker in the spleen of CD34+NSG mice of CD34+NSG mice after administration of circRNA (encoding anti-CD19 CAR), which depicted B cell depletion.

[0286] FIG. 20A is a graph of the percentage of anti-CD20 CAR positive cells as a percentage of CD3+ T cells in blood from cynomolgus monkeys. The panel shows a time course of CAR expression after each dose of 1 mg / kg, 2 mg / kg, or 3 mg / kg circRNA relative to a vehicle control.

[0287] FIG. 20B is a graph of the number of anti-CD20 CAR-positive CD3+ T cells present in each microliter (μL) of blood. The panel shows the number of CAR+ T cells generated and present in blood at each time depicted in the graph. Each cynomolgus monkey was dosed on day 0 and re-dosed on day 7. The number of CAR+ CD3+ T cells was measured by flow cytometry at baseline (e.g., 7 days prior to the dose administration), 6 hours, 24 hours after each dose, as well as 48 hours after the first dose.

[0288] FIG. 20C is a graph of the percentage of anti-CD20 CAR-positive CD3+ T cells present in the deep tissues (iliac lymph node and spleen). Each cynomolgus monkey was dosed on day 0 and day 7 at the 2 mg / kg and 3 mg / kg doses. The percentage of CAR+ CD3+ T cells was measured by flow cytometry at 24 hours after the second dose. The graphs represent the average of two cynomolgus monkeys per dose.

[0289] FIG. 20D is a graph of the percentage of remaining B cells in blood over time relative to a vehicle control. Each cynomolgus monkey in Cohort 1 was dosed on day 0 and day 7 at the 1 mg / kg, 2 mg / kg, or 3 mg / kg doses. The graphs represent the average of two cynomolgus monkeys per dose. The absolute count of B cells present in circulation at each time point was measured by flow cytometry and used to calculate the remaining B cells relative to the number of B cells relative to the vehicle control.

[0290] FIG. 20E is a graph of the percentage of remaining B cells in deep tissue (iliac lymph node and spleen) relative to a vehicle control. Each cynomolgus monkey was dosed on day 0 and day 7 at 1 mg / kg, 2 mg / kg, or 3 mg / kg doses. For the 1 mg / kg group, the panel shows the percentage of B cells present in the tissues at 8 days after the second dose. For the 2 mg / kg and 3 mg / kg groups, the panel shows the percentage of B cells present in the tissues at 24 hours after the second dose. Each data point indicates a different animal.

[0291] FIG. 20F is a graph of the percentage of anti-CD20 CAR-positive cells as a percentage of CD3+ T cells in blood from cynomolgus monkeys in Cohort 2. The panelATTORNEY DOCKET NO. ORB-019WO1shows a time course of CAR expression from both single-dose and double-dose groups. For the single-dose group, the monkeys were administered 0.75 mg / kg or 1 mg / kg doses, and CAR expression was measured at 6 hours, 24 hours, and 48 hours after the dose. For the double-dose groups, the monkeys were administered 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, or 1 mg / kg doses twice (on day 0 and day 7). CAR expression was measured at 6 hours, 24 hours after each dose, and after 48 hours as well as after five days for the single-dose groups.

[0292] FIG. 20G is a graph of the number of anti-CD20 CAR-positive CD3+ T cells present in each microliter (uL) of blood. The panel shows the number of CAR+ T cells generated and present in blood at each time point depicted in the graph. The number of CAR+ CD3+ T cells was measured by flow cytometry at baseline prior to the dose, at 6 hours, and 24 hours after each dose as well as 48 hours after the first dose.

[0293] FIG. 20H is a graph of the percentage of remaining B cells in the blood relative to a vehicle control. The panel shows a time course of B cell depletion for both single-dose and double-dose groups. For the single dose, the monkeys were administered circRNA (encoding anti-CD20 CAR) at 0.75 mg / kg or 1 mg / kg, and CAR expression was measured at 6 hours, 24 hours, and 48 hours after the dose. For the double-dose groups, the monkeys were administered twice at 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, or 1 mg / kg on day 0 and day 7. The number of B cells present in the blood was measured at 6 hours, 24 hours, and 48 hours after the first dose as well as five days later (120 hours) for the single-dose groups. In addition, the number of B cells was measured at 6 hours and 24 hours after the second dose.

[0294] FIG. 201 is a graph that shows the time course for the 0.25 mg / kg group in FIG. 20H.

[0295] FIG. 20 J is a graph that shows the time course for the 0.5 mg / kg group in FIG. 20H.

[0296] FIG. 20K is a graph that shows the time course for the 0.75 mg / kg group in FIG.20H.

[0297] FIG. 20L is a graph of the percentage of remaining B cells in the spleen relative to a vehicle control. For the single-dose groups, the monkeys were administered at 0.75 mg / kg or 1 mg / kg doses, and B cell depletion in the spleen was measured at day 5 after the dose. For the double-dose groups, the monkeys were administered twice each at 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, or 1 mg / kg on day 0 and day 7. On day 8, 24 hours after the second dose, the animals were sacrificed to quantify the number of B cells present in the tissue. B cellATTORNEY DOCKET NO. ORB-019WO1numbers were used to calculate the percentage of remaining B cell population relative to a vehicle control.

[0298] FIG. 20M is a graph of the percentage of remaining B cells in the iliac lymph node relative to a vehicle control. For the single-dose groups, the monkeys were administered 0.75 mg / kg or 1 mg / kg circRNA, and B cell depletion in the lymph node was measured at day 5 after the dose. For the double-dose groups, the monkeys were administered circRNA twice at 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, or 1 mg / kg each on day 0 and day 7. On day 8, 24 hours after the second dose, the monkeys were sacrificed to quantify the number of B cells present in the tissue. B cell numbers were used to calculate the % remaining B cell population relative to a vehicle control.

[0299] FIG. 20N is a graph of exemplary data from Cohorts 1 and 2 in the non-human primate study showing a liver enzyme (alanine aminotransferase, ALT) time course in blood following repeat administration of circRNA at day 0 and day 7 at doses of 0 mg / kg (vehicle control), 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, or 1.0 mg / kg each. This graph showed safety and tolerability of the circRNA in monkeys.

[0300] FIG. 200 is a graph of the percentage of remaining CD20+ B cells in the lymph node relative to a vehicle control in monkeys that were administered 0.25 mg / kg, 0.5 mg / kg, or 0.75 mg / kg circRNA twice on day 0 and day 7, and subsequently B cell depletion in the lymph node was measured 24 hours after the second dose (that is, on day 8). Viable lymphocytes isolated from monkeys were gated using a CD20 marker for quantifying a CD20+ B cell subpopulation by flow cytometry, and CD20+ B cell numbers were used to calculate the percentage of remaining CD20+ B cells relative to a vehicle control.

[0301] FIG. 20P is a graph showing the percentage of remaining CD20+ B cells in the spleen relative to a vehicle control (bottom panel). As part of the study, the monkeys in Cohort 1 were administered circRNA encoding anti-CD20 CAR at a single dose of 0.75 mg / kg or 1 mg / kg, and B cell depletion in the spleen was measured at day 5 after the dose (as illustrated in the top panel). CD20+ gated B cell numbers were used to calculate the percentage of remaining CD20+ B cells relative to a vehicle control.

[0302] FIG. 20Q is a graph showing the percentage of remaining CD20+ B cells in the spleen relative to a vehicle control. The monkeys (in Cohorts 1 and 2) were administered circRNA encoding anti-CD20 CAR twice each at 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1 mg / kg, 2 mg / kg or 3 mg / kg on day 0 and day 7. On day 8, 24 hours after the second dose, theATTORNEY DOCKET NO. ORB-019WO1animals were sacrificed to quantify the number of B cells present in the tissue. CD20+ gated B cell numbers were used to calculate the percentage of remaining CD20+ B cell population relative to a vehicle control. The graph combines the NHP data from Cohort 2 (for the 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, and 1 mg / kg doses) and Cohort 1 (for the 2 mg / kg and 3 mg / kg doses).

[0303] FIG. 20R depicts in situ hybridization (ISH) images that were generated 24 hours after the second dose of circRNA was administered to the monkeys. The ISH images showed that after two doses of circRNA, at 0.5 mg / kg or higher, formulated in targeted LNP, full depletion of B cells was seen in the spleen. The CD20 marker stained B cells, and DAPI stained cell nuclei.

[0304] FIG. 20S is a graph showing the percentage of remaining CD20+ B cells in the lymph node relative to a vehicle control (bottom panel). As part of the study, the monkeys were administered circRNA encoding anti-CD20 CAR as a single dose, and B cell depletion in the lymph node was measured at day 5 after the dose (as illustrated in the top panel).Representative data from the 0.75 mg / kg or 1 mg / kg doses are shown. CD20+ gated B cell numbers were used to calculate the percentage of remaining CD20+ B cells relative to a vehicle control.

[0305] FIG. 20T is a graph showing the percentage of remaining CD20+ B cells in the lymph node relative to a vehicle control. The monkeys (Cohorts 1 and 2) were administered circRNA encoding anti-CD20 CAR twice, on day 0 and day 7, each dose at 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, 1 mg / kg, 2 mg / kg or 3 mg / kg. On day 8, 24 hours after the second dose, the animals were sacrificed to quantify the number of B cells present in the tissue. CD20+ gated B cell numbers were used to calculate the percentage of remaining CD20+ B cells relative to a vehicle control. The graph combines the data from Cohort 2 (for the 0.25 mg / kg, 0.5 mg / kg, 0.75 mg / kg, and 1 mg / kg doses) and Cohort 1 (for the 2 mg / kg and 3 mg / kg doses).

[0306] FIG. 20U depicts ISH images in the lymph node that showed that after two doses of the circRNA encoding anti-CD20 CAR formulated in targeted LNP was administered to the cynomolgus monkeys, robust B cell depletion was observed at day 8, 24 hours after the second dose. The CD20 marker stained B cells, and DAPI stained cell nuclei.

[0307] FIG. 20V shows magnified versions of the ISH image for the 0.75 mg / kg dose from FIG. 20U. As shown in FIG. 20V (top right panel) although remaining B cells were detected, those B cells (CD20 stained) co-localized with the presence of circRNA (anti-CD20ATTORNEY DOCKET NO. ORB-019WO1CAR circRNA stained). Although not bound by any mechanism of action, the detection of the circRNA suggested that T cells transfected with the circRNA were in the process of depleting B cells, that is, that there was an ongoing B cell depletion phenomenon.

[0308] FIG. 21 A depicts ISH images of the spleen from day 1, day 5, and day 28 after a single administration of 1 mg / kg of the circRNA encoding anti-CD20 CAR formulated in targeted LNP to cynomolgus monkeys. The ISH images show T cells (stained with CD3 marker) and B cells (stained with CD20 marker) at day 1, day 5, and day 28 after administration of the circRNA, as well as after administration with a vehicle control at day 28.

[0309] FIG. 21B depicts ISH images of the lymph node from day 1, day 5, and day 28 after a single administration of 1 mg / kg of the circRNA encoding anti-CD20 CAR formulated in targeted LNP to cynomolgus monkeys. The ISH images show T cells (stained with CD3 marker) and B cells (stained with CD20 marker) at day 1, day 5, and day 28 after administration of the circRNA, as well as after administration with a vehicle control at day 28.DEFINITIONS

[0310] In order for the present disclosure to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification.[3H] About or approximately: As used herein, the term “about” or “approximately,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” is understood as within a range of normal tolerance in the art, for example, within two standard deviations of the mean, and refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value). In some embodiments, the term refers to a range of values that fall within 10% of the stated reference value. In some embodiments, the term refers to a range of values that fall within 5% of the stated refer. The term “between” includes the values of the specified boundaries and all intervening values and fractions.ATTORNEY DOCKET NO. ORB-019WO1

[0312] Adaptive immune response'. The adaptive immune response is typically understood to be antigen-specific. Antigen specificity allows for the generation of responses that are tailored to specific antigens, pathogens or pathogen-infected cells. The ability to mount these tailored responses is maintained in the body by “memory cells”. Should a pathogen infect the body more than once, these specific memory cells are used to quickly eliminate it. In this context, the first step of an adaptive immune response is the activation of naive antigenspecific T cells or different immune cells able to induce an antigen-specific immune response by antigen-presenting cells. This occurs in the lymphoid tissues and organs through which naive T cells are constantly passing. Cell types that can serve as antigen-presenting cells are inter alia dendritic cells, macrophages, and B cells. Each of these cells has a distinct function in eliciting immune responses. Dendritic cells take up antigens by phagocytosis and macropinocytosis and are stimulated by contact with e.g., a foreign antigen to migrate to the local lymphoid tissue, where they differentiate into mature dendritic cells. Macrophages ingest particulate antigens such as bacteria and are induced by infectious agents or other appropriate stimuli to express major histocompatibility complex (MHC) molecules. The unique ability of B cells to bind and internalize soluble protein antigens via their receptors may also be important to induce T cells. Presenting the antigen on MHC molecules leads to activation of T cells which induces their proliferation and differentiation into armed effector T cells. The most important function of effector T cells is the killing of infected cells by CD8+ cytotoxic T cells and the activation of macrophages by Thl cells which together make up cell-mediated immunity, and the activation of B cells by both Th2 and Thl cells to produce different classes of antibody, thus driving the humoral immune response. T cells recognize an antigen by their T cell receptors which do not recognize and bind antigen directly, but instead recognize short peptide fragments e.g., of pathogen-derived protein antigens, which are bound to MHC molecules on the surfaces of other cells.

[0313] Adenylation'. As used herein, “adenylation” or “AMPylation” is a process in which an adenosine monophosphate (AMP) molecule is covalently attached to the amino acid side chain of a protein. This covalent addition of AMP to a hydroxyl side chain of the protein is a posttranslational modification.

[0314] Adjuvant', the term “adjuvant” refers to agents which confer immunity by themselves. An adjuvant assists the immune system un-specifically to enhance the antigenspecific immune response by e.g., promoting presentation of an antigen to the immune system or induction of an unspecific innate immune response. Furthermore, an adjuvant mayATTORNEY DOCKET NO. ORB-019WO1preferably, e.g., modulate the antigen-specific immune response by e.g., shifting the dominating Th2 -based antigen specific response to a more Th 1 -based antigen specific response or vice versa. Accordingly, an adjuvant may favorably modulate cytokine expression / secretion, antigen presentation, type of immune response etc. An adjuvant or an adjuvant component in the broadest sense is typically a (e.g., pharmacological or immunological) agent or composition that may modify, e.g., enhance, the efficacy of other agents, such as a drug or vaccine. Conventionally the term refers in the context of the invention to a compound or composition that serves as a carrier or auxiliary substance for immunogens and / or other pharmaceutically active compounds. It is to be interpreted in a broad sense and refers to a broad spectrum of substances that are able to increase the immunogenicity of antigens incorporated into or co-administered with an adjuvant in question. In the context of the present invention an adjuvant will preferably enhance the specific immunogenic effect of the active agents of the present invention. Typically, “adjuvant” or “adjuvant component” has the same meaning and can be used interchangeably.

[0315] Antibody: As used herein, the term “antibody” is referred to in the broadest sense and specifically covers various embodiments including, but not limited to monoclonal antibodies, polyclonal antibodies, multi-specific antibodies (e.g., bispecific antibodies formed from at least two intact antibodies), and includes antibody fragments (e.g., diabodies) so long as they exhibit a desired biological activity (e.g., “functional”). Antibodies are primarily amino acid based molecules but may also comprise one or more modifications (including, but not limited to the addition of sugar moieties, fluorescent moieties, chemical tags, etc.). Nonlimiting examples of antibodies or fragments thereof include VH and VL domains, scFvs, Fab, Fab', F(ab')2, Fv fragment, diabodies, nanobodies, linear antibodies, single chain antibody molecules, multi-specific antibodies, bispecific antibodies, intrabodies, monoclonal antibodies, polyclonal antibodies, humanized antibodies, codon-optimized antibodies, tandem scFv antibodies, bispecific T-cell engagers, mAb2 antibodies, tetraval ent bi specific antibodies, biosynthetic antibodies, native antibodies, miniaturized antibodies, unibodies, maxibodies, antibodies to senescent cells, antibodies to conformers, antibodies to disease specific epitopes, or antibodies to innate defense molecules.

[0316] Antibody fragment or antigen-binding fragment: As used herein, “antibody fragment" or “antigen-binding fragment” refers to any portion of an intact antibody. In some embodiments, antibody fragments comprise antigen-binding regions from intact antibodies. Examples of antibodies or antigen binding fragments may include, but are not limited to Fab,ATTORNEY DOCKET NO. ORB-019WO1Fab', F(ab')2, Fv fragments; diabodies; single domain antibodies (sdAb) e.g., variable domain of heavy chain only (VHH) antibodies (also referred to as nanobodies) or Variable New Antigen Receptor (VNAR) antibodies); linear antibodies; single-chain antibody molecules (e.g., scFv); and multi-specific antibodies formed from antibody fragments. In some embodiments, the antibody or antigen-binding fragment is a VHH or an scFv. In some embodiments, the antibody or antigen-binding fragment is a VHH. In some embodiments, the antibody or antigen-binding fragment is an scFv.

[0317] Antibody variant: As used herein, “antibody variant” refers to a biomolecule resembling an antibody in structure and / or function comprising some differences in their amino acid sequence, composition or structure as compared to a native antibody.

[0318] Antigen'. The term “antigen” refers to a substance which may be recognized by the immune system, preferably by the adaptive immune system, and is capable of triggering an antigen-specific immune response, e.g., by formation of antibodies and / or antigen-specific T cells as part of an adaptive immune response. Typically, an antigen may be or may comprise a peptide or protein which may be presented by the MHC to T-cells. In the sense of the present invention an antigen may be the product of translation of a provided nucleic acid molecule, preferably an RNA as defined herein. In this context, fragments, variants and derivatives of peptides and proteins comprising at least one epitope are understood as antigen.

[0319] Associated: As used herein, the terms “associated with,” “conjugated,” “linked,” “attached,” and “tethered,” when used with respect to two or more moieties, means that the moieties are physically associated or connected with one another, either directly or via one or more additional moieties that serves as a linking agent, to form a structure that is sufficiently stable so that the moieties remain physically associated under the conditions in which the structure is used, e.g., physiological conditions. An “association” need not be strictly through direct covalent chemical bonding. It may also be through ionic or hydrogen bonding or a hybridization-based connectivity sufficiently stable such that the “associated” entities remain physically associated.

[0320] Biologic: As used herein, a “biologic” is a polypeptide-based molecule encoded by the linear and / or circular RNA provided herein which may be used to treat, cure, mitigate, prevent, or diagnose a disease or disorder.

[0321] Cellular immunity or cellular immune response: the phrase “cellular immunity”, which is used interchangeably with “cellular immune response”, relates typically to the activation of macrophages, natural killer cells (NK), antigen-specific cytotoxic TATTORNEY DOCKET NO. ORB-019WO1lymphocytes, and the release of various cytokines in response to an antigen. In a more general way, cellular immunity is not related to antibodies but to the activation of cells of the immune system. A cellular immune response is characterized e.g. by activating antigenspecific cytotoxic T lymphocytes that are able to induce apoptosis in body cells displaying epitopes of an antigen on their surface, such as virus-infected cells, cells with intracellular bacteria, and cancer cells displaying tumor antigens; activating macrophages and natural killer cells, enabling them to destroy pathogens; and stimulating cells to secrete a variety of cytokines that influence the function of other cells involved in adaptive immune responses and innate immune responses.

[0322] Circular RNA As used herein, the term “circular RNA” or “circRNA” refers to an RNA that forms a circular structure through covalent or non-covalent bonds. The terms “circRNA” or “circular polyribonucleotide” or “circular RNA” are used interchangeably. In some embodiments, circRNAs are covalently closed, single stranded RNA molecules. A circular RNA can be produced by various methods, including back-splicing of a linear precursor RNA, by chemical ligation and / or enzymatic ligation. Circular RNAs (circRNAs) can be endogenous or synthetic. Synthetically created and exogenously delivered circRNAs can be synthesized in vitro using self-splicing permuted introns (e.g., self-splicing Group I or Group II intron) from in vitro transcribed constructs. Unlike linear RNAs, circular RNAs are more resistant to the degradation by exonuclease and have a longer half-life than their corresponding linear counterparts. A circular RNA can be a circular RNA that encodes a polypeptide or synthetic protein (e.g., a chimeric antigen receptor). In some embodiments, circular RNAs disclosed herein have significantly improved drug-like properties compared to linear mRNA therapeutics, including enhanced longevity as protein production vectors.

[0323] Conjugation'. As used herein, the term "conjugation" refers to a process that creates a linkage between two moieties. In some embodiments, the conjugation is a chemical reaction. In some embodiments, the conjugation is an enzymatic, or enzyme-catalyzed, reaction.

[0324] Conjugation Handle. As used herein, the term “conjugation handle” refers to a short peptide containing one or more amino acids (e.g., 1-20 amino acids). In some embodiments, the conjugation handle includes at least one reactive group (e.g., azide, tetrazine, or thiol) that allows conjugation between two moieties via, e.g., chemical, enzymatic, or enzyme-catalyzed reaction. The reactive group in the conjugation handle may be a functional group of an amino acid on the “conjugation handle” or attached anywhere onATTORNEY DOCKET NO. ORB-019WO1the “conjugation handle” (e.g., at the N-terminus, C-terminus, or internal to the conjugation handle) directly or indirectly. In some embodiments, the reactive group described herein is suitable for a “click chemistry” reaction. In some embodiments, the conjugation handle comprises one or more glycine, alanine, cysteine, serine, and / or glutamine. A “conjugation handle” may contain, for example, 5-10 amino acids, such as 5, 6, 7, 8, 9, or 10 amino acids. A “conjugation handle” may be attached to (e.g., covalently attached to) a moiety (such as a targeting moiety) that is conjugated to, or is to be conjugated to, another moiety, such as a delivery vehicle. In such cases, a conjugation handle may include a naturally-occurring or modified amino acid containing a reactive group. In embodiments, a conjugation handle may be attached to a moiety (such as the targeting moiety), directly (e.g., by covalent attachment) or indirectly. As used herein, in some embodiments, “conjugation handle” also refers to a short peptide that is part of a conjugation product formed by joining two moieties. For instance, the conjugation handle may be joined to a targeting moiety and become part of a longer sequence, e.g., the conjugation handle may be attached to the sequence of a targeting moiety that comprises or consists of an antibody or antigen-binding fragment (e.g., a VHH). In some embodiments, the conjugation handle is attached to the C terminus of the antibody or antigen-binding fragment.

[0325] Corresponding to: As used herein, the term “corresponding to” refers to a nucleic acid sequence or an amino acid sequence at particular positions of an intron, or the corresponding positions in another intron. A sequence corresponding to the sequence at particular positions of an intron may comprise a corresponding substitution or a variant, e.g., the substituted nucleotides or amino acids do not naturally occur at the corresponding positions. The substituted nucleotides or amino acids may be the corresponding residues in another intron (e.g., Group I or II intron).

[0326] Delivery. As used herein, “delivery” refers to the act or manner of delivering a compound, substance, entity, moiety, cargo or payload.

[0327] DNA Nuclease: As used herein, “DNA nuclease” is an enzyme that catalyzes the cleavage of phosphodiester bonds. DNA nucleases play a role in DNA replication and various DNA repair processes, base excision repair, nucleotide excision repair, mismatch repair, and double strand break repair. Depending on whether a 5' or 3' end is required for substrate recognition and whether cleavage products are single or oligonucleotides, DNA nucleases are classified as exonucleases and endonucleases. For example, self-cleaving ribozymes cleave RNAs endonucleolytically, while exonucleases cleave one nucleotide at a time from one end,ATTORNEY DOCKET NO. ORB-019WO1either from 5' to 3' or from 3' to 5'. Some nucleases have both exonuclease and endonuclease activities, for example, Flap endonuclease 1 (FEN1) has 5' to 3' exonuclease activity in addition to endonuclease activity, and Mrel 1 has both endonuclease and 3' to 5' exonuclease activities.

[0328] DNase I: As used herein, “DNase I” or “deoxyribonuclease I” refers to enzymes that cleave single or double-stranded DNA and require divalent metal ions to hydrolyze DNA yielding 3'-hydroxyl and 5 '-phosphorylated products.

[0329] Downstream'. As used herein, the term “downstream” refers to sequence that is 3' to a particular sequence.

[0330] Encapsulate-. As used herein, the term “encapsulate” means to enclose, surround, or encase. As it relates to the formulation of the compositions of the disclosure, encapsulation may be substantial, complete or partial. The term “substantially encapsulated” means that at least greater than 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.9%, 99.9% or greater than 99.999% of the pharmaceutical composition of the disclosure may be enclosed, surrounded or encased within the delivery agent. “Partially encapsulated” means that less than 10%, 20%, 30%, 40%, 50%, or less of the pharmaceutical composition or compound of the disclosure may be enclosed, surrounded or encased within the delivery agent. In some embodiments, at least 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.9%, 99.99%, or greater than 99.99% of the pharmaceutical composition of the present disclosure are encapsulated in the delivery vehicle (e.g., a lipid nanoparticle (LNP)).

[0331] Encode-. As used herein, the term “encode” or “encoding” refers broadly to any process whereby the information in a polymeric macromolecule is used to direct the production of a second molecule that is different from the first. The second molecule may have a chemical structure that is different from the chemical nature of the first molecule. For example, DNA encodes RNA and RNA encodes a polypeptide or protein.

[0332] Enhance'. As used herein, the terms “enhance” and “enhancement” refers to an increase of at least about 5%, 10%, 20%, 25%, 50%, 75%, 100%, 150%, 200%, 300%, 400%, 500% or more of a reference; the reference may be a biological function of a nucleic acid or protein and a gene expression level, etc.

[0333] Expression: As used herein, “expression” of a nucleic acid sequence refers to one or more of the following events: (1) production of an RNA template from a DNA sequence (e.g., by transcription); (2) processing of an RNA transcript (e.g., by splicing, editing, 5' capATTORNEY DOCKET NO. ORB-019WO1formation, and / or 3' end processing) (RNA expression); (3) translation of an RNA into a polypeptide or protein; and (4) post-translational modification of a polypeptide or protein (Protein expression).

[0334] Feature: As used herein, a “feature” refers to a characteristic, a property, or a distinctive element. Features may refer to features of polynucleotides (e.g., circular polynucleotide of the present disclosure) such as sequence-based (nucleotides, modified nucleotides) or structural features (loops, folds, stem-loops, stems, hairpins, bulges, halfloops, etc.). Features may also refer to features of the polypeptides encoded by the present circular polynucleotide, such as surface manifestations, local conformational shape, folds, loops, half-loops, domains, half-domains, sites, termini or any combination thereof.

[0335] Formulation'. As used herein, a “formulation” includes one or more compound, substance, entity, moiety (e.g., targeting moiety), cargo, or payload (such as RNA) and a delivery vehicle (e.g., lipid nanoparticle or virus like particles).

[0336] Fragment'. A “fragment,” as used herein, refers to a portion. For example, an intron fragment may comprise a portion of the full intron sequence. Fragments of proteins may comprise polypeptides obtained by digesting full-length protein isolated from cultured cells.

[0337] Gap: As used therein, the term “gap” refers to a small region with the DNA splint comprising a stretch of nucleotides that are not complementary to any sequence of linear RNAs that are to be circularized. The gap region can be in the middle nucleotides of the splint, creating a situation where the splint ‘gap’ is single stranded, and the terminal 3' and 5' ends of the RNA molecule are also single stranded (depending on size of the gap).

[0338] Homology'. As used herein, the term “homology” refers to the overall relatedness between polymeric molecules, e.g., between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules). In some embodiments, polymeric molecules are considered to be “homologous” to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical or similar. The term “homologous” necessarily refers to a comparison between at least two polynucleotide sequences. In accordance with the disclosure, two polynucleotide sequences are considered to be homologous if the polypeptides they encode are at least about 50%, 60%, 70%, 80%, 90%, 95%, or even 99% identical for at least one stretch of at least about 20 amino acids. In some embodiments, homologous polynucleotide sequences are characterized by the ability to encode a stretch of at least 4-5 uniquely specified amino acids. For example, in someATTORNEY DOCKET NO. ORB-019WO1embodiments, for polynucleotide sequences less than 60 nucleotides in length, homology is determined by the ability to encode a stretch of at least 4-5 uniquely specified amino acids.

[0339] Hydropathy Index: As used herein, the term “hydropathy index” is a quantitative measure that refers to the hydrophilicity and hydrophobicity of a nucleobase sequence (e.g., an RNA sequence) by combining (i) experimentally measured nucleobase partition coefficients (log ) measuring distribution of the nucleobase between an immiscible organic solvent (e.g., n-Octanol) and water and (ii) normalization to length of the sequence, enabling comparison across sequences of different lengths. Typically, a more negative log P indicates a more hydrophilic nucleobase, while a less negative or positive log P indicates a more hydrophobic nucleobase. Experimental nucleobase log P values have been experimentally determined for canonical nucleobases and are as follows: cytosine: -1.7 (most hydrophilic), uracil: -1.1 (moderate hydrophilicity), guanine: -0.9 (moderate hydrophilicity) and adenine (least hydrophilic). (Zamora WJ et al., J. Phys. Chem. B, 2017, 13, 10908-10922). Poly A sequences, which have a higher hydropathy index, increasingly associate withionizable-lipid-rich regions at the LNP membrane surface remaining in contact with the membrane rather than remaining freely dispersed within the LNP. This heightened tendency to interact is undesirable, as it reflects stronger RNA-lipid association that can impede efficient delivery. In contrast, a sequence with a lower hydropathy index would be less likely to interact with ionizable lipids and thus support improved cellular uptake and cytoplasmic release. (Gilbert et al. 2024, Nanoscale, 2024, 16, 777-794). Protocol for Assigning a Residue’s Character on the Hydropathy Scale (PARCH) analysis has confirmed that base composition is the primary determinant of hydropathy as the phosphate-ribose backbone is uniformly highly hydrophilic and does not vary between sequences. Hydropathy of RNA sequences in the current disclosure is calculated by computing an additive hydropathy sum Hraw = Ei(nt * l°gPi) where n;- is the number of nucleobases of each type, e.g., nA, nc, no, nuand log P values are as determined by Zamora et al. Finally, the Hraw values are normalized to length of the RNA sequence so that hydropathy indices of sequences of different lengths may be directly compared, i.e., Hydropathy index =Further,normalizing to length provides a per nucleotide average hydrophilicity. As used herein, the hydropathy index refers to intrinsic hydropathy of the primary nucleic acid sequence without regard to folding, secondary structure or tertiary interactions.

[0340] Identity. As used herein, the term “identity” refers to the overall relatedness between polymeric molecules, e.g., between polynucleotide molecules (e.g., DNA moleculesATTORNEY DOCKET NO. ORB-019WO1and / or RNA molecules) and / or between polypeptide molecules. Calculation of the percent identity of two polynucleotide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second nucleic acid sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). In certain embodiments, the length of a sequence aligned for comparison purposes is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or 100% of the length of the reference sequence. The nucleotides at corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, considering the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, the percent identity between two nucleotide sequences can be determined using methods such as those described in Computational Molecular Biology, Lesk, A. M., ed., Oxford University Press, New York, 1988; Biocomputing:Informatics and Genome Projects, Smith, D. W., ed., Academic Press, New York, 1993; Sequence Analysis in Molecular Biology, von Heinje, G., Academic Press, 1987; Computer Analysis of Sequence Data, Part I, Griffin, A. M., and Griffin, H. G., eds., Humana Press, New Jersey, 1994; and Sequence Analysis Primer, Gribskov, M. and Devereux, J., eds., M Stockton Press, New York, 1991; each of which is incorporated herein by reference. For example, the percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (CAB IOS, 1989, 4:11-17), which has been incorporated into the ALIGN program (version 2.0) using a PAM120 weight residue table, a gap length penalty of 12 and a gap penalty of 4. The percent identity between two nucleotide sequences can, alternatively, be determined using the GAP program in the GCG software package using an NWSgapdna. CMP matrix. Methods commonly employed to determine percent identity between sequences include, but are not limited to those disclosed in Carillo, H. and Lipman, D., SIAM J Applied Math., 48:1073 (1988); incorporated herein by reference. Techniques for determining identity are codified in publicly available computer programs. Exemplary computer software to determine homology between two sequences include, but are not limited to, GCG program package, Devereux, J., et al., Nucleic Acids Research, 12(1), 387ATTORNEY DOCKET NO. ORB-019WO1(1984)), BLASTP, BLASTN, and FASTA Altschul, S. F. et al., J. Molec. Biol., 215, 403 (1990)).

[0341] Immune response'. The term “immune response” includes both a specific reaction of the adaptive immune system to a particular antigen (so called “specific” or “adaptive” immune response), and an unspecific reaction of the innate immune system (so called “unspecific” or “innate” immune response). In some aspects, the invention relates to eliciting specific reactions (adaptive immune responses) of the adaptive immune system. Furthermore, the invention also relates to eliciting an innate immune response, since the specific (adaptive) immune response can be supported by an additional unspecific reaction (innate immune response). Therefore, in some aspects, the invention also relates to simultaneous stimulation of the innate and the adaptive immune system to evoke an efficient immune response.

[0342] Innate immune response'. The term “innate immune response”, also known as nonspecific immune response, comprises the cells and mechanisms that defend the host from infection by other organisms in a non-specific manner. The cells of the innate system recognize and respond to pathogens in a generic way, but, unlike the adaptive immune system, the innate immune response does not confer long-lasting or protective immunity to the host. The innate immune system may be activated, for example, by ligands of pathogen-associated molecular patterns (PAMP) receptors, Toll-like receptors (TLRs), or other auxiliary substances such as lipopolysaccharides, TNF-alpha, CD40 ligand, or cytokines, monokines, lymphokines, interleukins or chemokines, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, IL-19, IL-20, IL-21, IL-22, IL-23, IL-24, IL-25, IL-26, IL-27, IL-28, IL-29, IL-30, IL-31, IL-32, IL-33, IFN-alpha, IFN-beta, IFN-gamma, GM-CSF, G-CSF, M-CSF, LT-beta, TNF-alpha, growth factors, and hGH, a ligand of human Toll-like receptor TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, a ligand of murine Toll-like receptor TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TLR11, TLR12 or TLR13, a ligand of aNOD-like receptor, a ligand of a RIG-1 like receptor, an immunostimulatory nucleic acid, an immunostimulatory RNA (isRNA), an antibacterial agent, or an anti-viral agent. Typically, a response of the innate immune system includes recruiting immune cells to sites of infection, through the production of chemical factors, including specialized chemical mediators, called cytokines; activation of the complement cascade; identification and removal of foreign substances present in organs, tissues, the blood and lymph, by specialized white blood cells;ATTORNEY DOCKET NO. ORB-019WO1activation of the adaptive immune system through, a process known as antigen presentation; and / or acting as a physical and chemical barrier to infectious agents.

[0343] Internal Homology Element: As used herein, the terms “internal homology region,” “internal homology element,” and “inner homology element (IHE)” are used interchangeably. The internal homology region refers to complementary sequences at the 5’ end and the 3’ end of precursor RNA (or DNA template from which the precursor RNA is transcribed), referred to as the 5’ internal homology element and the 3’ internal homology element, respectively. The 5’ internal homology element forms intramolecular Watson-Crick base pairs with the 3’ internal homology element, such that the homology elements have at least 75% sequence complementarity. These base-pairing interactions stabilize 5’ end to 3’ end association and promote circularization of the RNA. The internal homology region ranges from 5-50 nucleotides in length, more typically, between 5-20 nucleotides in length.

[0344] Intron: As used herein, the term “intron” means any sequence within a gene that is removed by RNA splicing during maturation of RNA transcripts from pre-mRNA transcripts. The term “intron” also refers to any sequence within a precursor linear RNA that is spliced out during circularization to form circular RNA. Introns in general are non-coding sequences of an RNA transcript. “Intron” can also refer to intron fragments.

[0345] Ionizable Lipid. As used herein, “ionizable lipid” refers to any of several lipid species that carry a net positive charge at a selected pH. For example, and without limitation, an ionizable lipid may carry a net positive charge at an acidic pH (e.g., pH of about 4.0), but remain neutrally charged at a physiological pH. Ionizable lipids typically comprise an ionizable head group such as amine linked to a hydrophobic tail region comprising aliphatic chain of varying degree of saturation.

[0346] Linker'. As used herein, “linker” refers to a short amino acid or polypeptide sequence that operably connects, joins or links two protein sequences, including without limitation, antibodies or domains, e.g., VH and VL. A linker can be fused to an N-terminus or a C-terminus of a polypeptide. A linker is of any length, for example, 3-100 amino acids long.

[0347] Lipid Nanoparticle'. As used herein “lipid nanoparticle” or “LNP” refers to a delivery vehicle comprising one or more lipids (e.g., cationic lipids, non-cationic lipids, Modified-PEG-lipids). An “ionizable lipid” refers to any of several lipid species that carry a net positive charge at a selected pH. A “non-cationic lipid” refers to any neutral, zwitterionic or anionic lipid.ATTORNEY DOCKET NO. ORB-019WO1

[0348] Liposome-. As used herein, “liposome” generally refers to a vesicle composed of lipids (e.g., amphiphilic lipids) arranged in one or more spherical bilayers or bilayers.

[0349] microRNA or miRNA or miR. As used herein, “microRNA” or “miRNA” or “miR” refers to small non-coding ribonucleic acids that regulates gene expression. A “miRNA” or “microRNA” is a single-stranded RNA of short length (for example, 18-25 nucleotides) that functions not to encode proteins, but to regulate biological processes such as cell proliferation or differentiation, immune response, and expression of other genes at the post-transcriptional level. The dysregulation of microRNA may be involved in various diseases, including autoimmune diseases and cancer.

[0350] Micro RNA (miRNA) binding sequence or miRNA binding site: As used herein, the term “microRNA binding sequence,” “miRNA binding sequence”, “microRNA binding site,” or “miRNA binding site” refers to a binding sequence (e.g.,, a short nucleic acid fragment) that binds to a miRNA. In some embodiments, the circular polynucleotide, such as circular RNA, comprise one or more miRNA binding sites. The number of miRNA binding sites in the circular polynucleotide is variable and relates to the length of the circular polynucleotide. As non-limiting examples, the circular polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more miRNA binding sites. The multiple miRNA binding sites may have the same nucleic acid sequences and bind to the same miRNA; or alternatively, the miRNA binding sites have different nucleic acid sequences and bind to different miRNAs, such as 2, 3, 4, 5, or more different miRNAs.

[0351] Modified. As used herein “modified” or, as appropriate, “modification” refers to a changed state or structure of a molecule. Molecules may be modified in many ways including chemically, structurally, and functionally. With respect to nucleic acid molecules (e.g., DNA and RNA), the modifications are modified version of A, G, C, U or T nucleotides. With respect to polypeptides, the term “modification” refers to a modification to the canonical set of 20 amino acids.

[0352] mRNA: As used herein, the term “messenger RNA” (mRNA) means a polynucleotide which encodes a polypeptide of interest or synthetic polypeptide, and which is capable of being translated to produce the encoded polypeptide of interest or synthetic polypeptide, such as a CAR construct, in vitro, in vivo, in situ or ex vivo.

[0353] Nucleic acid molecule '. As used herein, the term “ nucleic acid molecule" refers to a polymeric form of nucleotides, either deoxyribonucleotides (DNAs) or ribonucleotides (RNAs), or analogs thereof. The terms include single stranded or double-stranded moleculesATTORNEY DOCKET NO. ORB-019WO1comprised of nucleic acid bases. As such, the term includes, and may be used interchangeably with “plasmids”, “constructs”, or “vectors.” In the present disclosure, the terms “polynucleotide" and “nucleic acid' are used interchangeably. As is commonly understood, DNA is double-stranded. Any DNA sequences provided in the present disclosure may be sense or anti-sense sequences, and it is understood that the sequences of both DNA strands and the transcribed RNA product from either DNA strand are fully encompassed by the present disclosure. It is commonly understood that DNA includes nucleobases adenine, cytosine, thymine, and guanine, and RNA includes nucleobases adenine, cytosine, uracil, and guanine. Thymine in DNA is replaced by uracil in RNA; both thymine and uracil function as the complementary base to adenine. In the context of both a DNA and an RNA sequence, it is understood that A represents adenine, C represents cytosine, and G represents guanine. In the context of a DNA sequence, it is understood that T represents thymine. In the context of an RNA sequence, it is understood that U and T both represent uracil; throughout the present disclosure any reference to T in an RNA sequence means uracil.

[0354] Pharmaceutical composition'. As used herein the term “pharmaceutical composition” refers to compositions comprising at least one active ingredient or drug substance and optionally one or more pharmaceutically acceptable excipients. The pharmaceutical composition can comprise a circRNA as disclosed herein or a circRNA therapeutic as disclosed herein. In some embodiments, the pharmaceutical composition comprises a circRNA as disclosed herein as the active ingredient or drug substance. In some embodiments, the pharmaceutical composition comprises a targeted LNP as disclosed herein as an exemplary excipient.

[0355] Polydispersity Index: As used herein, the term “poly dispersity index” or “PDF’ refers to a range of value between 0 to 1 that defines the homogeneity or heterogeneity of lipid nanoparticle population. PDI value close to 0 indicates homogeneity of lipid nanoparticle population, whereas PDI value close to 1 indicates heterogeneity of the population. In some embodiments, the poly dispersity index (PDI) is about 0 to 0.1 (e.g., 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09 or 0.1).

[0356] Polypeptides'. As used herein, the term “polypeptides” refer to any polypeptide, both synthetic and naturally occurring, encoded in the linear and / or circular RNA of the present invention. As used herein, “polypeptide” means a polymer of amino acid residues (natural or unnatural) linked together, most often by peptide bonds. In some embodiments, the term, as used herein, refers to proteins (i.e., proteins of interest), polypeptides, andATTORNEY DOCKET NO. ORB-019WO1peptides of any size, structure, or function. In some embodiments, the polypeptide encoded is smaller than about 50 amino acids and the polypeptide is then termed a peptide. For example, a peptide is at least about 2, 3, 4, or at least 5 amino acid residues long. In some embodiments, polypeptides include gene products, naturally occurring polypeptides, synthetic polypeptides, homologs, orthologs, paralogs, fragments and other equivalents, variants, and analogs of the foregoing. In some embodiments, a polypeptide is a single molecule. In some embodiments, a polypeptide includes a multi-molecular complex such as a dimer, trimer or tetramer. In some embodiments, a polypeptide comprises single chain or multichain polypeptides such as antibodies or insulin and is associated or linked. In some embodiments, disulfide linkages are found in multichain polypeptides. In some embodiments, the term polypeptide applies to amino acid polymers in which one or more amino acid residues are an artificial chemical analogue of a corresponding naturally occurring amino acid. In some embodiments, the polypeptide (e.g., synthetic polypeptide) is a chimeric antigen receptor.

[0357] Polypeptide variant: As used herein, “polypeptide variant” refers to molecules which differ in their amino acid sequence from a native, wild-type or reference sequence. In some embodiments, the amino acid sequence variants possess substitutions, deletions, and / or insertions at certain positions within the amino acid sequence, as compared to a native or reference sequence. Typically, variants possess at least about 50% identity (homology) to a native or reference sequence. In some embodiments, they possess at least about 80%, at least about 90% identity to a native, wild-type or reference sequence.

[0358] Ribosome recruiting element'. As used herein, a “ribosome recruiting element” includes naturally occurring or engineered RNA sequences that directly or indirectly recruit the ribosome to initiate translation of RNA into protein. A ribosome recruiting element is an RNA sequence that recruits ribosomes to initiate translation of, for example, circRNA, in a cap-independent manner. In some embodiments, a ribosome recruiting element is an internal ribosome entry site (IRES). For circRNA, an IRES plays a critical role in translation initiation of the circRNA in the absence of a 5’ cap by directly recruiting ribosomes to an internal position of circRNA in a cap-independent manner to initiate translation. In other embodiments, a ribosome recruiting element is a branched cap structure (e.g., qRNA) added to circular RNA that initiates translation by the ribosome. In some embodiments, a ribosome recruiting element is an IRES transacting factor (ITAF) recruiting element. In some embodiments, IRES and ITAFs recruit the ribosome. In some embodiments, the ITAF isATTORNEY DOCKET NO. ORB-019WO1specific to an IRES or group of related IRES elements. In some embodiments, the ITAF is general (for example, polypyrimidine tract-binding protein, PTB).

[0359] Signal peptide: As used herein, a “signal sequence” or “signal peptide” is a polynucleotide or polypeptide, respectively, which is from about 9 to 200 nucleotides (3-60 amino acids) in length which is incorporated at the 5' terminus of the coding region or the N-terminus polypeptide encoded, respectively. In some embodiments, addition of these sequences results in trafficking of the encoded polypeptide to the endoplasmic reticulum through one or more secretory pathways. Some signal peptides are cleaved from the protein by signal peptidase after the proteins are transported.

[0360] Spacer'. As used herein, the term “spacer” refers to any contiguous nucleotide sequence (e.g., of one or more nucleotides) that provides distance or flexibility between two adjacent polynucleotide regions. The spacer can be a 5' or 3' spacer. In some embodiments, the nucleic acid for making a circular RNA (e.g., precursor RNA) of the present invention comprises a 5' spacer that is located between the upstream intron fragment and a sequence of interest ( / .<., a coding sequence to be included in an RNA to be circularized). In some embodiments, the 5' spacer is between the upstream intron fragment and the IRES. In some embodiments, the nucleic acid for making a circular RNA of the present invention comprises a 3' spacer that is located between a sequence of interest and the downstream intron fragment. The 5' and 3' spacer sequences may be 10 nucleotides to 100 nucleotides in length, or 20 nucleotides to 50 nucleotides in length. In some embodiments, the 5' spacer is at least 10 nucleotides in length. In some embodiments, the 5' spacer sequence is at least 15 nucleotides in length. In some embodiments, the 5' spacer is at least 20 nucleotides in length. In some embodiments, the 5' spacer sequence is at least 30 nucleotides in length. In some embodiments, a 3' spacer is located between a sequence to be circularized and the downstream intron sequence. In the precursor RNA and the resultant circular RNA following circularization, within the 5’ UTR or the 3’ UTR, more typically, the 3’ UTR, a microRNA binding site is flanked by an upstream spacer and a downstream spacer. The upstream spacer refers to a contiguous non-coding sequence 5’ of the microRNA binding site. The downstream spacer refers to a contiguous non-coding sequence 3 ’ of the microRNA binding site. Typically, the upstream spacer and / or downstream spacer comprises between 12-18 nucleotides and are of comparable length. In the present disclosure, the upstream spacer and / or downstream spacer comprise no more than five contiguous adenine sequences (for example, two, three, four or five contiguous adenine sequences are interrupted by a cytosine).ATTORNEY DOCKET NO. ORB-019WO1

[0361] Structural Lipid. As used herein, “structural lipid” refers to sterols and lipids containing sterol moieties.

[0362] Subject: As used herein, the terms “subject” and “patient” may be used interchangeably herein. As such, unless otherwise indicated, a “subject” refers to a human that is being treated for a disease, or prevention of a disease, such as a patient.

[0363] Transcription'. As used herein, the term “transcription” refers to the formation or synthesis of an RNA molecule by, e.g., an RNA polymerase using a DNA molecule as a template.

[0364] Translation'. As used herein, the term “translation” refers to the formation of a polypeptide molecule by a ribosome based upon an RNA template.

[0365] Treat and Prevent'. As used herein the terms “treat” or “prevent” as well as words stemming therefrom do not necessarily imply 100% or complete treatment or prevention. Rather, there are varying degrees of treatment or prevention of which one of ordinary skill in the art recognizes as having a benefit or therapeutic effect. Also, “prevention” can encompass delaying the onset of the disease, symptom or condition thereof.

[0366] Therapeutically effective amount'. As used herein, the term “therapeutically effective amount” means an amount effective, at dosages, frequency of administration, and for duration of time necessary to achieve the desired results such that one or more symptoms or biomarkers is improved after treatment.

[0367] Unmodified. As used herein, “unmodified” refers to any substance, compound or molecule prior to being changed in any way. Unmodified may, but does not always, refer to the wild type or native form of a biomolecule. Molecules may undergo a series of modifications whereby each modified molecule may serve as the “unmodified” starting molecule for a subsequent modification.

[0368] Untranslated Region '. As used herein, the term “untranslated region” or “UTR” refers to regions 5' or 3' of the coding sequence which are not translated. As use herein, “UTR” can include, for example, spacers.

[0369] Upstream'. As used herein, the term “upstream” refers to sequence that is 5' to a particular sequence.

[0370] Vector: As used herein, a “vector” is any molecule or moiety which transports, transduces or otherwise acts as a carrier of a heterologous molecule. Vectors of the present disclosure may be produced recombinantly and may be based on and / or may comprise viral parent or reference sequences. Such parent or reference viral sequences may serve as anATTORNEY DOCKET NO. ORB-019WO1original, second, third, or subsequent sequence for engineering vectors. In non-limiting examples, such parent or reference viral sequences may comprise any one or more of the following sequences: a polynucleotide sequence encoding a polypeptide or multi-polypeptide, which sequence may be wild-type or modified from wild-type and which sequence may encode full-length or partial sequence of a protein, protein domain, or one or more subunits of a protein; a polynucleotide comprising a modulatory or regulatory nucleic acid which sequence may be wild-type or modified from wild-type; and a transgene that may or may not be modified from wild-type sequence.

[0371] Vehicle'. The term “vehicle” refers to an agent, e.g., a carrier, that may typically be used within a pharmaceutical composition for facilitating administering of the components of the pharmaceutical composition to an individual.

[0372] Virus-like Particle: Virus-like particles (VLPs) are molecules that closely resemble viruses but are non-infectious because they contain no viral genetic material. As used herein, the term “virus-like particle” is used interchangeably with “viral like particles.” VLPs can be naturally occurring or synthesized through the individual expression of viral structural proteins, which can then self-assemble into the virus-like structure. As used herein, VLPs are used interchangeably with engineered viral particles.DETAILED DESCRIPTION

[0373] The present disclosure, provides, among other things, an improved in vivo CAR therapy for treating B cell mediated diseases based on circular ribonucleic acids (circRNA). In some aspects, the present disclosure provides targeted in vivo delivery of genes encoding CARs to select cell types (e.g., immune cells, or, more specifically, T cells) for treating B cell-mediated diseases e.g., autoimmune disease or cancer). The in vivo CAR therapy disclosed herein provides multiple benefits over current ex vivo CAR therapies, for which manufacturing is complex, laborious, and expensive.

[0374] Ex vivo CAR therapies to treat cancer or autoimmune diseases requires patients’ or donors’ T cells to be isolated from peripheral blood via apheresis, activated, and subsequently transduced by a lentivirus comprising a chimeric antigen receptor (CAR) construct to create an engineered CAR T cell. CAR T cells are further expanded before infusing engineered T cells into a patient’s bloodstream to attack target cells. Further, ex vivo therapy requires lymphodepletion prior to administration, and is also associated with serious side-effects including cytokine release syndrome (CRS), which can cause multi-organ failure and evenATTORNEY DOCKET NO. ORB-019WO1death, and immune effector cell-associated neurotoxicity syndrome (ICANS), which is a neuropsychiatric syndrome that can cause confusion and seizures (Bui, 2024; Mullard, 2024).

[0375] The in vivo CAR therapy of the present invention uses the body’s own immune system to manufacture the CAR T cells in vivo and bypasses the complex manufacturing steps required for ex vivo therapy. In addition, the in vivo CAR therapy does not require a step of lymphodepletion prior to administration of the therapy to the patient.

[0376] The present disclosure, provides, in part, a highly stable, durable, and targeted in vivo CAR therapy encoded by circRNA that is achieved through an unexpected and surprising synergy of a combination of precisely engineered features in the circRNA, delivery vehicle (e.g., lipid nanoparticle), and targeting moiety conjugated to the delivery vehicle. In some embodiments, the targeted in vivo CAR therapy in the present disclosure allows expression of the CAR therapy, and use of circRNA improves the duration and magnitude of CAR expression on T cells, which results in B cell depletion. As described herein, the targeted in vivo CAR therapy reaches the specific cell types and tissues, e.g., T cells, and allows for enhanced in vivo CAR T generation and improves potency for B cell depletion.

[0377] The circRNA therapeutics described in this disclosure provide multiple advantages compared to linear RNA. For example, circRNA therapeutics provides enhanced stability. Without wishing to be bound by any mechanism, since circRNA is a closed molecule with no free ends, it confers resistance to exonucleases, which leads to RNA durability. In addition, the absence of open 5’ and 3’ ends leads to potential reduction of an innate immune response by the patient. Moreover, circular RNA allows for efficient translation compared to linear RNA; IRES-mediated translation (cap-independent) leads to high synthesis of the protein encoded by the circular RNA.

[0378] The circRNAs and circRNA therapeutics of the present disclosure were precisely engineered to be efficacious and safe to treat diseases such as autoimmune diseases and cancer. In some embodiments, the circRNAs and circRNA therapeutics of the present disclosure: stably and durably express CAR to high levels, in part, due to increased stability of circRNA relative to a linear RNA; have specific Internal Ribosome Entry Site (IRES) elements that drive unexpectedly high expression of the CAR construct, including anti-CD19 CAR construct, in T cells; include variant 5' and / or 3' untranslated regions (UTR) with miRNA binding sites in the spacers to decrease off-target liver expression; and are codon-optimized for high expression of the CAR construct, including anti-CD19 CAR construct.ATTORNEY DOCKET NO. ORB-019WO1

[0379] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: (i) a circRNA comprising a ribosome recruiting element, and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) a targeting moiety, wherein the circRNA is encapsulated in a delivery vehicle, wherein the targeting moiety is conjugated to the delivery vehicle, and wherein the targeting moiety comprises an anti-CD5 antibody heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89.

[0380] In some aspects, provided herein is a circular ribonucleic acid (circRNA) for targeted delivery comprising: (i) a circRNA comprising: a ribosome recruiting element, and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) a targeting moiety, wherein the targeting moiety binds to a T cell, wherein the circRNA is in a delivery vehicle, wherein the targeting moiety that binds to a T cell is conjugated to the delivery vehicle, and wherein the targeting moiety is an anti-CD5 antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising complementarity determining region (CDR) sequencesGFTFSMYS (SEQ ID NO: 64), ISTGARDT (SEQ ID NO: 65) and GDLRYGPDGYDY (SEQ ID NO: 66).

[0381] Provided herein, among other things, is a circular ribonucleic acid (circRNA) for targeted delivery comprising: (i) a circRNA comprising a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (ii) a targeting moiety, and (iii) a delivery vehicle, wherein the targeting moiety is conjugated to the delivery vehicle (e.g., lipid nanoparticle, engineered viral particle, among others). In some embodiments, the targeting moiety comprises an anti-CD5 antibody or antigen-binding fragment thereof.

[0382] The present disclosure provides, among other things, a circRNA, comprising: a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA comprises an Internal Ribosome Entry Site (IRES) having at least 85% identity to a sequence comprising any one of SEQ ID NO: 1-11, and one or more untranslated region (UTR).

[0383] Among other things, provided herein is a circRNA comprising a codon-optimized coding sequence having at least 85% identity to SEQ ID NOS: 125-168 or 266.ATTORNEY DOCKET NO. ORB-019WO1

[0384] Provided herein, in part, is a circRNA comprising a sequence having at least 85% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, the sequence is selected from SEQ ID NOs: 175-233, 264, or 269.

[0385] Provided herein, among other things, is a circRNA therapeutic for targeted delivery, comprising: (i) a circRNA comprising a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) an anti-CD5 T cell targeting moiety; wherein the circRNA is encapsulated in a lipid nanoparticle; and wherein the T cell targeting moiety is conjugated to the lipid nanoparticle.

[0386] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: a circular RNA comprising a ribosome recruiting element and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA is encapsulated in an LNP, wherein a targeting moiety is conjugated to the LNP, wherein the targeting moiety is an anti-CD5 VHH, the anti-CD5 VHH comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89, wherein the LNP comprises one or more ionizable lipids at 30 to 49 mol% (e.g, at 30 to 45 mol%), one or more phospholipids at 5 to 40 mol% (e.g., at 15 to 35 mol%), one or more sterol lipids at 15 to 50 mol% (e.g., at 30 to 50 mol%), DMG-PEG 2000 at 0 to 10 mol% (e.g., at 0 to 5 mol%), and DSPE-PEG 2000-maleimide at 0.01 to 1 mol%, and wherein the VHH is conjugated to the LNP.

[0387] The present disclosure, provides among other things, a circRNA therapeutic for targeted delivery, comprising: (i) a circRNA comprising (a) a 5' untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES) having at least 85% identity to a nucleic acid sequence comprising SEQ ID NO: 1, (c) a coding sequence encoding: a CD8a signal peptide, and a Chimeric Antigen Receptor (CAR) comprising: an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a 4- IBB costimulatory domain, a CD3 zeta signaling domain, and (d) a 3' UTR, and (ii) an anti-CD5 T cell targeting moiety comprising the complementarity determining region (CDR) sequences of GFTFSMYS (SEQ ID NO:64), ISTGARDT (SEQ ID NO: 65) and GDLRYGPDGYDY (SEQ ID NO: 66), wherein the circRNA is encapsulated in a lipid nanoparticle (RNA-LNP), and wherein the anti-CD5 T cell targeting moiety is conjugated to the lipid nanoparticle.

[0388] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: a circular RNA comprising a ribosomeATTORNEY DOCKET NO. ORB-019WO1recruiting element and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA is encapsulated in an LNP, wherein a targeting moiety is conjugated to the LNP, wherein the targeting moiety is an anti-CD5 VHH, the anti-CD5 VHH comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89, wherein the LNP comprises one or more ionizable lipids at 30 to 49 mol%, one or more phospholipids at 5 to 40 mol%, one or more sterol lipids at 15 to 50 mol%, a DMG-PEG 2000 lipid at 0 to 10 mol%, and a DSPE-PEG 2000-maleimide lipid at 0.01 to 1 mol%, and wherein the VHH is conjugated to the LNP.

[0389] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: a circular RNA comprising a ribosome recruiting element and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA is encapsulated in an LNP, wherein a targeting moiety is conjugated to the LNP, wherein the targeting moiety is an anti-CD5 VHH, the anti-CD5 VHH comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89, wherein the LNP comprises one or more ionizable lipids at 30 to 45 mol%, one or more phospholipids at 15 to 35 mol%, one or more sterol lipids at 30 to 50 mol%, a DMG-PEG 2000 lipid at 0 to 5 mol%, and a DSPE-PEG 2000-maleimide lipid at 0.01 to 1 mol%, and wherein the VHH is conjugated to the LNP.

[0390] In some aspects, provided herein is a circRNA therapeutic, comprising: (i) a circRNA comprising in order: (a) a 5’ exon fragment and a 5' UTR, (b) an IRES having at least 85% identity to SEQ ID NO: 1, (c) a coding sequence encoding a chimeric antigen receptor (CAR), (d) a 3' UTR and a 3’ exon fragment, and (ii) an anti-CD5 T cell targeting moiety comprising the complementarity determining region (CDR) sequences CDR1, CDR2, and CDR3, wherein the CDR1, the CDR2, and the CDR3 amino acid sequences are GFTFSMYS (SEQ ID NO: 64), ISTGARDT (SEQ ID NO: 65), and GDLRYGPDGYDY (SEQ ID NO: 66), respectively, wherein the circRNA is encapsulated in a lipid nanoparticle, and wherein an anti-CD5 T cell targeting moiety is conjugated to the lipid nanoparticle.

[0391] In some aspects, provided herein is a circRNA therapeutic, comprising: (i) a circRNA comprising in order: (a) a 5’ exon fragment and a 5' UTR, (b) an IRES having at least 85% identity to SEQ ID NO: 1, (c) a coding sequence encoding a chimeric antigen receptor (CAR) that targets CD 19, (d) a 3' UTR and a 3’ exon fragment, and (ii) an anti-CD5 T cell targeting moiety comprising the complementarity determining region (CDR) sequences CDR1, CDR2, and CDR3, wherein the CDR1, the CDR2, and the CDR3 amino acidATTORNEY DOCKET NO. ORB-019WO1sequences are GFTFSMYS (SEQ ID NO: 64), ISTGARDT (SEQ ID NO: 65), and GDLRYGPDGYDY (SEQ ID NO: 66), respectively, wherein the circRNA is encapsulated in a lipid nanoparticle, and wherein an anti-CD5 T cell targeting moiety is conjugated to the lipid nanoparticle.

[0392] Provided herein is a circRNA therapeutic for targeted delivery, comprising: (i) a circRNA comprising (a) a 5' untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES) having 100% identity to a nucleic acid sequence comprising SEQ ID NO: 1, (c) a coding sequence encoding: a Chimeric Antigen Receptor (CAR) comprising: FMC63 (an anti-CD19 scFv) having a VL sequence of DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTSRLHSGVPSRF SGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEIT (SEQ ID NO: 54), linked by a linker having the amino acid sequence of GGGGSGGGGSGGGGS (SEQ ID NO: 56), and a VH sequence of EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGS ETTYYNSALKSRLTI IKDNSKSQVFLKMNSLQTDDTAI YYCAKHYYYGGSYAMDYWGQGTSV TVSS (SEQ ID NO: 55), a CD8a hinge, a CD8a transmembrane domain, a 4-1BB costimulatory domain, a CD3 zeta signaling domain, and a CD8 signal peptide; and (d) a 3' untranslated region (UTR) of SEQ ID NO: 46 (also known as SEQ ID NO: 272) comprising a miR-122 binding site, and (ii) an anti-CD5 T cell targeting moiety comprising the sequence of SEQ ID NO: 89, wherein the circRNA is encapsulated in a lipid nanoparticle, wherein the anti-CD5 T-cell targeting moiety is conjugated to the lipid nanoparticle via a thiol-maleimide conjugation. In some embodiments, the LNP comprises an ionizable lipid, a phospholipid, a sterol lipid, and one or more PEG-lipids.

[0393] Also provided by the present disclosure is a precursor RNA comprising, from 5' to 3' end: (i) a 3' Group I or Group II intron splicing fragment, (ii) a 5' untranslated region (UTR), (ii) an IRES, (iii) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (iv) a 3' untranslated region (UTR), and (v) a 5' Group I or Group II intron splicing fragment, wherein the 3' and 5' Group I intron splicing fragments are derived from a Group I or Group II intron.

[0394] Provided herein, among other things, is a precursor RNA comprising, from 5' to 3' end: (i) a 3' Group I intron splicing fragment, (ii) a 5' untranslated region (UTR), (iii) an IRES, (iv) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (v) a 3' untranslated region (UTR),ATTORNEY DOCKET NO. ORB-019WO1and (vi) a 5' Group I intron splicing fragment, wherein the 3' and 5' Group I intron splicing fragments are derived from a Twort-ORF142 Group I intron. As used herein, the terms “intron splicing fragment” and “intron fragment” are used interchangeably. The Twort-ORF142 intron sequence includes nucleotides 1055-1336 of the open reading frame Twort-ORF142 (unknown gene) in the genome of Staphylococcus phage Twort (GenBank:AF132670. 1) (Landthaler and Shub, PNAS, 1999, 96 (12), 7005-7010):AACTACTGAAAGCATAAATAATTGTGCCTTTATACAGTAATGTATATCGAAAAAT CCTCTAATTCAGGGAACACCTAAACAAACTAAGATGTAGGCAATCCTGAGCTAA GCTCTTAGTAATAAGAGAAAGTGCAACGACTATTCCGATAGGAAGTAGGGTCAA GTGACTCGAAATGGGGATTACCCTTCTAGGGTAGTGATATAGTCTGAACATATAT GGAAACATATAGAAGGATAGGAGTAACGAACCTATTCGTAACATAATTGAACTT TTAGTTATTT (SEQ ID NO: 273).

[0395] In some embodiments, the nucleic acid molecule for making a circular RNA comprises the upstream intron sequence comprising a sequence corresponding to positions 119 to 282 of SEQ ID NO: 273, and the downstream intron sequence comprising a sequence corresponding to positions 1 to 118 of SEQ ID NO: 273. In one non-limiting example, the upstream intron sequence corresponds to positions 119 to 282 of SEQ ID NO: 273, and the downstream intron sequence corresponds to positions 1 to 118 of SEQ ID NO: 273. In some embodiments, the upstream intron fragment comprises a sequence corresponding to positions 147 to 282 of SEQ ID NO: 273, and the downstream intron fragment comprises a sequence corresponding to positions 1 to 146 of SEQ ID NO: 273. In one non-limiting example, the upstream intron sequence corresponds to positions 147 to 282 of SEQ ID NO: 273, and the downstream intron sequence corresponds to positions 1 to 146 of SEQ ID NO: 273.Incorporated herein by reference in entirety is the disclosure in co-pending patent application US 63 / 660,303 by Applicants (entitled “Compositions and Methods for RNA Circularization”).Circular RNA for Targeted Delivery

[0396] The present disclosure provides, in part, a circular ribonucleic acid (circRNA) for targeted delivery comprising: (i) a circRNA comprising a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (ii) a targeting moiety, and (iii) a delivery vehicle, wherein the targetingATTORNEY DOCKET NO. ORB-019WO1moiety is conjugated to the delivery vehicle (e.g., lipid nanoparticle, viral particle, among others).

[0397] In some aspects, provided herein is a circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising: (i) a circRNA comprising a ribosome recruiting element, and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (ii) a targeting moiety, wherein the circRNA is encapsulated in a delivery vehicle, wherein the targeting moiety is conjugated to the delivery vehicle, and wherein the targeting moiety comprises an anti-CD5 antibody heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89.

[0398] In some embodiments, the CDR1 sequence comprises SEQ ID NO: 64, the CDR2 sequence comprises SEQ ID NO: 65, and the CDR3 sequence comprises SEQ ID NO: 66.Circular RNA

[0399] In some embodiments, the present invention provides a nucleic acid molecule for making a circRNA molecule that encodes an anti-CD19 CAR polypeptide. In some embodiments, the nucleic acid molecule is a DNA construct (e.g., a vector) that is transcribed into linear RNA, and the linear RNA circularizes into a circular RNA. In some embodiments, the nucleic acid molecule is a linear precursor RNA polynucleotide that circularizes into a circular RNA. The nucleic acid molecule described herein comprises different elements essential for the circular RNA synthesis and function. In some embodiments, the present invention provides a circular RNA encoding an anti-CD19 CAR polypeptide that is made from a nucleic acid molecule or by a method described herein.

[0400] In some embodiments, the nucleic acid molecule for making a circular RNA comprises self-splicing intron sequences and a coding sequence. A rational design of a synthetic circular RNA polynucleotide cassette includes at least two self-splicing intron sequences flanking the sequence to be circularized. The upstream and downstream intron sequences are self-spliced to generate a circular RNA comprising the coding sequence. In some embodiments, a 5' untranslated region and / or 3' untranslated region (UTR)) sequences are included between the intron sequences and the coding sequence (i.e., the sequence to be circularized). In some embodiments, one or more internal ribosome entry site (IRES) is added and operably linked to the coding sequence.ATTORNEY DOCKET NO. ORB-019WO1

[0401] In some embodiments, the circRNA encodes an amino acid sequence having at least 75% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 80% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 85% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 95% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 99% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence of any one of SEQ ID NOs: 169-174.

[0402] In some embodiments, provided herein is a circRNA comprising a sequence having at least 80% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, provided herein is a circRNA comprising a sequence having at least 85% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, provided herein is a circRNA comprising a sequence having at least 90% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, provided herein is a circRNA comprising a sequence having at least 95% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, the circRNA comprises a sequence having 100% identity to any one of SEQ ID NOs: 175-233, 264 or 269.

[0403] In some embodiments, the untranslated region (UTR) comprises a 5' UTR, a 3' UTR or both a 5' UTR and a 3' UTR. In some embodiments, the UTR comprises a 5' UTR. In some embodiments, the UTR comprises a 3' UTR.

[0404] In some embodiments, the UTR comprises a CAA40 comprising SEQ ID NO: 42 or a pAC18 comprises SEQ ID NO: 43 (also known as SEQ ID NO: 265). In some embodiments, the UTR comprises any one of SEQ ID NO: 43-45, 46 or 47. In some embodiments, the UTR comprises SEQ ID NO: 46 (also known as SEQ ID NO: 272) or SEQ ID NO: 47. In some embodiments, the UTR comprises one or more microRNA binding sites. In some embodiments, the one or more microRNA binding sites is selected from any one of SEQ ID NOs: 27-40.

[0405] In some embodiments, the circRNA further comprises one or more microRNA binding sites in the 5' UTR, the 3' UTR, or both. In some embodiments, the microRNA binding site is a miR-122 site. In some embodiments, the miR-122 binding site is in the 3' UTR. In some embodiments, the miR-122 binding site is in the 5' UTR and 3' UTR.ATTORNEY DOCKET NO. ORB-019WO1

[0406] In some aspects, provided herein is a circular ribonucleic acid (circRNA) comprising: (a) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (b) an Internal Ribosome Entry Site (IRES) comprising a sequence at least 85% identical to any one of SEQ ID NO: 1-11, (c) 3’ and 5’ untranslated region(s) (UTR), (d) 3’ and 5’ exon fragments, and (e) a single microRNA binding site.

[0407] In some aspects, provided herein isa circular ribonucleic acid (circRNA) comprising in order: (a) a 5’ exon fragment and 5’ untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES) comprising a sequence at least 85% identical to any one of SEQ ID NO: 1-11, (c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (d) a 3’ UTR and a 3’ exon fragment, and (e) a single microRNA binding site within the 5’ UTR or the 3’ UTR.

[0408] In some aspects, provided herein isa circular ribonucleic acid (circRNA) comprising in order: (a) a 5’ exon fragment and 5’ untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES) comprising a sequence at least 85% identical to any one of SEQ ID NO: 1-11, (c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (d) a 3’ UTR and a 3’ exon fragment, and (e) a single microRNA binding site within the 5’ UTR or the 3’ UTR.Ribosome Recruiting Element

[0409] In some embodiments, the circRNA comprises a ribosome recruiting element. As used here, a “ribosome recruiting element” includes naturally occurring or engineered RNA sequences that directly or indirectly recruit the ribosome to initiate translation of RNA into protein. In some embodiments, a ribosome recruiting element is an internal ribosome entry site (IRES). In other embodiments, a ribosome recruiting element is a branched cap structure (e.g., qRNA) added to circular RNA that initiates translation by the ribosome. In some embodiments, a ribosome recruiting element is an IRES transacting factor (ITAF) recruiting element. In some embodiments, IRES and ITAFs recruit the ribosome. In some embodiments, the ITAF is specific to an IRES or group of related IRES elements. In some embodiments, the ITAF is general (for example, polypyrimidine tract-binding protein, PTB).ATTORNEY DOCKET NO. ORB-019WO1Internal Ribosome Entry Site

[0410] In some embodiments, the ribosome recruiting element is an Internal Ribosome Entry Site (IRES). In some embodiments, the circRNA comprises an Internal Ribosome Entry site (IRES). Because circRNA is generated by spliceosome-mediated head-to-tail joining of pre-mRNAs, it does not contain the 5' cap that is commonly known to be required for cap-dependent translation. Thus, circRNA translation utilizes alternate mechanisms to initiate cap-independent translation, such as the use of an internal ribosome entry site (IRES) sequence that is recognized by ribosomes. Introduction of an IRES on synthetically generated circRNAs is sufficient to initiate translation of encoded circRNA proteins, thereby suggesting that endogenous circRNAs harboring IRES sequence may have translation potential as they are exported to the cytoplasm. As used herein, the term “internal ribosome entry site” or “IRES” refers to an RNA sequence or structural element ranging in size from 10 nucleotides to 1,000 nucleotides or more which is capable of initiating translation of a polypeptide in the absence of a normal RNA cap structure. Inclusion of an IRES element in circRNA engages a eukaryotic ribosome for translation, i.e., the IRES element initiates cap-independent translation and protein synthesis.

[0411] Notably, IRES function is cell-type dependent, i.e., the efficiency of IRES-mediated translation can vary significantly depending on the specific cell type due to the presence of different cellular transactivating factors, under stress conditions or developmental stage of the cell. In the present disclosure, IRES elements for optimal T cell function in activated and resting T cells and with breadth of function in different donors are carefully selected through experimentation.

[0412] In some embodiments, the IRES is a synthetic IRES. In some embodiments, the IRES is a viral IRES. In some embodiments, the IRES is a naturally-occurring viral IRES. In some embodiments, the IRES is a recombinant variant of a naturally-occurring viral IRES. In some embodiments, the IRES is a fragment of the full-length sequence of a naturally-occurring viral IRES. In some embodiments, the IRES includes but is not limited to an IRES derived from Apodemus agrarius picomavirus, Canine kobuvirus, Picomavirales / Parabovirus, Caprine kobuvirus, Mouse kobuvirus, Bejaponia (bovine) kobuvirus, Human Enterovirus C99, Bovine rhinitis A virus, Kobuvirus aichi, Human coxsackievirus A21, Falcon picomavirus, Aichivirus B, Rattus tanezumi hunnivirus, Human coxsackievirus A5, Human poliovirus 2, Enterovirus B69, Saffold vims 3, Salivirus FHB, or Human Echovirus E3, or variants or fragments derived from these viruses.ATTORNEY DOCKET NO. ORB-019WO1

[0413] In some embodiments, the ribosome recruiting element is an Internal Ribosome Entry Site (IRES), wherein the IRES comprises a sequence selected from Table 1.

[0414] In some embodiments, the IRES comprises a sequence selected from Table 1. In some embodiments, the IRES comprises any one of the sequences in Table 1. In some embodiments, the IRES comprises any one of the sequences selected from SEQ ID NO: 1-11 of Table 1. In some embodiments, the IRES comprises a sequence selected from SEQ ID NO: 1-11 of Table 1. In some embodiments, the IRES comprises a sequence having at least 90% identity to any one of SEQ ID NO: 1-11. In some embodiments, the IRES comprises a sequence having at least 95% identity to any one of SEQ ID NO: 1-11. In some embodiments, the IRES comprises a sequence of any one of SEQ ID NO: 1-11.

[0415] In some embodiments, the IRES comprises a sequence having at least 90% identity to any one of SEQ ID NO: 1-6. In some embodiments, the IRES comprises a sequence having at least 95% identity to any one of SEQ ID NO: 1-6. In some embodiments, the IRES comprises a sequence of any one of SEQ ID NO: 1-6. In some embodiments, IRES comprises a sequence having 95% identity to SEQ ID NO: 1. In some embodiments, the IRES comprises the sequence of SEQ ID NO: 1. In some embodiments, the IRES comprises SEQ ID NO: 1.

[0416] In some embodiments, the IRES comprises a sequence having at least 90% identity to SEQ ID NO: 354. In some embodiments, the IRES comprises a sequence having at least 95% identity to SEQ ID NO: 354. In some embodiments, the IRES comprises a sequence having 100% identity to SEQ ID NO: 354. In some embodiments, the IRES comprises a sequence of SEQ ID NO: 354.

[0417] In some embodiments, the IRES is associated with higher CAR expression in T cells in comparison to a reference Coxsackievirus B3 IRES (SEQ ID NO: 12). In some embodiments, the IRES comprising any one of SEQ ID NO: 1-11 is associated with higher CAR expression in T cells in comparison to a reference Coxsackievirus B3 IRES (SEQ ID NO: 12). In some embodiments, the IRES comprising SEQ ID NO: 1 is associated with higher CAR expression in T cells in comparison to a reference Coxsackievirus B3 IRES (SEQ ID NO: 12).

[0418] Table 1. Exemplary Internal Ribosome Entry Site (IRES) RNA Sequences*IRES263ATTORNEY DOCKET NO. ORB-019WO1TTACTCAGCGTAACTACTCCGGGTTACGTGATGAAGAAGAGGCTACGGAGATTCTCGGGCTACGGCCCTGGAG CCACTCCGGCTCCTAAAGATTTAGAAGTTTGAGCACACCCGCCCACTAGGGCCCCCCATCCAGGGGGGCAACG GGCAAGCACTTCTGTTTCCCCGGTATGATCTGATAGGCTGTAACCACGGCTGAAACAGAGATTATCGTTATCC GCTTCACTACTTCGAGAAGCCTAGTAATGATGGGTGAAATTGAATCCGTTGATCCGGTGTCTCCCCCACACCA GAAACTCATGATGAGGGTTGCCATCCCGGCTACGGCGACGTAGCGGGCATCCCTGCGCTGGCATGAGGCCTCT TAGGAGGACGGATGATATGGATCTTGTCGTGAAGAGCCTATTGAGCTAGTGTCGACTCCTCCGCCCCCGTGAA TGCGGCTAATCCTAACCCCGGAGCAGGTGGGTCCAATCCAGGGCCTGGCCTGTCGTAATGCGTAAGTCTGGGA CGGAACCGACTACTTTCGGGAAGGCGTGTTTCCATTTGTTCATTATTTGTGTGTTTATGGTGACAACTCTGGG T AAAC GTTCTATTGCGTTTATT GAGAGAT TGC C AAC AAT T GAAC AAAC GAGAAC TACCTGTTTTAT T AAAT T T ACACAGAGAAGAATTACA ( S EQ ID NO: 1 )IRES318 _ TTTCCCCTGTTCGTAACTAAGTGTGTGCCCAATCTCCTCACTCCTGCTGGCTTCACCGACCGGCAGTGTCCAA AATGCTAGGTGAATCCCCTCCCTTTCCTCTGGGCTTCTGCCCAGCTTCCTCCCCCCAGCCTGACGTGACACAG GCTGTGCAAAGACCCCGCGAAAGCTGCCAAAAGTGGCAATTGTGGGTCCCCCCTTTGTAAAGGCGTCGAGTCT TTCTCCCTCAAGGCTAGACCCGTCAGTGAATTCTGTCGGGCAACTAGTGACGCCACTGCACGCCTCTGACCTC GGCCGCGGAGTGCTGCCCCCCAAGTCGTGCCCCTGACCACAAGTTGTGCTGTCTGGCAAACATTGTCTGTGAG AATGTTCCGCTGTGGCTGCCAAGCCTGGCAACAGGCTGCCCCAGTGTGCGTAGTTCTCATCCAGACTTCGGTC TGGCAACTTGCTGTTAAGACACGGCGTAAGGGGCGTGTGCCAACGCCCTGGAACGAGTGTCCACTCTAATACC CCGAGGAATGCTACGCAGGTACCCCTGGTTCGCCAGGGATCTGAGCGTAGGCTAATTGTCTAAGGGTATTTTC ATTTCCCATTCTTTCTTTCTTGTTCATA ( SEQ ID NO: 2 )IRES303 _ TGAACCGTTACGCACCACTCAGTTGGTGTTTGGTGGCACCAATGATGGAACAAAAGGCTACACCACTTGGGCT ACGGCCCGCGCCACCTTGTGGCGCAAAGACATTAGAAGAATAGCATACCGCCCACTAGGGCCCTGCAGCCAGC AGGGTAACGGGCAAGCACTTCTGTCTCCCCGGTAGAACGGTATAGGCTGTACCCACGGCCGAAAACTGAACTA TCGTTACCCGACTCCGTACTTCGCAAAGCTTAGTAGGAAACTGGAAAGTTCGAGTTATTGACCCGGAGTGTTC CCCCCACTCCAGAAACGCGTGATGAGGGTTGCCACCCCGACCATGGCGACATGGTGGGCATCCCTGCGCTGGC ACGCGGCCTCTAAGAGGATAACTCGCTCCTACTGGTAACCGAAGAGCCCCGTGAGCTACGGTTTATTCCTCCG CCTCCCTGAATGCGGCTAATCCTAACCCATGAGCAGTTGCCATAGATCCATATGGTGGACTGTCGTAACGCGT AAGTTGTGGGCGGAACCGACTACTTTGGGATGGCGTGTTTCCTTGTTTTCTCCATTTGTTGTTGTATGGTGAC AAGTTATAGATCTCGATCTATAGCGTTTCTTGAGAGATTTCCAAACATTTATTCAAGTCGTACAATTCTTGTG TTTAAGCAGTACAGTGTAAGG ( SEQ ID NO: 3 ) _IRES 124 _ TTTAAGTGTTGTGCCCAATCTCTTGACTCCTGCTGGAACCACCGACCAGTAGTGTCCAAAATGCCAGGTGGAA AATCCTCCCTTCCCCTCTGGGCTTCATGCCCGGCATCCTCCCCCCAGCCTGACGTGCCACAGGCTGTGCAAAG ACCCCGCGAAAGCTGCCAAAAGTGGCAATTGTGGGTCCCCCCTTTGTCAAGGCGTCGAGTCTTTCTCCCTTAA GGCTAGTCCTGTCAGTGAACTCTGTCGGGCAACTAGTGACGCCACTGCATGCCTCCGACCTCGGCCGCGGAGT GCTGCCCCCCAAGTCATGCCCCTGACCACAAGTTGTGCTGTCTGGCAAACATTGTCTGTGAGAATGTTCCGCT GTGGCTGCCAAGCCTGGTAACAGGCTGCCCCAGTGTGCGTAATTCTCATCCAGACTTCGGTCTGGCAACTTGC TGTTAAGACATGGCGTAAGGGGCGTGTGCCAACGCCCTGGAACGAGTGTCCACTCTAATACCCCGAGGAATGC TACGCAGGTACCCCTGGCTCGCCAGGGATCTGAGCGTAGGCTAATTGTCTAAGGGTATTTTCATTTCCCACCC TCTTCTTCTTGTTCATA ( SEQ ID NO: 4 )IRES302 _ TTTGCTCAGCGTAACTTCTCCGGGTTACGTGGAGACCAAAAGGCTACGGAGACTCGGGCTACGGCCCTGGAGC ACCTAGGTGCTCCTAAAGACGTTAGAAGTTGTACAAACTCGCCCAATAGGGCCCCCCAACCAGGGGGGTAGCG GGCAAGCACTTCTGTTTCCCCGGTATGATCTCATAGGCTGTACCCACGGCTGAAAGAGAGATTATCGTTACCC GCCTCACTACTTCGAGAAGCCCAGTAATGGTTCATGAAGTTGATCTCGTTGACCCGGTGTTTCCCCCACACCA GAAACCTGTGATGGGGGTGGTCATCCCGGTCATGGCGACATGACGGACCTCCCCGCGCCGGCACAGGGCCTCT TCGGAGGACGAGTGACATGGATTCAACCGTGAAGAGCCTATTGAGCTAGTGTTGATTCCTCCGCCCCCGTGAA TGCGGCTAATCCCAACTCCGGAGCAGGCGGGCCCAAACCAGGGTCTGGCCTGTCGTAACGCGAAAGTCTGGAG CGGAACCGACTACTTTCGGGAAGGCGTGTTTCCTTTTGTTCCTTTTATCAAGTTTTATGGTGACAACTCCTGG TAGACGTTTTATTGCGTTTATT GAGAGAT T T C GAAC AAT T GAAC AGAC T AGAAC CACTTGTTTTAT C AAAC C C TCACAGAATAAGATAACA ( SEQ ID NO: 5 )IRES290ATTORNEY DOCKET NO. ORB-019WO1CCCCCTTTTCCCTAACCCCAACCGCCGTATCTGGTGGCGGCAAGACACACGGGTCTTTCCCTCTAAAGCACAA ATGTGTGTGTGTCCCAGGTCCTCCTGCGCTCGGTGCGGGAGTGCTCCCACCCAACTGTTGTAAGCCTGTCCAA CGTGTCGTCCTGGCAAGACTATGACGTCGCATGTTCCGCTGCGGATGCCGACCGGGTAACCGGTTCCCCAGTG TGTGTAGTGCGATCTTCCAGGTTCCCCTGGTTGGCGTTGTCCAGAAACTGCTTCAGGTAAGTGGGGTGTGCCC AATCCCTGCAAAGGTTGATTCTCTCACCACCTTAGGAATGCTCCGGAGGTACCCCAGCAACAGCTGGGATCTG ACCGGAGGCTAATTGTCTACGGGTGGTGTTTCCTTTTTCTTTTTCCACAACTTCATTGCTGACAACTCACTGA CTACCCATTTGCTCTCTTGTGCCTTTCTGCTCTGGTTCAAGCTCCTTGATTGTTTTTGATTGCTTTCTATTGC TTTCTTCCACTCAACCCTTGCTCAGTTCAAAGTC ( SEQ ID NO: 6) _IRES361 _ TTCAAGTGTGTGTGCTCGTAATCTTGACTCCTGCCGGAATGCCGCCCGGTTCAGTGAACAAACAGCTAGGCAA GTCCCTCCCTTCCCCTGTGGTCGGTTCTCACCGGCCACCATCCCTCCCCCAGCCTGACGTGTTACAGGCTGTG CAAAGCCCCCGCGAAAGCTGCTCACGTGGCAATTGTGGGTCCCCCCTTTGTCAAGACACCGAGTCTTTCTCCC TTAAGGCTAGCCCGGTCCCACGAACGTGGAACTGGCAACTAGTGGTGTCACTACACGCCTCCGACCTCGGACG CGGAGTGCTGTTCCCCAAGCTGTAACCCTGACCCAAGACTGTGCTGCCTGGCAAGCACCGTCTGGGAAGATGT TCCGCTGTGGCTGCCAAACCTGGTAACAGGTGCCCCAGTGTGTGTAGTCTTCCTCCAGTCTCCGGACTGGCAG TCTTGTGTAAAGATGCAGTGTAAGGTTCAAGTGCCAAATCCCTGGAAGGAGTGACCCTCTACTGCCCTAGGAA TGCTGTGCAGGTACCCCCAACTTCGGTTGGGGATCTGAGCACAGGCTAATTGTCTACGGGTAGTTTCATTTCC CATCCTCTCTTTTTTGGCATC ( SEQ ID NO: 7 ) _IRES289 _ GCCCGTCCCCCTCACCCTCTTTTCCGGTGGCCACGCCCGGGCCACCGATACTTCCCTTCACTCCCTCGGGACT GTTGGGGAGGAACACAACAGGGCTCCCCTGTATTTCCTCTTCCCATTCCCCCTTTCCTAACCCCAACCGCCGT ATCTGGTGGCGGTAAGACACACGGGTCTTTCCCTCTAAAGCACAATTGTGTGTGTGTCCCAGGTCCTCCTGCG TTCGGTGCGGGAGTGCTCCCACCCAACTGTTGTAAGCCTGTCCAACGTGTCGTCCTGGCAAGACTATGACGTC GCATGTTCCGCTGTGGATGCCGACCGGGTAACCGGTTCCCCAGTGTGTGTAGTGCGATCTTCCAGGTTCTCCT GGTTGGCGTTGTCCAGAAACTGCTTCGGGTAAGTGGGGTGTGCCCAATCCCTACAAGGGTTGATTCTTTCACC ACCTTAGGAATGCTCCGGAGGTACCCCAGCAACAGCTGGGATCTGACCGGAGGCTAATTGTCTACGGGTGGTG TTTCCATTTTCTTTTTCACACAACTTCATTGCTGACAACTCACTGACTAATCACTTGCTCTCTTGTGCCTTTC TGCTCTGGTTCAAGTTCCTTGATTGTTTGTTTGATTGCTTTTCACTGCTTTCTTCCCACAATCCTTGCTCAGT TCAAAGTC ( SEQ ID NO: 8 ) _IRES644 _ TTCCCCCTTTTCCCAACCCCAACCGCCGTATCTGGTGGCGGCAAGACACACGGGTCTTTCCCTCTAAAGCACA ATTGTGTGTGTGTCCCAGGTCCTCCTGCGTACGGTGCGGGAGTGCTCCCACCCAACTGTTGTAAGCCTGTCCA ACGCGTCGTCCTGGCAAGACTATGACGTCGCATGTTCCGCTGCGGATGCCGACCGGGTAACCGGTTCCCCAGT GTGTGTAGTGCGATCTTCCAGGTCCTCCTGGTTGGCGTTGTCCAGAAACTGCTTCAGGTAAGTGGGGTGTGCC CAATCCCTACAAAGGTTGATTCTTTCACCACCTTAGGAATGCTCCGGAGGTACCCCAGCAACAGCTGGGATCT GACCGGAGGCTAATTGTCTACGGGTGGTGTTTCCTTTTTCTTTTCACACAACTCTACTGCTGACAACTCACTG ACTATCCACTTGCTCTCTTGTGCCTTTCTGCTCTGGTTCAAGTTCCTTGATTGTTTTTGACTGCTTTTCACTG CTTTTCTTCTCACAATCCTTGCTCAGTTCAAAGTC ( SEQ ID NO: 9 ) _IRES596 _ GTAACTTCAAGTGTGTGTGCTCGTAATCTTGACTCCTGCCGGAATGCCGCCCGGTTCAGTGAACAAACAGCTA GGCAAGTCCCTCCCTTCCCCTGTGGTCGGTTCTCACCGGCCACCATCCCTCCCCCAGCCTGACGTGTTACAGG CTGTGCAAAGCCCCCGCGAAAGCTGCTCACGTGGCAATTGTGGGTCCCCCCTTTGTCAAGACACCGAGTCTTT CTCCCTTAAGGCTAGCCCGGTCCCACGAACGTGGAACTGGCAACTAGTGGTGTCACTACACGCCTCCGACCTC GGACGCGGAGTGCTGTTCCCCAAGCTGTAACCCTGACCCAAGACTGTGCTGCCTGGCAAGCACCGTCTGGGAA GATGTTCCGCTGTGGCTGCCAAACCTGGTAACAGGTGCCCCAGTGTGTGTAGTCTTCCTCCAGTCTCCGGACT GGCAGTCTTGTGTAAAGATGCAGTGTAAGGTTCAAGTGCCAAATCCCTGGAAGGAGTGACCCTCTACTGCCCT AGGAATGCTGTGCAGGTACCCCCAACTTCGGTTGGGGATCTGAGCACAGGCTAATTGTCTACGGGTAGTTTCA TTTCCCATCCTCTCTTTTTTGGCATCATGGCGAAC ( SEQ ID NO: 10 ) _IRES665 ~ CCCCCTCACCCTCTTTTCCGGCGGCGCATGTTCGCGTCGTCGTAAGTCTGGATTCCCAAGGCCCACTCGGTTC AACTTCGGTTTCCGGACAAATACAAAGAACCTCAGTCCTCTTGGTACTTTCTCGCCTGAGTCTACAAAGCGAG AAACCTGCCCCTCTAACGCCAGACGAGCGGCATAAAACTCGAACTTCTGGCACGTTCCACCACCCCTTCTCCT ATCCCAACCCCCATTGCGCTCTCAAGGTCGCGCTTTTCCGAGACTAGCTCGGATTCAAAAAGTTCCTGGCACC CTTTACCCCTTCAGGCCCTTAAGGTAGGAACTGACCTTGTGCTGTGATCTCGGTGCGGAAGTGCTACTGCGTA GTGATTGTAAGATCCTTTTGTGGTTCTGCCCTGGCAAGGCTACAGAGTGCTGTGATCCGCTGCGGATGCCATC CTGGTAACAGGACCCCCAGTGTGCGCAACAGTATGTTCACGGTCTTCCGTGTTCACCACATTCGGAACACTGC TTTCGTGAAACAGTGTGTGTCCAATCCCTGTGATCAGTATCAACCACACCACCTAGGAATGCTAGGAAGGTAC CCCGGTTCGCCGGGATCTGATCCTAGGCTAATTGTCTACGGTGGTGCTCCTTTTTATTTTCCACTTCAACTCA CTGGTTACAACTGCTTGATTTCTGTGTTTGCTGCTTTTCTCTGCTCTCACTGCCATTCTCAAGTGTTCACACTGTCCAAGCTCCTTTGGTTGTTCGCTTCCACTTGCCACTGTCAACTCTTGTC ( SEQ ID NO: 11 )ATTORNEY DOCKET NO. ORB-019WO1Coxsackievirus B3 (CVB3) Reference IRES _ TTAAAACAGCCTGTGGGTTGATCCCACCCACAGGCCCATTGGGCGCTAGCACTCTGGTATCACGGTACCTTTG TGCGCCTGTTTTATACCCCCTCCCCCAACTGTAACTTAGAAGTAACACACACCGATCAACAGTCAGCGTGGCA CACCAGCCACGTTTTGATCAAGCACTTCTGTTACCCCGGACTGAGTATCAATAGACTGCTCACGCGGTTGAAG GAGAAAGCGTTCGTTATCCGGCCAACTACTTCGAAAAACCTAGTAACACCGTGGAAGTTGCAGAGTGTTTCGC TCAGCACTACCCCAGTGTAGATCAGGTCGATGAGTCACCGCATTCCCCACGGGCGACCGTGGCGGTGGCTGCG TTGGCGGCCTGCCCATGGGGAAACCCATGGGACGCTCTAATACAGACATGGTGCGAAGAGTCTATTGAGCTAG TTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCTAACTGCGGAGCACACACCCTCAAGCCAGAGGGCAGTG TGTCGTAACGGGCAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCATTTTATTCCTATACTGGC TGCTTATGGTGACAATTGAGAGATCGTTACCATATAGCTATTGGATTGGCCATCCGGTGACTAATAGAGCTAT TATATATCCCTTTGTTGGGTTTATACCACTTAGCTTGAAAGAGGTTAAAACATTACAATTCATTGTTAAGTTG AATACAGCAAA ( SEQ ID NO: 12 )IRES004 Exemplary Control scrambled IRES _ TGTAGGTAGAAGGTATAAACGACCAGCCACACACTTCCTACCCAAAGCGCCAATAACAGGAGCCCGGATTGGT CTGGCACGCAAAGGAGTAGACATCCCCCAATGTTTTCCGAACCAAGCGGCAGCAGCGTCGGTTAGGATCCTCA TAATTCTGATCGAAGGACAGTACACGCGCTACTCAGGGTCTTCTATGATGATTACGATTTACTTACGACGGTG TGATGTAACGCCCTCGGCGACTTACTTGGACCTGTTCCCGAATTGAGAATGTGTAGGCCAGCGACCAATCGCC ACCACGAGGCCTAGACTGATCAACTTCAACTGTGAGCGACTCGCATCTGAGGAGTTGACCTAGCTAACACTTC CCATCCACTCTAATTTGGCTAACAAGGCTGCCACGTGATGATCCTTCTTCTAGGGTGTCTACCTGAGTAAGGG TACCCTTCTCGGAATAACGATAAGCGAAGGAGGACATGAACCTAGGCTGGATGTGATTAGCTCCGTAAGCTGC CAACCTGGAGCGCGTCAGATTCGGCCAACGGCAGATTGGGCTGATCGGGAGTGAGGAATCCCTGCTTACGCTC ACTGGGTGATTCCACGTAAAATGTCCAGTTATTATCTGTGTCCTCTTAACACATTCTTCTGTTTAAGCTTTTC GCCCGTTCTATAAACCTTGTCTGACTAAGTCCTTAGTCCGCTGCCATTACCGATTGGGTTGCGTAGGCAGCTT CTATTGAAACG ( SEQ ID NO: 263 )IRES642 GGCCTCGGCCCCCTCACCCTCTTTTCCGGTGGCCACGCCCGGGCCACCGATACTTCCCTTCACTCCTTCGGGA CTGTTGGGGAGGAACACAACAGGGCTCCCCTGTTTTCCCATTCCTTCCCCCTTTTCCCAACCCCAACCGCCGT ATCTGGTGGCGGCAAGACACACGGGTCTTTCCCTCTAAAGCACAATTGTGTGTGTGTCCCAGGTCCTCCTGCG TACGGTGCGGGAGTGCTCCCACCCAACTGTTGTAAGCCTGTCCAACGCGTCGTCCTGGCAAGACTATGACGTC GCATGTTCCGCTGCGGATGCCGACCGGGTAACCGGTTCCCCAGTGTGTGTAGTGCGATCTTCCAGGTCCTCCT GGTTGGCGTTGTCCAGAAACTGCTTCAGGTAAGTGGGGTGTGCCCAATCCCTACAAAGGTTGATTCTTTCACC ACCTTAGGAATGCTCCGGAGGTACCCCAGCAACAGCTGGGATCTGACCGGAGGCTAATTGTCTACGGGTGGTG TTTCCTTTTTCTTTTCACACAACTCTACTGCTGACAACTCACTGACTATCCACTTGCTCTCTTGTGCCTTTCT GCTCTGGTTCAAGTTCCTTGATTGTTTTTGACTGCTTTTCACTGCTTTTCTTCTCACAATCCTTGCTCAGTTC AAAGTC ( SEQ ID NO: 318 ) _IRES653 CCCCCTTTTCCCAACCCCAACCGCCGTATCTGGTGGCGGCAAGACACACGGGTCTTTCCCTCTAAAGCACAAT TGTGTGTGTGTCCCAGGTCCTCCTGCGTACGGTGCGGGAGTGCTCCCACCCAACTGTTGTAAGCCTGTCCAAC GCGTCGTCCTGGCAAGACTATGACGTCGCATGTTCCGCTGCGGATGCCGACCGGGTAACCGGTTCCCCAGTGT GTGTAGTGCGATCTTCCAGGTCCTCCTGGTTGGCGTTGTCCAGAAACTGCTTCAGGTAAGTGGGGTGTGCCCA ATCCCTACAAAGGTTGATTCTTTCACCACCTTAGGAATGCTCCGGAGGTACCCCAGCAACAGCTGGGATCTGA CCGGAGGCTAATTGTCTACGGGTGGTGTTTCCTTTTTCTTTTCACACAACTCTACTGCTGACAACTCACTGAC TATCCACTTGCTCTCTTGTGCCTTTCTGCTCTGGTTCAAGTTCCTTGATTGTTTTTGACTGCTTTTCACTGCT TTTCTTCTCACAATCCTTGCTCAGTTCAAAGTC (SEQ ID NO: 319 ) _IRES647 GCCTCGGCCCCCTCACCCTCTTTTCCGGTGGCCACGCCCGGGCCACCGATACTTCCCTTCACTCCTTCGGGAC TGTTGGGGAGGAACACAACAGGGCTCCCCTGTTTTCCCATTCCTTCCCCCTTTTCCCAACCCCAACCGCCGTA TCTGGTGGCGGCAAGACACACGGGTCTTTCCCTCTAAAGCACAATTGTGTGTGTGTCCCAGGTCCTCCTGCGT ACGGTGCGGGAGTGCTCCCACCCAACTGTTGTAAGCCTGTCCAACGCGTCGTCCTGGCAAGACTATGACGTCG CATGTTCCGCTGCGGATGCCGACCGGGTAACCGGTTCCCCAGTGTGTGTAGTGCGATCTTCCAGGTCCTCCTG GTTGGCGTTGTCCAGAAACTGCTTCAGGTAAGTGGGGTGTGCCCAATCCCTACAAAGGTTGATTCTTTCACCA CCTTAGGAATGCTCCGGAGGTACCCCAGCAACAGCTGGGATCTGACCGGAGGCTAATTGTCTACGGGTGGTGT TTCCTTTTTCTTTTCACACAACTCTACTGCTGACAACTCACTGACTATCCACTTGCTCTCTTGTGCCTTTCTG CTCTGGTTCAAGTTCCTTGATTGTTTTTGACTGCTTTTCACTGCTTTTCTTCTCACAATCCTTGCTCAGTTCAAAGTC ( SEQ ID NO: 320 )ATTORNEY DOCKET NO. ORB-019WO1IRES 147 TTAAAACAGCTCTGGGGTTGTTCCCACCCCAGAGGCCCACGTGGCGGCTAGTACTCTGGTATCACGGTACCTT TGTACGCCTGTTTTATATCCCTTCCCCCGTAACTTAGAAGCTTTGTACCAACAAGTTCAATAGAAGGGGTACA AACCAGTACCACCACGAACAAGCACTTCTGTTTCCCCGGTGCGATGACATAGACTGTACCCACGGTCAAAAGT CACCAATCCGTTACCCGCTTTGGTACTTCGAGACGCCTAGTATTGCCTTGGAATTCTCGATGCGTTGCGCTCA ACACTCGACCCTGGAGTGTAGCTTAGGCTGATGAGTCTGGGCATTCCCCACCGGTGACGGTGGCCCAGGCTGC GTTGGCGGCCTACCCATAGCTATTGCTATGGGATGCTAGAGTTGAACAAGGTGTGAAGAGCCAATTGAGCTAC TAAAGAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACCACGGATCAAGTGCGTGCAACCCAGCACGTAGCT TGTCGTAATGCGCAAGTCTGTGGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATTTTTATCATGGC TGCTTATGGTGACAATCTTGGATTGTTATCATATAGCGATTGGATTGGCCATCCGGTGAGAATAAAAGATATT GTTTATCTGTTTGTGGCTTTTACTCCATTAAACAAAAACACACTTTCCTTAATTAAAGTGGTACTATTGATCA GATCAGCATACCACA ( SEQ ID NO: 321 ) _IRES411 GTGTGTGTGCTCGTAATCTTGACTCCTGCCGGAATGCCGCCCGGTTCAGTGAACAAACAGCTAGGCAAGTCCC TCCCTTCCCCTGTGGTCGGTTCTCACCGGCCACCATCCCTCCCCCAGCCTGACGTGTTACAGGCTGTGCAAAG CCCCCGCGAAAGCTGCTCACGTGGCAATTGTGGGTCCCCCCTTTGTCAAGACACCGAGTCTTTCTCCCTTAAG GCTAGCCCGGTCCCACGAACGTGGAACTGGCAACTAGTGGTGTCACTACACGCCTCCGACCTCGGACGCGGAG TGCTGTTCCCCAAGCTGTAACCCTGACCCAAGACTGTGCTGCCTGGCAAGCACCGTCTGGGAAGATGTTCCGC TGTGGCTGCCAAACCTGGTAACAGGTGCCCCAGTGTGTGTAGTCTTCCTCCAGTCTCCGGACTGGCAGTCTTG TGTAAAGATGCAGTGTAAGGTTCAAGTGCCAAATCCCTGGAAGGAGTGACCCTCTACTGCCCTAGGAATGCTG TGCAGGTACCCCCAACTTCGGTTGGGGATCTGAGCACAGGCTAATTGTCTACGGGTAGTTTCATTTCCCATCC TCTCTTTTTTGGCATC ( SEQ ID NO: 322 ) _IRES 160 TTTTGCGGCTCTGCCGCCGTTCGGGTTTTACCTGTTTTCACAGAGCAAAACAGGACCTCTAGTTTCGTGCTTA AACGAGATCATGCTCGAACTAGAACTATAACGCTGGTCACTGGACCCGTGCCGCGCCTTGCGGATCTTTGCGG GAATGGTGGCTAGTGGGCTGTGGAAGTGACTCTAACCACACGCCCCTCAAGTGTGGGAAAACACGAACTGGTG TAGCGACGACGATAGGCCTTGGGACACCCTCTCCAGTGATGGAGACCCAAGGGGCCAAAAGCCACGCCTTGTG CCCTGTCGTTCACAACCCCAGTGCAGTTCGTGCCAGTACCTGCTTTTGGGAAGTGTGCTTTGGACAGCTGAAA ACAGTCCTAGTGGGAGACTAAGGATGCCCAGGAGGTACCCGGAGGTAACAAGTGACACTCTGGATCTGACTTG GGGAGAGCGGGTCTGCTTTACAGACGCCACTCTTTAAAAAACTTCTATGTCTCGTCAGGCACCGGAGGCCGGG CCTTTTCCTTTAAAACAATACACTTTATGAAGACAACAATA (SEQ ID NO: 323 ) _IRES217 CACCCATACACCCCCACCCCCTTTTCTGTAACTCAAGTATGTGTGCTCGTAATCTTGACTCCCACGGAATGGA TCGATCCGCTGGAGAACAAACTGCTAGATCCACATCCTCCCTCCCCTTGGGAGGACCTCGGTCCTCCCACATC CTCCCTCCAGCCTGACGTATCACAGGCTGTGTGAAGCCCCCGCGAAAGCTGCTCACGTGGCAATTGTGGGTCC CCCCTTCATCAAGACACCAGGTCTTTCCTCCTTAAGGCTAGCCCCGATGTGTGAATTCACATTGGGCAACTAG TGGTGTCACTGTGCGCTCCCAATCTCGGCCGCGGAGTGCTGTTCCCCAAGCCAAACCCCTGGCCCTTCACTAT GTGCCTGGCAAGCATATCTGAGAAGGTGTTCCGCTGTGGCTGCCAGCCTGGTAACAGGTGCCCCAGTGTGCGT AACCTTCTTCCGTCTCCGGACGGTAGTGATTGGTTAAGATTTGGTGTAAGGTTCATGTGCCAACGCCCTGTGC GGGATGAAACCTCTACTGCCCTAGGAATGCCAGGCAGGTACCCCACCTTCGGGTGGGATCTGAGCCTGGGCTA ATTGTCTACGGGTAGTTTCATTTCCAATTCTTTTATGCTGGAGTC (SEQ ID NO: 324 ) _ IRES288 CACCCACTTTTCCGGCGGCGCACGTTCGCGTCGTCGTAAGTCTGGACTCCCAAGGCCAACTCGGTTCAACTTC GGTTTCCGGACAAATACAAGAACCTCAGTCCTCTTGGTACTTTCTCGCCTGAGTCAACAAAGCGAGAAACCTG CCCCTCTAACGCCAGACGAGCGGCATGAAACTTGAACTTCTGGCATGTTCCACTTTCCCTCTCCCTCACCCCA CCCCACCGCGCTCTCAAGGTCGCGCTCTTCCGGGACTAGCCCGGTTTTAAAAGTTCCTGGCACCCTTTGCCCC TCTAGGCCCTTAAGGTAGGAACTGACCTTGTGCTGTGATCTCGGTGCGGAAGTGCTACTGCGTAGTGATTGTA AGATCCCTTTGTGGTTCTGCCCTGGCAAGGCTACAGAGTACTGTGATCCGCTGCGGACGCCATCCTGGTAACA GGACCCCCAGTGTGCGCAACAGTATGTCCACGGTCTTCCGTGTTCACCACATTCGGAACACTGCTCTCGTGAA ACAGTGTGTGTCCAATCCCTGTGACCAGTATTAACCACACCACCTAGGAATGCTAGGAAGGTACCCCGGTTCG CCGGGATCTGATCCTAGGCTAATTGTCTACAGTGGTGCTCCTTTTATTTTCCCCTCACTTCATTGACTACAAC TGCTTGATTTCCGTGTTTGCTGCTCTTCTTAGCTCTCACTGCCATTCTCAAGTGTTCACACTGTCCAAGCTCCTTTGGTTGTTTGCCTCCACTTGCCACTGTCAACTCTTGTC (SEQ ID NO: 325 )ATTORNEY DOCKET NO. ORB-019WO1IRES655 TTAAAACAGCTCTGGGGTTGCTCCCACCCCAGAGGCCCACGTGGCGGCTAGTAATCTGGTATCAGGTACCTTT GTACGCCTGTTTTATATCCCTTCCCCCGTAACTTTAGAAGCTTATCAAAAGTTCAATAGCAGGGGTACAAGCC AGTACCTCTACGAACAAGCACTTCTGTTTCCCCGGTGAAATCATATAGACTGTACCCACGGTCAAAAGTGATT GATCCGTTATCCGCTTGAGTACTTCGAGAAGCCTAGTATCGCCTTGGAATCTTCGACGCGTTGCGCTCAACAC TCTGCCCCGAGTGTAGCTTAGGCTGATGAGTCTGGGCACTCCCCACCGGCGACGGTGGCCCAGGCTGCGTTGG CGGCCTACCCATGGCTGATGCCGTGGGACGCTAGTTGTGAACAAGGTGTGAAGAGCCTATTGAGCTACTCAAG AGTCCTCCGGCCCCTGAATGCGGCTAATCCTAACCACGGAGCAATCGCTCACGACCCAGTGAGTAGGTTGTCG TAATGCGTAAGTCTGTGGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCCTTTATATTCATACTGGCTGCTT ATGGTGACAATTTACGAATTGTTACCATATAGCTATTGGATTGGCCACCCAGTGCTGTGCAATATATTTGAGT GCTTCTTTCATAGGTGTTACCAACATCACATTTAAACCACAATAGTCAGTGCAA ( SEQ ID NO: 326) IRES726 CCTAGCCCCTTTCTGTAACGGTCTGTGTGAGACCTAAAACCACACATAATATCGTGCCTGATGGTTAACGCTT GTTAGCCGTCTGTAAACTAAGAACCTACCAACACACTATGCACCTCTTGGCGAAGCCGCTTGGAATAAGTTGA GAGGAATTATGCATGCTAGTTGTGTTTGTTTACAACTAATTGTTCTAATCCAAGTGAAGCTCTTCGCTTGGGG CGGCACGACACTTGCCGTAATTCTTCTACCGTCCCTCCACACCTTGTGGATGAAGGGCCGGATGTGTGGCCTC TGGCTAACCCCTCTCTCTGGGGTGATGCTACTGGATGTTTTACTCCTAGACCAAATCACATGAACTCCTCTTG ATCCACTTCGGTGGGGCTATGAGCCTGCGGATTAATAGCTGGCGACAGCTACCCCAGGGGCCAAAAGCCACGG TGTTAGCAGCACCCTCATAGTCTGATGCCCAAGGGCTGATGTTGGGAGCTAGTAGTGTGTGTCTGGCCTATGT TTAGGACTTCTGGCCAAGCGCAGAGGAGTGGGGCTGAAGGATGCCCAGAAGGTACCCGTAGGTAACCTTAAGA GACTATGGATCTGATCTGGGGCCCCCTCACGTGGCTTTACCACGTGTTGGGGGTTAAAAAACGTCTAGGCCCC ACCAGCCCACGGGAGTGGGCTTTCCCTTAAAAAGCCCAACAATATTTATGGTGACAATTCACTGTTTCTTCTT TGCAATTTTTGTATTCTTGGACTCCTTATTTTTTGTTTGCTTGATTTTAGTGGACATTCAGATTCAAATACAA AATGGCAGGA ( SEQ ID NO: 327 ) _IRES639 ATTCCCAAGGCCCACTCGGTTCAACTTCGGTTTCCGGACAAATACAAAGAACCTCAGTCCTCTTGGTACTTTC TCGCCTGAGTCTACAAAGCGAGAAACCTGCCCCTCTAACGCCAGACGAGCGGCATAAAACTCGAACTTCTGGC ACGTTCCACCACCCCTTCTCCTATCCCAACCCCCATTGCGCTCTCAAGGTCGCGCTTTTCCGAGACTAGCTCG GATTCAAAAAGTTCCTGGCACCCTTTACCCCTTCAGGCCCTTAAGGTAGGAACTGACCTTGTGCTGTGATCTC GGTGCGGAAGTGCTACTGCGTAGTGATTGTAAGATCCTTTTGTGGTTCTGCCCTGGCAAGGCTACAGAGTGCT GTGATCCGCTGCGGATGCCATCCTGGTAACAGGACCCCCAGTGTGCGCAACAGTATGTTCACGGTCTTCCGTG TTCACCACATTCGGAACACTGCTTTCGTGAAACAGTGTGTGTCCAATCCCTGTGATCAGTATCAACCACACCA CCTAGGAATGCTAGGAAGGTACCCCGGTTCGCCGGGATCTGATCCTAGGCTAATTGTCTACGGTGGTGCTCCT TTTTATTTTCCACTTCAACTCACTGGTTACAACTGCTTGATTTCTGTGTTTGCTGCTTTTCTCTGCTCTCACT GCCATTCTCAAGTGTTCACACTGTCCAAGCTCCTTTGGTTGTTCGCTTCCACTTGCCACTGTCAACTCTTGTC( SEQ ID NO: 328 ) _IRES307 CCCCCCTCCCCCCCTTCCCTTCCCTTTGCAACGCAACAATTGTAAGTGCCCTCACCTGTCAATTGGGACCACC ACTTTCAGTGACCCCATGCGAAGTGCTGAGAGAAAGGAAGCTTTCTTACCCTTCATTTGTGAACCCACTGGTC TAAGCCGCTTGGAATACGATGAGTGGAAAAGTTCATTCTTAATGGAGTGAAACATGCTTAAATTTCCAGCTCG TGCTGGTCTTTCCAGTACGGGGCGGCCCTGTCTGGCCGTAATTCTTCAGAGTGTCACGCCACACTTGTGGATC TCACGTGCCACATGACAGCGCTACAGCTGGAACTGGGTGCTTGGTGCCCATGGAGTAACAGCGAAAAGTGTTA GATCAAGCCTTGCTTGGGCTATGAGCCTGCGGAACAACAACTGGTAACAGTTGCCTCAGGGGCCGAAAGCCAC GGTGTTAACAGCACCCTCATAGTTTGATCCACCTCAGGGTGGTGATGTTTAGCAGTTAGTAGTTGCCAATCTG TGTTCACTGAAATCTCGGCATACCGTGTAGTGTACAGGGGTGAAGGATGCCCAGAAGGTACCCGTAGGTAACC TTAAGAGACTATGGATCTGATCTGGGGCCTTGTCCGGAGTGCTTTACACACGGCTCAAGGTTAAAAAACGTCT AGCCCCACAGAGCCCGAGGGATTCGGGTTTTCCCTTTAAAAACCCGACTAGAGCTTATGGTGACAATTATTGC TGTTCAGACGAACAGTGTAATTGTTGTCTATTCACAGCAGTTCTATCAGAGCTTTTCCCACAACGGATCTTCTTGGCAAGCAAATACAGCAGGAGTCAAT ( SEQ ID NO: 329 )ATTORNEY DOCKET NO. ORB-019WO1IRES389 TTAAAACAGCCTGTGGGTTGCACCCACCCACAGGGCCCACTGGGCGCCAGCACTCTGGTATCGCGGTACCTTT GTGCGCCTGTTTTATGACCCCTCCCCAACCGAAACTTAGAAGTTACACATACCGATCAATAGTGGGCATAGCA CGCCAGCTGTGTCTTGATCAAGCACTTCTGTTTCCCCGGACCGAGTATCAATAGACTGCTCACGCGGTTGAAG GAGAAAGCGTTCGTTATCCGGCTAACTACTTCGAGAAACCTAGTAACACCATGAAAGTTGCGGAGTGTTTCGC TCAGCACTTCCCCAGTGTAGATCAGGTCGATGAGTCACCGCGTTCCCCACGGGCGACCGTGGCGGTGGCTGCG CTGGCGGCCTGCCTATGGGATGACCCATAGGACGCTTCAACACAGACATGGTGTGAAGAGTCTATTGAGCTAG TTAGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCTAACTGCGGAGCACGTGCCCTCAAGCCAGAGGGTGGCG TGTCGTAACGGGCAACTCTGCAGCGGAACCGACTACTTTGGGAGTCCGTGTTTCCTTTTATTCTTACACTGGC TGCTTATGGTGACAATTGAGAGATTGTTACCATATAGCTATTGGATTGGCCATCCGGTGAGCAACAGAGCGAT TGTATACTCTTTTGTGGGTTTTGTGCCATTAAATCTTACTGTGATTACCACACTGAAACTCATCTTGCAGTTT AATACAGCAAA ( SEQ ID NO: 330 ) _IRES325 TTAAAACAGCTCTGGGGTTGTTCCCACCCCAGAGGCCCACGTGGCGGCTAGTACACTGGTATTACGGTACCCT TGTACGCCTGTTTTATACTCCCTTCCCCGTAACTTAGAAGCTATGTAATCCAAGTTCAATAGAAGGGGGTACA AACCAGTACCACCACGAACGAGCACTTCTGTTTCCCCGGTGAGATTGTATAAGCTGTTCCCACGGCCGAAAAC GATCGATCCGTTACCCGCTCTTGCACTTCGAGAAGCCTAGTATCATCTTGGGATCTTCGATGCGTTGCGCTCA GCACTCTACCCCAGAGTGTAGCTTAGGTCGATGAGTCTGGACATTCCTCACCGGCGACGGTGGTCCAGGCTGC GTTGGCGGCCTACCTGTGGTCCAAAGCCACAGGACGCTAGTTGTAAACAAGGTGTGAAGAGCCTATTGAGCTG CAAGAGAGTCCTCCGGCCCCTGAATGCGGCTAATCCCAACCACGGAGCAGGTGGTTGCAACCCAGTAACTGGC CTGTCGTAACGCGTAAGTCTGTGGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATTTCTATAATGG CTGCTTATGGTGACAATCATAGATTGTTATCATAAAGCGAATTGGATTGGCCATCCGGTGAGAGTTAAGACAA TTATCCACTTGTTTGTTGGATTTACCCCTCTGACTAATTACACACTCAACTTTATTAGGATTGTCATTCTAGT GAGATATTACCATCACA ( SEQ ID NO: 331 ) _IRES327 TTAAAACAGCTCTGGGGTTGTTCCCACCCCAGAGGCCCACGTGGCGGCTAGTACTCCGGTATCACGGTACCCT TGTACGCCTGTTTTACACTCCCTTCCCCCGTAACTTAGAAGAAACAAACAAAGTTCAATAGAAGGGGGTACAA ACCAGTACCACCACGAACAAGCACTTCTGTTTCCCCGGTGACGTTGTATAGACTGTACCCACGGTCGAAAACG ATTGATCCGTTATCCGCTTTTGTACTTCGAGAAGCCTAGTATCATCTTGGAATCTTCGATGCGTTGCGCTCAG CACTCAATCCCAGAGTGTAGCTTAGGTCGATGAGTCTGGACGTTCCTCACCGGCGACGGTGGTCCAGGCTGCG TTGGCGGCCTACCTGTGGCCCAAAGCCACAGGACGCTAGTTGTGAACAAGGTGTGAAGAGCCTATTGAGCTAC AAGAGAGTCCTCCGGCCCCTGAATGCGGCTAATCCTAACCACGGAGCAAGGGTGTGTGAGCCAACACATACCT TGTCGTAATGCGTAAGTTCGTGGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATTCTTGTATTGGC TGCTTATGGTGACAATCATAGATTGTTATCATAAAGCGACTTGGATTGGCCATCCAGTGGAAGTGAAATACCT TGTCTACCTGTCTGTTGGTTTTACTCCATTTGACTAATTCACTTACAAGCTTATTGCTATAATTTTTATTGAC TAAACAACAATACTACA ( SEQ ID NO: 332 ) _IRES659 CACCCTCTTTTCCGGCGGCGCATGTTCGCGTCGTCGTAAGTCTGGATTCCCAAGGCCCACTCGGTTCAACTTC GGTTTCCGGACAAATACAAAGAACCTCAGTCCTCTTGGTACTTTCTCGCCTGAGTCTACAAAGCGAGAAACCT GCCCCTCTAACGCCAGACGAGCGGCATAAAACTCGAACTTCTGGCACGTTCCACCACCCCTTCTCCTATCCCA ACCCCCATTGCGCTCTCAAGGTCGCGCTTTTCCGAGACTAGCTCGGATTCAAAAAGTTCCTGGCACCCTTTAC CCCTTCAGGCCCTTAAGGTAGGAACTGACCTTGTGCTGTGATCTCGGTGCGGAAGTGCTACTGCGTAGTGATT GTAAGATCCTTTTGTGGTTCTGCCCTGGCAAGGCTACAGAGTGCTGTGATCCGCTGCGGATGCCATCCTGGTA ACAGGACCCCCAGTGTGCGCAACAGTATGTTCACGGTCTTCCGTGTTCACCACATTCGGAACACTGCTTTCGT GAAACAGTGTGTGTCCAATCCCTGTGATCAGTATCAACCACACCACCTAGGAATGCTAGGAAGGTACCCCGGT TCGCCGGGATCTGATCCTAGGCTAATTGTCTACGGTGGTGCTCCTTTTTATTTTCCACTTCAACTCACTGGTT ACAACTGCTTGATTTCTGTGTTTGCTGCTTTTCTCTGCTCTCACTGCCATTCTCAAGTGTTCACACTGTCCAAGCTCCTTTGGTTGTTCGCTTCCACTTGCCACTGTCAACTCTTGTC (SEQ ID NO: 333 )ATTORNEY DOCKET NO. ORB-019WO1IRES693 AACAGCCTGTGGGTTGTACCCACCCACAGGGCCCACTGGGCGCTAGCACACTGGTATCACGGTACCTTTGTGC GCCTGTTTTATACCCCCTACCCCAACTTTAAACTTAGAAGCAAAGCAAACCCGATCAATAGCAGATGTAGCAA GCCAGTTGCATCTTGATCAAGTACTTCTGTCTCCCCGGACTGAGTATCAATAGACTGCTCACGCGGTTGAAGG AGAAAACGTTCGTTACCCGACCAACTACTTCGAGAAACCTAGTAACACCATGAAAGTAGCAGGGTGTTTCGAT CAGCACAACCCCAGTGTAGATCAGGCTGATGAGTCACCGCATCCCCCACGGGCGACCGTGGCGGTGGCTGCGC TGGCGGCCTGCCCATGGGGCAACCCATGGGACGCTCTAATATGGACATGGTGTGAAGAGTCTATTGAGCTAGT TGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCTAACTGCGGAGCACATACCCCAAAGCCAAGGGGCAGTGT GTCGTAACGGGCAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATTCTCAAACTGGCT GCTTATGGTGACAATCTCAGAGTTGTTACCATATAGCTATTGGATTGGCCATCCGGTGAGCAACAGAGCAATT GTCTACCAATTTGTTGGATTTATTCCACTCAATCTAGCGGTTTTCAGAACATTGAATTATATTCTAACCCTCA ACAAGACGAA ( SEQ ID NO: 334 ) _IRES772 TGGAATTAACATCATTTCCGACGAAAGTGCTATCATGCCTCCCCGATTGTGTGATGTTTTCTGCCCTGCTGGG CGGAGCATTCTCGGGTTGAGAAACCTTGAATCTTTTCCTTTGGAACCTTGGTTCCCCCGGTCTAAGCCGCTTG GAATATGACAGGGTTATTTTCCTTATCTTACTTCTACTTTCACGGGTTCTATCCGTGAAAAGGGTACGTGTTG CCCCTTCCTTCTTCGGAGAATTCACACGGCGGTCTTTCCGTCTCTCGACAGGTGTGAATGCAACATGCCGGAA ACGGTGAAGAAAACAGTTTTCTGCGGAAGTCTTGAGTGCGCATCGAAACAGCTGTAGCGACCTCACAGTAGCA GCGGACTCCCCTCTTGGTGACAAGAGCCTCTGCGGCCAAAAGCCCCGTGGATAAGATCCACTGCTGTGAGCGG TGCAACCCCAGCACCCTGATTCGATGAACATTCTTTACGGAACCAGAAGATGGTTTTCTTAAGCCCTCCGGTA GAGAAGCCAAGAATGTCCTGAAGGTACCCCGCGTGCGGGATCTGATCAGGAGACCAATTGACTGTGCTTTACA CTGTCACTTTGGTTTAAAAACTGTCACAGCTTCTCCAAACCAAGTGGTCTTGGTTTTCCAATTTTACTAACTG GCAAT ( SEQ ID NO: 335 ) _IRES537 TTACCCGTATTCCTTCCCTTCCCCCGCAACCACCACGCTTACTCGCGCACGTGTTGAGTGGCACGTGCGTTGT CCAAACAGCTACACCCACACCCTTCGGGGCGGGTTTGTCCCGCCCTCGGGTTCCTCGCGGAACCCCCCCCTCC CTCTCTCTCTTTCTATCCGCCCTCACTTCCCATAACTACAGTGCTTTGGTAGGTGAGCACCCTGACCCCCCGC GGAAGCTGCTAACGTGGCAACTGTGGGGATCCAGGCAGGTTATCAAAGGCACCCGGTCTTTCCGCCTTCAGGA GTATCTCTGCCGGTGAATTCCGGTAGGGCTCTGCTTGGTGCCAACCTCCCCCAAATGCGCGCTGCGGGAGTGC TCTTCCCCAACTCATCTTAGTAACCTCTCATGTGTGTGCTTGGTCAGCATATCTGAGGCGACGTTCCGCTGTC CCAGACCAGTCCAGCAATGGACGGGCCAGTGTGCGTAGTCGCTTTCCGGTTTTCCGGCGCATGTTTGGCGAAA CGCTGAGGTAAGGTTGGTGTGCCCAACGCCCGTAATTTGGTGATACCTCAAGACCACCCAGGAATGCCAGGGA GGTACCCCACTTCGGTGGGATCTGACCCTGGGCTAATTGTCTACGGTGGTTCTTCTTGCTTCCACTTCTCTTT TTTCTGGC ( SEQ ID NO: 336) _IRES703 AACAGCCTGTGGGTTGTACCCACCCACAGGGCCCACTGGGCGCCAGCACTCTGGTATTACGGTACCCTTGTGC GCCTGTTTTATATCCCTTCCCCAAGGTAAACGTTAGAAGCAATGCACTCCCGATCAATAGAAGGTGCGACGCG CCAGTCGTATCTCGATCAAGCACTTCTGTTTCCCCGGACCGAGTATCAATAGACTGCTAGCGCGGTTGAAGGA GAAAACGTTCGTTACCCGACTAACTACTTCGAGAAGCCCAGTAGCGCCATGAAAGTTGCAGAGTGTTTCGCTC AGCACTTCCCCCGTGTAGATCAGGCCGATGAGTCACCGCAATCCCCACGGGCGACCGTGGCGGTGGCTGCGTT GGCGGCCTGCCTATGGGGAAACCCATAGGACGCTCAAATACAGACATGGTGCGAAGAGTCTATTGAGCTAGTT AGTACTCCTCCGGCCCCTGAATGCGGCTAATCCTAACTGCGGAGCAAGTACTCACAATCCAGTGGGCAGCTTG TCGTAACGGGTAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATTCCTACATTGGCTG CTTATGGTGACAATCAAAGAGTTGTTACCATATAGCTATTGGATTGGCCATCCGGTGACTAATAAAGCAATTA TATTTCTTTTTGTTGGGTTTGTACCCCTTAATACAACCAGCTTCACCACATTGTGTTACATCATTAGACTGAA CAAGGCAAA ( SEQ ID NO: 337 ) _IRES758 TCTTTTTGACGTGGTTGGAATTAACATCATTTCCGACGAAAGTGCTATCATGCCTCCCCGATTGTGTGATGTT TTCTGCCCTGCTGGGCGGAGCATTCTCGGGTTGAGAAACCTTGAATCTTTTCCTTTGGAACCTTGGTTCCCCC GGTCTAAGCCGCTTGGAATATGACAGGGTTATTTTCCTTATCTTACTTCTACTTTCACGGGTTCTATCCGTGA AAAGGGTACGTGTTGCCCCTTCCTTCTTCGGAGAATTCACACGGCGGTCTTTCCGTCTCTCGACAGGTGTGAA TGCAACATGCCGGAAACGGTGAAGAAAACAGTTTTCTGCGGAAGTCTTGAGTGCGCATCGAAACAGCTGTAGC GACCTCACAGTAGCAGCGGACTCCCCTCTTGGTGACAAGAGCCTCTGCGGCCAAAAGCCCCGTGGATAAGATC CACTGCTGTGAGCGGTGCAACCCCAGCACCCTGATTCGATGAACATTCTTTACGGAACCAGAAGATGGTTTTC TTAAGCCCTCCGGTAGAGAAGCCAAGAATGTCCTGAAGGTACCCCGCGTGCGGGATCTGATCAGGAGACCAAT TGACTGTGCTTTACACTGTCACTTTGGTTTAAAAACTGTCACAGCTTCTCCAAACCAAGTGGTCTTGGTTTTCCAATTTTACTAACTGGCAAT ( SEQ ID NO: 338 )ATTORNEY DOCKET NO. ORB-019WO1IRES753 TTTGAAAAGGGGGTGGGGGGGCTTCGGCCCCCTCACCCTCTTTTCCGGCGGCGCATGTTCGCGTCGTCGTAAG TCTGGATTCCCAAGGCCCACTCGGTTCAACTTCGGTTTCCGGACAAATACAAAGAACCTCAGTCCTCTTGGTA CTTTCTCGCCTGAGTCTACAAAGCGAGAAACCTGCCCCTCTAACGCCAGACGAGCGGCATAAAACTCGAACTT CTGGCACGTTCCACCACCCCTTCTCCTATCCCAACCCCCATTGCGCTCTCAAGGTCGCGCTTTTCCGAGACTA GCTCGGATTCAAAAAGTTCCTGGCACCCTTTACCCCTTCAGGCCCTTAAGGTAGGAACTGACCTTGTGCTGTG ATCTCGGTGCGGAAGTGCTACTGCGTAGTGATTGTAAGATCCTTTTGTGGTTCTGCCCTGGCAAGGCTACAGA GTGCTGTGATCCGCTGCGGATGCCATCCTGGTAACAGGACCCCCAGTGTGCGCAACAGTATGTTCACGGTCTT CCGTGTTCACCACATTCGGAACACTGCTTTCGTGAAACAGTGTGTGTCCAATCCCTGTGATCAGTATCAACCA CACCACCTAGGAATGCTAGGAAGGTACCCCGGTTCGCCGGGATCTGATCCTAGGCTAATTGTCTACGGTGGTG CTCCTTTTTATTTTCCACTTCAACTCACTGGTTACAACTGCTTGATTTCTGTGTTTGCTGCTTTTCTCTGCTC TCACTGCCATTCTCAAGTGTTCACACTGTCCAAGCTCCTTTGGTTGTTCGCTTCCACTTGCCACTGTCAACTC TTGTCATGGACCCCTCAAAC ( SEQ ID NO: 339 ) _IRES650 TTAAAACAGCCTGTGGGTTGTACCCACCCACAGGGCCCACTGGGCGCTAGCACACTGGTATCACGGTACCTTT GTGCGCCTGTTTTATACCCCCTACCCCAACTTTAAACTTAGAAGCAAAGCAAACCCGATCAATAGCAGATGTA GCAAGCCAGTTGCATCTTGATCAAGTACTTCTGTCTCCCCGGACTGAGTATCAATAGACTGCTCACGCGGTTG AAGGAGAAAACGTTCGTTACCCGACCAACTACTTCGAGAAACCTAGTAACACCATGAAAGTAGCAGGGTGTTT CGATCAGCACAACCCCAGTGTAGATCAGGCTGATGAGTCACCGCATCCCCCACGGGCGACCGTGGCGGTGGCT GCGCTGGCGGCCTGCCCATGGGGCAACCCATGGGACGCTCTAATATGGACATGGTGTGAAGAGTCTATTGAGC TAGTTGGTAGTCCTCCGGCCCCTGAATGCGGCTAATCCTAACTGCGGAGCACATACCCCAAAGCCAAGGGGCA GTGTGTCGTAACGGGCAACTCTGCAGCGGAACCGACTACTTTGGGTGTCCGTGTTTCCTTTTATTCTCAAACT GGCTGCTTATGGTGACAATCTCAGAGTTGTTACCATATAGCTATTGGATTGGCCATCCGGTGAGCAACAGAGC AATTGTCTACCAATTTGTTGGATTTATTCCACTCAATCTAGCGGTTTTCAGAACATTGAATTATATTCTAACCCTCAACAAGACGAA ( SEQ ID NO: 340 ) _*Because the sequences are RNA sequences, it is understood that the T' s represent uracil in the sequences of this table.Untranslated Region

[0419] In some embodiments, the circRNA comprises one or more untranslated regions (UTR). In some embodiments, the circRNA comprises a 5' UTR, a 3' UTR or both a 5' UTR and a 3' UTR.

[0420] In some embodiments, the one or more UTR comprises a CAA40 comprising SEQ ID NO: 41 or a pAC18 comprising SEQ ID NO: 42.

[0421] In some embodiments, one or more UTR comprises any one of SEQ ID NO: 41-45, 46, 47 or 272. In some embodiments, the one or more UTR comprises SEQ ID NO: 47 or SEQ ID NO: 272.

[0422] As shown in Table 2A below, as used herein, SEQ ID NO: 43 and SEQ ID NO: 265 refer to the same sequence and are interchangeable throughout the specification. As used herein, SEQ ID NO: 46 and SEQ ID NO: 272 refer to the same 3’ UTR sequence and are interchangeable throughout the specification.

[0423] In some embodiments, the 3’ UTR comprises a 3’ exon fragment. In some embodiments, the 3’ UTR comprises a 3’ exon fragment as shown in FIG. 3 A. In some embodiments, the 3’ UTR comprises a 3’ exon fragment from Twort. In some embodiments, the 3’ UTR comprises a 3’ exon fragment from a Group I intron. In some embodiments, the 3’ UTR comprises a 3’ exon fragment from a Group II intron.ATTORNEY DOCKET NO. ORB-019WO1

[0424] In some embodiments, the 3’ UTR comprises SEQ ID NO: 268 or SEQ ID NO: 272. In some embodiments, the 3’ UTR comprises SEQ ID NO: 358 or SEQ ID NO: 362. It is understood that SEQ ID NO: 358 is identical to SEQ ID NO: 268 with Ts replaced by Us. It is understood that SEQ ID NO: 362 is identical to SEQ ID NO: 272 with Ts replaced by Us. It is understood that in the context of circRNA, SEQ ID NO: 268 is equivalent to SEQ ID NO: 358 and comprises uracils in place of thymines, and may be interchangeably used herein. Similarly, in the context of RNA, SEQ ID NO: 362 is equivalent to SEQ ID NO: 272 and comprises uracils in place of thymines, and may be interchangeably used herein.

[0425] In some embodiments, the 3’ UTR does not comprise a 3’ exon fragment. In some embodiments, the 3’ UTR does not comprise a 3’ exon fragment as shown in FIG. 3 A. In some embodiments, the 3’ UTR does not comprise a 3’ exon fragment from Twort. In some embodiments, the 3’ UTR does not comprise a 3’ exon fragment from a Group I intron. In some embodiments, the 3’ UTR does not comprise a 3’ exon fragment from a Group II intron.

[0426] In some embodiments, the 3’ UTR comprises TAATAGaacaaacacaaacCAAACACCATTGTCACACTCCAaacaacaaacaaacaACCACACA AATGGTCGCCGA (SEQ ID NO: 349).

[0427] In some embodiments, the 3’ UTR comprises TAATAGaaaaacacaaacCAAACACCATTGTCACACTCCAaaaaaaacaaacaACCACACAAA TGGTCGCCGA (SEQ ID NO: 350).

[0428] In some embodiments, the 3’ UTR comprises SEQ ID NO: 359. In the context of RNA, it is understood that SEQ ID NO: 359 is equivalent to SEQ ID NO: 349 and comprises uracils in place of thymines, and may be interchangeably used herein.

[0429] In some embodiments, the 3’ UTR comprises SEQ ID NO: 363. In the context of RNA, it is understood that SEQ ID NO: 363 is equivalent to SEQ ID NO: 350 and comprises uracils in place of thymines, and may be interchangeably used herein.

[0430] In some embodiments, the 3’ UTR comprises a single microRNA binding site.

[0431] In some embodiments, the 5’ UTR comprises a 5’ exon fragment. In some embodiments, the 5’ UTR comprises a 5’ exon fragment as shown in FIG. 3 A. In some embodiments, the 5’ UTR comprises a 5’ exon fragment from Twort. In some embodiments, the 5’ UTR comprises a 5’ exon fragment from a Group I intron. In some embodiments, the 5’ UTR comprises a 5’ exon fragment from a Group II intron.

[0432] In some embodiments, the 5’ UTR comprises SEQ ID NO: 265.ATTORNEY DOCKET NO. ORB-019WO1

[0433] In some embodiments, the 5’ UTR comprises SEQ ID NO: 352. In the context of RNA, it is understood that SEQ ID NO: 352 is equivalent to SEQ ID NO: 265 and comprises uracils in place of thymines, and may be interchangeably used herein.

[0434] In some embodiments, the 5’ UTR does not comprise a 5’ exon fragment. In some embodiments, the 5’ UTR does not comprise a 5’ exon fragment as shown in FIG. 3 A. In some embodiments, the 5’ UTR does not comprise a 5’ exon fragment from Twort. In some embodiments, the 5’ UTR does not comprise a 5’ exon fragment from a Group I intron. In some embodiments, the 5’ UTR does not comprise a 5’ exon fragment from a Group II intron.

[0435] In some embodiments, the 5’ UTR comprises TCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCAT ATGACCTCTGCGTGTGCCTGGTAATTGACTAAGATATC (SEQ ID NO: 348).

[0436] In some embodiments, the 5’ UTR comprises SEQ ID NO: 353. In the context of RNA, it is understood that SEQ ID NO: 353 is equivalent to SEQ ID NO: 348 and comprises uracils in place of thymines, and may be interchangeably used herein.

[0437] In some embodiments, the 5’ UTR comprises SEQ ID NO: 348 and the 3’ UTR comprises SEQ ID NO: 349. In some embodiments, the 5’ exon fragment comprises SEQ ID NO: 382 and a 3’ exon fragment comprises SEQ ID NO: 389.

[0438] In some embodiments, the 5’ UTR comprises SEQ ID NO: 353 and the 3’ UTR comprises SEQ ID NO: 359.

[0439] In some embodiments, the 5’ UTR comprises SEQ ID NO: 348 and the 3’ UTR comprises SEQ ID NO: 350. In some embodiments, the 5’ UTR comprises SEQ ID NO: 353 and the 3’ UTR comprises SEQ ID NO: 363.

[0440] Table 2A. Exemplary UTR Sequences*CAA40ACAACAACAAACAACAAAACAACAAACAACACAAAACCAA ( SEQ ID NO: 41 )pAC18AAAAACAAAAAACAAAAACCACACAAATGGTCGCCGA ( SEQ ID NO: 42 )Exemplary 5' UTR5’ UTR_1 AACTTTTAGTTATTTTCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCATATG ACCTCTGCGTGTGCCTGGTAATTGACTAAGATATC (SEQ ID NO: 43 ) AACTTTTAGTTATTTTCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCATATG ACCTCTGCGTGTGCCTGGTAATTGACTAAGATATC (SEQ ID NO: 265 )5’ UTR_2ATTORNEY DOCKET NO. ORB-019WO1TCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCATATGACCTCTGCGTGTGCC TGGTAATTGACTAAGATATC ( SEQ ID NO: 348 )Exemplary 3' UTR3' UTR_1TAATAGAAAAACAAAAAACAAAAACCACACAAATGGTCGCCGAAACTACTGAAAGCAT ( SEQ ID NO: 44 )3' UTR_2TGATAATAGAAAAACAAAAAACAAAAACCACACAAATGGTCGCCGAAACTACTGAAAGCAT ( SEQ ID NO: 45 )3' UTR_3T AAT AGAAAAAC AC AAAC C AAAC AC CAT T GT C AC AC T C C AAAAAAAAC AAAC AAC C AC ACAAAT GGTCGCCGA AACTACTGAAAGCAT ( SEQ ID NO: 46)T AAT AGAAAAAC AC AAAC C AAAC AC CAT T GT C AC AC T C C AAAAAAAAC AAAC AAC C AC ACAAAT GGTCGCCGA AACTACTGAAAGCAT ( SEQ ID NO: 272 )3' UTR_4T AAT AGAAAAACACAAAC CAAACAC CAT T GT CACACT C CAACT CAT GAT CAGT ACAGAACAAACAC CAT T GT C ACACT CCAAGCT GACATACATAT CATAACT CTACTATAGACAAACACCATT GT CACACT CCAAAAAAAACAAA CAACCACACAAATGGTCGCCGAAACTACTGAAAGCAT ( SEQ ID NO: 47 )3’ UTR_5 TAATAGaacaaacacaaacCAAACACCATTGTCACACTCCAaacaacaaacaaacaACCACACAAATGGTCGCC GAaactactgaaagcat (SEQ ID NO: 268 )3' UTR_6T AAT AGAAC AAAC AC AAAC CAAACAC CAT T GT C AC AC T C C AAAC AAC AAAC AAAC AAC C AC AC AAAT G GT C G C CGAAACTACTGAAAGCAT (SEQ ID NO: 271 )3' UTR_7 TAATAGaacaaacacaaacCAAACACCATTGTCACACTCCAaacaacaaacaaacaACCACACAAATGGTCGC CGA ( SEQ ID NO: 349 ).3' UTR_8 TAATAGaaaaacacaaacCAAACACCATTGTCACACTCCAaaaaaaacaaacaACCACACAAATGGTCGCCGA ( SEQ ID NO: 350 ).*Because the sequences are RNA sequences, it is understood that the T' s represent uracil in the sequences of this table.5' and 3’ Spacers

[0441] In some embodiments, the nucleic acid molecule for making a circular RNA may comprise a 5’ UTR comprising a 5' spacer.

[0442] The 5' spacer sequence comprises a random sequence that increases circularization efficiency. The 5' spacer sequence may be of any length (e.g., 10 to 100 nucleotides, 10 to 90 nucleotides, 10 to 80 nucleotides, 10 to 70 nucleotides, 10 to 60 nucleotides, 10 to 50 nucleotides, 10 to 40 nucleotides, 10 to 30 nucleotides, 10 to 20 nucleotides, 20 to 100 nucleotides, 20 to 90 nucleotides, 20 to 80 nucleotides, 20 to 70 nucleotides, 20 to 60 nucleotides, 20 to 50 nucleotides, 20 to 40 nucleotides, 20 to 30 nucleotides, 30 to 100 nucleotides, 30 to 90 nucleotides, 30 to 80 nucleotides, 30 to 70 nucleotides, 30 to 60ATTORNEY DOCKET NO. ORB-019WO1nucleotides, 30 to 50 nucleotides, 30 to 40 nucleotides, 40 to 100 nucleotides, 40 to 90 nucleotides, 40 to 80 nucleotides, 40 to 70 nucleotides, 40 to 60 nucleotides, 40 to 50 nucleotides, 50 to 100 nucleotides, 50 to 90 nucleotides, 50 to 80 nucleotides, 50 to 70 nucleotides, 50 to 60 nucleotides, 60 to 100 nucleotides, 60 to 90 nucleotides, 60 to 80 nucleotides, 60 to 70 nucleotides, or 50 nucleotides). For example, in some embodiments, the length of the 5' spacer is selected to optimize translation of the protein-coding nucleic acid sequence.

[0443] In some embodiments, the 5' spacer sequence is between 20 and 50 nucleotides in length. In some embodiments, the 5' spacer sequence is between 30 and 100 nucleotides in length. In certain embodiments, the 5' spacer sequence is 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49,50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 nucleotides in length. In some embodiments, the 5' spacer sequence includes a 5' inner homology element. As used herein, the terms “internal homology region” and “inner homology element (IHE)” are used interchangeably. In some embodiments, the internal homology element is about 5-50 nucleotides in length. In some embodiments, the internal homology element is about 5-30 nucleotides in length. In some embodiments, the internal homology region is about 10-25 nucleotides in length. In some embodiments, the internal homology element is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides in length. In some embodiments, the 5' internal homology element is located at the 5' end of the 5' spacer sequence. The internal homology element forms base-pairing with an internal homology region, e.g., at the 3' end (3' inner homology element). In some embodiments, the nucleic acid described herein comprising 5' and 3' inner homology elements that are 75%, 80%, 85%, 90%, 95%, or 100% complementary to each other.

[0444] In some embodiments, the 5' spacer sequence comprises a polyA sequence. The polyA sequence may comprise 15-30 As. In some embodiments, the polyA sequence comprises 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 or 30 As. In other embodiments, the 5' spacer sequence comprises a polyAC sequence.

[0445] In some embodiments, the nucleic acid molecule for making a circular RNA comprises a 3’ UTR comprising a 3' spacer. In some embodiments, the 3' spacer comprises a microRNA binding site. In some embodiments, the circRNA comprises UTRs comprising a spacer sequence, or variants thereof, selected from Table 2B.ATTORNEY DOCKET NO. ORB-019WO1

[0446] In some aspects, provided herein is a circular ribonucleic acid (circRNA) comprising: (a) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, (b) an Internal Ribosome Entry Site (IRES) comprising a sequence at least 85% identical to any one of SEQ ID NO: 1-11, (c) 3’ and 5’ untranslated region(s) (UTR), (d) 3’ and 5’ exon fragments, and (e) a single microRNA binding site.

[0447] In some embodiments, the 3’ UTR comprises the single microRNA binding site.

[0448] In some embodiments, the microRNA binding site comprises SEQ ID NO: 40.

[0449] In some embodiments, the single microRNA binding site is flanked by an upstream spacer and a downstream spacer, and wherein the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3.

[0450] In some embodiments, the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3. In some embodiments, the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.4. In some embodiments, the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.5.

[0451] In some embodiments, the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.4. In some embodiments, the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.5.

[0452] In some embodiments, the miR-122_2 upstream spacer has a hydropathy index of -0.59. In some embodiments, the miR-122_2 downstream spacer has a hydropathy index of -0.53. In some embodiments, the miR-122_l upstream spacer has a hydropathy index of -0.5. In some embodiments, the miR-122_l downstream spacer has a hydropathy index of -0.346. In contrast, a 15 nucleotide poly A sequence of consecutive adenines has a hydropathy index of -0.1, while a 15 nucleotide poly C sequence of consecutive cytosines has a hydropathy index of -1.7. A 16 nucleotide sequence of alternating cytosines and adenines has a hydropathy index of -0.9.

[0453] In some aspects, provided herein is a circular ribonucleic acid (circRNA) comprising: (a) a 5’ exon fragment and a 5’ untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES), (c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (d) a 3’ UTR comprising an upstream spacer, a microRNA binding site, and a downstream spacer, and a 3’ exon fragment, wherein each of the upstream spacer and downstream spacerATTORNEY DOCKET NO. ORB-019WO1comprise multiple instances of consecutive adenines (As) but do not comprise more than five (5) consecutive adenines (As).

[0454] In some aspects, provided herein is a circular ribonucleic acid (circRNA) comprising, in order: (a) a 5’ exon fragment and a 5’ untranslated region (UTR), (b) an Internal Ribosome Entry Site (IRES), (c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and (d) a 3’ UTR comprising an upstream spacer, a microRNA binding site, and a downstream spacer, and a 3’ exon fragment, wherein each of the upstream spacer and downstream spacer comprise multiple instances of consecutive adenines (As) but do not comprise more than five (5) consecutive adenines (As).

[0455] In some embodiments, each of the upstream spacer and downstream spacer include consecutive adenines but do not comprise more than five (5) consecutive adenines (As). In some embodiments, each of the upstream spacer and the downstream spacer comprise 2, 3, 4 or 5 consecutive adenines.

[0456] In some embodiments, each of the upstream spacer and downstream spacer include consecutive adenines but do not comprise more than four (4) consecutive adenines (As). In some embodiments, each of the upstream spacer and the downstream spacer comprise 2, 3 or 4 consecutive adenines.

[0457] In some embodiments, each of the upstream spacer and downstream spacer include consecutive adenines but do not comprise more than three (3) consecutive adenines (As). In some embodiments, each of the upstream spacer and the downstream spacer comprise 2 or 3 consecutive adenines.

[0458] In some embodiments, the upstream spacer comprises 2, 3, 4 or 5 consecutive adenines. In some embodiments, the upstream spacer comprises two consecutive adenines. In some embodiments, the upstream spacer comprises three consecutive adenines. In some embodiments, the upstream spacer comprises four consecutive adenines. In some embodiments, the upstream spacer comprises five consecutive adenines.

[0459] In some embodiments, the downstream spacer comprises 2, 3, 4 or 5 consecutive adenines. In some embodiments, the downstream spacer comprises two consecutive adenines. In some embodiments, the downstream spacer comprises three consecutive adenines. In some embodiments, the downstream spacer comprises four consecutive adenines. In some embodiments, the downstream spacer comprises five consecutive adenines.ATTORNEY DOCKET NO. ORB-019WO1

[0460] In some embodiments, each of the upstream spacer and downstream spacer comprise one or more instances of a cytosine (C) interrupting consecutive As.

[0461] In some embodiments, the upstream spacer and downstream spacer facilitate circularization. In some embodiments, the circularization is increased by 5%, 6%, 7%, 8%, 9%, 10% or more by interrupting a poly A sequence comprising greater than five consecutive adenines in the upstream and / or downstream spacer by inserting a cytosine.

[0462] As a non-limiting example, the miR-122_2 redesigned spacer where poly A sequences were interrupted by inserting a cytosine increased circularization by about 8.2 % relative to a miR-122_l spacer (FIG. 4G and FIG. 4H).

[0463] In some aspects, provided herein is a circular ribonucleic acid (circRNA) comprising:a 5’ untranslated region (UTR), an Internal Ribosome Entry Site (IRES), a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigenbinding fragment thereof, and a 3’ untranslated region (UTR) comprising a microRNA binding site flanked by an upstream spacer and a downstream spacer, wherein each of the upstream spacer and downstream spacer comprise multiple instances of consecutive adenines (As) but do not comprise more than five (5) consecutive adenines (As). In some embodiments, each of the upstream spacer and the downstream spacer comprise 2, 3, 4 or 5 consecutive adenines.

[0464] In some embodiments, each of the upstream spacer and downstream spacer comprise no uracil.

[0465] In some embodiments, the upstream spacer sequence and the downstream spacer sequence consist only of As and Cs.

[0466] In some embodiments, each of the upstream spacer and downstream spacer have a length of not more than 18 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 12 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 13 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 14 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 15 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 16 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 17 nucleotides. In someATTORNEY DOCKET NO. ORB-019WO1embodiments, each of the upstream spacer and downstream spacer have a length of 18 nucleotides.

[0467] In some embodiments, each of the upstream spacer and downstream spacer have a length of 12-18 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 12, 13, 14, 15, 16, 17 or 18 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 12 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 13 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 14 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 15 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 16 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 17 nucleotides. In some embodiments, each of the upstream spacer and downstream spacer have a length of 18 nucleotides.

[0468] In some embodiments, each of the upstream spacer and downstream spacer have a length of 12-15 nucleotides.

[0469] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 4 nucleotides.

[0470] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 3 nucleotides.

[0471] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 2 nucleotides.

[0472] In some embodiments, the upstream spacer and the downstream spacer do not differ in length by more than 1 nucleotide.

[0473] In some embodiments, the microRNA binding site is a microRNA-122 (miR-122) binding site.

[0474] Table 2B. Exemplary Spacer sequences*Human p -actin 3 ' UTR taaACCGGACTGTTACCAACACCCACACCCCTGTGATGAAACAAAACCCATAAATGC ( SEQ ID NO: 396 ) Chicken p -actin 3 ' UTRAAGTTCTACAATGCATCTGGGACTTTGATTGTACATTGT ( SEQ ID NO: 397 )Internal Deletion from Human p -actin 3 ' UTR taaACCGGACTGTaaaaaAACCCATAAATGC ( SEQ ID NO: 398 )huRRBPl 5 ' UTR (nucleotide 111-219), " IRES" D3ATTORNEY DOCKET NO. ORB-019WO1GGCGGCGTCAGGGTCGCAGCGTCTACAGCTGCTCGGGGGCGGTTTCTTGGCGGAGGCTTGGCCGGCTCCTCTCTCC CGGCTCCGCGGCGGCTGCGAAGGCGGCGGCTCC ( SEQ ID NO: 399 )huRRBPl 5 ' UTR (nucleotide 1-69), " IRES" DI CGGCGATCCCGGCGGAGGGGGCCGTTCGCCAGCTCTGAGGCAGAAAGTGCCACGACTCCACACGCGCGCCGGCGAT CCCGGCGGAGGGGGCCGTTCGCCAGCTCTGAGGCAGAAAGTGCCACGACTCCACACGCGCGC ( SEQ ID NO: 400 )huRRBPl 5 ' UTR (nucleotide 70-110), " IRES" D2a ACGCAGCCAGCGAGCGGCCGGAGCGGACGGCAGACGGGGCG ( SEQ ID NO: 401 )huRRBPl 5 ' UTR (nucleotide 220-277), " IRES" D2b TGCCCTCTCGCTTTCCCTCTCGCGTCTCTGGCTGCAGGTGAAAGGAAAGCAAGCCAGG ( SEQ ID NO: 402 ) HCV DIV AGACCGTGCACCATGAGCACGAATCCA ( SEQ ID NO: 403 )22-mer structRNACACCTATATAGTTATATAATAA ( SEQ ID NO: 404 )IgMl-2 pre-mRNAcaagTggacagcaaTTcacacTgTcTcTgg ( SEQ ID NO: 405 )IgM-MS2 substratecaagTggacagcaaTTcacacTgTcTcTgTcaccTgcagg ( SEQ ID NO: 406 )dsx-MS2 substrateTgTaTaaTaaaaTgccgaaTgTcTcTcgaTcTgaTcaaaccag ( SEQ ID NO: 407 )longARFlgagtgccagaagctgcctcTTcggaggcagtttctggtactc ( SEQ ID NO: 408 )shortARFlggcagaagcTgccTcTTcggaggcagTTTcTgcc ( SEQ ID NO: 409 )Scpl60 binding motifTGAAAAATTTTaaaaaGCGATGAGATaaaaaGTTAATTTG ( SEQ ID NO: 410 )Mpt5 5 ' UTR. l CTCCTGTTTCATTATTTGCGTGAGGCATTCTTGTATCTCTTGCAAAGTTTTCTTTTTTCTTTCACTTTCGCACTG( SEQ ID NO: 411 )Mpt5 5 ' UTR. 2 AGACTCTGTGTCTGTGTGTGCATTAGTATTTCGTCTTATTCCTTTCCTTTTCTTGCTGTACCATTGTATTTATTC( SEQ ID NO: 412 )Mpt5 5 ' UTR. 3 AAGGCCAGCTCAGCCATATACTCTTACCACATACTCAAATTCTGCTGCTCTGCTCTGCTTTTGCGTTGCTGAGTT( SEQ ID NO: 413 )Mpt5 5 ' UTR. 4 TCTTGTTTGTCTTCTAAACACTACTATAAAGCGTCTGCGTTTTTCTCTTCGTGTGTATTTCTCTTTTTTTTCGCA( SEQ ID NO: 414 )Whi3 binding motifATTORNEY DOCKET NO. ORB-019WO1GCATaaaaaGCATaaaaaGCAT ( SEQ ID NO: 415 )ASH1 El AAGACTATGTTAAAATACGCGAAGAAGTGGCTCATTTCAAGCCATTAAGTATACCCAACTTAACTAATAATCAAAA TAA ( SEQ ID NO: 416 )ASH1 E3ATGGATAACTGAATCTCTTTCAACTAATAAGAGACATTATCACGAAACAAT ( SEQ ID NO: 299 ) Vgl. VgLE. 1TATTTCTACTTTATTTCTACACTGTTA ( SEQ ID NO: 300 )Vgl. VgLE. 2TTTGCCTTGACTGTTAGCTGTTACCTGTTA ( SEQ ID NO: 301 )Vgl. VgLE. 3CTATTTCACTAAAATTAGATGAGGTGCTTGCACTATTCACTGCACAGAG ( SEQ ID NO: 302 )PTB. 1CTCCCTCTCTCCCTCTCTTCCTTCCTCTCTCTCTTTCTCT ( SEQ ID NO: 303 )PTB. 2CTCCCTCTCTCCCTCTCTTC ( SEQ ID NO: 304 )PTB. 3TTCTCCTCCTTTCTTTCCTC ( SEQ ID NO: 305 )PCBP. 1CCTTACCCTTTCCCCTTCCTCCAATCCCTTTCCCCTTCCA ( SEQ ID NO: 306 )PCBP. 2CCTTACCCTTTCCCCTTCCT ( SEQ ID NO: 307 )PCBP. 3CCAATCCCTTTCCCCTTCCA ( SEQ ID NO: 308 )ALPP 3 ' UTR pA signal CAAAAATAAACAAATAAATTTTAAAAATAAATAAATAATAAAAGGAAGTGTTAGACAATGTAA ( SEQ ID NO: 309 )CALR 3 ' UTR pA signal GAGAATGTAAGAACTACAAACAAAATTTCTATTAAATTAAATTTTGTGTCTC ( SEQ ID NO: 310 ) Randl9 / 35. 1ACTCATGATCAGTACAGAA ( SEQ ID NO: 311 )Rand32 / 30. 1AGCTGACATACATATCATAACTCTACTATAGA ( SEQ ID NO: 312 )Rand50 / 30. 1GTAAGCCTCATTTAATTACACTGTTGTAATCGTTATTATACTTTTAAGTA ( SEQ ID NO: 313 ) Rand50 / 70. 1GGTGCGGCCCGAAGCGAGTATCACCTCACGATCCAAACGACCTCTAAGAC ( SEQ ID NO: 314 )pAC, 2 OntACCAAACAACAACACAAAAA ( SEQ ID NO: 315 )ATTORNEY DOCKET NO. ORB-019WO1pAC, 25ntCAACAAAAAACAAACAAAAAAAACA ( SEQ ID NO: 316)pAC, 40ntACAACAACAAACAACAAAACAACAAACAACACAAAACCAA ( SEQ ID NO: 41 )pAC18_lAAAAACAAAAAACAAAA ( SEQ ID NO: 317 )pAC18AAAAACAAAAAACAAAAACCACACAAATGGTCGCCGA ( SEQ ID NO: 42 )*Because the sequences are RNA sequences, it is understood that T' s represent uracil in the sequences of this table.MicroRNA Binding Sites

[0475] In some embodiments, the UTR of the circRNA constructs described herein comprises one or more microRNA (miRNA) binding sites. In some embodiments, the circRNA is engineered to minimize off-target expression, e.g., through the use of miRNA binding sites (engineered within spacers in the 5’ and / or 3’ UTR of the circRNA). For example, miR-122 is highly expressed in the liver, and is the most abundant miRNA in the adult liver. The use of miRNA binding sites, such as miR-122 binding sites, in the circRNA (encoding for, e.g., the CAR protein) decreases CAR expression in the liver (CAR expression in the liver is an undesirable off-target effect). The miRNA binding site in the circRNA binds to the miRNA in the liver, which leads to translation inhibition, thus reducing expression of the CAR in the liver.

[0476] In some embodiments, the number of miRNA binding sites in the circular polynucleotide is variable and relates to the length of the circular polynucleotide. As nonlimiting examples, the circular polynucleotide comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more miRNA binding sites. In some other embodiments, the multiple miRNA binding sites has the same nucleic acid sequences and bind to the same miRNA. In other embodiments, the miRNA binding sites have different nucleic acid sequences and bind to different miRNAs, such as 2, 3, 4, 5, or more different miRNAs.

[0477] In some embodiments, the one or more microRNA binding sites is selected from any one of SEQ ID NOs: 27-40.

[0478] In some embodiments, the UTR comprises one microRNA binding site. In some embodiments, the 5' UTR comprises one microRNA binding site. In preferred embodiments, the 3' UTR comprises one microRNA binding site. In some embodiments, the circRNA comprises a single microRNA binding site. As used herein, a “single microRNA binding site” means only one microRNA binding site. In some embodiments, the one microRNA bindingATTORNEY DOCKET NO. ORB-019WO1site is selected from any one of SEQ ID NOs: 27-40. In some embodiments, the one microRNA binding site is a miR-122 binding site. In some embodiments, the circRNA comprises a single miR-122 binding site.

[0479] In some embodiments, the single microRNA binding site is SEQ ID NO: 40.

[0480] In some embodiments, the single microRNA binding site is SEQ ID NO: 356. In the context of RNA, it is understood that SEQ ID NO: 356 is equivalent to SEQ ID NO: 40.

[0481] In some embodiments, the UTR comprises two microRNA binding sites. In some embodiments, the 5' UTR comprises two microRNA binding sites. In some embodiments, the 3' UTR comprises two microRNA binding sites. In some embodiments, the 5' UTR comprises one microRNA binding site and the 3' UTR comprises one microRNA binding site. In some embodiments, each of the two microRNA binding sites are selected from any one of SEQ ID NOs: 27-40.

[0482] In some embodiments, the UTR comprises three microRNA binding sites. In some embodiments, the 5' UTR comprises three microRNA binding sites. In some embodiments, the 3' UTR comprises three microRNA binding sites. In some embodiments, the 5' UTR comprises two microRNA binding sites and the 3' UTR comprises one microRNA binding site. In some embodiments, the 5' UTR comprises one microRNA binding site and the 3' UTR comprises two microRNA binding sites. In some embodiments, each of the three microRNA binding sites are selected from any one of SEQ ID NOs: 27-40.

[0483] In some embodiments, the circRNA further comprises one or more miR-122 binding sites in the 5' UTR, the 3' UTR, or both. In some embodiments, the microRNA binding site is a miR-122 site. In some embodiments, the miR-122 binding site is in the 3' UTR. In some embodiments, the circRNAs described herein comprises one or more exemplary microRNA binding sites, or variants thereof, selected from Table 3.

[0484] Table 3. Exemplary microRNA and microRNA Binding Site Sequences* microRNA Reverse Complementary Sequence (miRNA (miRNA) microRNA (miRNA) Sequence binding site sequence)TGAGGAGATCGTCGAGGTTGG ( SEQ ID CCAACCTCGACGATCTCCTCA ( SEQ ID miR-3150b-3p NO: 13 ) NO: 27 )TCATCCTCGTCTCCCTCCCAG ( SEQ ID CTGGGAGGGAGACGAGGATGA ( SEQ ID miR-5196-3p NO: 14 ) NO: 28 )GTAGATAAAATATTGGTACCTG ( SEQ CAGGTACCAATATTTTATCTAC ( SEQ miR-577-5p ID NO: 15 ) ID NO: 29 )CGGGGCAGCTCAGTACAGGAT ( SEQ ID ATCCTGTACTGAGCTGCCCCG ( SEQ ID miR-486-3p NO: 16) NO: 30 )TAGATAAAATATTGGTACCTG ( SEQ ID CAGGTACCAATATTTTATCTA ( SEQ ID miR-577 NO: 17 ) NO: 31 )AGAGTTAACTCAAAATGGACTA ( SEQ TAGTCCATTTTGAGTTAACTCT ( SEQmiR-4424 ID NO: 18 ) ID NO: 32 )ATTORNEY DOCKET NO. ORB-019WO1GCCCGGAGAGCTGGGAGCCAG ( SEQ ID CTGGCTCCCAGCTCTCCGGGC ( SEQ ID miR-10398-3p NO: 19 ) NO: 33 )TGTAAACATCCTCGACTGGAAG ( SEQ CTTCCAGTCGAGGATGTTTACA ( SEQ miR-30a-5p ID NO: 20 ) ID NO: 34 )AAGGTTTGGATAGATGCAATA ( SEQ ID TATTGCATCTATCCAAACCTT ( SEQ ID miR-4464-5p NO: 21 ) NO: 35 )CTAGACTGAAGCTCCTTGAGG ( SEQ ID CCTCAAGGAGCTTCAGTCTAG ( SEQ ID miR-151a-3p NO: 22 ) NO: 36)CATAAAGTAGAAAGCACTACT ( SEQ ID AGTAGTGCTTTCTACTTTATG ( SEQ ID miR-142-3p NO: 23 ) NO: 37 )ACAGGTGAGGTTCTTGGGAGCC ( SEQ GGCTCCCAAGAACCTCACCTGT ( SEQ miR-125a-3p ID NO: 24 ) ID NO: 38 )AGGCAAGATGCTGGCATAGCT ( SEQ ID AGCTATGCCAGCATCTTGCCT ( SEQ ID miR-31-5p NO: 25 ) NO: 39 )TGGAGTGTGACAATGGTGTTTG ( SEQ CAAACACCATTGTCACACTCCA ( SEQmiR-122-5p ID NO: 26) ID NO: 40 )*Because the sequences are RNA sequences, it is understood that T' s represent uracil in the sequences of this table.Codon-optimized Coding Sequences

[0485] The present invention provides, among other things, circRNA molecules comprising coding sequences encoding for anti -CD 19 CAR. The coding sequence, also known as open reading frame (ORF) sequence, is defined herein as the region that encodes the protein of interest.

[0486] In some embodiments, the coding sequence includes a region to initiate translation. In some embodiments, the region includes any translation initiation sequence or signal including a start codon. As a non-limiting example, the coding sequence includes one or more start codon(s). In some embodiments, as related to a DNA sequence, the start codon is “ATG,” “ACG,” “AGG,” “ATA,” “ATT,” “CTG,” “GTG,” or “TTG.” In some embodiments, as related to an RNA sequence, the start codon is “AUG,” “AU A,” “AUU,” “CUG,” “GUG,” or “UUG.”

[0487] In some embodiments, the coding sequence further includes a region to stop translation. This region may include any translation termination sequence or signal including a stop codon. As a non-limiting example, the region includes one or more stop codon(s). In some embodiments, as related to a DNA sequence, the stop codon is “TGA,” “TAA,” “TGA,” or “TAG.” In some embodiments, as related to an RNA sequence, the stop codon is “UGA,” “UAA,” “UGA” or “UAG.”

[0488] In some embodiments, the circRNA molecule of the present disclosure comprises a coding sequence. In some embodiments, the circRNA molecule of the present disclosure comprises a codon-optimized coding sequence. In some embodiments, the CDS comprises a nucleic acid sequence or variants thereof selected from Table 4, which are exemplary codon-optimized codon sequences comprising anti-CD19 antigen-binding domain.ATTORNEY DOCKET NO. ORB-019WO1

[0489] In some aspects, provided herein is a circRNA comprising a codon-optimized sequence having at least 75% identity to SEQ ID NOS: 125-168 or 266. In some aspects, provided herein is a circRNA comprising a codon-optimized sequence having at least 80% identity to SEQ ID NOS: 125-168 or 266. In some aspects, provided herein is a circRNA comprising a codon-optimized sequence having at least 85% identity to SEQ ID NOS: 125-168 or 266. In some aspects, provided herein is a circRNA comprising a codon-optimized sequence having at least 90% identity to SEQ ID NOS: 125-168 or 266. In some aspects, provided herein is a circRNA comprising a codon-optimized sequence having at least 95% identity to SEQ ID NOS: 125-168 or 266. In some embodiments, the circRNA comprises a codon-optimized sequence having 100% identity to SEQ ID NOS: 125-168 or 266.

[0490] Table 4. Exemplary Codon-Optimized Coding Sequence* Comprising Anti-CD19 Antigen-Binding Domain ATGGCTCTGCCTGTGACAGCTCTGCTGCTGCCTCTGGCTCTGCTTCTGCATGCCGCCAGACCTGACATCCAGATGA CCCAGACAACCAGCAGCCTGTCTGCCAGCCTGGGCGACAGAGTGACCATCAGCTGTAGAGCCAGCCAGGACATCAG CAAGTACCTGAACTGGTATCAGCAGAAACCCGACGGCACCGTGAAGCTGCTGATCTACCACACCAGCAGACTGCAC AGCGGCGTGCCATCTAGATTTTCTGGCAGCGGCTCTGGCACCGACTACAGCCTGACAATCAGCAACCTGGAACAAG AGGATATCGCTACCTACTTCTGCCAGCAAGGCAACACCCTGCCTTACACATTCGGCGGAGGCACCAAGCTGGAAAT CACAGGTGGCGGAGGATCTGGCGGAGGTGGAAGCGGAGGCGGTGGATCTGAAGTGAAACTGCAAGAGTCTGGCCCT GGCCTGGTGGCTCCATCTCAGTCTCTGAGCGTGACCTGTACCGTCAGCGGAGTGTCTCTGCCTGATTACGGCGTGT CCTGGATCAGACAGCCTCCTAGAAAAGGCCTGGAATGGCTGGGCGTGATCTGGGGCAGCGAGACAACCTACTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACTCCAAGAGCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGATTACTGGG GCCAGGGAACCAGCGTGACCGTGTCTAGCACAACAACCCCTGCTCCTAGACCTCCTACACCAGCTCCTACAATCGC CAGCCAGCCTCTGTCTCTGAGGCCAGAGGCTTGTAGACCTGCTGCTGGCGGAGCTGTGCACACAAGAGGCCTGGAT TTCGCCTGCGACATCTACATCTGGGCCCCTCTGGCTGGAACATGTGGCGTGCTGCTGCTGAGCCTGGTCATCACCC TGTATTGCAAGAGAGGCCGGAAGAAGCTGCTCTACATCTTCAAGCAGCCCTTCATGAGGCCCGTGCAGACCACACA AGAAGAGGACGGCTGCTCCTGCAGATTCCCCGAGGAAGAGGAAGGCGGCTGCGAGCTGAGAGTGAAGTTCAGCAGA AGCGCTGACGCCCCTGCTTACCAGCAGGGACAGAACCAGCTGTACAACGAGCTGAACCTGGGGAGAAGAGAAGAGT ACGACGTGCTGGACAAGCGGAGAGGCAGAGATCCTGAGATGGGCGGCAAGCCCAGAAGAAAGAACCCTCAAGAGGG CCTGTATAATGAGCTGCAGAAAGACAAGATGGCCGAGGCCTACAGCGAGATCGGAATGAAGGGCGAGCGCAGAAGA GGCAAGGGACACGATGGACTGTACCAGGGCCTGAGCACAGCCACCAAGGACACATATGACGCCCTGCACATGCAGG CCCTGCCTCCAAGA ( SEQ ID NO: 125 ) ATGGCACTACCCGTTACAGCATTACTATTACCTCTCGCCCTCCTCCTCCACGCCGCCAGACCAGATATCCAGATGA CACAGACCACCTCTAGCCTCAGCGCCAGCCTCGGCGATAGAGTGACAATCTCTTGCAGAGCCTCCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTCCTGATCTACCACACCAGCAGGCTGCAC AGCGGCGTGCCATCTAGATTCTCTGGATCTGGATCTGGCACCGATTACAGCCTGACCATCAGCAATCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAATACCCTGCCCTACACCTTTGGCGGCGGCACAAAGCTGGAGAT TACAGGTGGTGGTGGTTCTGGGGGAGGAGGGTCTGGTGGTGGTGGTAGTGAAGTGAAACTGCAGGAATCTGGACCT GGCCTGGTGGCCCCTTCTCAGTCGCTGAGCGTGACATGCACCGTGTCTGGCGTGAGCCTGCCGGATTACGGCGTGA GCTGGATTCGGCAGCCTCCTCGGAAGGGCCTGGAATGGCTGGGCGTGATCTGGGGCTCCGAGACCACCTACTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGTCCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGATGACACGGCCATCTACTACTGCGCCAAGCACTACTACTACGGAGGCTCCTACGCCATGGATTACTGGG GCCAGGGAACCTCCGTGACAGTGTCTTCTACAACAACACCAGCTCCTAGGCCTCCTACACCTGCTCCTACAATTGC TTCTCAGCCTCTGTCTCTGAGACCAGAAGCTTGTAGACCAGCTGCTGGAGGAGCTGTGCATACAAGAGGCCTGGAT TTCGCCTGTGACATTTACATCTGGGCTCCACTGGCCGGCACCTGTGGCGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGAGGGGCAGAAAGAAGCTGCTGTACATCTTCAAGCAGCCTTTCATGCGCCCCGTGCAGACAACACA GGAGGAAGATGGCTGCAGCTGTAGATTTCCTGAAGAAGAGGAGGGCGGCTGCGAGCTGAGAGTGAAATTTTCTAGAAGCGCCGACGCCCCTGCCTACCAGCAGGGCCAGAATCAGTTATATAACGAACTGAATCTGGGCCGGCGGGAGGAGTATTORNEY DOCKET NO. ORB-019WO1ACGACGTGCTGGATAAGAGAAGAGGCAGAGATCCTGAAATGGGAGGAAAACCTAGAAGAAAGAATCCGCAGGAGGG CTTATATAATGAACTGCAGAAGGACAAGATGGCGGAGGCCTACAGCGAGATCGGCATGAAGGGCGAGAGAAGAAGA GGCAAGGGCCACGATGGCCTGTACCAGGGCCTGAGCACGGCCACCAAGGATACCTACGATGCCCTGCACATGCAGG CCCTGCCTCCTAGA ( SEQ ID NO: 126) ATGGCATTACCAGTTACAGCCCTCCTTCTTCCTCTCGCCCTCTTGCTCCACGCCGCCAGGCCAGATATCCAGATGA CACAGACCACCAGCAGCCTCAGCGCCAGCCTCGGCGATAGAGTGACAATCTCTTGCAGAGCCTCCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGTTGCTGATCTACCACACCAGCAGACTGCAC AGCGGAGTGCCTTCTAGATTCTCTGGAAGCGGCTCCGGCACAGACTACTCGCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGCAATACCCTGCCCTACACCTTCGGCGGCGGCACAAAACTGGAGAT TACAGGTGGTGGTGGTAGTGGTGGTGGGGGAAGTGGTGGTGGTGGTTCTGAAGTGAAACTGCAGGAATCAGGGCCT GGCCTGGTGGCCCCTTCTCAGTCGCTGAGCGTGACCTGTACCGTGTCAGGCGTGAGCCTGCCAGATTACGGAGTGA GCTGGATTAGACAGCCCCCTCGGAAGGGCCTGGAATGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGTCCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGATTACTGGG GCCAGGGCACCTCTGTGACGGTGTCTTCTACAACAACACCTGCTCCTAGGCCTCCTACACCAGCTCCTACAATTGC TTCTCAGCCGCTGAGTCTGCGGCCAGAAGCTTGTAGGCCTGCTGCAGGAGGAGCTGTGCATACAAGAGGACTGGAC TTTGCCTGTGATATCTACATCTGGGCCCCTCTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGCGCGGCCGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGAGGCCCGTGCAGACAACACA GGAGGAGGATGGCTGCTCCTGTAGATTTCCTGAAGAGGAGGAGGGCGGCTGCGAGCTGAGAGTGAAATTTTCTAGA AGCGCCGATGCCCCTGCCTACCAGCAGGGCCAGAACCAGTTATATAACGAACTGAACCTGGGCAGAAGAGAGGAGT ACGACGTGCTGGATAAGAGAAGAGGCAGAGATCCAGAAATGGGAGGAAAGCCTAGAAGAAAGAATCCGCAGGAGGG CCTATATAATGAACTGCAGAAGGACAAGATGGCCGAGGCCTACAGCGAGATCGGCATGAAGGGCGAGAGAAGAAGA GGCAAGGGCCACGATGGACTGTATCAGGGGCTGAGCACCGCCACCAAGGATACCTACGATGCCCTGCACATGCAGG CGCTGCCTCCTAGA ( SEQ ID NO: 127 ) ATGGCATTACCAGTTACAGCCCTCCTCCTCCCTCTCGCCCTCCTTCTCCACGCCGCCAGACCAGATATCCAGATGA CCCAGACCACCTCTAGCCTCAGCGCCAGCCTCGGCGATAGAGTGACAATCTCTTGCAGAGCCTCCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTCCTGATCTACCACACCAGCAGGCTGCAC TCTGGCGTGCCATCTAGATTTTCTGGATCTGGCAGCGGCACCGACTACAGCCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAATACCCTGCCCTACACCTTTGGCGGCGGCACAAAGCTGGAGAT TACAGGTGGTGGTGGTTCTGGTGGTGGTGGTAGTGGTGGTGGGGGATCTGAAGTGAAACTGCAGGAATCTGGACCT GGCCTGGTGGCCCCTTCTCAGTCTCTGAGCGTGACATGCACCGTGTCCGGCGTGAGCCTGCCGGATTACGGCGTGA GCTGGATTCGGCAGCCTCCTAGAAAGGGCCTGGAATGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCGCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGTCCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGATGACACCGCCATCTACTACTGTGCGAAGCACTACTACTACGGCGGCTCCTACGCCATGGATTACTGGG GCCAGGGCACCAGCGTGACGGTGTCTTCTACAACAACACCTGCTCCTAGACCTCCTACACCAGCTCCTACAATTGC TTCTCAGCCTCTGTCGCTGAGACCTGAAGCATGTAGACCTGCTGCTGGAGGAGCTGTGCATACAAGAGGACTGGAT TTTGCCTGCGACATCTACATCTGGGCCCCGCTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGAGGGGCAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGAGGCCTGTGCAGACCACCCA GGAGGAGGATGGCTGCAGCTGCAGATTTCCTGAAGAAGAGGAGGGAGGCTGCGAGCTGAGAGTGAAATTTTCCAGG AGCGCCGACGCTCCTGCCTACCAGCAGGGCCAGAATCAGTTATATAATGAACTGAACCTGGGCCGGCGGGAGGAGT ACGACGTGCTGGATAAGAGAAGAGGCAGAGATCCTGAAATGGGAGGAAAGCCTAGAAGAAAGAATCCGCAGGAGGG CCTATATAACGAACTGCAGAAGGACAAGATGGCCGAGGCCTATTCCGAGATCGGAATGAAGGGCGAGAGAAGAAGA GGCAAGGGCCACGATGGCCTGTACCAGGGCCTGAGCACGGCCACCAAGGATACCTACGACGCCCTGCACATGCAGG CCCTGCCTCCTAGA ( SEQ ID NO: 128 ) ATGGCCCTACCCGTTACAGCTCTACTGCTGCCGCTGGCACTCTTGCTGCACGCAGCCAGACCTGACATTCAGATGA CGCAGACTACGTCGTCCCTGTCAGCGTCCCTTGGGGATCGGGTTACTATAAGTTGCCGCGCCAGCCAGGATATAAG TAAATACCTCAATTGGTATCAGCAAAAGCCGGATGGTACCGTGAAGCTCCTCATCTACCATACGTCCCGGCTCCAC TCTGGTGTGCCGAGCAGGTTCTCCGGGAGTGGGAGTGGGACAGACTACAGCCTCACGATCAGCAATCTAGAGCAGG AGGACATCGCTACTTACTTTTGCCAGCAGGGGAATACCCTTCCCTACACTTTCGGTGGCGGCACGAAGTTGGAGAT AACTGGAGGCGGCGGGTCTGGCGGCGGTGGGTCAGGCGGTGGGGGCTCGGAGGTGAAGCTGCAAGAAAGCGGTCCA GGGCTGGTGGCCCCTTCACAGTCCCTTAGTGTGACCTGCACTGTCAGCGGAGTCTCACTGCCCGATTACGGGGTGA GCTGGATCAGGCAACCGCCGAGGAAGGGGCTGGAATGGTTAGGTGTCATCTGGGGGTCAGAGACTACTTACTATAA CAGCGCTTTGAAGTCTAGACTGACTATCATTAAGGATAACTCCAAATCCCAAGTCTTCCTCAAGATGAACAGTTTA CAGACAGATGACACAGCCATCTACTACTGTGCCAAGCACTACTACTACGGAGGCTCATATGCGATGGACTATTGGGGCCAAGGTACGTCGGTTACAGTAAGCTCGACGACCACTCCCGCACCTCGGCCTCCCACCCCTGCTCCTACCATCGCATTORNEY DOCKET NO. ORB-019WO1CAGTCAACCCCTGTCCCTGCGGCCTGAGGCCTGTCGTCCCGCGGCCGGGGGTGCGGTCCATACGCGCGGACTTGAC TTTGCGTGTGATATCTACATTTGGGCCCCCTTAGCTGGCACATGTGGTGTTCTCCTGCTCTCCCTGGTCATCACCT TGTACTGCAAGCGTGGTCGCAAGAAGTTGCTATATATATTTAAGCAGCCCTTCATGCGGCCAGTCCAGACTACTCA GGAAGAAGATGGCTGTAGTTGCCGGTTCCCCGAGGAAGAGGAAGGCGGCTGCGAGCTGCGGGTTAAGTTCTCTCGC AGTGCCGATGCCCCTGCGTATCAGCAGGGGCAGAATCAACTGTATAACGAGTTGAATCTTGGCCGTCGGGAAGAGT ACGATGTTCTCGATAAACGGCGGGGTCGAGATCCCGAGATGGGAGGGAAGCCCCGGAGAAAGAACCCACAAGAGGG GCTTTATAACGAGCTGCAGAAAGACAAGATGGCCGAGGCCTACTCGGAGATCGGGATGAAGGGAGAACGGCGCCGG GGTAAGGGCCACGACGGCCTTTACCAAGGGTTAAGTACCGCGACAAAAGATACTTATGACGCACTGCACATGCAGG CGTTGCCGCCAAGG ( SEQ ID NO: 129) ATGGCCTTGCCGGTCACTGCATTGCTGCTGCCTCTCGCACTGCTGCTTCATGCCGCAAGGCCTGACATTCAGATGA CTCAGACAACCAGCTCGCTCAGCGCATCGCTGGGGGATCGCGTGACGATCAGTTGCCGTGCATCCCAGGATATTTC CAAGTACCTCAACTGGTACCAGCAGAAGCCAGATGGTACAGTAAAATTGCTGATATACCATACATCCAGATTACAC TCAGGCGTACCCTCGCGCTTTAGTGGCTCCGGAAGTGGCACCGATTACTCTTTGACCATCAGTAATCTCGAGCAAG AAGACATTGCAACGTACTTCTGCCAACAGGGGAACACTCTCCCATACACCTTTGGGGGAGGTACTAAGCTCGAGAT TACGGGTGGCGGGGGGTCGGGCGGTGGCGGCTCTGGAGGCGGAGGGAGCGAGGTTAAACTGCAGGAGTCAGGCCCT GGGCTTGTAGCTCCCAGTCAGTCACTGTCTGTGACTTGTACTGTGTCAGGTGTTTCCTTGCCAGACTATGGGGTAT CCTGGATCCGCCAGCCACCCCGTAAAGGTCTGGAGTGGCTCGGGGTCATATGGGGCTCCGAAACTACTTATTACAA CTCGGCATTGAAGAGCAGGCTGACAATTATTAAAGATAATAGTAAGTCGCAGGTGTTCCTTAAAATGAACTCCCTG CAAACTGATGATACGGCCATATACTATTGTGCCAAACACTATTATTACGGCGGCAGCTATGCTATGGACTATTGGG GTCAGGGCACATCTGTTACCGTTAGTTCGACGACAACGCCCGCACCGCGCCCTCCAACCCCAGCTCCGACGATTGC ATCGCAGCCCCTCTCACTGCGCCCCGAGGCCTGTCGGCCAGCCGCAGGTGGCGCTGTGCATACACGGGGGCTGGAC TTCGCATGCGATATTTATATCTGGGCTCCACTGGCCGGGACGTGCGGAGTTCTTCTCCTGTCACTGGTGATCACCC TGTATTGTAAGCGCGGGCGGAAGAAGCTGCTATATATTTTTAAGCAGCCCTTTATGAGGCCTGTGCAGACGACACA GGAAGAAGACGGGTGCAGTTGCCGATTCCCTGAGGAAGAGGAGGGGGGCTGCGAGCTGCGCGTGAAATTTTCGCGC TCCGCTGACGCACCTGCGTACCAGCAAGGCCAGAATCAACTCTACAATGAGTTGAATTTGGGGAGACGCGAAGAGT ACGACGTCCTCGACAAGCGACGAGGGAGGGATCCTGAAATGGGAGGAAAGCCTCGCAGAAAGAACCCTCAGGAGGG GTTGTACAATGAGTTACAGAAGGACAAGATGGCGGAAGCCTACTCCGAGATAGGGATGAAGGGCGAGCGGAGGCGG GGGAAGGGCCACGACGGGCTTTATCAAGGTCTGTCCACAGCCACCAAGGATACCTATGACGCTCTGCATATGCAGG CCCTGCCCCCACGC ( SEQ ID NO: 130) ATGGCCCTTCCAGTGACAGCATTGTTGCTTCCTTTGGCACTGCTGTTACATGCCGCCCGCCCCGACATTCAGATGA CACAGACAACTTCCTCACTGAGTGCCAGTCTCGGAGACAGAGTGACCATCAGCTGTCGGGCCTCCCAAGATATCAG TAAATACCTTAACTGGTACCAGCAGAAGCCCGATGGGACAGTTAAACTTCTCATCTATCATACCAGCAGGCTGCAC AGCGGCGTTCCTAGCAGATTTAGTGGTAGCGGGAGCGGCACTGATTACAGCCTGACAATTAGCAATCTCGAGCAGG AGGATATCGCTACCTACTTTTGCCAGCAGGGTAACACCCTGCCCTACACTTTCGGGGGGGGAACTAAACTGGAGAT AACAGGGGGTGGCGGGAGCGGGGGAGGTGGCAGTGGGGGGGGCGGGAGTGAAGTGAAACTCCAGGAGAGCGGGCCA GGTCTCGTGGCCCCCTCTCAATCTTTGTCAGTGACTTGCACAGTCTCTGGCGTTTCCCTGCCCGACTATGGCGTCT CATGGATTCGACAGCCCCCTCGAAAGGGGTTAGAGTGGCTCGGAGTGATATGGGGCTCCGAAACCACCTATTACAA TTCCGCTTTAAAGAGCAGATTGACAATCATCAAGGATAATTCCAAGTCTCAGGTGTTTTTGAAAATGAACTCCCTT CAGACTGATGACACTGCAATCTACTACTGTGCCAAACATTACTATTACGGGGGGAGTTACGCCATGGACTACTGGG GGCAGGGCACAAGTGTGACTGTCTCCTCCACCACCACTCCTGCTCCCCGACCTCCTACACCTGCTCCAACTATCGC CTCTCAGCCACTGAGCCTGCGACCCGAGGCTTGCAGGCCTGCAGCTGGAGGCGCTGTCCATACCAGAGGTCTGGAC TTCGCCTGCGATATCTACATCTGGGCCCCACTAGCCGGTACCTGCGGCGTCCTCTTATTGAGCCTCGTGATCACTC TTTACTGCAAGAGAGGCCGCAAGAAGCTGTTGTACATATTCAAACAGCCTTTTATGCGGCCTGTGCAGACTACACA GGAGGAAGACGGGTGCTCATGTAGATTCCCTGAGGAAGAGGAGGGTGGATGCGAGCTTCGGGTGAAGTTCAGCCGA AGCGCTGACGCCCCCGCATACCAACAGGGCCAGAACCAACTCTACAATGAGTTGAATTTGGGCCGCAGGGAAGAAT ACGACGTGCTGGATAAACGAAGGGGCCGAGACCCCGAAATGGGGGGCAAGCCTCGGCGGAAGAATCCCCAGGAAGG CTTGTACAATGAGTTGCAGAAAGATAAAATGGCCGAGGCCTACTCCGAGATTGGTATGAAGGGCGAAAGAAGGCGG GGAAAAGGGCACGACGGGTTGTATCAGGGGCTGAGTACCGCAACAAAGGACACCTATGATGCCCTACATATGCAGG CTCTACCTCCCAGA ( SEQ ID NO: 131) ATGGCTCTTCCCGTGACAGCCTTGCTGCTCCCATTGGCTCTCCTACTGCACGCAGCACGACCCGATATTCAGATGA CCCAGACTACCTCTTCCCTCAGCGCCTCCCTCGGGGACCGAGTGACTATCTCCTGTCGGGCATCCCAGGATATTAG TAAGTACCTCAATTGGTACCAGCAAAAGCCCGACGGGACAGTCAAACTATTAATATACCACACCTCTAGACTACAT AGTGGTGTTCCTTCTCGGTTCTCCGGGAGCGGCTCTGGGACTGACTACAGTCTCACCATAAGCAACCTGGAGCAGG AGGACATAGCCACATACTTCTGTCAGCAGGGCAACACCCTGCCCTATACCTTCGGAGGGGGCACCAAACTAGAGATTACTGGCGGAGGGGGTTCAGGAGGAGGCGGGAGCGGGGGCGGTGGAAGTGAGGTGAAGTTACAGGAGTCCGGGCCTATTORNEY DOCKET NO. ORB-019WO1GGGCTCGTCGCCCCTTCCCAGAGCCTGTCAGTTACCTGTACTGTGAGCGGAGTCTCATTACCCGACTACGGAGTCA GCTGGATCCGCCAGCCCCCCAGGAAGGGACTGGAGTGGCTGGGCGTGATCTGGGGTTCTGAGACCACATATTATAA TTCTGCTCTAAAATCTAGGCTGACAATCATCAAAGACAACAGCAAGAGCCAGGTGTTTCTGAAGATGAACTCATTA CAAACAGATGACACTGCTATCTACTACTGCGCAAAGCACTACTATTATGGTGGGAGCTATGCCATGGATTATTGGG GGCAGGGGACCAGCGTCACTGTCTCCAGTACAACAACCCCCGCTCCACGACCTCCAACCCCAGCACCCACTATTGC TTCACAGCCCCTGAGTTTACGGCCCGAGGCATGTCGCCCTGCCGCTGGAGGGGCTGTTCATACCAGAGGACTGGAC TTCGCCTGTGATATCTACATTTGGGCCCCACTGGCCGGAACATGTGGCGTTCTCCTGCTCTCCCTCGTCATAACAC TCTATTGCAAGAGAGGCAGGAAGAAACTTCTCTATATCTTTAAACAGCCTTTTATGAGGCCTGTGCAGACTACACA GGAAGAGGATGGATGTTCCTGCCGATTCCCAGAGGAGGAGGAGGGCGGGTGTGAGCTACGAGTGAAGTTCTCACGC TCTGCAGATGCCCCCGCTTACCAGCAGGGGCAGAATCAGCTCTACAACGAGCTGAATCTCGGACGCCGAGAAGAAT ACGACGTGCTGGACAAACGCAGGGGTAGAGACCCCGAGATGGGGGGAAAACCTAGGCGCAAAAACCCCCAGGAGGG CCTGTATAACGAGCTGCAAAAGGATAAGATGGCCGAAGCTTACAGTGAGATCGGCATGAAGGGGGAGCGCCGCCGC GGTAAAGGGCACGACGGTCTGTACCAGGGACTTAGCACAGCTACCAAGGACACCTACGATGCTCTCCACATGCAGG CATTACCACCCAGG ( SEQ ID NO: 132 ) ATGGCCTTGCCCGTCACTGCCCTATTACTTCCTCTCGCCCTGTTGCTCCACGCCGCTCGCCCCGACATCCAAATGA CACAGACCACCTCTTCCCTATCCGCATCCTTGGGAGATAGGGTTACCATCTCCTGTAGGGCATCCCAGGATATAAG CAAATATTTGAATTGGTATCAGCAAAAACCTGATGGTACAGTGAAGCTACTCATTTATCACACATCCCGGTTGCAT TCCGGCGTGCCCTCCCGCTTCAGCGGAAGCGGTTCCGGGACTGACTATTCACTTACTATAAGTAATCTAGAGCAAG AGGACATTGCAACTTATTTCTGCCAGCAAGGAAACACCCTACCCTACACATTTGGAGGAGGAACAAAATTGGAAAT CACCGGGGGCGGGGGTTCAGGTGGTGGTGGAAGCGGTGGTGGTGGATCAGAGGTTAAGTTGCAAGAGAGCGGCCCC GGCCTTGTGGCTCCCAGTCAGAGTCTATCAGTGACCTGTACCGTGTCTGGCGTTAGTCTCCCAGACTATGGAGTTA GCTGGATCAGACAACCACCACGCAAGGGACTGGAGTGGCTTGGAGTTATATGGGGATCTGAGACCACATATTACAA TTCTGCCCTTAAGTCCCGACTAACAATCATTAAAGACAACTCTAAGAGCCAGGTTTTCCTCAAAATGAACTCTTTG CAGACCGATGACACCGCTATTTACTATTGTGCCAAGCACTACTATTACGGTGGGTCATACGCTATGGATTACTGGG GGCAGGGCACATCAGTCACTGTGAGCTCTACAACCACACCCGCTCCCAGGCCTCCAACCCCCGCCCCCACTATCGC CTCTCAACCTCTCAGCTTAAGGCCAGAGGCTTGTCGACCTGCTGCCGGCGGAGCTGTTCATACCCGCGGTTTGGAC TTCGCTTGCGACATATACATTTGGGCACCACTCGCCGGCACCTGCGGTGTGCTCCTCCTTTCTCTCGTGATTACCC TGTATTGCAAGCGAGGCAGAAAAAAATTATTATATATTTTTAAACAGCCCTTCATGCGGCCTGTGCAGACTACACA AGAAGAAGATGGGTGCAGCTGCAGGTTCCCTGAGGAGGAGGAAGGCGGCTGTGAACTTCGGGTCAAATTTTCCCGC AGTGCCGATGCTCCTGCATACCAACAAGGTCAAAACCAGCTGTACAATGAGTTAAATCTTGGACGGAGAGAAGAGT ATGATGTCTTGGACAAGCGGCGAGGTCGGGATCCTGAGATGGGCGGAAAGCCACGGCGCAAGAACCCACAAGAAGG TCTCTACAACGAGCTTCAGAAAGATAAAATGGCTGAAGCTTATAGCGAAATTGGGATGAAAGGAGAAAGACGCAGG GGAAAGGGACACGACGGACTTTATCAAGGTCTATCTACCGCTACCAAAGATACTTATGACGCCCTCCACATGCAGG CACTTCCTCCACGC ( SEQ ID NO: 133) ATGGCCTTGCCAGTAACCGCACTTCTGCTGCCACTAGCACTGCTGCTGCATGCAGCCCGACCAGATATACAAATGA CCCAGACAACCTCGAGTTTGAGTGCCTCCCTTGGAGATCGCGTAACTATCAGCTGTCGGGCTAGTCAGGACATTTC AAAGTACCTCAACTGGTATCAGCAGAAACCTGATGGAACTGTGAAGCTGCTGATTTACCATACATCACGCTTACAC TCCGGAGTGCCCTCACGTTTCTCCGGATCCGGTTCCGGAACTGATTATAGTCTTACCATTAGCAACCTTGAGCAGG AAGACATAGCCACCTATTTCTGCCAACAAGGAAATACTTTGCCTTATACCTTTGGGGGCGGCACTAAGCTGGAAAT CACAGGTGGCGGGGGAAGCGGGGGCGGGGGAAGTGGCGGTGGGGGGTCTGAAGTGAAGCTCCAAGAATCTGGACCA GGTTTAGTTGCACCGAGCCAAAGCCTATCCGTGACCTGCACTGTTTCCGGGGTGTCTCTTCCAGATTACGGAGTGT CCTGGATACGCCAGCCGCCGCGAAAAGGGCTCGAGTGGTTGGGCGTCATATGGGGCAGCGAAACGACCTATTATAA CAGTGCCCTGAAAAGCCGGCTAACGATCATTAAAGATAACTCTAAGTCACAAGTATTCCTCAAGATGAACTCCCTT CAGACGGATGATACGGCCATATACTACTGTGCTAAGCATTATTATTATGGCGGGTCATACGCGATGGACTACTGGG GCCAGGGGACATCCGTCACCGTATCCTCTACGACGACACCGGCACCAAGACCTCCTACTCCTGCACCCACCATAGC CAGCCAGCCTTTGTCACTGAGACCCGAGGCTTGTAGACCAGCTGCCGGTGGCGCAGTGCACACCCGAGGCTTGGAC TTTGCCTGTGACATTTACATCTGGGCACCACTTGCAGGGACTTGCGGGGTACTCTTATTGAGCCTCGTAATCACAC TGTACTGCAAGAGAGGACGCAAAAAATTGCTATATATTTTTAAACAGCCTTTTATGCGCCCTGTTCAGACTACGCA GGAGGAAGATGGATGTTCTTGTAGGTTTCCCGAAGAAGAGGAGGGCGGGTGTGAGCTAAGAGTAAAATTCAGTAGA TCTGCGGATGCACCGGCCTACCAGCAGGGTCAGAACCAGTTATACAACGAGCTAAACTTAGGACGCAGAGAAGAGT ACGATGTATTAGATAAACGCCGTGGTCGCGATCCTGAAATGGGTGGTAAGCCTCGTCGGAAGAATCCTCAGGAGGG ACTGTACAACGAACTACAGAAAGATAAGATGGCTGAGGCTTACTCCGAGATTGGAATGAAGGGAGAGCGCAGAAGG GGAAAGGGACATGACGGCCTGTACCAAGGACTGAGCACTGCAACTAAGGATACCTACGACGCTCTGCACATGCAGG CCCTGCCCCCAAGA ( SEQ ID NO: 134 )ATGGCTCTACCAGTCACAGCACTACTCCTCCCTCTTGCCCTCCTTCTTCACGCCGCCAGGCCAGATATCCAGATGACACAGACCACCTCCTCCCTTTCCGCCTCTCTCGGGGACCGAGTCACTATTTCTTGTCGGGCTTCTCAGGATATCAGATTORNEY DOCKET NO. ORB-019WO1TAAGTACCTTAATTGGTACCAGCAGAAGCCCGACGGGACAGTGAAGCTCCTGATCTACCACACCAGTAGGCTCCAC TCTGGTGTCCCCTCCAGATTTAGTGGGTCAGGGAGTGGGACCGACTACTCCCTGACCATCTCCAATCTGGAGCAGG AGGATATCGCCACCTACTTCTGTCAGCAGGGGAATACCCTGCCATATACATTTGGAGGGGGCACCAAGCTGGAGAT TACTGGTGGTGGTGGATCAGGAGGAGGTGGAAGTGGAGGAGGTGGTTCTGAAGTGAAACTTCAGGAATCTGGACCT GGCCTGGTGGCTCCCAGCCAGTCCCTGAGTGTGACTTGCACCGTGTCTGGCGTGTCTCTGCCAGACTACGGTGTGA GCTGGATCCGCCAGCCACCCAGGAAAGGCCTGGAGTGGCTGGGCGTGATCTGGGGCAGTGAGACCACCTACTATAA TAGTGCTTTGAAGTCCAGGCTGACCATCATCAAGGACAACTCCAAGAGTCAGGTGTTCTTGAAGATGAACAGCCTG CAGACTGATGATACCGCAATCTACTATTGCGCAAAGCACTATTATTATGGGGGGTCTTACGCCATGGATTACTGGG GGCAGGGGACTAGTGTTACTGTGAGTAGCACTACTACCCCTGCCCCTAGACCTCCAACACCAGCTCCTACCATTGC AAGTCAGCCTCTGTCTCTGAGACCAGAGGCTTGTAGACCTGCAGCTGGTGGAGCTGTGCATACTCGGGGACTGGAT TTTGCATGTGACATTTACATCTGGGCTCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGTCTGGTGATCACTC TGTACTGCAAGCGCGGAAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCAGTGCAGACCACTCA GGAGGAGGACGGCTGCAGCTGCCGGTTTCCAGAGGAGGAAGAAGGAGGGTGTGAGCTGAGGGTGAAGTTTTCACGC TCAGCTGACGCCCCTGCTTATCAGCAGGGCCAGAATCAGTTATATAATGAACTGAATCTGGGCAGGAGGGAGGAGT ACGATGTGCTGGATAAACGGAGAGGCAGGGATCCCGAAATGGGAGGTAAGCCTCGGCGGAAGAATCCTCAGGAGGG TCTATATAACGAACTCCAGAAGGATAAGATGGCCGAGGCTTACTCCGAGATCGGGATGAAAGGGGAGAGGCGCCGG GGTAAAGGCCACGATGGCCTTTACCAGGGCCTCTCCACCGCAACAAAGGATACCTATGATGCCCTGCATATGCAGG CCCTGCCTCCCCGG ( SEQ ID NO: 135) ATGGCACTACCCGTTACCGCTCTCTTGCTCCCTTTGGCCCTCCTCCTCCACGCCGCCAGACCCGATATCCAGATGA CTCAGACTACATCTAGTTTGAGTGCATCTCTCGGAGATCGGGTCACCATTTCTTGTCGGGCTTCTCAGGATATCAG TAAGTACCTTAATTGGTACCAGCAGAAGCCCGACGGGACAGTGAAGCTCCTGATCTACCACACCAGTAGGCTCCAC TCTGGTGTCCCCTCCAGATTTAGTGGGTCAGGGAGTGGGACCGACTACTCCCTGACCATCTCCAATCTGGAGCAGG AGGATATCGCCACCTACTTCTGCCAGCAGGGGAATACCCTGCCATATACATTTGGAGGGGGCACCAAGCTGGAGAT TACTGGTGGTGGTGGATCAGGAGGTGGTGGTAGTGGAGGAGGAGGGTCAGAGGTGAAACTGCAGGAGAGCGGTCCT GGTCTCGTTGCCCCAAGTCAGTCCCTGTCCGTGACATGTACCGTGTCTGGCGTGTCTCTGCCAGACTACGGAGTGT CATGGATCAGACAGCCACCCAGGAAAGGCCTGGAGTGGCTGGGAGTGATTTGGGGCAGCGAGACCACATATTATAA CTCCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACTCCAAGAGTCAGGTGTTCTTGAAGATGAATAGCCTG CAGACAGATGACACGGCCATCTACTACTGTGCGAAGCACTACTACTATGGCGGCAGCTACGCCATGGATTACTGGG GGCAGGGGACATCTGTGACTGTGAGTTCAACAACCACTCCAGCTCCACGGCCTCCTACTCCAGCTCCAACCATTGC AAGTCAGCCTCTGAGTCTGAGACCAGAGGCTTGTAGACCTGCAGCTGGTGGAGCTGTTCATACTCGCGGGCTGGAT TTCGCCTGCGATATCTACATCTGGGCTCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGCCTGGTGATCACTC TGTACTGCAAGCGCGGAAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCAGTGCAGACCACTCA GGAGGAGGACGGCTGCAGCTGCCGGTTTCCAGAGGAGGAGGAAGGAGGCTGTGAGCTGAGAGTGAAGTTTTCTCGC AGCGCAGATGCCCCTGCCTACCAGCAGGGCCAGAATCAGTTATATAATGAACTGAATCTGGGCCGCAGGGAGGAGT ATGATGTGCTGGATAAGCGGAGGGGCAGGGATCCCGAAATGGGGGGTAAGCCTCGGCGGAAGAATCCTCAGGAGGG TCTATATAACGAACTCCAGAAGGATAAGATGGCCGAGGCTTACTCCGAGATCGGGATGAAAGGGGAGAGGCGCCGG GGTAAAGGCCACGATGGCCTTTACCAGGGCCTCTCCACCGCAACAAAGGATACCTATGATGCCCTGCATATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 136) ATGGCCCTACCAGTGACAGCCCTCCTCCTCCCTTTGGCCTTGTTGCTCCACGCAGCAAGGCCAGATATTCAGATGA CACAGACAACCAGCAGCCTCTCCGCCTCTCTTGGAGACAGGGTGACCATTTCTTGTCGGGCTTCTCAGGATATCAG TAAGTACCTTAATTGGTACCAGCAGAAGCCCGACGGGACAGTGAAGTTGCTGATCTACCACACTAGCAGGCTGCAT AGCGGTGTCCCTTCCAGATTCTCGGGATCTGGAAGCGGCACCGACTACAGCCTGACTATCAGCAACCTGGAGCAGG AGGACATCGCCACATATTTCTGCCAGCAGGGGAATACCCTGCCCTATACATTTGGCGGGGGCACAAAGCTGGAGAT TACAGGTGGCGGAGGCTCTGGAGGAGGAGGATCAGGCGGGGGCGGATCTGAAGTGAAACTTCAGGAATCCGGCCCC GGCCTGGTCGCTCCTTCCCAGAGCCTGTCCGTCACCTGTACAGTCTCCGGCGTGTCCCTGCCAGATTATGGGGTGA GTTGGATTCGCCAGCCCCCTAGAAAGGGGCTGGAATGGTTAGGCGTGATTTGGGGGTCAGAGACCACCTACTACAA CTCTGCCCTCAAGTCACGCCTGACCATTATCAAGGACAATAGTAAGAGCCAGGTGTTTCTGAAGATGAATAGTCTT CAGACCGATGACACCGCTATTTACTATTGTGCCAAGCACTACTACTATGGCGGCAGCTACGCCATGGATTACTGGG GGCAGGGGACTAGTGTTACTGTGAGTAGCACTACTACCCCTGCCCCTAGACCTCCAACTCCAGCTCCAACCATTGC AAGTCAGCCTCTGAGTCTGAGACCAGAGGCTTGTAGACCTGCAGCTGGTGGAGCTGTTCATACAAGGGGCCTGGAT TTTGCCTGTGACATTTACATCTGGGCCCCTCTGGCAGGAACATGTGGCGTGCTCCTCCTGTCTCTGGTGATCACCC T GTACT GTAAGAGAGGCAGGAAGAAGCT GOT GTACAT CTT CAAGCAGCCATT CAT GAGACCT GT GCAGACAACACA GGAAGAGGATGGCTGCTCCTGCAGGTTCCCTGAGGAGGAGGAGGGGGGCTGTGAACTGCGCGTCAAATTCTCCCGG TCTGCCGATGCCCCTGCTTATCAGCAGGGGCAGAATCAGTTATATAATGAACTGAATCTCGGCAGACGGGAGGAGT ATGACGTGCTGGATAAGCGGAGGGGCAGGGATCCCGAAATGGGGGGTAAGCCTCGGCGGAAGAATCCTCAGGAGGGTCTATATAACGAACTCCAGAAGGATAAGATGGCCGAGGCTTACTCCGAGATCGGGATGAAAGGGGAGAGGCGCCGGATTORNEY DOCKET NO. ORB-019WO1GGTAAAGGCCACGATGGCCTTTACCAGGGCCTCTCCACCGCAACAAAGGATACCTATGATGCCCTGCATATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 137 ) ATGGCATTACCAGTTACCGCCCTCCTCCTCCCCCTCGCCCTCCTCCTCCACGCCGCCCGGCCCGACATCCAGATGA CCCAGACCACCAGCAGCCTCAGCGCCAGCCTCGGCGACCGGGTGACCATCTCTTGCCGGGCCAGCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCGGACGGCACCGTGAAGCTCCTGATCTACCACACCAGCCGGCTGCAC TCCGGGGTGCCCAGCCGGTTCAGCGGCAGCGGCAGCGGCACCGACTACTCCCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAACACCCTGCCCTACACCTTCGGCGGCGGCACCAAGCTGGAGAT CACCGGCGGCGGCGGCAGCGGCGGCGGCGGCAGCGGCGGGGGCGGCAGCGAGGTGAAGCTGCAGGAGAGCGGCCCC GGCCTGGTGGCCCCCAGCCAGAGCCTGTCCGTGACCTGCACCGTGTCCGGCGTGAGCCTGCCCGACTACGGCGTGT CCTGGATCCGCCAGCCCCCCAGGAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCGCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGAGCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGACTACTGGG GCCAGGGCACCAGCGTGACCGTGTCCAGCACCACCACCCCCGCCCCCCGGCCCCCCACCCCCGCCCCCACCATCGC CAGCCAGCCCCTGAGCCTGCGGCCGGAGGCCTGCAGGCCCGCCGCCGGCGGCGCCGTGCACACCCGGGGCCTGGAC TTCGCCTGCGACATCTACATCTGGGCCCCCCTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGCGGGGCCGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCCGTGCAGACCACCCA GGAGGAGGACGGCTGCAGCTGCCGCTTCCCCGAGGAGGAGGAGGGCGGCTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCCGACGCCCCTGCGTACCAGCAGGGCCAGAACCAGCTGTACAACGAGCTGAACCTGGGCCGGCGCGAGGAGT ACGACGTGCTGGACAAGCGCCGGGGCCGGGACCCCGAGATGGGCGGCAAGCCCCGGCGCAAGAACCCCCAGGAGGG CCTGTACAACGAGCTGCAGAAGGACAAGATGGCCGAGGCCTACAGCGAGATCGGCATGAAGGGCGAGCGGCGGCGG GGCAAGGGCCACGACGGCCTGTACCAGGGCCTGTCCACCGCCACCAAGGACACCTACGACGCCCTGCACATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 138 ) ATGGCCCTGCCCGTGACCGCCCTGCTGCTCCCCCTGGCCCTGCTGCTGCACGCAGCAAGGCCAGACATCCAGATGA CCCAGACTACGAGCAGCCTGAGCGCCTCCCTGGGCGACAGGGTGACAATCAGCTGCCGGGCCAGTCAGGACATCAG CAAGTACCTGAACTGGTACCAGCAGAAGCCTGACGGCACCGTGAAGCTGCTGATTTATCACACCTCCAGGCTGCAC AGCGGCGTGCCCTCCAGATTCTCCGGCAGCGGCTCCGGCACCGACTACAGCCTGACCATCTCCAACCTGGAGCAGG AGGATATCGCCACCTACTTCTGCCAGCAGGGGAACACCCTGCCCTATACCTTTGGGGGCGGAACCAAGCTGGAGAT CACCGGCGGTGGCGGATCCGGCGGCGGCGGGAGCGGCGGCGGAGGATCTGAGGTCAAGCTGCAGGAGTCTGGCCCT GGGCTGGTGGCCCCAAGCCAGAGCCTGTCCGTGACATGTACCGTGTCCGGCGTGTCTCTGCCTGACTACGGCGTGT CATGGATCAGGCAGCCTCCTCGCAAGGGGCTGGAGTGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGTGCCCTGAAGTCCCGGCTGACCATCATCAAGGACAACAGCAAGAGCCAGGTGTTCTTGAAGATGAACAGCCTC CAGACCGACGACACCGCCATTTACTACTGTGCCAAGCACTATTACTATGGCGGGTCCTACGCCATGGATTACTGGG GCCAGGGCACCAGTGTGACCGTGTCCAGCACCACGACTCCTGCCCCACGCCCACCCACTCCAGCCCCTACCATCGC CAGCCAGCCACTGAGCCTGCGGCCTGAGGCCTGCAGGCCAGCGGCTGGCGGGGCGGTGCACACCCGCGGGCTGGAC TTCGCCTGCGACATCTACATCTGGGCCCCCCTCGCCGGCACCTGCGGCGTCCTCCTCCTGAGCCTGGTGATCACTC TGTACTGCAAGAGGGGCCGCAAGAAGCTGCTGTATATCTTCAAGCAGCCTTTCATGCGGCCCGTGCAGACCACTCA GGAGGAGGACGGCTGCAGCTGCCGGTTTCCTGAGGAGGAGGAGGGGGGCTGTGAGCTGCGGGTGAAGTTCAGCCGC TCCGCCGACGCCCCCGCCTACCAGCAGGGCCAGAATCAGCTGTACAACGAGCTGAATCTGGGCCGCAGGGAGGAGT ACGACGTGCTCGACAAGCGGCGGGGGCGGGACCCCGAGATGGGCGGCAAGCCCAGGCGGAAAAACCCCCAGGAGGG GCTGTACAATGAGCTCCAGAAGGACAAAATGGCCGAGGCTTACAGCGAGATCGGCATGAAGGGGGAGAGGCGGAGG GGCAAGGGGCATGACGGCCTGTACCAGGGGCTGTCCACAGCCACCAAGGACACCTACGACGCCCTGCACATGCAGG CCCTGCCTCCTAGG ( SEQ ID NO: 139) ATGGCCCTGCCCGTGACCGCCCTGCTGCTTCCCCTGGCCCTGCTGCTGCACGCCGCAAGGCCAGACATCCAGATGA CCCAGACTACCAGTTCTCTGTCCGCCTCTCTGGGCGACAGAGTGACGATCAGTTGCCGGGCCAGTCAGGACATCAG CAAGTACCTGAACTGGTACCAGCAGAAGCCTGACGGCACCGTGAAGCTGCTGATCTACCACACCAGCCGGCTGCAC AGCGGAGTGCCCAGCCGGTTCAGCGGCTCCGGGTCCGGCACTGATTACAGCCTGACCATCTCAAACCTGGAGCAGG AGGACATCGCCACTTACTTCTGCCAGCAGGGCAATACCCTGCCCTACACCTTCGGAGGCGGCACCAAGCTGGAGAT CACCGGCGGCGGCGGCTCCGGCGGCGGTGGCTCCGGCGGGGGCGGCTCCGAGGTGAAGCTGCAGGAGAGTGGCCCA GGCCTGGTTGCCCCCAGCCAGAGCCTGAGCGTGACATGCACCGTGTCAGGCGTGAGTCTTCCTGACTACGGGGTGA GCTGGATCAGGCAGCCCCCTAGGAAGGGCCTGGAGTGGCTGGGGGTGATCTGGGGCTCAGAGACCACATACTACAA CTCTGCCCTCAAGAGCAGGCTGACCATCATCAAGGACAACTCCAAGTCCCAGGTGTTCTTGAAGATGAACAGCCTG CAGACCGACGATACCGCCATTTACTACTGTGCCAAGCACTACTACTATGGCGGGTCCTACGCCATGGACTATTGGG GGCAGGGGACTAGTGTGACTGTGAGCTCCACCACCACCCCAGCCCCCAGGCCCCCTACTCCTGCCCCTACCATCGC CAGCCAGCCCCTGAGCCTGCGGCCTGAGGCCTGCAGGCCAGCCGCCGGGGGCGCCGTGCACACCAGGGGCCTGGAC TTTGCCTGTGACATTTACATCTGGGCCCCTCTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCCTGTACTGCAAGAGGGGCAGGAAGAAGCTGCTGTATATCTTCAAGCAGCCCTTCATGCGGCCAGTGCAGACCACTCAATTORNEY DOCKET NO. ORB-019WO1GGAGGAGGACGGCTGCAGCTGCCGCTTCCCTGAGGAGGAGGAGGGCGGCTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCCGATGCCCCTGCCTACCAGCAGGGCCAGAATCAGCTGTACAATGAGCTGAATCTGGGCCGTCGGGAGGAGT ACGACGTGCTGGATAAGCGCAGGGGCAGAGATCCTGAGATGGGCGGCAAGCCCCGCAGGAAGAATCCTCAGGAGGG TCTGTACAATGAGCTGCAGAAGGACAAGATGGCTGAGGCTTATTCAGAGATCGGCATGAAGGGCGAGAGGCGGCGG GGCAAGGGCCACGATGGCCTCTATCAGGGGCTGAGCACGGCCACCAAGGATACATACGACGCCCTGCACATGCAGG CGCTGCCTCCCCGG ( SEQ ID NO: 140) ATGGCCCTCCCCGTCACCGCCCTGCTCCTCCCCCTGGCACTCCTGCTGCACGCCGCCAGGCCCGACATCCAGATGA CCCAGACCACCAGCAGCCTCAGTGCCTCCCTGGGAGATAGGGTGACTATCTCCTGCAGGGCCAGTCAGGACATCAG CAAGTACCTGAATTGGTACCAGCAGAAGCCTGACGGCACTGTGAAGCTGCTGATCTACCACACCTCCCGCCTGCAC AGCGGTGTGCCCAGCCGGTTCTCCGGCTCTGGGTCCGGGACTGACTACAGCCTGACCATCTCCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAACACCCTGCCATACACCTTTGGTGGCGGGACCAAGCTGGAGAT CACCGGCGGAGGCGGGAGCGGCGGGGGCGGCTCCGGCGGAGGCGGGAGCGAGGTCAAGCTGCAGGAGTCTGGCCCC GGCCTGGTGGCCCCCAGCCAGAGCCTGAGCGTGACCTGCACCGTCTCCGGCGTGAGCCTGCCCGACTATGGCGTGA GCTGGATCAGGCAGCCACCCAGGAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGGTCTGAGACCACCTACTATAA CAGCGCCCTCAAGTCACGCCTGACCATCATCAAGGACAACAGCAAGTCCCAGGTGTTCCTGAAGATGAACAGTCTG CAGACAGACGACACCGCCATCTACTACTGTGCCAAGCACTACTACTATGGCGGGTCTTACGCCATGGACTACTGGG GGCAGGGGACGAGTGTGACTGTGTCCAGCACAACCACACCCGCCCCTAGGCCCCCAACACCCGCCCCCACTATCGC AAGCCAGCCCCTGAGTCTGAGGCCCGAGGCCTGCCGGCCCGCCGCCGGCGGCGCCGTGCACACCAGGGGCCTGGAC TTCGCTTGCGATATCTACATCTGGGCTCCTCTGGCCGGAACCTGCGGCGTGCTGCTGTTGTCCTTGGTGATCACTC TGTACTGCAAGAGGGGCCGCAAGAAGCTGCTGTACATCTTTAAGCAGCCTTTCATGCGGCCCGTGCAGACCACCCA GGAGGAGGACGGCTGCAGCTGCAGGTTCCCAGAGGAGGAGGAGGGCGGGTGTGAGCTGAGGGTGAAGTTCTCCAGG TCTGCCGACGCCCCCGCCTACCAGCAGGGCCAGAATCAGCTCTACAATGAGCTGAATCTGGGCCGCCGGGAGGAGT ACGATGTGCTCGACAAGCGGCGGGGGCGCGACCCAGAGATGGGCGGCAAGCCTAGGAGAAAGAACCCTCAGGAGGG GCT CTACAAT GAGCT GCAGAAGGACAAGATGGCT GAGGCGTATT CAGAGAT CGGGAT GAAAGGGGAGAGGAGGAGG GGCAAGGGGCACGACGGCCTGTATCAGGGGCTGAGCACCGCCACCAAGGACACCTATGACGCACTGCACATGCAGG CCCTGCCCCCGCGG ( SEQ ID NO: 141) ATGGCCCTGCCCGTGACCGCCCTGCTTCTGCCCCTGGCCCTGCTGCTCCACGCCGCCAGGCCAGACATCCAGATGA CCCAGACAACCTCCAGCCTGAGCGCCAGCCTGGGCGACAGAGTGACCATCAGTTGCCGGGCCAGCCAGGACATCAG CAAGTACCTGAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTGCTGATCTACCACACCAGCCGCCTGCAC AGCGGCGTGCCCAGCAGGTTCAGCGGCAGCGGCTCCGGCACCGACTACAGCCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGCAATACCCTGCCCTACACCTTTGGCGGCGGCACCAAGCTGGAGAT CACCGGCGGCGGCGGAAGCGGCGGCGGCGGCTCCGGCGGGGGCGGCAGCGAGGTGAAGCTGCAGGAGAGCGGCCCA GGCCTGGTGGCCCCCAGCCAGAGCCTGAGCGTGACCTGCACCGTGAGCGGCGTGTCCCTGCCTGACTACGGGGTGA GCTGGATCAGGCAGCCCCCCAGAAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA TTCCGCCCTGAAGTCCCGGCTGACCATCATCAAGGACAATTCCAAGAGCCAGGTGTTCCTGAAGATGAACTCCCTG CAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCTCCTACGCCATGGACTACTGGG GCCAGGGCACCTCCGTGACCGTGAGCTCTACCACCACCCCCGCCCCTCGGCCCCCAACACCTGCCCCCACCATCGC TAGCCAGCCCCTGTCTCTGAGACCCGAGGCTTGCAGACCAGCCGCCGGCGGCGCCGTGCACACCAGAGGCCTGGAC TTCGCCTGCGACATCTACATCTGGGCCCCCCTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGAGGGGAAGGAAGAAGCTGCTGTACATCTTTAAGCAGCCTTTCATGAGACCCGTCCAGACCACACA GGAGGAGGACGGCTGTAGCTGCAGGTTCCCAGAGGAGGAGGAGGGCGGCTGCGAGCTGCGGGTGAAGTTCAGCAGG TCCGCTGACGCCCCCGCCTACCAGCAGGGCCAGAACCAGCTGTACAATGAGCTGAACCTGGGCCGGAGAGAGGAGT ACGACGTGCTGGACAAGAGGAGAGGCAGGGACCCCGAGATGGGCGGCAAGCCTAGAAGGAAGAACCCCCAGGAGGG ACTGTACAACGAGCTGCAGAAGGACAAGATGGCCGAGGCCTACTCTGAGATCGGCATGAAGGGCGAGAGGAGAAGA GGCAAGGGCCACGATGGCCTGTACCAGGGCCTGAGCACCGCCACCAAGGACACCTACGACGCCCTGCACATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 142 ) ATGGCATTACCAGTTACCGCCCTCCTCCTCCCCCTCGCCCTCCTCCTCCACGCCGCCCGGCCAGATATCCAGATGA CCCAGACCACCTCCTCCCTCAGCGCCTCCCTCGGCGACCGGGTGACCATCTCTTGCCGGGCCAGCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCGGACGGCACCGTGAAGCTCCTGATCTACCACACCTCCCGCCTGCAC AGCGGGGTGCCCAGCAGGTTCAGCGGCAGCGGCAGCGGCACCGACTACTCCCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAACACCCTGCCCTACACCTTCGGCGGCGGCACCAAGCTGGAGAT CACCGGGGGCGGGGGCAGCGGGGGCGGGGGCAGCGGCGGGGGCGGCAGCGAGGTGAAGCTGCAGGAGAGCGGCCCA GGCCTGGTGGCCCCCAGCCAGAGCCTGTCCGTGACCTGCACCGTGAGCGGCGTGTCCCTGCCCGACTATGGCGTGA GCTGGATCCGCCAGCCACCCAGGAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGAGCCAGGTGTTCCTGAAGATGAACAGCCTGCAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGACTACTGGGATTORNEY DOCKET NO. ORB-019WO1GCCAGGGCACCTCCGTGACCGTGAGCAGCACCACCACCCCCGCCCCCCGCCCCCCCACCCCCGCCCCCACCATCGC CAGCCAGCCGCTGAGCCTGCGGCCGGAGGCCTGCAGGCCCGCCGCCGGAGGCGCCGTGCACACCCGGGGCCTGGAC TTCGCCTGCGACATCTACATCTGGGCCCCCCTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGCGGGGCAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGGCCCGTGCAGACCACCCA GGAGGAGGATGGCTGCAGCTGCAGGTTCCCTGAGGAGGAGGAGGGCGGCTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCCGACGCCCCCGCCTACCAGCAGGGCCAGAATCAGCTGTACAACGAGCTGAATCTGGGCCGGCGGGAGGAGT ACGACGTGCTGGACAAGCGCCGGGGCCGGGACCCCGAGATGGGCGGCAAGCCCCGGCGCAAGAACCCCCAGGAGGG CCTGTACAACGAGCTGCAGAAGGATAAGATGGCCGAGGCCTACAGCGAGATCGGCATGAAGGGCGAGCGGCGGAGG GGGAAGGGCCACGACGGCCTGTATCAGGGCCTGTCCACCGCCACCAAGGACACCTACGACGCCCTGCACATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 143) ATGGCATTACCCGTGACGGCCCTCCTACTCCCCTTAGCTCTCTTGCTCCACGCCGCCCGCCCGGACATCCAGATGA CCCAGACGACCTCGAGCTTGAGTGCGTCACTCGGGGACCGAGTGACGATCTCTTGCCGAGCGTCGCAGGACATCTC GAAGTACTTGAATTGGTACCAGCAGAAGCCCGACGGGACCGTTAAGCTATTAATCTACCACACGAGTAGATTGCAT AGCGGCGTTCCCAGTCGGTTCTCTGGTAGCGGTTCAGGTACTGACTACTCGCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGTAACACGCTTCCCTACACCTTCGGTGGGGGGACCAAGCTGGAGAT CACTGGTGGCGGAGGTAGCGGTGGCGGTGGTTCGGGCGGGGGCGGGAGCGAGGTGAAGCTGCAGGAGAGTGGGCCT GGCCTCGTCGCGCCGTCACAGAGCCTCTCTGTGACGTGCACCGTGAGCGGGGTCAGCCTGCCGGACTACGGAGTGT CCTGGATCCGGCAGCCGCCCCGCAAAGGCCTCGAGTGGCTCGGGGTCATTTGGGGGAGCGAGACAACGTACTACAA TAGTGCGTTGAAGTCGCGACTTACTATTATCAAGGATAATAGTAAGTCGCAGGTCTTTCTGAAGATGAACTCACTT CAGACTGACGACACCGCAATATACTATTGCGCCAAGCACTACTACTATGGAGGCAGTTACGCGATGGACTACTGGG GGCAGGGGACATCCGTGACTGTCTCCAGCACCACCACGCCGGCCCCGCGGCCCCCCACCCCCGCGCCGACGATCGC GTCTCAGCCCCTCAGCCTGAGGCCTGAGGCGTGTCGTCCAGCGGCGGGTGGGGCCGTGCACACACGGGGACTAGAC TTTGCATGCGACATCTACATCTGGGCCCCGCTGGCGGGGACCTGTGGAGTGTTGCTGCTGAGTCTAGTCATCACCT TGTACTGCAAGCGCGGCCGTAAGAAGCTCCTGTACATCTTCAAGCAGCCCTTCATGAGGCCTGTTCAAACGACGCA GGAGGAGGACGGGTGCAGCTGCCGCTTCCCTGAGGAGGAGGAGGGCGGCTGCGAGCTGAGAGTGAAATTCTCACGC TCGGCGGACGCCCCAGCTTATCAGCAGGGCCAGAACCAGCTGTACAATGAGCTGAACTTAGGCCGCAGGGAGGAGT ACGACGTGCTCGACAAGCGCCGGGGTCGTGACCCGGAGATGGGTGGCAAGCCCCGGCGCAAGAATCCGCAGGAGGG GCTCTACAACGAGCTCCAGAAGGACAAGATGGCGGAGGCCTACTCCGAGATTGGCATGAAGGGGGAGCGCCGTCGT GGTAAGGGCCACGACGGCCTGTACCAGGGCCTGTCGACGGCCACTAAGGACACTTACGACGCCCTGCACATGCAGG CGTTGCCGCCCCGG ( SEQ ID NO: 144 ) ATGGCATTACCAGTTACAGCCCTGTTGCTGCCCCTGGCCCTGCTGCTGCACGCCGCCCGGCCAGATATCCAGATGA CGCAGACCACCAGCTCCCTGTCAGCCTCCCTGGGGGACAGGGTGACCATCAGCTGCCGGGCCAGCCAGGACATCAG CAAGTACCTGAACTGGTACCAGCAGAAGCCTGATGGCACCGTGAAGCTGCTGATCTACCACACCTCCCGCCTGCAC AGCGGGGTGCCCAGCAGGTTCAGCGGCAGCGGGTCAGGCACCGACTACAGCCTGACCATCTCCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAACACCCTGCCCTACACCTTCGGCGGGGGCACCAAGCTGGAGAT CACTGGCGGCGGGGGGAGCGGGGGCGGGGGGTCTGGGGGCGGGGGTAGTGAGGTGAAGCTGCAGGAGAGCGGACCA GGCCTGGTGGCCCCCAGCCAGAGCCTGTCCGTGACCTGCACCGTGTCTGGCGTGAGCCTGCCAGACTACGGCGTGT CCTGGATCCGCCAGCCACCCAGGAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCGCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGAGCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGTGCGAAGCACTACTACTATGGCGGGTCCTACGCCATGGATTACTGGG GCCAGGGGACCTCAGTGACCGTGAGCAGCACCACTACCCCCGCCCCCAGACCCCCCACCCCCGCTCCCACCATCGC CAGCCAGCCTCTGAGCCTGCGGCCAGAGGCCTGCAGGCCTGCCGCCGGGGGGGCCGTGCACACCCGCGGGCTGGAC TTCGCCTGCGACATCTACATCTGGGCCCCCCTGGCCGGCACCTGCGGGGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGCGGGGCCGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCCGTGCAGACCACCCA GGAGGAGGACGGCTGCAGCTGCCGCTTCCCTGAGGAGGAGGAGGGCGGCTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCCGACGCCCCTGCGTACCAGCAGGGCCAGAATCAGCTGTACAACGAGCTGAATCTGGGCAGGCGGGAGGAGT ACGACGTGCTGGACAAGCGCCGGGGCCGGGACCCCGAGATGGGGGGCAAGCCCCGGCGCAAGAACCCCCAGGAGGG GCTGTACAACGAGCTGCAGAAGGATAAGATGGCCGAGGCCTATTCCGAGATCGGAATGAAGGGCGAGCGGCGGCGG GGCAAGGGGCACGACGGGCTGTACCAGGGCCTGAGCACCGCCACCAAGGACACCTACGATGCCCTGCACATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 145) ATGGCTCTACCAGTGACAGCCCTCCTCCTCCCACTCGCCCTCCTCTTGCACGCCGCGCGCCCAGATATACAGATGA CCCAGACGACGTCCAGCTTGAGTGCGTCACTCGGGGACCGAGTGACGATCTCTTGCCGGGCGTCGCAGGACATCTC CAAGTACCTCAATTGGTACCAGCAGAAGCCAGACGGGACGGTGAAGCTCCTGATCTATCACACGTCCCGTCTGCAT TCTGGTGTCCCGAGCCGGTTCTCCGGCTCGGGAAGTGGGACTGACTACAGTCTCACGATATCCAACCTGGAGCAGG AGGATATCGCAACGTATTTCTGCCAGCAGGGAAATACGTTGCCGTACACCTTCGGGGGAGGGACAAAGCTGGAGATCACTGGTGGGGGCGGGAGTGGGGGGGGGGGGAGCGGGGGAGGGGGCAGCGAGGTCAAGCTCCAGGAGTCTGGGCCGATTORNEY DOCKET NO. ORB-019WO1GGCCTCGTTGCCCCCTCCCAGTCACTCTCAGTCACTTGCACCGTTTCCGGGGTGTCGCTGCCGGACTACGGGGTGT CGTGGATCCGGCAGCCACCCCGGAAAGGGCTGGAGTGGCTGGGAGTGATCTGGGGGTCCGAGACTACGTACTACAA TAGTGCGTTGAAGTCTCGGCTCACCATAATAAAGGACAATAGTAAGTCGCAGGTCTTTCTGAAGATGAACAGTCTT CAGACGGACGACACTGCGATTTACTATTGTGCCAAGCACTATTATTATGGTGGGTCGTACGCCATGGATTACTGGG GTCAGGGGACCTCAGTGACCGTGAGCAGTACGACCACCCCCGCTCCCCGCCCCCCCACTCCCGCCCCCACCATAGC ATCCCAGCCTTTGTCCCTCCGCCCGGAGGCGTGCCGCCCCGCCGCGGGCGGGGCGGTGCACACGCGAGGGCTGGAC TTCGCGTGTGACATCTATATCTGGGCGCCGCTGGCGGGGACGTGCGGAGTTCTTCTGCTCTCGCTTGTAATCACGC TGTATTGCAAGCGAGGGCGGAAGAAGCTCCTGTACATCTTCAAGCAGCCGTTCATGCGGCCGGTCCAGACGACCCA AGAGGAGGACGGGTGCTCCTGCCGGTTCCCGGAGGAGGAGGAGGGGGGCTGTGAGCTGCGGGTCAAGTTCAGCCGT TCAGCTGATGCCCCTGCGTACCAGCAGGGGCAGAATCAGCTGTACAACGAGCTGAACTTGGGCCGTCGAGAGGAGT ACGACGTCCTCGACAAGCGTCGAGGACGTGATCCCGAGATGGGGGGTAAGCCCCGCCGTAAGAATCCTCAGGAGGG GCTCTACAACGAGCTCCAGAAGGATAAGATGGCGGAGGCTTACTCCGAAATCGGGATGAAGGGAGAGCGCCGTCGT GGCAAGGGCCACGACGGGCTCTACCAGGGCCTCTCGACGGCCACCAAGGACACCTACGACGCCCTGCACATGCAGG CGTTGCCGCCGCGG ( SEQ ID NO: 146) ATGGCCCTACCCGTGACCGCTCTGCTGCTACCCCTGGCTCTGTTACTGCACGCCGCCCGCCCTGACATTCAGATGA CTCAGACTACCAGCAGCCTGAGCGCCTCGTTGGGAGACCGCGTTACCATCTCCTGTCGCGCTAGCCAGGACATATC TAAATATCTCAATTGGTACCAACAGAAGCCTGACGGCACAGTTAAACTTCTCATTTATCATACCTCACGCCTCCAT TCCGGAGTGCCCAGTAGGTTCAGTGGAAGCGGCAGTGGAACCGACTATAGCCTCACTATTAGTAACTTGGAACAAG AGGACATAGCAACATATTTTTGCCAACAGGGGAATACCTTGCCATATACATTCGGTGGGGGAACTAAACTCGAGAT CACCGGCGGAGGTGGTAGCGGGGGAGGAGGATCAGGGGGAGGAGGTAGTGAGGTGAAGTTGCAGGAGTCAGGTCCG GGACTGGTCGCGCCATCCCAGTCACTTTCAGTGACATGTACAGTGAGTGGTGTTTCACTGCCCGACTACGGAGTCT CCTGGATCCGCCAGCCTCCCCGAAAGGGATTGGAGTGGCTTGGGGTAATATGGGGGAGCGAAACGACCTACTATAA CTCAGCCCTCAAAAGTAGGCTGACAATTATAAAGGATAACAGTAAAAGTCAAGTTTTCCTTAAAATGAACAGCCTG CAGACAGATGATACGGCCATCTACTACTGTGCGAAACATTACTACTACGGGGGCTCCTATGCCATGGATTATTGGG GACAGGGAACCTCAGTCACTGTGTCATCTACAACCACTCCGGCTCCACGGCCTCCTACACCCGCACCCACTATTGC CTCACAGCCTCTCAGCCTGAGACCTGAAGCCTGCAGACCCGCTGCGGGCGGCGCCGTACACACCCGAGGTCTGGAT TTTGCCTGTGACATTTATATCTGGGCCCCGTTGGCTGGAACATGCGGTGTATTATTGCTGTCCCTGGTGATAACTT TGTACTGTAAGCGGGGAAGGAAGAAGCTACTGTACATCTTCAAGCAGCCCTTTATGCGCCCTGTGCAAACAACTCA AGAGGAGGATGGTTGTTCGTGCAGGTTCCCAGAGGAGGAGGAAGGCGGGTGTGAGTTGCGTGTTAAGTTTTCGAGA TCAGCTGACGCCCCTGCTTACCAGCAAGGGCAGAACCAGCTCTACAACGAGCTGAATCTTGGACGTCGAGAAGAGT ACGATGTTTTGGATAAAAGGCGTGGGCGCGATCCAGAGATGGGCGGTAAACCAAGGCGTAAGAATCCTCAGGAGGG TCTCTATAACGAGTTGCAGAAGGATAAGATGGCTGAGGCTTACAGCGAAATCGGAATGAAAGGAGAGAGACGGCGA GGTAAAGGTCACGATGGTCTCTATCAGGGATTGAGCACAGCGACCAAGGACACGTACGACGCCCTTCACATGCAAG CGCTGCCTCCGAGG ( SEQ ID NO: 147 ) ATGGCTCTGCCAGTAACCGCGCTTCTACTGCCATTAGCCTTGCTGCTCCACGCAGCCCGACCCGACATTCAGATGA CTCAGACTACTAGCTCTCTGAGTGCATCGCTAGGTGATCGTGTTACGATTTCCTGCCGGGCCTCACAGGATATTTC TAAATACCTGAACTGGTACCAACAGAAGCCCGATGGGACGGTGAAGTTATTGATCTACCACACCAGTCGGTTGCAC TCCGGCGTCCCATCCAGGTTTTCCGGGTCAGGCAGCGGTACTGACTATAGCCTTACCATCAGCAACCTGGAACAGG AAGACATAGCCACCTACTTCTGCCAGCAAGGCAACACACTGCCCTACACCTTTGGTGGCGGAACCAAGCTGGAGAT TACTGGGGGCGGGGGTTCTGGTGGTGGTGGTTCGGGCGGGGGAGGGAGTGAGGTCAAGCTTCAGGAATCGGGACCT GGTCTAGTGGCTCCATCTCAGTCCCTGTCCGTGACTTGCACTGTGTCCGGCGTGTCGTTGCCGGACTACGGTGTCT CGTGGATCAGGCAACCCCCCCGCAAGGGCCTCGAGTGGCTAGGGGTGATTTGGGGCTCCGAAACTACCTACTACAA CTCCGCGCTCAAGTCTAGGCTGACGATCATCAAGGACAACTCTAAATCCCAAGTGTTCTTAAAGATGAACAGTCTG CAAACAGACGATACCGCGATATATTATTGCGCTAAGCATTATTATTATGGTGGCTCCTACGCCATGGATTACTGGG GTCAAGGCACTTCCGTTACAGTCAGTTCAACAACAACCCCAGCTCCACGTCCTCCAACACCAGCCCCGACCATCGC CTCTCAGCCACTGTCCCTCAGGCCTGAGGCCTGCCGGCCTGCGGCCGGAGGGGCAGTGCATACGCGGGGGTTGGAC TTTGCTTGCGACATTTATATCTGGGCGCCCCTGGCCGGTACATGTGGAGTGCTCCTTTTGTCCCTGGTGATCACCT TGTACTGCAAGAGAGGGAGAAAGAAGCTACTCTACATCTTTAAGCAGCCGTTCATGCGACCGGTCCAGACCACCCA GGAAGAGGACGGCTGCAGCTGCCGGTTTCCTGAAGAGGAAGAAGGCGGCTGCGAATTACGCGTGAAGTTTTCGCGC AGCGCCGATGCTCCCGCTTATCAGCAAGGCCAAAACCAGCTCTACAATGAGTTGAACCTGGGGCGACGTGAGGAGT ATGATGTTCTTGATAAGCGGCGGGGCCGAGACCCTGAGATGGGCGGGAAACCGCGACGTAAGAATCCTCAGGAGGG GCTGTACAACGAGCTCCAGAAAGATAAGATGGCGGAAGCGTATAGCGAGATCGGTATGAAGGGGGAGCGCCGGCGC GGGAAGGGTCACGATGGACTGTACCAGGGCCTCAGTACCGCGACAAAGGATACCTATGACGCGTTGCATATGCAGG CCCTGCCCCCAAGG ( SEQ ID NO: 148 )ATGGCCCTGCCTGTGACCGCCCTGCTGCTGCCACTGGCCCTGCTGCTGCACGCCGCCCGGCCAGATATCCAGATGACACAGACAACCTCCAGCCTGAGCGCCAGCCTGGGGGACAGAGTGACCATCAGCTGCCGGGCCTCCCAAGACATCAGATTORNEY DOCKET NO. ORB-019WO1CAAATACCTGAACTGGTACCAGCAGAAACCCGATGGCACCGTGAAGCTGCTGATCTACCACACTTCCAGGCTGCAC AGTGGGGTGCCCTCCCGGTTCTCCGGCAGCGGCAGCGGCACCGACTACAGCCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTTTGCCAGCAGGGCAACACCCTGCCCTACACCTTCGGCGGCGGCACCAAGCTGGAAAT CACCGGGGGCGGGGGATCTGGGGGCGGCGGCTCCGGCGGCGGAGGATCCGAGGTGAAGCTGCAGGAGTCCGGCCCA GGCCTGGTGGCCCCCAGCCAAAGTCTGTCCGTGACCTGTACCGTGTCCGGCGTGTCCCTGCCCGACTACGGCGTGA GCTGGATTCGCCAGCCCCCCCGGAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTATTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATTAAAGATAATAGCAAGAGCCAAGTGTTCCTCAAGATGAATAGCCTG CAGACCGACGATACCGCCATCTACTACTGTGCCAAACACTACTACTACGGCGGGAGCTACGCCATGGACTACTGGG GCCAAGGCACCTCCGTGACCGTGAGCAGCACCACCACCCCAGCCCCCAGGCCCCCTACCCCCGCTCCTACAATTGC CAGCCAGCCACTGTCCCTGCGCCCTGAGGCCTGCCGGCCCGCCGCCGGCGGCGCCGTGCATACCCGGGGCCTGGAC TTCGCCTGTGACATCTACATCTGGGCCCCCCTGGCTGGGACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACAC TGTACTGCAAAAGGGGCCGGAAAAAACTGCTGTACATCTTCAAGCAGCCCTTCATGAGACCCGTGCAGACCACCCA GGAGGAGGACGGATGCTCCTGCCGCTTCCCAGAGGAGGAGGAGGGCGGGTGCGAGCTGAGAGTGAAGTTTAGCCGC AGCGCCGATGCTCCCGCCTACCAGCAGGGCCAGAATCAGCTGTATAACGAACTCAACCTGGGCAGAAGAGAAGAGT ACGATGTGCTGGATAAAAGACGGGGCAGGGACCCCGAGATGGGCGGCAAGCCAAGAAGAAAGAATCCCCAGGAGGG CCTGTACAACGAGCTGCAGAAGGATAAGATGGCCGAAGCCTACAGCGAGATTGGCATGAAGGGAGAGCGGAGGCGC GGCAAGGGCCACGACGGCCTCTACCAGGGCCTGTCCACCGCCACCAAGGATACCTACGACGCTCTGCACATGCAAG CTCTGCCCCCCCGC ( SEQ ID NO: 149) ATGGCCCTGCCCGTCACCGCCCTGCTGCTGCCCCTGGCCCTGCTGCTGCACGCCGCCCGGCCCGATATCCAAATGA CACAGACCACCAGCTCCCTGAGCGCCAGCCTGGGCGACAGAGTGACCATCTCCTGCAGGGCTAGCCAGGACATCAG CAAGTATCTGAACTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTCCTGATCTATCACACCTCCAGGCTGCAC AGCGGCGTGCCCAGCAGATTCTCCGGCAGCGGCAGCGGCACCGACTACTCCCTGACCATTAGCAACCTGGAGCAAG AGGACATCGCCACCTACTTCTGCCAGCAGGGCAACACACTGCCCTATACCTTTGGCGGCGGCACCAAGCTGGAGAT CACCGGCGGCGGCGGCAGCGGCGGCGGCGGCAGCGGCGGCGGCGGCTCCGAGGTGAAACTGCAGGAAAGCGGCCCC GGCCTGGTGGCCCCCTCCCAGAGCCTGAGCGTGACCTGCACCGTGAGCGGCGTGAGCCTGCCTGACTACGGCGTGT CCTGGATTAGGCAACCCCCCAGGAAGGGACTGGAGTGGCTCGGCGTGATTTGGGGAAGCGAGACCACCTACTACAA CTCCGCCCTGAAGTCCAGACTCACCATCATTAAAGATAACAGCAAATCCCAAGTGTTCCTGAAGATGAACTCCCTG CAAACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGAGGCAGCTACGCCATGGACTATTGGG GCCAGGGCACCAGCGTGACCGTGAGCAGCACTACAACCCCCGCCCCCAGACCTCCTACCCCTGCCCCCACCATCGC CTCACAGCCTCTGAGCCTGAGACCCGAGGCCTGCAGGCCCGCCGCCGGCGGCGCCGTGCACACCCGGGGACTGGAC TTCGCCTGCGACATTTACATCTGGGCCCCCCTGGCAGGCACCTGCGGCGTGCTCCTGCTGAGCCTGGTGATTACCC TGTACTGCAAGAGAGGAAGAAAGAAACTGCTGTACATCTTCAAGCAGCCCTTCATGAGGCCCGTGCAGACCACCCA GGAAGAGGACGGCTGCTCCTGTAGGTTCCCCGAGGAGGAGGAGGGCGGCTGCGAGCTCAGAGTGAAGTTTAGCAGG AGCGCCGACGCCCCCGCTTACCAGCAAGGCCAGAACCAGCTGTACAATGAACTGAACCTGGGCAGACGGGAGGAAT ACGACGTGCTGGACAAAAGGAGAGGAAGAGACCCCGAGATGGGCGGAAAGCCCCGGCGCAAGAACCCACAGGAGGG ACTGTACAACGAGCTGCAAAAAGACAAGATGGCCGAAGCTTACAGCGAGATCGGAATGAAGGGCGAGAGGAGACGC GGCAAGGGACACGATGGCCTCTACCAGGGCCTGAGCACCGCCACCAAAGACACCTATGATGCCCTCCACATGCAAG CCCTGCCCCCTAGG ( SEQ ID NO: 150) ATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCCAGGCCGGACATCCAGATGA CACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAG TAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACAC TCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAG AAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGAT CACAGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAGGTGAAACTGCAGGAGTCAGGACCT GGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAA GCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAA TTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTG CAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGG GCCAAGGAACCTCAGTCACCGTCTCCTCAACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACCATCGC GTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGAC TTCGCCTGTGATATCTACATCTGGGCGCCCTTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTGGTTATCACCC TTTACTGCAAACGGGGCAGAAAGAAACTCCTGTATATATTCAAACAACCATTTATGAGACCAGTACAAACTACTCA AGAGGAAGATGGCTGTAGCTGCCGATTTCCAGAAGAAGAAGAAGGAGGATGTGAACTGAGAGTGAAGTTCAGCAGG AGCGCAGACGCCCCCGCGTACCAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGT ACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGATTORNEY DOCKET NO. ORB-019WO1GGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGG CCCTGCCCCCTCGC ( SEQ ID NO: 151) ATGGCACTTCCCGTTACCGCCCTCCTCCTTCCACTCGCCCTCCTCCTCCACGCCGCCCGCCCAGATATACAGATGA CACAGACCACCTCCAGCTTGAGTGCCTCCCTCGGGGACAGGGTGACCATCTCTTGTCGGGCTTCTCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGGACCGTGAAGCTCCTGATCTACCACACCAGCAGGCTGCAC AGCGGCGTGCCCTCCAGGTTTAGCGGGTCAGGGAGTGGGACCGACTACTCCCTGACCATCTCAAACCTGGAGCAGG AGGACATCGCCACCTACTTTTGCCAGCAGGGCAACACCCTGCCCTACACCTTCGGGGGAGGCACTAAGCTGGAGAT CACTGGCGGTGGTGGGAGCGGGGGTGGGGGGTCTGGGGGCGGGGGTAGCGAGGTGAAGCTGCAGGAGTCCGGACCC GGCCTGGTCGCCCCCAGCCAGTCCCTGTCCGTGACATGCACCGTGTCTGGCGTGAGCCTGCCAGACTACGGCGTGT CATGGATCAGGCAGCCCCCCCGGAAGGGGCTGGAGTGGCTGGGGGTGATCTGGGGGTCCGAGACTACCTACTACAA CAGCGCGCTGAAGTCCAGGCTGACCATCATCAAGGACAACTCCAAGAGTCAGGTGTTCTTGAAGATGAACAGCCTG CAGACCGATGACACCGCCATATACTACTGTGCGAAGCACTACTACTATGGCGGGTCATACGCCATGGATTACTGGG GCCAGGGGACCTCAGTGACCGTGAGCAGCACCACCACCCCCGCCCCCAGACCCCCCACCCCCGCTCCCACCATCGC CAGCCAGCCCCTGAGCCTGCGCCCCGAGGCCTGCCGGCCCGCGGCCGGCGGCGCCGTGCACACTCGGGGGCTGGAC TTCGCCTGTGACATCTATATCTGGGCCCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGTCTGGTGATCACCC TGTACTGCAAGCGCGGAAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCAGTGCAGACCACTCA GGAGGAGGACGGCTGCAGCTGCCGGTTCCCAGAGGAGGAGGAGGGCGGGTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCCGATGCCCCTGCGTACCAGCAGGGGCAGAATCAGCTCTACAACGAGCTGAACCTGGGCCGCCGCGAGGAGT ACGACGTGCTCGACAAGCGGCGGGGCCGGGATCCCGAGATGGGGGGTAAGCCTCGGCGGAAGAATCCTCAGGAGGG GCTCTACAACGAGCTCCAGAAGGATAAGATGGCCGAGGCTTACTCCGAGATCGGGATGAAGGGGGAGCGGCGGCGG GGCAAGGGGCACGACGGCCTGTATCAGGGCCTGAGTACCGCCACCAAGGATACTTACGACGCCCTGCACATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 152 ) ATGGCCCTACCAGTCACCGCCCTCCTCCTCCCCTTGGCCCTCCTCCTCCACGCCGCCCGGCCAGATATTCAGATGA CACAGACCACCTCCAGCCTTAGTGCCTCCCTCGGAGATAGGGTGACCATTTCTTGTCGGGCTTCTCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGGACAGTGAAGTTGCTGATCTACCACACTAGCAGGCTGCAT AGTGGCGTGCCCTCCCGCTTCTCTGGAAGCGGGAGCGGCACCGACTACAGCCTGACTATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAATACCCTGCCCTATACCTTCGGGGGAGGCACTAAGCTGGAGAT TACTGGCGGAGGCGGCAGTGGAGGAGGAGGGTCAGGGGGAGGAGGATCTGAAGTGAAACTTCAGGAGAGTGGGCCC GGCCTTGTTGCTCCTTCTCAGAGCCTGTCCGTGACATGTACCGTGTCTGGCGTGTCTCTGCCAGACTACGGCGTGT CATGGATCAGGCAGCCTCCTAGGAAGGGGCTGGAGTGGCTGGGCGTGATCTGGGGTAGTGAGACTACTTACTACAA TAGTGCCCTGAAGTCCCGGCTGACTATTATCAAGGATAATAGCAAGTCTCAGGTGTTCCTGAAGATGAACAGCCTG CAGACAGATGACACGGCCATCTACTACTGTGCGAAGCACTACTACTATGGCGGCAGCTACGCCATGGATTACTGGG GCCAGGGCACTAGTGTGACAGTGAGTAGTACCACTACCCCAGCTCCACGCCCACCTACTCCAGCCCCTACAATTGC CTCCCAGCCTCTGTCTCTGCGCCCTGAAGCTTGTCGCCCTGCTGCTGGTGGAGCTGTGCACACCAGAGGGCTGGAC TTCGCCTGCGACATCTATATCTGGGCTCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGTCTGGTGATCACCC TGTACTGCAAGCGCGGAAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCAGTGCAGACCACTCA GGAGGAGGACGGCTGCAGCTGCCGGTTTCCAGAGGAGGAAGAGGGTGGATGTGAGCTGCGGGTGAAGTTCAGCCGC TCTGCTGATGCACCAGCCTACCAGCAGGGCCAGAATCAGCTCTACAACGAGCTGAATCTGGGCCGCAGGGAGGAGT ACGACGTGCTCGACAAGCGCAGGGGCAGAGATCCAGAGATGGGAGGCAAACCTAGGAGGAAGAATCCCCAGGAGGG GCTCTACAACGAGCTGCAGAAGGACAAGATGGCCGAGGCCTACTCTGAGATCGGAATGAAGGGCGAGAGGAGGCGG GGAAAAGGCCACGATGGCCTGTATCAGGGGCTGTCCACCGCCACCAAGGACACCTATGATGCCCTGCACATGCAGG CCCTGCCTCCTCGG ( SEQ ID NO: 153) ATGGCTCTCCCAGTGACCGCCCTCCTCCTTCCCCTTGCCCTCCTCTTGCACGCCGCTAGACCCGACATCCAGATGA CTCAGACTACCTCTAGTTTGAGTGCATCCTTGGGTGATCGGGTCACCATCTCTTGTCGGGCTTCTCAGGATATCAG TAAGTACCTTAATTGGTATCAGCAGAAGCCCGACGGGACCGTGAAGCTCTTGATCTACCATACATCAAGGCTTCAC AGCGGCGTGCCCTCCAGGTTTAGCGGGTCAGGGAGTGGGACCGACTACTCCCTGACCATCTCAAACCTGGAGCAGG AGGACATCGCTACTTACTTCTGCCAGCAGGGTAACACCCTGCCCTACACCTTTGGGGGAGGGACAAAGCTGGAGAT CACTGGTGGGGGCGGGAGTGGGGGGGGGGGGAGCGGGGGTGGGGGCAGCGAGGTCAAGCTCCAGGAGTCTGGGCCT GGCCTCGTTGCCCCATCCCAGAGTCTGTCAGTGACATGCACTGTGAGTGGCGTGTCACTGCCAGACTATGGGGTGT CCTGGATCCGCCAGCCACCCAGGAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGGAGTGAGACCACCTACTATAA TAGTGCTTTGAAGTCTAGACTTACTATTATCAAGGATAATAGTAAGTCTCAGGTCTTTCTGAAGATGAACAGTCTT CAGACAGACGACACCGCAATATACTATTGCGCAAAGCACTATTATTATGGGGGGTCTTACGCCATGGACTACTGGG GTCAGGGAACGTCCGTGACCGTCAGTAGTACCACCACCCCCGCTCCCCGCCCCCCCACTCCCGCCCCCACCATTGC ATCCCAGCCTTTGTCCCTCCGCCCAGAGGCCTGTAGGCCAGCCGCCGGGGGTGCGGTGCACACCCGGGGGCTGGAC TTCGCATGTGACATCTACATATGGGCTCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGTCTGGTGATCACCCTGTACTGCAAGCGCGGAAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCAGTGCAGACCACTCAATTORNEY DOCKET NO. ORB-019WO1GGAGGAGGACGGCTGCAGCTGCCGGTTCCCAGAGGAGGAGGAGGGGGGGTGTGAGCTGAGGGTGAAGTTTTCACGC TCAGCTGACGCCCCCGCCTACCAGCAGGGCCAGAATCAGCTCTACAACGAGCTGAATCTGGGCCGGCGGGAGGAGT ACGATGTCCTCGACAAGAGGAGGGGGAGGGATCCCGAGATGGGGGGTAAGCCTCGGCGGAAGAATCCTCAGGAGGG GCTCTACAACGAGCTCCAGAAGGATAAGATGGCCGAGGCTTACTCCGAGATCGGGATGAAGGGGGAGCGGCGGAGG GGGAAGGGCCACGATGGACTGTATCAGGGGTTGTCCACTGCCACAAAGGACACCTATGATGCACTCCATATGCAGG CCCTGCCCCCTCGG ( SEQ ID NO: 154 ) ATGGCTCTCCCAGTTACAGCCCTCTTGTTGCCTTTGGCCTTGCTCTTGCACGCCGCAAGGCCCGATATCCAGATGA CCCAGACCACCAGTTCCCTAAGCGCCAGCTTGGGGGACCGGGTGACCATCTCTTGTCGGGCCAGTCAGGATATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCGGACGGCACCGTGAAGTTGCTGATCTATCACACCAGTAGGCTGCAC TCCGGCGTGCCCTCCCGGTTCTCCGGGAGCGGCAGCGGGACAGACTACAGTCTGACCATCAGCAACCTGGAGCAGG AGGACATTGCCACCTACTTCTGCCAGCAGGGGAACACCCTGCCCTATACATTTGGCGGGGGCACCAAGCTGGAGAT TACAGGCGGGGGAGGCTCTGGCGGCGGGGGCAGCGGGGGGGGCGGCTCTGAGGTGAAGCTTCAGGAGTCCGGCCCC GGCCTCGTTGCCCCCAGCCAGAGCCTCTCCGTGACCTGTACCGTCTCCGGCGTGTCCCTGCCAGATTATGGGGTGT CCTGGATCCGCCAGCCACCCCGGAAGGGGCTGGAGTGGCTGGGCGTGATCTGGGGCTCAGAGACAACCTACTACAA CTCTGCCCTGAAGAGTAGACTTACTATTATCAAGGATAATAGTAAGTCTCAGGTGTTTCTGAAGATGAATAGCCTG CAGACAGATGACACGGCCATCTACTACTGCGCTAAGCACTACTACTATGGCGGCAGCTATGCCATGGACTACTGGG GCCAGGGGACTAGCGTGACAGTGTCCTCCACCACCACCCCTGCTCCCAGGCCCCCCACGCCTGCCCCCACCATCGC GAGCCAGCCACTGTCCCTCCGGCCAGAGGCCTGCCGGCCTGCTGCCGGAGGGGCAGTGCACACAAGAGGGCTGGAT TTCGCCTGTGACATTTACATCTGGGCTCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGCCTGGTGATCACCC TCTACTGCAAGAGGGGCAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCTTTCATGAGGCCCGTGCAGACCACCCA GGAGGAGGACGGCTGCAGCTGCCGGTTCCCAGAGGAGGAGGAGGGCGGCTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCAGACGCCCCCGCCTACCAGCAGGGCCAGAATCAGCTGTACAACGAGCTGAATCTGGGCCGGCGGGAGGAGT ACGACGTGCTGGACAAGCGCCGGGGCCGGGACCCCGAGATGGGGGGCAAGCCCCGGCGCAAGAATCCTCAGGAGGG GCTGTACAACGAGCTCCAGAAGGATAAGATGGCGGAGGCCTACAGCGAGATCGGGATGAAGGGCGAGCGCAGGCGG GGCAAGGGGCATGACGGGCTGTACCAGGGCCTGTCAACTGCCACCAAGGACACCTACGACGCCCTGCACATGCAGG CCCTGCCTCCCCGG ( SEQ ID NO: 155) ATGGCCTTACCAGTTACGGCGCTATTGCTCCCGTTGGCGCTATTGCTCCACGCCGCGCGCCCGGACATACAGATGA CGCAGACGACGTCTTCGTTATCGGCGAGTTTGGGCGACCGGGTTACGATATCGTGCAGGGCTTCGCAGGACATAAG TAAGTACCTTAATTGGTACCAGCAGAAGCCCGACGGTACGGTAAAACTACTCATCTATCATACGTCGCGTTTGCAT AGTGGAGTACCCTCCCGTTTTTCGGGGAGTGGGTCGGGTACCGACTATTCGCTAACGATATCGAACTTGGAGCAAG AAGATATCGCGACGTACTTTTGCCAGCAAGGGAATACGTTACCGTACACGTTTGGTGGAGGTACGAAACTAGAGAT CACGGGTGGTGGTGGTTCGGGTGGTGGTGGTTCGGGTGGGGGCGGCTCGGAAGTGAAACTTCAAGAGTCGGGTCCC GGTCTAGTGGCGCCGAGCCAATCGCTATCGGTAACGTGTACGGTTTCGGGTGTATCGCTACCGGATTACGGTGTAT CGTGGATACGCCAACCGCCGCGTAAGGGCCTAGAATGGCTAGGGGTTATATGGGGTAGTGAGACTACCTATTATAA CTCGGCGTTGAAATCGCGTCTAACGATAATAAAAGATAATTCAAAATCGCAGGTTTTTCTAAAAATGAATTCGCTA CAAACGGATGATACGGCGATATATTATTGTGCGAAGCACTATTATTATGGTGGTTCCTATGCGATGGATTATTGGG GTCAAGGTACGTCGGTAACCGTGTCGTCGACGACGACGCCGGCGCCGCGCCCGCCGACGCCGGCGCCGACGATAGC GTCGCAACCGCTATCGTTGCGGCCGGAGGCGTGCCGCCCGGCGGCGGGCGGGGCGGTGCATACGCGCGGCCTCGAT TTTGCGTGTGACATTTACATATGGGCGCCGCTAGCGGGCACGTGTGGTGTCCTACTACTATCGTTGGTGATCACCC TATATTGTAAGCGTGGCAGGAAGAAGCTTCTTTATATATTTAAACAGCCGTTTATGCGGCCGGTACAAACGACGCA AGAAGAAGATGGTTGTTCGTGCCGCTTCCCGGAAGAAGAAGAAGGTGGCTGTGAGCTACGCGTAAAGTTTTCGCGT TCGGCGGATGCTCCGGCGTATCAACAAGGTCAAAATCAGTTGTATAACGAACTGAATCTAGGGCGTCGTGAAGAAT ACGACGTCCTAGATAAACGTCGTGGTCGGGACCCGGAAATGGGTGGTAAGCCGCGTCGTAAAAACCCGCAAGAGGG GTTATATAATGAACTCCAAAAGGATAAAATGGCGGAGGCGTATTCGGAGATCGGGATGAAAGGCGAGCGTCGCCGT GGTAAGGGCCACGACGGCCTTTATCAAGGTCTCTCGACGGCGACGAAGGATACGTATGATGCGTTGCATATGCAAG CGTTGCCGCCGCGT ( SEQ ID NO: 156) ATGGCACTACCGGTAACGGCGCTACTACTACCGCTAGCGTTACTATTGCATGCAGCACGTCCGGATATACAAATGA CGCAAACAACTTCGAGTTTATCGGCATCTTTAGGAGACCGTGTAACCATATCGTGTCGAGCGTCTCAAGACATATC TAAGTATTTAAATTGGTATCAACAAAAACCTGACGGCACGGTAAAATTACTAATATACCATACGTCGCGACTCCAT TCGGGTGTACCATCGCGTTTTTCGGGATCGGGTTCGGGTACTGATTACTCGCTTACTATATCGAATTTAGAACAAG AAGATATCGCGACTTATTTTTGCCAACAAGGGAACACGCTACCTTATACCTTTGGTGGAGGGACCAAATTGGAAAT AACGGGGGGGGGCGGGAGTGGAGGAGGAGGTTCAGGGGGGGGGGGTTCAGAAGTTAAATTACAAGAATCAGGACCA GGATTAGTAGCGCCGTCGCAATCGTTATCAGTTACGTGTACGGTCTCGGGTGTAAGTCTACCGGATTACGGAGTAT CGTGGATTAGGCAACCCCCGCGCAAGGGGCTTGAATGGCTTGGCGTTATCTGGGGCTCGGAAACTACGTATTATAA CTCGGCTCTAAAATCACGTTTGACGATAATAAAAGACAATTCGAAATCGCAAGTATTCTTAAAAATGAACAGCCTACAAACGGACGACACGGCGATATACTATTGTGCGAAACATTATTATTATGGTGGTAGTTATGCGATGGATTACTGGGATTORNEY DOCKET NO. ORB-019WO1GCCAAGGAACAAGCGTTACCGTATCGTCCACTACGACGCCTGCGCCGCGACCGCCCACGCCGGCGCCTACCATAGC TTCACAACCATTAAGCCTCCGTCCAGAGGCGTGCCGTCCTGCGGCGGGTGGTGCTGTACATACGAGGGGTCTAGAC TTTGCGTGTGATATTTATATATGGGCTCCATTAGCGGGGACATGCGGGGTTCTACTACTATCGCTAGTAATAACTC TATACTGCAAACGTGGACGTAAAAAGTTACTATATATATTTAAACAGCCGTTCATGAGACCGGTACAAACTACGCA AGAGGAAGATGGGTGCTCCTGTAGATTTCCGGAGGAAGAAGAAGGTGGTTGCGAATTAAGAGTTAAATTCAGTCGA AGTGCGGACGCGCCAGCGTATCAACAAGGGCAAAATCAACTATACAATGAACTAAATCTTGGTCGAAGAGAAGAAT ATGACGTATTAGACAAACGGCGTGGACGAGACCCTGAGATGGGTGGAAAACCGCGAAGAAAAAATCCACAAGAAGG TTTATACAATGAATTACAAAAAGATAAAATGGCGGAGGCATATAGTGAAATAGGTATGAAAGGAGAACGAAGACGT GGTAAAGGGCATGACGGATTATATCAAGGATTGTCGACGGCAACGAAAGATACTTACGACGCGTTACATATGCAGG CCCTACCGCCGCGA ( SEQ ID NO: 157 ) ATGGCCTTACCAGTAACAGCCTTATTACTTCCACTTGCCCTCCTTCTACATGCCGCTAGACCAGATATCCAAATGA CACAAACGACCTCAAGCCTTTCCGCGTCTCTCGGTGACAGGGTAACTATTAGTTGTCGGGCCTCCCAAGATATTTC AAAATACTTAAATTGGTATCAACAGAAGCCAGATGGTACCGTGAAGCTCCTCATCTACCACACTAGCCGTCTTCAC TCTGGAGTACCAAGCAGGTTTTCCGGCTCCGGAAGTGGTACTGACTACAGTTTGACTATAAGCAACCTAGAGCAAG AGGATATTGCCACATACTTCTGTCAACAAGGAAACACTCTGCCATATACATTCGGTGGTGGTACAAAACTGGAAAT AACTGGAGGGGGAGGTTCAGGCGGCGGCGGATCTGGGGGGGGTGGCAGTGAGGTCAAATTGCAGGAGTCTGGACCC GGTCTGGTTGCTCCATCCCAATCACTTTCAGTGACCTGCACTGTATCCGGAGTTTCTCTGCCAGATTATGGAGTGA GTTGGATCCGCCAGCCTCCACGCAAGGGCCTTGAATGGCTCGGGGTTATATGGGGTAGTGAAACTACATACTATAA TAGCGCTCTGAAATCTCGACTCACAATCATTAAGGATAATAGCAAAAGCCAGGTGTTTTTGAAGATGAATAGCCTG CAAACCGATGATACCGCCATATATTACTGTGCCAAACATTACTATTACGGAGGCTCCTACGCTATGGATTATTGGG GACAGGGGACAAGCGTGACCGTTAGCTCTACTACTACCCCCGCCCCTCGCCCTCCAACTCCAGCTCCCACCATTGC CAGTCAACCACTATCCTTAAGACCCGAGGCATGCAGACCCGCCGCAGGGGGCGCCGTACATACCCGAGGTTTGGAT TTTGCCTGCGACATCTATATCTGGGCCCCCCTTGCTGGGACATGTGGAGTGTTGCTACTCTCGTTGGTTATCACTC TGTATTGCAAGAGAGGAAGAAAGAAACTATTATATATTTTTAAGCAGCCATTCATGAGACCGGTGCAGACTACCCA AGAAGAAGACGGATGTTCATGTAGGTTCCCTGAAGAAGAGGAAGGAGGCTGCGAACTGCGAGTGAAATTTTCCAGA TCTGCCGATGCACCAGCTTATCAACAAGGACAAAACCAGCTATATAATGAACTTAATTTAGGGAGACGAGAGGAGT ACGACGTCCTGGACAAACGCCGCGGAAGAGACCCTGAGATGGGAGGTAAGCCCAGAAGAAAAAACCCACAAGAAGG ACTGTACAATGAACTCCAAAAAGACAAAATGGCTGAAGCGTATTCCGAAATTGGCATGAAAGGAGAACGGAGAAGA GGTAAGGGTCACGACGGCTTATATCAGGGACTCTCTACAGCAACCAAAGACACATATGATGCACTACACATGCAGG CACTACCACCCAGG ( SEQ ID NO: 158 ) ATGGCTCTACCAGTCACAGCACTACTATTGCCGTTGGCTCTATTGCTTCACGCGGCGCGACCGGATATACAGATGA CCCAAACGACGTCGAGTCTCTCGGCGTCGTTGGGTGATCGTGTAACGATATCGTGTCGTGCGTCGCAAGATATCTC GAAATATCTAAATTGGTATCAACAAAAACCGGACGGGACGGTGAAATTATTAATATATCACACGTCCCGTCTACAT TCTGGTGTTCCGAGTCGATTTAGTGGGAGTGGTTCGGGTACGGACTACTCCCTAACTATATCGAACTTGGAACAAG AGGATATAGCGACGTACTTCTGCCAACAAGGTAATACGCTACCGTATACCTTTGGCGGAGGTACGAAGCTAGAAAT AACTGGTGGGGGTGGGAGTGGTGGTGGTGGTTCGGGTGGTGGTGGTTCTGAAGTGAAACTTCAAGAATCGGGACCG GGGCTGGTAGCCCCGTCCCAATCGTTATCGGTAACGTGCACCGTATCGGGTGTATCGTTACCGGATTATGGTGTAT CGTGGATACGCCAACCGCCCCGTAAAGGCCTAGAATGGCTAGGTGTTATATGGGGTTCGGAAACGACGTACTACAA TAGTGCGTTGAAATCGCGACTTACTATTATCAAGGATAATAGTAAGTCGCAAGTATTTTTAAAGATGAATTCGTTA CAGACGGACGATACCGCGATATACTATTGCGCGAAGCACTATTATTATGGTGGTTCGTATGCGATGGATTATTGGG GTCAAGGTACGTCTGTAACGGTTTCATCTACTACTACTCCGGCACCACGACCACCAACTCCTGCTCCTACTATAGC GTCGCAACCGTTATCGTTGCGACCTGAAGCATGTAGACCAGCGGCAGGTGGTGCTGTTCATACGCGAGGTCTAGAC TTCGCGTGTGATATATATATATGGGCCCCGCTTGCGGGTACGTGTGGTGTACTATTACTATCGTTAGTAATAACGC TATACTGCAAGCGGGGCCGAAAAAAATTATTATATATATTTAAACAACCGTTTATGCGGCCGGTACAAACGACGCA AGAGGAAGATGGGTGTTCTTGTCGTTTTCCTGAAGAGGAAGAAGGTGGATGCGAACTACGTGTAAAATTTTCGCGT AGTGCGGATGCGCCGGCCTATCAACAAGGCCAAAATCAACTCTATAACGAGTTGAATCTAGGACGTCGTGAAGAGT ACGACGTCCTAGATAAACGACGTGGTCGTGACCCGGAAATGGGTGGCAAACCGCGTCGTAAAAACCCGCAAGAGGG TTTATATAATGAGCTACAAAAAGATAAAATGGCTGAAGCGTACTCGGAAATTGGTATGAAAGGTGAACGTCGTCGT GGTAAAGGCCATGACGGTTTATATCAAGGTCTGAGTACGGCGACGAAGGACACCTATGATGCTCTACATATGCAAG CGTTACCGCCGCGT ( SEQ ID NO: 159) ATGGCTCTACCCGTCACGGCGTTATTACTTCCGCTCGCGTTGCTATTACACGCAGCGCGACCGGATATACAAATGA CGCAAACGACGTCGTCGTTATCGGCGTCGCTAGGAGATCGTGTGACGATCTCTTGCCGAGCGTCGCAAGATATCTC GAAATATCTAAATTGGTATCAACAAAAACCGGACGGTACGGTTAAATTGTTGATATATCATACGTCGAGGCTACAT TCTGGTGTTCCGAGTCGATTTAGTGGGAGTGGTTCGGGTACGGACTACTCCCTAACTATATCGAACTTGGAACAAG AGGATATAGCCACGTATTTTTGCCAACAGGGCAATACCCTCCCCTACACGTTTGGGGGAGGGACGAAACTAGAAATTACAGGTGGTGGTGGTTCTGGGGGTGGTGGATCAGGTGGTGGTGGTTCTGAAGTGAAACTTCAAGAATCAGGACCGATTORNEY DOCKET NO. ORB-019WO1GGTCTAGTCGCCCCCAGTCAATCGCTATCGGTAACCTGTACGGTTTCTGGTGTCAGCTTACCGGATTATGGTGTAA GCTGGATACGTCAACCGCCCCGTAAAGGCCTAGAATGGCTAGGTGTTATATGGGGTTCGGAGACGACGTACTACAA TAGTGCGTTAAAATCGCGACTTACTATTATCAAAGATAATAGTAAGTCGCAGGTTTTTTTAAAGATGAACTCTTTA CAAACGGATGATACGGCGATATACTATTGCGCGAAGCACTATTATTATGGTGGTTCGTATGCGATGGATTACTGGG GGCAGGGTACGTCGGTAACTGTGAGTTCGACTACGACTCCTGCTCCTAGACCGCCGACTCCGGCGCCAACTATAGC ATCGCAGCCTCTATCGCTTCGTCCGGAAGCGTGTAGACCAGCTGCAGGTGGTGCTGTACATACGCGAGGTCTAGAC TTCGCGTGTGATATATATATATGGGCGCCCCTAGCGGGTACGTGTGGGGTGCTATTACTATCGTTAGTAATAACGC TATATTGCAAACGTGGTAGGAAAAAGCTACTATATATATTTAAACAACCTTTTATGCGTCCGGTACAAACGACGCA AGAAGAGGATGGTTGTAGTTGTCGATTTCCGGAAGAAGAAGAGGGAGGTTGCGAACTACGAGTAAAATTTAGTCGA TCGGCGGATGCGCCGGCGTATCAACAAGGTCAAAATCAATTATATAATGAATTAAATCTAGGACGTCGTGAAGAAT ATGACGTCCTAGATAAACGACGTGGTCGTGACCCAGAAATGGGTGGTAAACCGCGGCGTAAAAACCCGCAAGAAGG TTTATATAATGAGCTTCAAAAAGATAAAATGGCGGAAGCTTATTCGGAAATAGGTATGAAAGGTGAACGCCGTCGT GGTAAAGGGCATGACGGTCTTTACCAAGGTTTGTCGACGGCGACGAAAGATACGTATGATGCGTTACACATGCAAG CGTTACCGCCGCGT ( SEQ ID NO: 160) ATGGCATTACCCGTTACCGCCCTCCTCTTGCCCCTCGCCCTCTTGCTCCACGCCGCCCGGCCAGATATCCAGATGA CCCAGACCACCTCCTCCCTCAGCGCCTCCCTCGGCGACCGGGTGACCATCTCTTGCCGGGCCTCACAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTTCTGATCTACCACACCTCCCGGCTGCAC AGCGGGGTGCCCAGCAGGTTCAGCGGCAGCGGGTCAGGCACCGACTACAGCCTGACCATCTCCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGGAACACCCTGCCCTACACCTTCGGCGGGGGCACCAAGCTGGAGAT CACTGGCGGCGGGGGGAGCGGGGGCGGGGGGTCTGGGGGCGGGGGTAGTGAGGTGAAGCTGCAGGAGAGCGGACCA GGCCTGGTGGCCCCCAGCCAGAGCCTGTCCGTGACCTGCACCGTGTCTGGCGTGAGCCTGCCAGACTACGGCGTGT CCTGGATCCGCCAGCCACCCAGGAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCGCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGAGCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGTGCGAAGCACTACTACTATGGCGGGTCCTACGCCATGGATTACTGGG GCCAGGGGACCTCAGTGACCGTGAGCAGCACCACTACCCCCGCCCCCAGACCCCCCACCCCCGCTCCCACCATCGC CAGCCAGCCTCTGAGCCTGCGGCCAGAGGCCTGCAGGCCTGCCGCCGGGGGGGCCGTGCACACCCGCGGGCTGGAC TTCGCCTGCGACATCTACATCTGGGCCCCCCTGGCCGGCACCTGCGGGGTGCTGCTGCTGAGCCTGGTGATCACCC TGTACTGCAAGCGCGGGAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGAGGCCCGTGCAGACCACCCA GGAGGAGGATGGCTGCAGCTGCAGGTTCCCTGAGGAGGAGGAGGGCGGCTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCAGACGCCCCCGCCTACCAGCAGGGCCAGAATCAGCTGTACAACGAGCTGAATCTGGGCCGGCGGGAGGAGT ACGACGTGCTGGACAAGCGCCGGGGCCGGGACCCCGAGATGGGGGGCAAGCCCCGGCGCAAGAATCCTCAGGAGGG CCTGTACAACGAGCTGCAGAAGGATAAGATGGCCGAGGCCTACAGCGAGATCGGGATGAAGGGCGAGAGGAGGCGC GGCAAGGGGCACGACGGCCTGTATCAGGGCCTGTCCACCGCCACCAAGGACACCTACGACGCCCTGCACATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 161) ATGGCTCTACCAGTTACAGCCCTCTTGCTCCCTTTGGCCTTGTTGCTCCACGCAGCAAGGCCAGACATCCAGATGA CTCAGACTACCTCTAGTTTGAGTGCATCCTTGGGAGACCGGGTGACTATCAGTTGCCGGGCCTCCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCGGACGGCACCGTGAAGTTGCTGATCTACCACACCAGCAGGCTGCAC AGCGGCGTGCCCTCCCGGTTCTCCGGGAGCGGGTCAGGGACCGACTACTCCCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCTACTTACTTCTGCCAGCAGGGGAACACCCTGCCCTACACCTTTGGCGGAGGGACAAAGCTGGAGAT CACTGGTGGGGGCGGGAGTGGGGGGGGGGGGAGCGGGGGTGGGGGGTCTGAGGTGAAGCTGCAGGAGTCCGGCCCC GGGCTGGTCGCCCCCTCACAGAGCCTGTCGGTGACATGCACTGTGAGCGGGGTGAGCCTGCCCGACTACGGGGTGA GCTGGATCCGCCAGCCCCCCCGGAAGGGCCTGGAGTGGCTCGGAGTGATCTGGGGCTCCGAGACCACTTACTACAA CAGCGCCCTGAAGTCCAGGTTGACTATTATCAAGGATAATAGCAAGTCTCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGATGACACCGCCATATACTACTGTGCGAAGCACTACTACTATGGCGGGTCATACGCCATGGACTACTGGG GCCAGGGCACCTCAGTGACCGTCAGCTCCACCACCACCCCCGCTCCCCGCCCCCCCACTCCCGCCCCCACCATCGC ATCCCAGCCCCTGTCCCTCCGGCCAGAGGCCTGCCGGCCCGCCGCGGGCGGCGCCGTGCACACCCGCGGGCTGGAC TTCGCCTGCGACATCTACATCTGGGCTCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGCCTGGTGATCACTC TGTACTGCAAGCGCGGAAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCAGTGCAGACCACCCA GGAGGAGGACGGCTGCAGCTGCCGGTTCCCAGAGGAGGAGGAGGGCGGGTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCCGACGCACCCGCCTACCAGCAGGGCCAGAATCAGCTCTACAACGAGCTGAATCTGGGCCGGCGGGAGGAGT ACGATGTGCTGGACAAGCGGAGGGGCAGGGATCCCGAGATGGGGGGTAAGCCTCGGCGGAAGAATCCTCAGGAGGG GCTCTACAACGAGCTCCAGAAGGATAAGATGGCCGAGGCTTACTCCGAGATCGGGATGAAGGGGGAGAGGCGCCGG GGTAAGGGCCACGATGGCCTTTACCAGGGCCTCTCCACCGCAACAAAGGACACCTATGATGCCCTGCACATGCAGG CCCTGCCCCCCCGG ( SEQ ID NO: 162 )ATGGCTTTACCCGTTACAGCCCTGTTGCTGCCCCTTGCCCTGCTGCTGCACGCCGCCCGGCCAGATATCCAGATGACGCAGACCACCAGCTCATTGTCAGCCTCCCTGGGGGATCGGGTGACCATCAGCTGCCGGGCCAGTCAGGACATCAGATTORNEY DOCKET NO. ORB-019WO1CAAGTACCTGAACTGGTACCAGCAGAAGCCTGATGGCACCGTGAAGCTGCTGATCTACCACACCAGCAGGCTTCAC TCCGGCGTGCCTTCCCGCTTCTCTGGGTCCGGCTCCGGCACAGACTACAGTCTGACCATCTCCAACCTGGAGCAGG AGGACATCGCTACTTACTTCTGCCAGCAGGGGAACACCCTGCCCTACACCTTCGGCGGGGGCACCAAGCTGGAGAT CACTGGCGGTGGTGGGAGCGGGGGTGGGGGGTCTGGGGGCGGGGGTAGCGAGGTGAAGCTGCAGGAGTCCGGACCC GGCCTGGTCGCCCCCAGCCAGTCCCTGTCCGTGACATGCACCGTGTCTGGCGTGAGCCTGCCAGACTACGGCGTGT CATGGATCAGGCAGCCCCCCCGGAAGGGGCTGGAGTGGCTGGGGGTGATCTGGGGGTCCGAGACTACCTACTACAA CAGCGCGCTGAAGTCCAGGCTGACCATCATCAAGGACAACTCCAAGAGTCAGGTGTTCTTGAAGATGAACAGCCTG CAGACCGATGACACCGCCATATACTACTGTGCGAAGCACTACTACTATGGCGGGTCATACGCCATGGATTACTGGG GCCAGGGGACCTCAGTGACCGTGAGCAGCACCACCACCCCCGCCCCCAGACCCCCCACCCCCGCTCCCACCATCGC CAGCCAGCCCCTGAGCCTGCGCCCCGAGGCCTGCCGGCCCGCGGCCGGCGGCGCCGTGCACACTCGGGGGCTGGAC TTCGCCTGCGATATCTATATCTGGGCCCCTCTGGCCGGCACCTGCGGCGTCCTCCTCCTGAGTCTGGTGATCACCC TGTACTGCAAGCGCGGAAGGAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGCGCCCAGTGCAGACCACTCA GGAGGAGGACGGCTGCAGCTGCCGGTTCCCAGAGGAGGAGGAGGGGGGCTGCGAGCTGCGGGTGAAGTTCTCCCGC AGCGCAGACGCCCCCGCCTACCAGCAGGGCCAGAATCAGCTCTACAACGAGCTGAATCTGGGCCGGCGGGAGGAGT ACGATGTCCTGGACAAGCGCCGGGGCCGGGACCCAGAGATGGGCGGGAAGCCACGCCGGAAGAACCCCCAGGAGGG GCTGTACAATGAGCTGCAGAAGGATAAGATGGCCGAGGCCTATTCCGAGATCGGAATGAAGGGCGAGCGGCGGCGG GGCAAGGGGCACGACGGGCTGTACCAGGGGCTGAGCACCGCAACAAAGGACACCTATGATGCCCTGCACATGCAGG CTCTGCCCCCCCGG ( SEQ ID NO: 163) ATGGCTCTGCCTGTGACAGCTCTGCTGCTGCCTCTGGCTCTGCTTCTGCATGCCGCCAGACCTGACATCCAGATGA CCCAGACAACCAGCAGCCTGTCTGCCAGCCTGGGCGACAGAGTGACCATCAGCTGTAGAGCCAGCCAGGACATCAG CAAGTACCTGAACTGGTATCAGCAGAAACCCGACGGCACCGTGAAGCTGCTGATCTACCACACCAGCAGACTGCAC AGCGGCGTGCCATCTAGATTTTCTGGCAGCGGCTCTGGCACCGACTACAGCCTGACAATCAGCAACCTGGAACAAG AGGATATCGCTACCTACTTCTGCCAGCAAGGCAACACCCTGCCTTACACATTCGGCGGAGGCACCAAGCTGGAAAT CACAGGTGGCGGAGGATCTGGCGGAGGTGGAAGCGGAGGCGGTGGATCTGAAGTGAAACTGCAAGAGTCTGGCCCT GGCCTGGTGGCTCCATCTCAGTCTCTGAGCGTGACCTGTACCGTCAGCGGAGTGTCTCTGCCTGATTACGGCGTGT CCTGGATCAGACAGCCTCCTAGAAAAGGCCTGGAATGGCTGGGCGTGATCTGGGGCAGCGAGACAACCTACTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACTCCAAGAGCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGATTACTGGG GCCAGGGAACCAGCGTGACCGTGTCTAGCACAACAACCCCTGCTCCTAGACCTCCTACACCAGCTCCTACAATCGC CAGCCAGCCTCTGTCTCTGAGGCCAGAGGCTTGTAGACCTGCTGCTGGCGGAGCTGTGCACACAAGAGGCCTGGAT TTCGCCTGCGACATCTACATCTGGGCCCCTCTGGCTGGAACATGTGGCGTGCTGCTGCTGAGCCTGGTCATCACCC TGTATTGCAGAGTGAAGTTCAGCAGAAGCGCTGACGCCCCTGCTTACCAGCAGGGACAGAACCAGCTGTTTAACGA GCTGAACCTGGGGAGAAGAGAAGAGTTCGACGTGCTGGACAAGCGGAGAGGCAGAGATCCTGAGATGGGCGGCAAG CCCAGAAGAAAGAACCCTCAAGAGGGCCTGTTTAATGAGCTGCAGAAAGACAAGATGGCCGAGGCCTTCAGCGAGA TCGGAATGAAGGGCGAGCGCAGAAGAGGCAAGGGACACGATGGACTGTTCCAGGGCCTGAGCACAGCCACCAAGGA CACATTCGACGCCCTGCACATGCAGGCCCTGCCTCCAAGA ( SEQ ID NO: 164 ) ATGCTGCTGCTGGTGACAAGCCTGCTGCTGTGCGAGCTGCCCCACCCCGCCTTTCTGCTGATCCCCGACATCCAGA TGACCCAGACCACCAGCAGCCTGAGCGCCAGCCTGGGCGACAGAGTGACCATCAGCTGCCGGGCCAGCCAGGACAT CAGCAAGTACCTGAACTGGTATCAGCAGAAACCCGACGGCACCGTGAAGCTGCTGATCTACCACACCTCCCGGCTG CACAGCGGCGTGCCCAGCAGATTTTCTGGCAGCGGCAGCGGCACCGACTACAGCCTGACCATCTCCAACCTGGAAC AAGAAGATATCGCTACCTATTTTTGTCAACAAGGCAACACCCTGCCCTACACCTTCGGCGGAGGCACCAAACTGGA AATCACCGGCAGCACCAGCGGCTCCGGCAAGCCTGGATCTGGCGAGGGCAGCACCAAGGGCGAAGTGAAGCTGCAA GAAAGCGGCCCTGGCCTGGTGGCCCCTAGCCAGAGCCTGTCCGTGACCTGTACCGTGTCCGGCGTGTCCCTGCCCG ACTACGGCGTGTCCTGGATCCGGCAGCCCCCCAGAAAGGGCCTGGAATGGCTGGGCGTGATCTGGGGCAGCGAGAC AACCTACTACAACAGCGCCCTGAAGTCCCGGCTGACCATCATCAAGGACAACAGCAAGAGCCAGGTGTTCCTGAAG ATGAACAGCCTGCAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCA TGGACTACTGGGGCCAGGGCACCAGCGTGACCGTGTCCAGCTTCGTGCCCGTGTTTCTGCCCGCCAAGCCTACCAC CACCCCTGCCCCTAGACCTCCCACCCCAGCCCCAACAATCGCCAGCCAGCCTCTGTCCCTGCGGCCCGAAGCCTGT AGACCAGCTGCCGGCGGAGCCGTGCACACCAGAGGCCTGGACTTCGCCTGCGACATCTACATCTGGGCCCCTCTGG CCGGCACCTGTGGCGTGCTGCTGCTGAGCCTGGTGATCACCCTGTACTGCAACCACCGGAACAGAAGCAAGCGGAG CCGGCTGCTGCACAGCGACTACATGAACATGACCCCAAGACGGCCTGGCCCCACCCGGAAGCACTACCAGCCTTAC GCCCCTCCCAGAGACTTCGCCGCCTACCGGTCCAGAGTGAAGTTCAGCAGATCCGCCGACGCCCCTGCCTACCAGC AGGGACAGAACCAGCTGTACAACGAGCTGAACCTGGGCAGACGGGAAGAGTACGACGTGCTGGACAAGCGGAGAGG CCGGGACCCCGAGATGGGCGGAAAGCCCAGACGGAAGAACCCCCAGGAAGGCCTGTATAACGAACTGCAGAAAGACAAGATGGCCGAGGCCTACAGCGAGATCGGCATGAAGGGCGAGCGGAGGCGCGGCAAGGGCCACGATGGCCTGTACCATTORNEY DOCKET NO. ORB-019WO1AGGGCCTGAGCACCGCCACCAAGGACACCTACGACGCCCTGCACATGCAGGCCCTGCCCCCCAGA ( SEQ ID NO: 165 ) ATGGCATTACCAGTTACAGCCCTCCTCCTCCCTCTCGCTCTCCTCCTCCACGCCGCCAGGCCAGATATCCAGATGA CACAGACCACCAGCAGCCTCAGCGCCAGCCTCGGCGATAGAGTGACAATCTCTTGCAGAGCCTCCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTCCTGATCTACCACACCAGCAGACTGCAC AGCGGCGTGCCTTCTAGATTCTCTGGAAGCGGCAGCGGCACCGACTACAGCCTGACCATCAGCAACCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGCAATACCCTGCCCTACACCTTCGGCGGCGGCACAAAACTGGAAAT TACAGGTGGTGGTGGTTCTGGGGGAGGAGGGTCTGGTGGTGGTGGTAGTGAAGTGAAACTGCAGGAATCTGGACCA GGACTGGTGGCCCCATCTCAGTCTCTGAGCGTGACATGTACAGTGTCTGGCGTGTCTCTGCCTGATTACGGAGTGT CTTGGATTAGACAGCCTCCTCGGAAGGGCCTGGAGTGGCTGGGAGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGTCCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGATTACTGGG GCCAGGGCACCAGCGTGACGGTGTCTTCTACAACAACACCTGCTCCTAGGCCTCCTACACCAGCTCCTACAATTGC TTCTCAGCCGCTGTCTCTGCGGCCAGAAGCTTGTAGGCCTGCTGCAGGAGGAGCGGTGCATACAAGAGGACTGGAC TTTGCCTGTGATATCTACATCTGGGCCCCTCTGGCCGGCACCTGCGGCGTGCTGCTGCTGAGCCTGGTGATCACCC T GTACT GCAACCACAGAAACAGAAGCAAGAGAAGCAGGCT GCT GCACAGCGACTACAT GAATAT GACACCTAGGAG ACCTGGCCCTACAAGAAAACATTATCAGCCTTATGCCCCTCCTAGAGATTTTGCCGCCTATAGAAGCCGGGTGAAG TTTTCTAGATCTGCTGATGCCCCAGCCTATCAGCAGGGCCAGAACCAGTTATATAATGAACTGAACCTGGGCCGGC GGGAGGAGTACGAT GT GCT GGATAAGAGAAGAGGCAGAGAT GCT GAAAT GGGAGGAAAACCTAGAAGAAAGAAT CC CCAGGAGGGCCTATATAACGAACTGCAGAAGGACAAGATGGCCGAGGCCTACAGCGAGATCGGCATGAAGGGCGAG AGAAGAAGAGGCAAGGGCCACGATGGACTGTATCAGGGGCTGAGCACCGCCACCAAGGATACCTACGATGCCCTGC ACATGCAGGCCCTGCCTCCTAGA ( SEQ ID NO: 166) ATGCTATTATTAGTCACCAGCCTCCTCCTTTGCGAGCTCCCCCACCCCGCCTTCCTCCTCATCCCCGACATCCAGA TGACCCAGACCACCAGCAGCCTCAGCGCCAGCCTCGGCGATAGAGTGACAATCTCTTGCAGAGCCTCCCAGGACAT CAGCAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTGCTGATCTACCACACCAGCAGACTG CACAGCGGCGTGCCTTCTAGATTCTCTGGAAGCGGCAGCGGCACCGACTACAGCCTGACCATCAGCAACCTGGAGC AGGAGGACATCGCCACCTACTTCTGCCAGCAGGGCAATACCTTGCCCTACACCTTCGGCGGCGGCACAAAGCTGGA AATTACAGGTGGTGGTGGTAGTGGTGGTGGTGGTTCTGGTGGTGGTGGATCTGAAGTGAAACTGCAGGAATCTGGA CCTGGCCTGGTGGCCCCTTCTCAGTCGCTGAGCGTGACATGTACAGTGAGCGGCGTGAGCCTGCCTGATTATGGAG TGAGCTGGATCAGACAGCCTCCAAGAAAGGGCCTGGAGTGGCTGGGCGTGATCTGGGGCTCCGAGACCACCTACTA CAACAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGAGCCAGGTGTTCCTGAAGATGAACAGC CTGCAGACCGATGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGATTACT GGGGCCAGGGCACATCTGTGACAGTGTCTTCTGCTGCTGCTCCAGCTCCTAGACCTCCTACACCTGCTCCTACAAT TGCTTCTCAGCCGCTGAGCCTGAGACCTGAAGCTTGTAGACCTGCTGCTGGAGGAGCTGTGCATACAAGAGGACTG GATTTTGATACCGCCCTGGCCGCCGTGATCTGTTCTGCCCTGGCCACAGTGCTGCTGGCCCTGCTGATCCTGTGCG TGATCTACTGCAAGAGACAGCCCAGAAGAAAGAAGCTGCTGTACATCTTCAAGCAGCCCTTCATGAGACCTGTGCA GACAACACAGGAGGAGGATGGCTGTTCCTGTAGATTTCCAGAGGAAGAGGAGGGCGGCTGCGAGCTGAGAGTGAAA TTTTCTAGATCTGCCGATGCCCCTGCCTACCAGCAGGGCCAGAACCAGCTATATAATGAACTGAACCTGGGCAGAA GAGAGGAGTACGACGTGCTGGATAAGAGAAGAGGCAGAGATCCTGAAATGGGAGGAAAACCTAGAAGAAAGAATCC GCAGGAGGGCCTATATAACGAACTGCAGAAGGACAAGATGGCCGAGGCCTACAGCGAGATCGGCATGAAGGGCGAG AGAAGAAGAGGCAAGGGCCACGATGGACTGTATCAGGGGCTGAGCACAGCCACCAAGGATACCTACGATGCCCTGC ACATGCAGGCCCTGCCTCCTAGA ( SEQ ID NO: 167 ) ATGGCTCTACCCGTTACAGCATTACTACTACCTCTTGCCCTCCTCCTCCACGCCGCCAGACCAGATATCCAGATGA CCCAGACCACCTCTAGCCTCAGCGCCAGCCTCGGCGATAGAGTGACAATCTCTTGCAGAGCCTCCCAGGACATCAG CAAGTACCTCAATTGGTACCAGCAGAAGCCCGACGGCACCGTGAAGCTCCTGATCTACCACACCAGCAGGCTGCAC TCTGGCGTGCCATCTAGATTTTCTGGAAGCGGCAGCGGCACAGATTACAGCCTGACCATCAGCAATCTGGAGCAGG AGGACATCGCCACCTACTTCTGCCAGCAGGGCAATACCCTGCCCTACACCTTCGGCGGCGGCACAAAACTGGAAAT TACAGGTGGTGGTGGTTCAGGGGGAGGAGGGTCTGGTGGTGGAGGTTCTGAAGTGAAACTGCAGGAATCTGGACCT GGCCTGGTGGCCCCTTCTCAGTCGCTGAGCGTGACATGTACCGTGAGCGGCGTGAGCCTGCCTGATTACGGGGTGA GCTGGATCAGACAGCCCCCTAGAAAGGGCCTGGAATGGCTGGGCGTGATCTGGGGCAGCGAGACCACCTACTACAA CAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACAGCAAGTCCCAGGTGTTCCTGAAGATGAACAGCCTG CAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGGCAGCTACGCCATGGATTACTGGG GCCAGGGCACCTCTGTGACAGTGAGCAGCGCCGCTGCCATTGAGGTGATGTACCCTCCCCCCTACCTGGACAACGA GAAGTCCAACGGCACCATCATCCACGTGAAGGGCAAGCACCTGTGCCCTAGCCCCCTGTTTCCAGGCCCATCTAAG CCTTTTTGGGTGCTGGTGGTGGTGGGCGGCGTGCTGGCCTGCTACAGCCTGCTGGTGACCGTGGCCTTCATCATCTTCTGGGTGAGAAGCAAGAGGTCCCGGCTGCTGCACAGCGACTACATGAATATGACACCCAGGAGACCTGGCCCTACATTORNEY DOCKET NO. ORB-019WO1AAGAAAACATTATCAGCCTTATGCCCCTCCTAGAGATTTTGCCGCCTATAGAAGCCGGGTGAAGTTTTCTAGAAGC GCTGATGCCCCAGCCTACCAGCAGGGCCAGAACCAGCTGTACAACGAGCTGAACCTGGGCAGAAGAGAGGAGTACG ACGTGCTGGATAAGAGAAGAGGCAGAGATCCGGAAATGGGAGGAAAGCCTAGAAGAAAGAATCCTCAGGAGGGCCT GTACAAT GAGCT GCAGAAGGACAAGAT GGCCGAGGCCTACT CT GAGAT CGGCAT GAAGGGCGAGAGAAGAAGAGGC AAGGGCCACGATGGCCTGTACCAGGGCCTGAGCACGGCCACCAAGGATACCTACGATGCCCTGCACATGCAGGCCC TGCCTCCTAGA ( SEQ ID NO: 168 ) ATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCCAGGCCGGACATCCAGATGA CACAGACTACATCCTCCCTGTCTGCCTCTCTGGGAGACAGAGTCACCATCAGTTGCAGGGCAAGTCAGGACATTAG TAAATATTTAAATTGGTATCAGCAGAAACCAGATGGAACTGTTAAACTCCTGATCTACCATACATCAAGATTACAC TCAGGAGTCCCATCAAGGTTCAGTGGCAGTGGGTCTGGAACAGATTATTCTCTCACCATTAGCAACCTGGAGCAAG AAGATATTGCCACTTACTTTTGCCAACAGGGTAATACGCTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAGAT CACAGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAGGTGAAACTGCAGGAGTCAGGACCT GGCCTGGTGGCGCCCTCACAGAGCCTGTCCGTCACATGCACTGTCTCAGGGGTCTCATTACCCGACTATGGTGTAA GCTGGATTCGCCAGCCTCCACGAAAGGGTCTGGAGTGGCTGGGAGTAATATGGGGTAGTGAAACCACATACTATAA TTCAGCTCTCAAATCCAGACTGACCATCATCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTG CAAACTGATGACACAGCCATTTACTACTGTGCCAAACATTATTACTACGGTGGTAGCTATGCTATGGACTACTGGG GCCAAGGAACCTCAGTCACCGTCTCCTCAACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACCATCGC GTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGAC TTCGCCTGTGATATCTACATCTGGGCGCCCTTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTGGTTATCACCC TTTACTGCAAACGGGGCAGAAAGAAACTCCTGTATATATTCAAACAACCATTTATGAGACCAGTACAAACTACTCA AGAGGAAGATGGCTGTAGCTGCCGATTTCCAGAAGAAGAAGAAGGAGGATGTGAACTGAGAGTGAAGTTCAGCAGG AGCGCAGACGCCCCCGCGTACAAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGT ACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGAAGG CCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGG GGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCTAA ( SEQ ID NO: 346)*Because the sequences are RNA sequences, it is understood that T' s represent uracil in the sequences of this table.

[0491] In some embodiments, the circRNA molecule comprises one or more modified nucleotides. In some embodiments, the circRNA molecule is unmodified.

[0492] In some embodiments, the circRNA molecules of the present invention may include one, two, three, or more modifications. In some embodiments, the modified nucleotides are located in coding region(s). In some embodiments, the modified nucleotides are in the untranslated region(s).

[0493] In some embodiments, the modifications stabilize the circRNA molecule and enhance resistance to degradation as compared to unmodified nucleotides. In some embodiments, modified nucleotides enhance biological functions of nucleic acid molecules, for example, increase binding to a RNA binding protein or increasing translation.

[0494] In some embodiments, the modified nucleotide is one or more of Nl-methylpseudouridine, 5-methoxyuridine, N6-methyladenosine, pseudouridine or 5-methylcytosine.

[0495] In some embodiments, the modified nucleotide is N1 -methylpseudouridine. In some embodiments, the modified nucleotide is 5-methoxyuridine. In some embodiments, theATTORNEY DOCKET NO. ORB-019WO1modified nucleotide is N6-methyladenosine. In some embodiments, the modified nucleotide is pseudouridine. In some embodiments, the modified nucleotide is 5-methylcytosine.

[0496] In some embodiments, the encoded protein is an anti-CD19 antibody or antigen binding domain. In some embodiments, the encoded protein is any one of SEQ ID Nos: 169-174 selected from Table 5. In some embodiments, the circRNA encodes an amino acid sequence having at least 75% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 80% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 85% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 90% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 95% identity to any one of SEQ ID NOs: 169-174. In some embodiments, the circRNA encodes an amino acid sequence having at least 100% identity to any one of SEQ ID NOs: 169-174.

[0497] Table 5. Exemplary Encoded Anti-CD19 Antibody or Antigen Binding Domain (Amino Acid Sequences)Encoded Anti-CD19 Amino Acid Sequences MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTS RLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVK LQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKS QVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRP AAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFP EEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQK DKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO: 169 ) _ MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTS RLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVK LQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKS QVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRP AAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCRVKFSRSADAPAYQQGQNQLFNELNLGRREEFD VLDKRRGRDPEMGGKPRRKNPQEGLFNELQKDKMAEAFSEIGMKGERRRGKGHDGLFQGLSTATKDTFDALHM QALPPR ( SEQ ID NO: 170 ) _ MLLLVTSLLLCELPHPAFLLIPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHT SRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGSTSGSGKPGSGEGSTK GEVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKD NSKSQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSFVPVFLPAKPTTTPAPRPPTPAPTI ASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNM TPRRPGPTRKHYQPYAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGG KPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO: 171 ) _ MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTS RLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVK LQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKS QVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRP AAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCNHRNRSKRSRLLHSDYMNMTPRRPGPTRKHYQP YAPPRDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO: 172 )ATTORNEY DOCKET NO. ORB-019WO1MLLLVTSLLLCELPHPAFLLIPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHT SRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEV KLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSK SQVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSAAAPAPRPPTPAPTIASQPLSLRPEACR PAAGGAVHTRGLDFDTALAAVICSALATVLLALLILCVIYCKRQPRRKKLLYIFKQPFMRPVQTTQEEDGCSC RFPEEEEGGCELRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNE LQKDKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO: 173 ) _ MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTS RLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVK LQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKS QVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKH LCPSPLFPGPSKPFWVLVWGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPP RDFAAYRSRVKFSRSADAPAYQQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQKD KMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO: 174 ) _ MALPVTALLLPLALLLHAARPDIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHTS RLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTKLEITGGGGSGGGGSGGGGSEVK LQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVIWGSETTYYNSALKSRLTIIKDNSKS QVFLKMNSLQTDDTAIYYCAKHYYYGGSYAMDYWGQGTSVTVSSTTTPAPRPPTPAPTIASQPLSLRPEACRP AAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFP EEEEGGCELRVKFSRSADAPAYKQGQNQLYNELNLGRREEYDVLDKRRGRDPEMGGKPRRKNPQEGLYNELQK DKMAEAYSEIGMKGERRRGKGHDGLYQGLSTATKDTYDALHMQALPPR ( SEQ ID NO: 347 )

[0498] In some aspects, provided herein is a full-length circRNA sequence selected from Table 6. As discussed here, the circRNA sequences comprise features, including, for example, the 5’ UTR, ribosome recruiting element (such as IRES), coding sequence, and 3’ UTR. In some aspects, provided herein is a full-length circRNA comprising a sequence selected from any one of SEQ ID NOs: 175-233 in Table 6 or SEQ ID NOs: 264 or 269 in Table 7B. In some aspects, provided herein is a circRNA comprising a sequence having at least 80% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, provided herein is a circRNA comprising a sequence having at least 85% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, provided herein is a circRNA comprising a sequence having at least 90% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, provided herein is a circRNA comprising a sequence having at least 95% identity to any one of SEQ ID NOs: 175-233, 264 or 269. In some embodiments, provided herein is a circRNA comprising a sequence having at least 100% identity to any one of SEQ ID NOs: 175-233, 264 or 269.

[0499] Table 6. Exemplary Full-length CircRNA Sequences* AACTTTTAGTTATTTTCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCATATGACC TCTGCGTGTGCCTGGTAATTGACTAAGATATCTTACTCAGCGTAACTACTCCGGGTTACGTGATGAAGAAGAGGCT ACGGAGATTCTCGGGCTACGGCCCTGGAGCCACTCCGGCTCCTAAAGATTTAGAAGTTTGAGCACACCCGCCCACT AGGGCCCCCCATCCAGGGGGGCAACGGGCAAGCACTTCTGTTTCCCCGGTATGATCTGATAGGCTGTAACCACGGC TGAAACAGAGATTATCGTTATCCGCTTCACTACTTCGAGAAGCCTAGTAATGATGGGTGAAATTGAATCCGTTGAT CCGGTGTCTCCCCCACACCAGAAACTCATGATGAGGGTTGCCATCCCGGCTACGGCGACGTAGCGGGCATCCCTGC GCTGGCATGAGGCCTCTTAGGAGGACGGATGATATGGATCTTGTCGTGAAGAGCCTATTGAGCTAGTGTCGACTCC TCCGCCCCCGTGAATGCGGCTAATCCTAACCCCGGAGCAGGTGGGTCCAATCCAGGGCCTGGCCTGTCGTAATGCGTAAGTCTGGGACGGAACCGACTACTTTCGGGAAGGCGTGTTTCCATTTGTTCATTATTTGTGTGTTTATGGTGACAATTORNEY DOCKET NO. ORB-019WO1ACTCTGGGTAAACGTTCTATTGCGTTTATTGAGAGATTCCCAACAATTGAACAAACGAGAACTACCTGTTTTATTA AATTTACACAGAGAAGAATTACAATGGCTCTGCCTGTGACAGCTCTGCTGCTGCCTCTGGCTCTGCTTCTGCATGC CGCCAGACCTGACATCCAGATGACCCAGACAACCAGCAGCCTGTCTGCCAGCCTGGGCGACAGAGTGACCATCAGC TGTAGAGCCAGCCAGGACATCAGCAAGTACCTGAACTGGTATCAGCAGAAACCCGACGGCACCGTGAAGCTGCTGA TCTACCACACCAGCAGACTGCACAGCGGCGTGCCATCTAGATTTTCTGGCAGCGGCTCTGGCACCGACTACAGCCT GACAATCAGCAACCTGGAACAAGAGGATATCGCTACCTACTTCTGCCAGCAAGGCAACACCCTGCCTTACACATTC GGCGGAGGCACCAAGCTGGAAATCACAGGTGGCGGAGGATCTGGCGGAGGTGGAAGCGGAGGCGGTGGATCTGAAG TGAAACTGCAAGAGTCTGGCCCTGGCCTGGTGGCTCCATCTCAGTCTCTGAGCGTGACCTGTACCGTCAGCGGAGT GTCTCTGCCTGATTACGGCGTGTCCTGGATCAGACAGCCTCCTAGAAAAGGCCTGGAATGGCTGGGCGTGATCTGG GGCAGCGAGACAACCTACTACAACAGCGCCCTGAAGTCCAGGCTGACCATCATCAAGGACAACTCCAAGAGCCAGG TGTTCCTGAAGATGAACAGCCTGCAGACCGACGACACCGCCATCTACTACTGCGCCAAGCACTACTACTACGGCGG CAGCTACGCCATGGATTACTGGGGCCAGGGAACCAGCGTGACCGTGTCTAGCACAACAACCCCTGCTCCTAGACCT CCTACACCAGCTCCTACAATCGCCAGCCAGCCTCTGTCTCTGAGGCCAGAGGCTTGTAGACCTGCTGCTGGCGGAG CTGTGCACACAAGAGGCCTGGATTTCGCCTGCGACATCTACATCTGGGCCCCTCTGGCTGGAACATGTGGCGTGCT GCTGCTGAGCCTGGTCATCACCCTGTATTGCAAGAGAGGCCGGAAGAAGCTGCTCTACATCTTCAAGCAGCCCTTC ATGAGGCCCGTGCAGACCACACAAGAAGAGGACGGCTGCTCCTGCAGATTCCCCGAGGAAGAGGAAGGCGGCTGCG AGCTGAGAGTGAAGTTCAGCAGAAGCGCTGACGCCCCTGCTTACCAGCAGGGACAGAACCAGCTGTACAACGAGCT GAACCTGGGGAGAAGAGAAGAGTACGACGTGCTGGACAAGCGGAGAGGCAGAGATCCTGAGATGGGCGGCAAGCCC AGAAGAAAGAACCCTCAAGAGGGCCTGTATAATGAGCTGCAGAAAGACAAGATGGCCGAGGCCTACAGCGAGATCG GAATGAAGGGCGAGCGCAGAAGAGGCAAGGGACACGATGGACTGTACCAGGGCCTGAGCACAGCCACCAAGGACAC ATATGACGCCCTGCACATGCAGGCCCTGCCTCCAAGATAATAGAAAAACAAAAAACAAAAACCACACAAATGGTCG CCGAAACTACTGAAAGCAT ( SEQ ID NO: 175 ) _ AACTTTTAGTTATTTTCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCATATGACC TCTGCGTGTGCCTGGTAATTGACTAAGATATCTTACTCAGCGTAACTACTCCGGGTTACGTGATGAAGAAGAGGCT ACGGAGATTCTCGGGCTACGGCCCTGGAGCCACTCCGGCTCCTAAAGATTTAGAAGTTTGAGCACACCCGCCCACT AGGGCCCCCCATCCAGGGGGGCAACGGGCAAGCACTTCTGTTTCCCCGGTATGATCTGATAGGCTGTAACCACGGC TGAAACAGAGATTATCGTTATCCGCTTCACTACTTCGAGAAGCCTAGTAATGATG...

Claims

ATTORNEY DOCKET NO. ORB-019WO1CLAIMSWe claim:

1. A circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising:(i) a circRNA comprising a ribosome recruiting element, and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and(ii) a targeting moiety,wherein the circRNA is encapsulated in a delivery vehicle, and wherein the targeting moiety is conjugated to the delivery vehicle, andwherein the targeting moiety comprises an anti-CD5 antibody heavy chain variable region comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89.

2. The circRNA therapeutic of claim 1, wherein the CDR1 sequence comprises SEQ ID NO: 64, the CDR2 sequence comprises SEQ ID NO: 65, and the CDR3 sequence comprises SEQ ID NO: 66.

3. A circular ribonucleic acid (circRNA) for targeted delivery comprising:(i) a circRNA comprising:a ribosome recruiting element, anda coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and(ii) a targeting moiety, wherein the targeting moiety binds to a T cell,wherein the circRNA is in a delivery vehicle,wherein the targeting moiety that binds to a T cell is conjugated to the delivery vehicle, and wherein the targeting moiety is an anti-CD5 antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising complementarity determining region (CDR) sequences GFTFSMYS (SEQ ID NO: 64), ISTGARDT (SEQ ID NO: 65) and GDLRYGPDGYDY (SEQ ID NO: 66).

4. The circRNA therapeutic of any one of claims 1 to 3, wherein the delivery vehicle is a lipid nanoparticle (LNP).ATTORNEY DOCKET NO. ORB-019WO15. The circRNA therapeutic of any one of claims 1 to 4, wherein the anti-CD5 antibody or antigen-binding fragment comprises a sequence having at least 95% identity to SEQ ID NO: 89.

6. The circRNA therapeutic of any one of claims 1 to 5, wherein the anti-CD5 antibody or antigen-binding fragment comprises SEQ ID NO: 89.

7. The circRNA therapeutic of any one of claims 1 to 6, wherein the anti-CD5 antibody or antigen-binding fragment is a VHH.

8. The circRNA therapeutic of any one of the preceding claims, wherein the anti-CD5 antibody or antigen-binding fragment is conjugated to a conjugating PEG-lipid in the delivery vehicle.

9. The circRNA therapeutic of claims 1 to 8, wherein the anti-CD5 antibody or antigenbinding fragment is covalently attached to a conjugation handle.

10. The circRNA therapeutic of claim 9, wherein the conjugation handle is covalently attached to the C-terminus of the heavy chain of the anti-CD5 antibody or antigen-binding fragment.

11. The circRNA therapeutic of claims 9 or 10, wherein the conjugation handle is 5-10 amino acids in length.

12. The circRNA therapeutic of any one of claims 9 to 11, wherein the conjugation handle comprises SEQ ID NO: 120.

13. The circRNA therapeutic of any one of claims 9 to 12, wherein the conjugation handle consists of SEQ ID NO: 120.

14. The circRNA therapeutic of any one of claims 9 to 13, wherein the anti-CD5 antibody or antigen-binding fragment is covalently attached to the conjugation handle, and the sequence of the anti-CD5 antibody or antigen-binding fragment with attached conjugation handle comprises SEQ ID NO: 122.

15. The circRNA therapeutic of claim 8, wherein the anti-CD5 antibody or antigenbinding fragment is conjugated to the conjugating PEG-lipid in the delivery vehicle via aATTORNEY DOCKET NO. ORB-019WO1thiol-maleimide conjugation.

16. The circRNA therapeutic of any one of claims 9 to 15, wherein the anti-CD5 antibody or antigen-binding fragment is conjugated to the conjugating PEG-lipid in the delivery vehicle via a thiol-maleimide conjugation between the conjugating PEG-lipid and the conjugation handle.

17. The circRNA therapeutic of any one of claims 8, 15 and 16, wherein the delivery vehicle is an LNP comprising two PEG-lipids, one of which is the conjugating PEG-lipid.

18. The circRNA therapeutic of any one of claims 8 and 15 to 17, wherein the conjugating PEG-lipid comprises a long chain fatty acid.

19. The circRNA therapeutic of claim 18, wherein the conjugating PEG-lipid is DSPE-PEG 2000-maleimide.

20. The circRNA therapeutic of any one of claims 17 to 19, wherein the LNP further comprises one or more ionizable lipids, one or more phospholipids, and one or more sterol lipids.

21. The circRNA therapeutic of claim 20, wherein the LNP comprises the one or more ionizable lipids at 30 to 49 mol% (e.g., at 30 to 45 mol%), the one or more phospholipids at 5 to 40 mol% (e.g., at 15 to 35 mol%), the one or more sterol lipids at 15 to 50 mol% (e.g., at 30 to 50 mol%), the PEG lipid (e.g., the DMG-PEG 2000) at 0 to 10 mol% (e.g., at 0 to 5 mol%), and the conjugating PEG-lipid (e.g., the DSPE-PEG 2000-maleimide) at 0.01-1 mol%.

22. The circRNA therapeutic of claim 20, wherein the LNP comprises the one or more ionizable lipids, the one or more phospholipids, the one or more sterol lipids, the DMG-PEG 2000 lipid, and the DSPE-PEG 2000-maleimide lipid at a ratio of 39: 20: 39: 1.9: 0.1 mol%.

23. The circRNA therapeutic of any one of claims 20 to 22, wherein the one or more ionizable lipids isATTORNEY DOCKET NO. ORB-019WO1(Compound24. The circRNA therapeutic of any one of claims 20 to 23, wherein the one or more phospholipids is DSPC.

25. The circRNA therapeutic of any one of claims 20 to 24, wherein the sterol lipid is cholesterol.

26. The circRNA therapeutic of any one of claims 1 to 25, wherein the ribosome recruiting element is an Internal Ribosome Entry Site (IRES), wherein the IRES comprises a sequence selected from Table 1.

27. The circRNA therapeutic of claim 26, wherein the IRES comprises a sequence selected from SEQ ID NO: 1-11 of Table 1.

28. The circRNA therapeutic of claim 26 or 27, wherein the IRES comprises the sequence of SEQ ID NO: 1.

29. The circRNA therapeutic of any one of the preceding claims, wherein:the coding sequence further encodes a CD8 signal peptide, andthe CAR comprises a CD8 hinge, a CD8 transmembrane domain, a 4-1BB costimulatory domain, and a CD3 zeta signaling domain.

30. The circRNA therapeutic of any one of the preceding claims, wherein the anti-CD19 antibody or antigen-binding fragment thereof comprises a VL sequence of SEQ ID NO: 54 and a VH sequence of SEQ ID NO: 55.

31. The circRNA therapeutic of any one of the preceding claims, wherein the coding sequence encodes an amino acid sequence of SEQ ID NO: 58.ATTORNEY DOCKET NO. ORB-019WO132. A circular ribonucleic acid (circRNA) therapeutic for targeted delivery comprising:a circular RNA comprising a ribosome recruiting element and a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, wherein the circRNA is encapsulated in an LNP,wherein a targeting moiety is conjugated to the LNP,wherein the targeting moiety is an anti-CD5 VHH, the anti-CD5 VHH comprising CDR1, CDR2, and CDR3 sequences from SEQ ID NO: 89,wherein the LNP comprises one or more ionizable lipids at 30 to 49 mol%, one or more phospholipids at 5 to 40 mol%, one or more sterol lipids at 15 to 50 mol%, a DMG-PEG 2000 lipid at 0 to 10 mol%, and a DSPE-PEG 2000-mal eimide lipid at 0.01 to 1 mol%, andwherein the VHH is conjugated to the LNP.

33. The circRNA therapeutic of claim 32, wherein the VHH has a length of 110 to 130 (e.g., 115 to 125) amino acids.

34. The circRNA therapeutic of claim 32 or 33, wherein the VHH is conjugated to the LNP via a thiol-maleimide conjugation, and the thiol-maleimide conjugation is between the DSPE-PEG 2000-maleimide and a conjugation handle that is covalently attached to the VHH.

35. The circRNA therapeutic of claim 34, wherein the N-terminus of the conjugation handle is covalently attached to the C-terminus of the VHH.

36. The circRNA therapeutic of claim 34 or 35, wherein the VHH with conjugation handle has a length of 120 to 140 amino acids.

37. The circRNA therapeutic of claim 36, wherein the VHH with attached conjugation handle has a length of 125 to 135 amino acids.ATTORNEY DOCKET NO. ORB-019WO138. The circRNA therapeutic of any one of claims 32 to 37, wherein the VHH comprises SEQ ID NO: 89.

39. The circRNA therapeutic of any one of claims 32 to 38, wherein the conjugation handle comprises SEQ ID NO: 120.

40. The circRNA therapeutic of any one of claims 32 to 39, wherein the sequence of the VHH with the attached conjugation handle comprises SEQ ID NO: 122.

41. The circRNA therapeutic of any one of claims 32 to 40, wherein the sequence of the VHH with the attached conjugation handle is SEQ ID NO: 122.

42. The circRNA therapeutic of any one of claims 32 to 41, wherein the LNP comprises the one or more ionizable lipids, the one or more phospholipids, the one or more sterol lipids, the DMG-PEG 2000 lipid, and the DSPE-PEG 2000-maleimide lipid at a ratio of 39: 20: 39: 1.9: 0.1 mol%.

43. The circRNA therapeutic of any one of claims 32 to 42, wherein the one or more ionizable lipids is Compound 7.

44. The circRNA therapeutic of any one of claims 32 to 43, wherein the one or more phospholipids is l,2-distearoyl- wglycero-3 -phosphocholine (DSPC).

45. The circRNA therapeutic of any one of claims 32 to 44, wherein the one or more sterol lipids is cholesterol.

46. The circRNA therapeutic of any one of claims 1 to 45, comprising a circRNA sequence having at least 85% identity to SEQ ID NO: 264.

47. The circRNA therapeutic of any one of claims 1 to 45, comprising a circRNA sequence according to SEQ ID NO: 264.

48. A circRNA therapeutic comprising(i) a circular RNA (circRNA) comprising a coding sequence encoding an anti-CD19 CAR, wherein the circRNA is encapsulated in an LNP,ATTORNEY DOCKET NO. ORB-019WO1(ii) the LNP, wherein the LNP comprises Compound 7 at 39 mol%, DSPC at 20 mol%, cholesterol at 39 mol%, DMG-PEG 2000 lipid at 1.9 mol%, and DSPE-PEG 2000-maleimide lipid at 0.1 mol%, and(iii) a VHH with a conjugation handle comprising the sequence of SEQ ID NO: 122, wherein the VHH with the conjugation handle is attached to the LNP via thiol-maleimide conjugation of the cysteine in the conjugation handle with the DSPE-PEG 2000-maleimide lipid.

49. The circRNA therapeutic of claim 48, wherein the circRNA comprises a sequence according to SEQ ID NO: 264.

50. A circRNA therapeutic, comprising:(i) a circRNA comprising, in order:(a) a 5’ exon fragment and a 5' UTR,(b) an IRES having at least 85% identity to SEQ ID NO: 1,(c) a coding sequence encoding a chimeric antigen receptor (CAR) that targets CD 19,(d) a 3 ' UTR and a 3 ’ exon fragment, and(ii) an anti-CD5 T cell targeting moiety comprising the complementarity determining region (CDR) sequences CDR1, CDR2, and CDR3, wherein the CDR1, the CDR2, and the CDR3 amino acid sequences are GFTFSMYS (SEQ ID NO: 64), ISTGARDT (SEQ ID NO: 65), and GDLRYGPDGYDY (SEQ ID NO: 66), respectively,wherein the circRNA is encapsulated in a lipid nanoparticle, andwherein the anti-CD5 T cell targeting moiety is conjugated to the lipid nanoparticle.

51. The circRNA therapeutic of claim 50, wherein the coding sequence encoding the CAR that targets CD19 comprises SEQ ID NO: 125.ATTORNEY DOCKET NO. ORB-019WO152. The circRNA therapeutic of claim 51, wherein the CAR comprises an anti-CD19 scFv, a CD8a hinge, a CD8a transmembrane domain, a 4-1BB costimulatory domain, and a CD3 zeta signaling domain.

53. The circRNA therapeutic of claim 52, wherein the CAR comprises an amino acid sequence of SEQ ID NO: 58.

54. The circRNA therapeutic of claim 52, wherein the anti-CD19 scFv comprisesa VL sequence of DIQMTQTTSSLSASLGDRVTISCRASQDISKYLNWYQQKPDGTVKLLIYHT SRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCQQGNTLPYTFGGGTK LEIT (SEQ ID NO: 54),linked by a linker having the amino acid sequence of GGGGSGGGGSGGGGS (SEQ ID NO: 56), anda VH sequence of EVKLQESGPGLVAPSQSLSVTCTVSGVSLPDYGVSWIRQPPRKGLEWLGVI WGSETTYYNSALKSRLTIIKDNSKSQVFLKMNSLQTDDTAIYYCAKHYYY GGSYAMDYWGQGTSVTVSS (SEQ ID NO: 55).

55. A circular ribonucleic acid (circRNA) comprising in order:(a) a 5’ exon fragment and a 5’ untranslated region (UTR),(b) an Internal Ribosome Entry Site (IRES) comprising a sequence at least 85% identical to any one of SEQ ID NO: 1-11,(c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof,(d) a 3 ’ UTR and a 3 ’ exon fragment, and(e) a single microRNA binding site within the 5’ UTR or the 3’ UTR.

56. The circRNA of claim 55, wherein the 3’ UTR comprises the single microRNA binding site.ATTORNEY DOCKET NO. ORB-019WO157. The circRNA of claim 55 or 56, wherein the microRNA binding site comprises SEQ ID NO: 40.

58. The circRNA of any one of claims 55 to 57, wherein the single microRNA binding site is flanked by an upstream spacer and a downstream spacer, and wherein the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3.

59. The circRNA of claim 58, wherein the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.4.

60. The circRNA of claim 58 or 59, wherein each of the upstream spacer and downstream spacer comprise consecutive adenines but do not comprise more than five (5) consecutive adenines (As).

61. The circRNA of claim 60, wherein each of the upstream spacer and downstream spacer comprise consecutive adenines but do not comprise more than four (4) consecutive adenines (As).

62. The circRNA of claim 61, wherein each of the upstream spacer and downstream spacer include consecutive adenines but do not comprise more than three (3) consecutive adenines (As).

63. A circular ribonucleic acid (circRNA) comprising, in order:(a) a 5’ exon fragment and a 5’ untranslated region (UTR),(b) an Internal Ribosome Entry Site (IRES),(c) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof, and(d) a 3’ UTR comprising an upstream spacer, a microRNA binding site, and a downstream spacer, and a 3 ’ exon fragment,ATTORNEY DOCKET NO. ORB-019WO1wherein each of the upstream spacer and downstream spacer comprise multiple instances of consecutive adenines (As) but do not comprise more than five (5) consecutive adenines (As).

64. The circRNA of claim 63, wherein each of the upstream spacer and the downstream spacer do not comprise more than four (4) consecutive adenines (As).

65. The circRNA of claim 64, wherein each of the upstream spacer and the downstream spacer do not comprise more than three (3) consecutive adenines (As).

66. The circRNA of any one of claims 63-65, wherein the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3.

67. The circRNA of claim 66, wherein the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.4.

68. The circRNA of any one of claims 63-67, wherein each of the upstream spacer and downstream spacer comprise one or more instances of a cytosine (C) interrupting consecutive As.

69. The circRNA of any one of claims 63-68, wherein each of the upstream spacer and downstream spacer comprise no uracil.

70. The circRNA of any one of claims 63-69, wherein the upstream spacer sequence and the downstream spacer sequence consist only of As and Cs.

71. The circRNA of any one of claims 63-70, wherein each of the upstream spacer and downstream spacer have a length of not more than 18 nucleotides.

72. The circRNA of claim 71, wherein each of the upstream spacer and downstream spacer have a length of 12-18 nucleotides.

73. The circRNA of claim 72, wherein each of the upstream spacer and downstream spacer have a length of 12-15 nucleotides.

74. The circRNA of any one of claims 63-73, wherein the upstream spacer and the downstream spacer do not differ in length by more than 4 nucleotides.ATTORNEY DOCKET NO. ORB-019WO175. The circRNA of claim 74, wherein the upstream spacer and the downstream spacer do not differ in length by more than 3 nucleotides.

76. The circRNA of claim 75, wherein the upstream spacer and the downstream spacer do not differ in length by more than 2 nucleotides.

77. The circRNA of any one of claims 63-76, where the microRNA binding site is a microRNA-122 (miR-122) binding site.

78. The circRNA of any one of claims 63-77, wherein the 3’ UTR comprises TAATAGaacaaacacaaacCAAACACCATTGTCACACTCCAaacaacaaacaaacaACCACACA AATGGTCGCCGA (SEQ ID NO: 349).

79. The circRNA of any one of claims 63-78, wherein the 5’ UTR comprises TCGGCGACCATTTGTGTGGTGAGCTCTAAGTGTTGATCAGCGGGCGCAGCTGCAT ATGACCTCTGCGTGTGCCTGGTAATTGACTAAGATATC (SEQ ID NO: 348).

80. The circRNA of any one of claims 63-79, wherein the 5’ UTR comprises SEQ ID NO: 348, and the 3’ UTR comprises SEQ ID NO: 349.

81. The circRNA of any one of claims 63-79, wherein the 5’ exon fragment comprises SEQ ID NO: 382, and a 3’ exon fragment comprises SEQ ID NO: 389.

82. The circRNA of any one of claims 63-81, wherein the IRES comprises a sequence having at least 90% identity to any one of SEQ ID NO: 1-6.

83. The circRNA of claim 82, wherein the IRES comprises a sequence of SEQ ID NO: 1.

84. The circRNA of any one of claims 63-83, wherein:the coding sequence further encodes:a CD8 signal peptide, andthe anti-CD19 CAR further comprises a CD8 hinge, a CD8 transmembrane domain, a 4-1BB costimulatory domain, and a CD3 zeta signaling domain.ATTORNEY DOCKET NO. ORB-019WO185. The circRNA of any one of claims 63-84, wherein the anti-CD19 antibody or antigenbinding fragment thereof comprises a VL sequence of SEQ ID NO: 54, and a VH sequence of SEQ ID NO: 55.

86. The circRNA of claim 85, wherein the VL and the VH are joined by a linker.

87. The circRNA of claim 86, wherein the linker comprises SEQ ID NO: 56.

88. The circRNA of any one of claims 63-87, wherein the coding sequence encodes an amino acid sequence having at least 90% identity to SEQ ID NO: 58.

89. The circRNA of claim 88, wherein the coding sequence encodes an amino acid sequence having at least 95% identity to SEQ ID NO: 58.

90. The circRNA of claim 89, wherein the coding sequence encodes an amino acid sequence of SEQ ID NO: 58.

91. A circRNA comprising a sequence having at least 85% identity to SEQ ID NO: 264.

92. The circRNA of claim 91, comprising a sequence having at least 90% identity to SEQ ID NO: 264.

93. The circRNA of claim 92, comprising a sequence having at least 95% identity to SEQ ID NO: 264.

94. The circRNA of claim 93, comprising a sequence having at least 98% identity to SEQ ID NO: 264.

95. The circRNA of claim 94, comprising a sequence of SEQ ID NO: 264.

96. The circRNA therapeutic of any one of claims 32 to 44, comprising circRNA according to any one of claims 55-95.

97. The circRNA therapeutic of claim 48, wherein the circRNA is according to any one of claims 55-94.

98. A circRNA therapeutic comprising the circRNA of any one of claims 55-95 encapsulated in an LNP.ATTORNEY DOCKET NO. ORB-019WO199. The circRNA therapeutic of claim 98, wherein the LNP is conjugated to a targeting moiety, and wherein the targeting moiety binds to T cells.

100. The circRNA therapeutic of claim 99, wherein the targeting moiety comprises an anti-CD5 antibody or antigen-binding fragment thereof.

101. A precursor RNA comprising, from 5 ' to 3 ' end:(i) a 5' intron fragment and a 5’ exon fragment,(ii) a 5' untranslated region (UTR),(iii) an IRES,(iv) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof,(v) a 3' untranslated region (UTR), and(vi) a 3 ' exon fragment and a 3 ’ intron fragment,wherein the 5' and 3' intron fragments are derived from a Twort- ORF142 Group I intron, andwherein the 3’ UTR comprises a single microRNA binding site.

102. The precursor RNA of claim 101, wherein the single microRNA binding site is flanked by an upstream spacer and a downstream spacer.

103. The precursor RNA of claim 102, wherein each of the upstream spacer and the downstream spacer comprise no more than five (5) consecutive adenines (As).

104. The precursor RNA of claim 103, wherein each of the upstream spacer and the downstream spacer do not comprise more than four (4) consecutive adenines (As).

105. The precursor RNA of claim 104, wherein each of the upstream spacer and the downstream spacer do not comprise more than three (3) consecutive adenines (As).ATTORNEY DOCKET NO. ORB-019WO1106. The precursor RNA of any one of claims 101-105, wherein the sequence of each of the upstream spacer and the downstream spacer has a hydropathy index between -0.6 to -0.3.

107. The precursor RNA of claim 106, wherein the sequence of the upstream spacer has a hydropathy index of -0.6 to -0.4.

108. The precursor RNA of any one of claims 101-107, wherein each of the upstream spacer and downstream spacer comprise one or more instances of a cytosine (C) interrupting consecutive As.

109. The precursor RNA of any one of claims 101-108, wherein each of the upstream spacer and downstream spacer comprise no uracil.

110. The precursor RNA of any one of claims 101-109, wherein the upstream spacer sequence and the downstream spacer sequence consist only of As and Cs.

111. The precursor RNA of any one of claims 101-110, wherein each of the upstream spacer and downstream spacer have a length of not more than 18 nucleotides.

112. The precursor RNA of claim 111, wherein each of the upstream spacer and downstream spacer have a length of 12-18 nucleotides.

113. The precursor RNA of claim 112, wherein each of the upstream spacer and downstream spacer have a length of 12-15 nucleotides.

114. The precursor RNA of any one of claims 101-113, wherein the upstream spacer and the downstream spacer do not differ in length by more than 4 nucleotides.

115. The precursor RNA of claim 114, wherein the upstream spacer and the downstream spacer do not differ in length by more than 3 nucleotides.

116. The precursor RNA of claim 115, wherein the upstream spacer and the downstream spacer do not differ in length by more than 2 nucleotides.

117. The precursor RNA of any one of claims 101-116, where the microRNA binding site is a microRNA-122 (miR-122) binding site.ATTORNEY DOCKET NO. ORB-019WO1118. A precursor RNA comprising, from 5' to 3' end:(i) a 5' intron fragment and a 5’ exon fragment,(ii) a 5' untranslated region (UTR),(iii) an IRES,(iv) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof,(v) a 3' untranslated region (UTR), and(vi) a 3 ' exon fragment and 3 ’ intron fragment,wherein the 5' and the 3' intron fragments are derived from a Twort- ORF142 Group I intron,wherein the 3’ UTR comprises a microRNA binding site flanked by an upstream spacer and a downstream spacer, andwherein each of the upstream spacer and the downstream spacer comprise no more than five (5) consecutive adenines (As).

119. The precursor RNA of claim 118, wherein each of the upstream spacer and the downstream spacer comprise no more than four (4) consecutive adenines (As).

120. The precursor RNA of claim 119, wherein each of the upstream spacer and the downstream spacer comprise no more than three (3) consecutive adenines (As).

121. The precursor RNA of any one of claims 118-120, wherein each of the upstream spacer and the downstream spacer comprise one or more instances of a cytosine (C) interrupting consecutive As.

122. The precursor RNA of any one of claims 118-120, wherein each of the upstream spacer and the downstream spacer comprise no uracil.ATTORNEY DOCKET NO. ORB-019WO1123. The precursor RNA of any one of claims 118-121, wherein each of the upstream spacer and the downstream spacer comprise 12-18 nucleotides.

124. The precursor RNA of claim 123, wherein each of the upstream spacer and downstream spacer have a length of 12-15 nucleotides.

125. The precursor RNA of any one of claims 118-124, wherein the upstream spacer and the downstream spacer do not differ in length by more than 4 nucleotides.

126. The precursor RNA of claim 125, wherein the upstream spacer and the downstream spacer do not differ in length by more than 3 nucleotides.

127. The precursor RNA of claim 126, wherein the upstream spacer and the downstream spacer do not differ in length by more than 2 nucleotides.

128. The precursor RNA of any one of claims 118-127, where the microRNA binding site is a microRNA-122 (miR-122) binding site.

129. The precursor RNA of any one of claims 101-117 and 118-128, wherein the 5’ intron fragment comprises SEQ ID NO: 381.

130. The precursor RNA of any one of claims 101-117 and 118-129, wherein the 5’ exon fragment comprises SEQ ID NO: 382.

131. The precursor RNA of any one of claims 101-117 and 118-130, wherein the 5’ UTR comprises SEQ ID NO: 348.

132. The precursor RNA of any one of claims 101-117 and 118-131, wherein the IRES comprises a ribonucleic acid (RNA) sequence having at least 90% identity to any one of SEQ ID NO: 1-6.

133. The precursor RNA of claim 132, wherein the IRES comprises the sequence of SEQ ID NO: 1.

134. The precursor RNA of any one of claims 101-117 and 118-133, wherein the coding sequence encoding the CAR comprising the anti-CD19 antibody or antigen-binding fragment thereof comprises the sequence of SEQ ID NO: 355.ATTORNEY DOCKET NO. ORB-019WO1135. The precursor RNA of any one of claims 101-117 and 118-134, wherein the 3’ UTR comprises SEQ ID NO: 349 or 350.

136. The precursor RNA of any one of claims 101-117 and 118-135, wherein the 3’ exon fragment comprises SEQ ID NO: 389.

137. The precursor RNA of any one of claims 101-117 and 118-136, wherein the 3’ intron fragment comprises SEQ ID NO: 390.

138. The precursor RNA of any one of claims 101-117 and 118-137, wherein the 5’ UTR comprises SEQ ID NO: 348, and the 3’ UTR comprises SEQ ID NO: 349.

139. The precursor RNA of any one of claims 101-117 and 118-138, wherein the 5’ intron fragment and the 5’ exon fragment together comprises SEQ ID NO: 276, and the 3’ exon fragment and the 3’ intron fragment together comprises SEQ ID NO: 277.

140. A precursor RNA comprising, from 5' to 3' end:(i) a 5' intron fragment and a 5’ exon fragment together comprising SEQ ID NO: 276,(ii) a 5' untranslated region (UTR),(iii) an IRES,(iv) a coding sequence encoding a chimeric antigen receptor (CAR) comprising an anti-CD19 antibody or antigen-binding fragment thereof,(v) a 3' untranslated region (UTR), and(vi) a 3’ exon fragment and a 3' intron fragment together comprising SEQ ID NO: 277.

141. A circular RNA circularized from the precursor RNA of any one of claims 100-140.

142. A circular RNA (circRNA) therapeutic comprising the circular RNA of claim 141.ATTORNEY DOCKET NO. ORB-019WO1143. A method of depleting B cells comprising administering to a human patient in need thereof the circRNA therapeutic of any one of claims 1-54 or 95-99 and 141-142.

144. A method of treating a B cell-mediated disease comprising administering to a human patient in need thereof the circRNA therapeutic of claim 143.

145. The method of claim 143 or 144, wherein the B cell-mediated disease is an autoimmune disease.

146. The method of claim 143 or 144, wherein the B cell-mediated disease is cancer.

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