Modified immune cells having enhanced Anti-neoplasia activity and immunosuppression resistance

Abstract

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Description

INCOPORATION BY REFERENCE 5

[0001] This application claims the benefit of U.S. Provisional Application No. 62 / 793,277 filed on January 16, 2019 and U.S. Provisional Application No. 62 / 839,870 filed on April 29, 2019. BACKGROUND OF THE INVENTION

[0002] Autologous and allogeneic immunotherapies are neoplasia treatment approaches in 10 which immune cells expressing chimeric antigen receptors are administered to a subject. To generate an immune cell that expresses a chimeric antigen receptor (CAR), the immune cell is first collected from the subject (autologous) or a donor separate from the subject receiving treatment (allogeneic) and genetically modified to express the chimeric antigen receptor. The resulting cell expresses the chimeric antigen receptor on its cell surface (e.g., CAR T-cell), and 15 upon administration to the subject, the chimeric antigen receptor binds to the marker expressed by the neoplastic cell. This interaction with the neoplasia marker activates the CAR-T cell, which then cell kills the neoplastic cell. But for autologous or allogeneic cell therapy to be effective and efficient, significant conditions and cellular responses, such as T cell signaling inhibition, must be overcome or avoided. For allogeneic cell therapy, graft versus host disease 20 and host rejection of CAR-T cells may provide additional challenges. Editing genes involved in these processes can enhance CAR-T cell function and resistance to immunosuppression or inhibition, but current methodologies for making such edits have the potential to induce large, genomic rearrangements in the CAR-T cell, thereby negatively impacting its efficacy. Thus, there is a significant need for techniques to more precisely modify immune cells, especially 25 CAR-T cells. This application is directed to this and other important needs. SUMMARY OF THE INVENTION

[0003] As described below, the present invention features genetically modified immune cells having enhanced anti-neoplasia activity, resistance to immune suppression, and decreased risk of 30 eliciting a graft versus host reaction, or host versus graft reaction where host CD8+ T cells recognize a graft as non-self (e.g., where a transplant recipient generates an immune response 2026204466   11 Jun 2026 against the transplanted organ), or a combination thereof. In one embodiment, a subject having or having a propensity to develop graft versus host disease (GVHD) is administered a CAR-T cell that lacks or has reduced levels of functional TRAC. In one embodiment, a subject having or having a propensity to develop host versus graft disease (HVGD) is administered a CAR-T 5 cell that lacks or has reduced levels of functional beta2 microglobulin (B2M). The present invention also features methods for producing and using these modified immune cells.

[0004] In one aspect, provided herein is a method for producing a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity by multiplexed editing, the method comprising: modifying at least four gene sequences or regulatory elements thereof, at a 10 single target nucleobase in each thereof in an immune cell, thereby generating the modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity.

[0005] In another aspect, provided herein is a method for producing a population of modified immune cells with reduced immunogenicity and / or increased anti-neoplasia activity by multiplexed editing, the method comprising: modifying at least four gene sequences or 15 regulatory elements thereof at a single target nucleobase in each thereof in a population of immune cells, thereby generating the population of modified immune cells with reduced immunogenicity and / or increased anti-neoplasia activity.

[0006] In some embodiments, the at least one of the at least four gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an 20 immunogenic gene sequence.

[0007] In some embodiments, the modifying reduces expression of at least one of the at least four gene sequences.

[0008] In some embodiments, the expression of at least one of the at least four genes is reduced by at least 80% as compared to a control cell without the modification. 25

[0009] In some embodiments, the expression of each one of the at least four genes is reduced by at least 80% as compared to a control cell without the modification.

[0010] In some embodiments, the expression of at least one of the at least four genes is reduced in at least 50% of the population of immune cells.

[0011] In some embodiments, the expression of each one of the at least four genes is reduced 30 in at least 50% of the population of immune cells. 2026204466   11 Jun 2026

[0012] In some embodiments, the at least four gene sequences comprise a TRAC gene sequence.

[0013] In some embodiments, the at least four gene sequences comprise a check point inhibitor gene sequence. 5

[0014] In some embodiments, the at least four gene sequences comprise a PDCD1 gene sequence.

[0015] In some embodiments, the at least four gene sequences comprise a T cell marker gene sequence.

[0016] In some embodiments, the at least four gene sequences comprise a CD52 gene 10 sequence.

[0017] In some embodiments, the at least four gene sequences comprises a CD7 gene sequence.

[0018] In some embodiments, the at least four gene sequences comprise a TRAC gene sequence, a PDCD1 gene sequence, a CD52 gene sequence, or a CD7 gene sequence. 15

[0019] In some embodiments, the at least four sequences comprise a TCR complex gene sequence, a CD7 gene sequence, a CD52 gene sequence ,and a gene sequence selected from the group consisting of CIITA a CD2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence 20

[0020] In some embodiments, the at least four gene sequences comprise a gene sequence selected from the group consisting of a CD2 gene sequence, a TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 25 gene sequence, a B2M gene sequence, and a CIITA gene sequence.

[0021] The method of some embodiments described herein comprises modifying five gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the immune cell.

[0022] The method of some embodiments described herein comprises modifying six gene 30 sequences or regulatory elements thereof at a single target nucleobase in each thereof in the immune cell. 2026204466   11 Jun 2026

[0023] The method of some embodiments described herein comprises modifying seven gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the immune cell.

[0024] The method of some embodiments described herein comprises modifying eight gene 5 sequences or regulatory elements thereof at a single target nucleobase in each thereof in the immune cell.

[0025] The method of some embodiments described herein comprises modifying five gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the population of immune cells. 10

[0026] The method of some embodiments described herein comprises modifying six gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the population of immune cells.

[0027] The method of some embodiments described herein comprises modifying seven gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the 15 population of immune cells.

[0028] The method of some embodiments described herein modifying eight gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the population of immune cells.

[0029] In some embodiments, the five, six, seven, or eight gene sequences or regulatory 20 elements thereof are selected from the group consisting of a CD2 gene sequence, a TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence. 25

[0030] In some embodiments, the five, six, seven, or eight gene sequences or regulatory elements thereof at comprises a CD3 gene sequence, a CD7 gene sequence, a CD2 gene sequence, a CD5 gene sequence, and a CD52 gene sequence.

[0031] In some embodiments, the modifying comprises deaminating the single target nucleobase. 30

[0032] In some embodiments, the deaminating is performed by a polypeptide comprising a deaminase. 2026204466   11 Jun 2026

[0033] In some embodiments, the deaminase is associated with a nucleic acid programmable DNA binding protein (napDNAbp) to form a base editor.

[0034] In some embodiments, the deaminase is fused to the nucleic acid programmable DNA binding protein (napDNAbp). 5

[0035] In some embodiments, the napDNAbp comprises a Cas9 polypeptide or a portion thereof.

[0036] In some embodiments, the napDNAbp comprises a Cas9 nickase or nuclease dead Cas9.

[0037] In some embodiments, the deaminase is a cytidine deaminase.

[0038] In some embodiments, the single target nucleobase is a cytosine (C) and wherein the 10 modification comprises conversion of the C to a thymine (T).

[0039] In some embodiments, the base editor further comprises a uracil glycosylase inhibitor.

[0040] In some embodiments, the deaminase is an adenosine deaminase.

[0041] In some embodiments, the single target nucleobase is a adenosine (A) and wherein the modification comprises conversion of the A to a guanine (G). 15

[0042] In some embodiments, the modifying comprises contacting the immune cell with a guide nucleic acid sequences.

[0043] In some embodiments, the modifying comprises contacting the immune cell with at least four guide nucleic acid sequences, wherein each guide nucleic acid sequence targets the napDNAbp to one of the at least four gene sequences or regulatory elements thereof. 20

[0044] In some embodiments, the guide nucleic acid sequence comprises a sequence selected from guide RNA sequences of table 8 A, table 8B, or table 8C.

[0045] In some embodiments, the guide nucleic acid sequence comprises a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC, 25 CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

[0046] In some embodiments, the modifying comprises replacing the single target nucleobase with a different nucleobase by target-primed reverse transcription with a reverse transcriptase 30 and an extended guide nucleic acid sequence. 2026204466   11 Jun 2026

[0047] In some embodiments, the extended guide nucleic acid sequence comprises a reverse transcription template sequence, a reverse transcription primer binding site, or a combination thereof.

[0048] In some embodiments, the single target nucleobase is in an exon. 5

[0049] In some embodiments, modifying generates a premature stop codon in the exon.

[0050] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of the TRAC gene sequence.

[0051] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 5 of the PCDC1 gene sequence. 10

[0052] In some embodiments, the single target nucleobase is within an exon 1 or an exon 2 of the CD52 gene sequence.

[0053] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of the CD7 gene sequence.

[0054] In some embodiments, the single target nucleobase is within an exon 1 or an exon 2 of 15   the B2M gene sequence.

[0055] In some embodiments, the single target nucleobase is within an exon 2, an exon 3, an exon 4, an exon 5, an exon 6, an exon 7, or an exon 8 of the CD5 gene sequence.

[0056] In some embodiments, the single target nucleobase is within an exon 2, an exon 3, an exon 4, or an exon 5 of the CD2 gene sequence. 20

[0057] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, an exon 4, an exon 7, an exon 8, an exon 9, an exon 10, an exon 11, an exon 12, an exon 14, an exon 15, an exon 18, or an exon 19 of the CIITA gene sequence.

[0058] In some embodiments, the single target nucleobase is in a splice donor site or a splice acceptor site. 25

[0059] In some embodiments, the single target nucleobase is in an exon 1 splice acceptor site, an exon 1 splice donor site, or an exon 3 splice acceptor site of the TRAC gene sequence.

[0060] In some embodiments, the single target nucleobase is in an exon 1 splice acceptor site, an exon 1 splice donor site, an exon 2 splice acceptor site, an exon 3 splice donor site, an exon 4 splice acceptor site, an exon 4 splice donor site, or an exon 5 splice acceptor site of the PDCD1 30 gene sequence. 2026204466   11 Jun 2026

[0061] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, or an exon 2 splice acceptor site of the CD52 gene sequence.

[0062] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, an exon 2 splice donor site, an exon 2 splice acceptor site, or an exon 3 splice acceptor site of the 5 CD7 gene sequence.

[0063] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, an exon 2 splice donor site, an exon 2 splice acceptor site, or an exon 3 splice acceptor site of the B2M gene sequence.

[0064] In some embodiments, the single target nucleobase is in an exon 3 splice donor site of 10 the CD2 gene sequence.

[0065] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, an exon 1 splice acceptor site, an exon 3 splice acceptor site, an exon 3 splice donor site, an exon 4 splice acceptor site, an exon 5 splice donor site, an exon 6 splice acceptor site, an exon 9 splice donor site, an exon 10 splice acceptor site of the CD5 gene sequence. 15

[0066] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, an exon 7 splice donor site, an exon 8 splice acceptor site, an exon 9 slice donor site, an exon 10 splice acceptor site, an exon 11 splice acceptor site, an exon 14 splice acceptor site, an exon 14 splice donor site, an exon 15 splice donor site, an exon 16 splice acceptor site, an exon 16 splice donor site, an exon 17 splice acceptor site, an exon 17 splice donor site, or an exon 19 splice 20 acceptor site of the CIITACIITA gene sequence.

[0067] In some embodiments, the immune cell is a human cell. In some embodiments, the immune cell is a cytotoxic T cell, a regulatory T cell, a T helper cell, a dendritic cell, a B cell, or aNK cell.

[0068] In some embodiments, the population of immune cells are human cells. 25

[0069] In some embodiments, the population of immune cells are cytotoxic T cells, regulatory T cells, T helper cells, dendritic cells, B cells, or NK cells.

[0070] In some embodiments, the modifying is ex vivo.

[0071] In some embodiments, the immune cell or the population of immune cells are derived from a single human donor. 2026204466   11 Jun 2026

[0072] In some embodiments, the method further comprising contacting the immune cell or the population of immune cells with a polynucleotide that encodes an exogenous functional chimeric antigen receptor (CAR) or a functional fragment thereof.

[0073] In some embodiments, contacting the immune cell or the population of immune cells 5 with a lentivirus comprising the polynucleotide that encodes the CAR.

[0074] In some embodiments, contacting the immune cell or the population of immune cells with a napDNAbp and a donor DNA sequence comprising the polynucleotide that encodes the CAR.

[0075] In some embodiments, the napDNAbp is a Cas 12b. 10

[0076] In some embodiments, the CAR specifically binds a marker associated with neoplasia.

[0077] In some embodiments, the neoplasia is a T cell cancer, a B cell cancer, a lymphoma, a leukemia, or a multiple myeloma.

[0078] In some embodiments the CAR specifically binds CD7.

[0079] In some embodiments, the CAR specifically binds BCMA. 15

[0080] In some embodiments, the immune cell or the population of immune cells comprises no detectable translocation. In some embodiments, at least 50% of the population of immune cells express the CAR. In some embodiments, at least 50% of the population of immune cells are viable. In some embodiments, at least 50% of the population of immune cells expand at least 80% of expansion rate of a population of control cells of a same type without the modification. 20

[0081] In the method of some embodiments described herein, the modifying generates less than 1% of indels in the immune cell. In some embodiments, the modifying generates less than 5% of non-target edits in the immune cell. In some embodiments, the modifying generates less than 5% of off-target edits in the immune cell.

[0082] In one aspect, provided herein is a modified immune cell produced according to some 25 embodiments described in the preceding paragraphs.

[0083] In one aspect, provided herein is a population of modified immune cells produced according to some embodiments described in the preceding paragraphs.

[0084] In another aspect, provided herein is a modified immune cell with reduced immunogenicity or increased anti-neoplasia activity, wherein the modified immune cell 30 comprises a single target nucleobase modification in each one of at least four gene sequences or regulatory elements thereof. In some embodiments, in the modified immune cell described 2026204466   11 Jun 2026 above, each one of the at least four gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.

[0085] In the modified immune cell of the preceding embodiments the at least four gene sequences comprise a TCR complex gene sequence. 5

[0086] In some embodiments, the at least four gene sequences comprise a TRAC gene sequence. In some embodiments, the at least four gene sequences comprise a check point inhibitor gene sequence. In some embodiments, the at least four gene sequences comprise a PDCD1 gene sequence.

[0087] In some embodiments, the at least four gene sequences comprise a T cell marker gene 10 sequence.

[0088] In some embodiments, the at least four gene sequences comprise CD52 gene sequence.

[0089] In some embodiments, the at least four gene sequences comprises a CD7 gene sequence.

[0090] In some embodiments, the expression of one of the at least four genes is reduced by at 15 least 80% as compared to a control cell without the modification.

[0091] In some embodiments, the expression of each one of the at least four genes is reduced by at least 90% as compared to a control cell without the modification.

[0092] In some embodiments, the immune cell comprises a modification at a single target nucleobase in each one of five gene sequences or regulatory elements thereof, wherein each one 20   of the five gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.

[0093] In some embodiments, the immune cell comprises a modification at a single target nucleobase in each one of six gene sequences or regulatory elements thereof, wherein each one of the six gene sequences is a checkpoint inhibitor gene sequence, an immune response 25 regulation gene sequence, or an immunogenic gene sequence.

[0094] In some embodiments, the immune cell comprises a modification at a single target nucleobase in each one of seven gene sequences or regulatory elements thereof, wherein each one of the seven gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence or an immunogenic gene sequence. 30

[0095] In some embodiments, the immune cell comprises a modification at a single target nucleobase in each one of eight gene sequences or regulatory elements thereof, wherein each one 2026204466   11 Jun 2026 of the eight gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.

[0096] In some embodiments, the expression of at least one of the five, six, seven or eight genes is reduced by at least 90% as compared to a control cell without the modification. 5

[0097] In some embodiments, the expression of each one of the five, six, seven, or eight genes is reduced by at least 90% as compared to a control cell without the modification.

[0098] In some embodiments, the five, six, seven, or eight gene sequences or regulatory elements thereof comprise a sequence selected from the group consisting of a CD2 gene sequence, a TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, 10 a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence.

[0099] In one aspect, provided herein is a modified immune cell comprising a single target 15 nucleobase modification in each one of a CD3 gene sequence, a CD5 gene sequence, a CD52 gene sequence, and a CD7 gene sequence, wherein the modified immune cell exhibits reduced immunogenicity or increased anti-neoplasia activity as compared to a control cell of a same type without the modification.

[00100] In some embodiments, the modified immune cell further comprises a single target 20 nucleobase modification in a CD2 gene sequence, CIITA or a regulatory element of each thereof.

[00101] In some embodiments, the modified immune cell comprises a single target nucleobase modification in a TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, or a TRBC2 gene sequence further comprises a single target nucleobase modification in a gene sequence a CD4 gene 25 sequence, a CD30 gene sequence, a CD33 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence or a regulatory element of each thereof.

[00102] In some embodiments, the modified immune cell comprises a single nucleobase modification in each one of a TRAC gene sequence, a PDCD1 gene sequence, a CD52 gene sequence, a CD7 gene sequence, a CD2 gene sequence, a CD5 gene sequence, a CIITA gene 30 sequence, and a B2M gene sequence. 2026204466   11 Jun 2026

[00103] In some embodiments, the modified immune cell comprises no detectable translocation.

[00104] In some embodiments, the modified immune cell comprises less than 1% of indels.

[00105] In some embodiments, the modified immune cell comprises less than 5% of non-target 5 edits.

[00106] In some embodiments, the modified immune cell comprises less than 5% of off-target edits.

[00107] In some embodiments, the modified immune has increased growth or viability compared to a reference cell. In some embodiments, the reference cell is an immune cell 10 modified with a Cas9 nuclease.

[00108] In some embodiments, the modified immune cell is a mammalian cell.

[00109] In some embodiments, the modified immune cell is a human cell.

[00110] In some embodiments, the modified immune cell is a cytotoxic T cell, a regulatory T cell, a T helper cell, a dendritic cell, a B cell, or a NK cell. 15

[00111] In some embodiments, the modified the immune cell is in an ex vivo culture.

[00112] In some embodiments, the modified the immune cell is derived from a single human donor.

[00113] In some embodiments, the modified the immune cell further comprises a polynucleotide that encodes an exogenous functional chimeric antigen receptor (CAR) or a 20 functional fragment thereof.

[00114] In some embodiments, the polynucleotide that encodes the CAR is integrated in the genome of the immune cell.

[00115] In some embodiments, the CAR specifically binds a marker associated with neoplasia.

[00116] In some embodiments, the neoplasia is a T cell cancer, a B cell cancer, a lymphoma, a 25 leukemia, or a multiple myeloma.

[00117] In some embodiments, the CAR specifically binds CD7.

[00118] In some embodiments, the CAR specifically binds BCMA.

[00119] In some embodiments, the single target nucleobase is in an exon.

[00120] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or 30 an exon 3 of the TRAC gene sequence. 2026204466   11 Jun 2026

[00121] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 5 of the PCDC1 gene sequence.

[00122] In some embodiments, the single target nucleobase is within an exon 1 or an exon 2 of the CD52 gene sequence. 5

[00123] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of a CD7 gene sequence.

[00124] In some embodiments, the single target nucleobase is in a splice donor site or a splice acceptor site.

[00125] In some embodiments, the single target nucleobase is in an exon 1 splice acceptor site, 10 an exon 1 splice donor site, or an exon 3 splice acceptor site of the TRAC gene sequence.

[00126] In some embodiments, the single target nucleobase is in an exon 1 splice acceptor site, an exon 1 splice donor site, an exon 2 splice acceptor site, an exon 3 splice donor site, an exon 4 splice acceptor site, an exon 4 splice donor site, or an exon 5 splice acceptor site of the PDCD1 gene sequence. 15

[00127] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, or an exon 2 splice acceptor site of the CD52 gene sequence.

[00128] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, an exon 2 splice donor site, an exon 2 splice acceptor site, or an exon 3 splice acceptor site of the CD7 gene sequence. 20

[00129] In one aspect, provided herein is a population of modified immune cells, wherein a plurality of the population of cells comprise a single target nucleobase modification in each one of at least four gene sequences or regulatory elements thereof, and wherein the plurality of the population of cells having the modification exhibit reduced immunogenicity or increased antineoplasia activity as compared to a plurality of control cells of a same type without the 25 modification.

[00130] In some embodiments, the plurality of cells comprises at least 50% of the population.

[00131] In some embodiments, each one of the at least four gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence. 30

[00132] In some embodiments, the at least four gene sequences comprise a TCR component gene sequence, a check point inhibitor gene sequence, or a T cell marker gene sequence. 2026204466   11 Jun 2026

[00133] In some embodiments, the at least four gene sequences comprise a TRAC gene sequence.

[00134] In some embodiments, the at least four gene sequences comprise a PDCD1 gene sequence. 5

[00135] In some embodiments, the at least four gene sequences comprise CD52 gene sequence.

[00136] In some embodiments, the at least four gene sequences comprises a CD7 gene sequence.

[00137] In the population of some embodiments, expression of at least one of the at least four genes is reduced by at least 80% in the plurality of cells having the modification as compared to 10 a control cell without the modification

[00138] In the population of some embodiments, expression of each one of the at least four genes is reduced by at least 80% in the plurality of cells having the modification as compared to a control cell without the modification.

[00139] In some embodiments, the plurality of the population comprises a modification at a 15 single target nucleobase in each one of five gene sequences or regulatory elements thereof, wherein each one of the five gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.

[00140] In some embodiments, the plurality of the population comprises a modification at a single target nucleobase in each one of six gene sequences or regulatory elements thereof, 20 wherein each one of the six sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence

[00141] In some embodiments, the plurality of the population comprises a modification at a single target nucleobase in each one of seven gene sequences or regulatory elements thereof, wherein each one of the seven gene sequences is a checkpoint inhibitor gene sequence, an 25 immune response regulation gene sequence, or an immunogenic gene sequence.

[00142] In some embodiments, the plurality of the population comprises a modification at a single target nucleobase in each one of eight gene sequences or regulatory elements thereof, wherein each one of the eight gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence. 2026204466   11 Jun 2026

[00143] In the population of some embodiments, the expression of at least one of the five, six, seven, or eight genes is reduced by at least 90% in the plurality of cells having the modification as compared to a control cell without the modification.

[00144] In the population of some embodiments, the expression of each one of the five, six, 5 seven, or eight genes is reduced by at least 90% in the plurality of cells having the modification as compared to a control cell without the modification.

[00145] In the population of some embodiments, the expression of at least one of the five, six, seven, or eight genes is reduced by at least 90% in the plurality of cells having the modification as compared to a control cell without the modification. 10

[00146] In some embodiments, the expression of each one of the five, six, seven, or eight genes is reduced by at least 90% in the plurality of cells having the modification as compared to a control cell without the modification.

[00147] In some embodiments, the five, six, seven, or eight gene sequences or regulatory elements thereof are selected from the group consisting of a CD2 gene sequence, a TRAC gene 15 sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence.

[00148] In one aspect, provided herein is a population of modified immune cells, wherein a 20 plurality of the population comprise a single target nucleobase modification in each one of a TRAC gene sequence, a PDCD1 gene sequence, a CD52 gene sequence, and a CD7 gene sequence, and wherein the plurality of the population having the modification exhibit reduced immunogenicity or increased anti-neoplasia activity as compared to a plurality of control cells of a same type without the modification. 25

[00149] In some embodiments, the plurality of the population further comprises a single target nucleobase modification in a CD2 gene sequence, a CD5 gene sequence, a CIITA gene sequence, a B2M gene sequence, or a regulatory element of each thereof. In some embodiments, the plurality of the population further comprises a single target nucleobase modification in a gene sequence of a gene selected from the group consisting of a CD2 gene sequence, a TRAC gene 30 sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene 2026204466   11 Jun 2026 sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence or a regulatory element of each thereof. In some embodiments, the plurality of the population comprises a single nucleobase modification in each one of a TRAC gene sequence, a PDCD1 5 gene sequence, a CD52 gene sequence, a CD7 gene sequence, a CD2 gene sequence, a CD5 gene sequence, a CIITA gene sequence, and a B2M gene sequence.

[00150] In the population of modified immune cells of some embodiments, the plurality of the population comprises no detectable translocation.

[00151] In the population of modified immune cells of some embodiments, the at least 60% of 10 the population of immune cells are viable. In the population of modified immune cells of some embodiments, the at least 60% of the population of immune cells expand at least 80% of expansion rate of a population of control cells of a same type without the modification. In the population of modified immune cells of some embodiments, the population of immune cells are human cells. In the population of modified immune cells of some embodiments, the population 15    of immune cells are cytotoxic T cells, regulatory T cells, T helper cells, dendritic cells, B cells, or NK cells. In the population of modified immune cells of some embodiments, the population of immune cells are derived from a single human donor. In the population of modified immune cells of some embodiments, the plurality of cells having the modification further comprises a polynucleotide that encodes an exogenous functional chimeric antigen receptor (CAR) or a 20 functional fragment thereof.

[00152] In some embodiments, the at least 50% of the population of immune cells express the CAR.

[00153] In some embodiments, the the CAR specifically binds a marker associated with neoplasia. 25

[00154] In some embodiments, the neoplasia is a T cell cancer, a B cell cancer, a lymphoma, a leukemia, or a multiple myeloma.

[00155] In some embodiments, the CAR specifically binds CD7.

[00156] In some embodiments, the CAR specifically binds BCMA.

[00157] In some embodiments, the single target nucleobase is in an exon. 30

[00158] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of the TRAC gene sequence. 2026204466   11 Jun 2026

[00159] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 5 of the PCDC1 gene sequence.

[00160] In some embodiments, the single target nucleobase is within an exon 1 or an exon 2 of the CD52 gene sequence. 5

[00161] In some embodiments, the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of a CD7 gene sequence.

[00162] In the population of modified immune cells of some embodiments, the single target nucleobase is in a splice donor site or a splice acceptor site.

[00163] In some embodiments, the single target nucleobase is in an exon 1 splice acceptor site, 10 an exon 1 splice donor site, or an exon 3 splice acceptor site of the TRAC gene sequence.

[00164] In some embodiments, the single target nucleobase is in an exon 1 splice acceptor site, an exon 1 splice donor site, an exon 2 splice acceptor site, an exon 3 splice donor site, an exon 4 splice acceptor site, an exon 4 splice donor site, or an exon 5 splice acceptor site of the PDCD1 gene sequence. 15

[00165] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, or an exon 2 splice acceptor site of the CD52 gene sequence.

[00166] In some embodiments, the single target nucleobase is in an exon 1 splice donor site, an exon 2 splice donor site, an exon 2 splice acceptor site, or an exon 3 splice acceptor site of the CD7 gene sequence. 20

[00167] In one aspect, provided herein is a composition comprising deaminase and a nucleic acid sequence, wherein the guide nucleic acid sequence comprises a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC, CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, 25 CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

[00168] In some embodiments, the deaminase is associated with a nucleic acid programmable DNA binding protein (napDNAbp) to form a base editor.

[00169] In some embodiments, the napDNAbp comprises a Cas9 nickase or nuclease dead Cas9 and wherein the deaminase is a cytidine deaminase. 30

[00170] In some embodiments, the base editor further comprises a uracil glycosylase inhibitor. 2026204466   11 Jun 2026

[00171] In some embodiments, the napDNAbp comprises a Cas9 nickase or nuclease dead Cas9 and wherein the deaminase is a adenosine deaminase.

[00172] In one aspect, provided herein is a composition comprising a polymerase and a guide nucleic acid sequence, wherein the guide nucleic acid sequence comprises a sequence selected 5 from the group consisting of the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC, CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC. 10

[00173] In some embodiments, the polymerase is a reverse transcriptase and wherein the guide nucleic acid sequence is an extended guide nucleic acid sequence comprising a reverse transcription template sequence, a reverse transcription primer binding site, or a combination thereof.

[00174] In one aspect, provided herein is a method for producing a modified immune cell with 15 reduced immunogenicity and / or increased anti-neoplasia activity, the method comprising: a) modifying a single target nucleobase in a first gene sequence or a regulatory element thereof in an immune cell; and b) modifying a second gene sequence or a regulatory element thereof in the immune cell with a Cas 12 polypeptide, wherein the Cas 12 polypeptide generates a site-specific cleavage in the second gene sequence; wherein each of the first gene and the second gene is a 20 immunogenic gene, a checkpoint inhibitor gene, or an immune response regulation gene, thereby generating a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity.

[00175] In some embodiments, the method further comprises expressing an exogenous functional chimeric antigen receptor (CAR) or a functional fragment thereof in the immune cell. 25

[00176] In some embodiments, a polynucleotide encoding the CAR or the functional fragment thereof is inserted into the site specific cleavage generated by the Cas 12 polypeptide.

[00177] In some embodiments, the Cas 12 polypeptide is a Cas 12b polypeptide.

[00178] In one aspect, provided herein is a method for producing a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity, the method comprising: 30

[00179] a) modifying a single target nucleobase in a first gene sequence or a regulatory element thereof in an immune cell; and b) modifying a second gene sequence or a regulatory 2026204466   11 Jun 2026 element thereof in the immune cell by inserting an exogenous functional chimeric antigen receptor (CAR) or a functional fragment thereof or an exogenous functional T cell receptor or a functional fragment thereof in the second gene; wherein each of the first gene and the second gene is a immunogenic gene, a checkpoint inhibitor gene, or an immune response regulation 5 gene, thereby generating a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity.

[00180] In some embodiments, the step b) further comprises generating a site-specific cleavage in the second gene sequence with a nucleic acid programmable DNA binding protein (napDNAbp). 10

[00181] In some embodiments, the napDNAbp is a Cas 12b.

[00182] In some embodiments, the expression of the first gene is reduced by at least 60% or wherein expression of the second gene is reduced by at least 60% as compared to a control cell of a same type without the modification.

[00183] In some embodiments, the first gene is selected from the group consisting of CD3 15 epsilon, CD3 gamma, CD3 delta, CD4, TRAC, TRBC1, TRBC2, PDCD1, CD30, CD33, CD7, CD52, B2M, CD70, CIITA, CD2, and CD5.

[00184] In some embodiments, the first gene or the second gene is selected from the group consisting of TRAC, CIITA, CD2, CD5, CD7, and CD52.

[00185] In some embodiments, the second gene is TRAC. 20

[00186] In some embodiments, the step a) further comprises modifying a single target nucleobase in two other gene sequences or regulatory elements thereof.

[00187] In some embodiments, the step a) further comprises modifying a single target nucleobase in three other gene sequences or regulatory elements thereof.

[00188] In some embodiments, the step a) further comprises modifying a single target 25 nucleobase in four other gene sequences or regulatory elements thereof.

[00189] In some embodiments, the step a) further comprises modifying a single target nucleobase in five other gene sequences or regulatory elements thereof.

[00190] In some embodiments, the step a) further comprises modifying a single target nucleobase in six other gene sequences or regulatory elements thereof. 30

[00191] In some embodiments, the step a) further comprises modifying a single target nucleobase in seven other gene sequences or regulatory elements thereof. 2026204466   11 Jun 2026

[00192] In some embodiments, the modifying in step a) comprises deaminating the single target nucleobase with a base editor comprising a deaminase and a nucleic acid programmable DNA binding protein (napDNAbp).

[00193] In some embodiments, the napDNAbp comprises a Cas9 nickase or nuclease dead 5 Cas9.

[00194] In some embodiments, the deaminase is a cytidine deaminase and wherein the modification comprises conversion of a cytidine (C) to a thymine (T).

[00195] In some embodiments, the deaminase is an adenosine deaminase and wherein the modification comprises conversion of an adenine (A) to a guanine (G). 10

[00196] In some embodiments, the modifying in a) comprises contacting the immune cell with a guide nucleic acid sequence.

[00197] In some embodiments, the guide nucleic acid sequence comprises a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC, 15 CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

[00198] In some embodiments, the modifying in b) comprises contacting the immune cell with a guide nucleic acid sequence. 20

[00199] In some embodiments, the guide nucleic acid sequence comprises a sequence selected from sequences in Table 1.

[00200] In some embodiments, the modifying in a) comprises replacing the single target nucleobase with a different nucleobase by target-primed reverse transcription with a reverse transcriptase and an extended guide nucleic acid sequence, wherein the extended guide nucleic 25 acid sequence comprises a reverse transcription template sequence, a reverse transcription primer binding site, or a combination thereof.

[00201] In some embodiments, wherein the modifying in a) and b) generates less than 1% indels in the immune cell.

[00202] In some embodiments, the modifying in a) and b) generates less than 5% off target 30 modification in the immune cell. 2026204466   11 Jun 2026

[00203] In some embodiments, the modifying in a) and b) generate less than 5% non-target modification in the immune cell.

[00204] In some embodiments, the immune cell is a human cell.

[00205] In some embodiments, the immune cell is a cytotoxic T cell, a regulatory T cell, a T 5 helper cell, a dendritic cell, a B cell, or a NK cell.

[00206] In some embodiments, the CAR specifically binds a marker associated with neoplasia.

[00207] In some embodiments, the CAR specifically binds CD7.

[00208] In one aspect, provided herein is a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity, wherein the modified immune cell 10 comprises:

[00209] a) a single target nucleobase modification in a first gene sequence or a regulatory element thereof; and b) a modification in a second gene sequence or a regulatory element thereof, wherein the modification is a Cas 12 polypeptide generated site-specific cleavage; wherein each of the first gene and the second gene is a immunogenic gene, a checkpoint inhibitor 15   gene, or an immune response regulation gene. In one embodiment, the immune cell further comprises an exogenous functional chimeric antigen receptor (CAR) or a functional fragment thereof.

[00210] In some embodiments, a polynucleotide encoding the CAR or the functional fragment thereof is inserted into the site specific cleavage generated by the Cas 12 polypeptide. 20

[00211] In one aspect, provided herein is a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity, the modified immune cell comprising: a) a single target nucleobase modification in a first gene sequence or a regulatory element thereof in an immune cell; and b) a modification in a second gene sequence or a regulatory element thereof, wherein the modification is an insertion of an exogenous chimeric antigen receptor 25 (CAR) or a functional fragment thereof or an exogenous T cell receptor or a functional fragment thereof; wherein each of the first gene and the second gene is a immunogenic gene, a checkpoint inhibitor gene, or immune response regulation gene.

[00212] In some embodiments, the modification in b) is generated by a site-specific cleavage with a Cas 12b. 2026204466   11 Jun 2026

[00213] In some embodiments, expression of the first gene is reduced by at least 60% or wherein expression of the second gene is reduced by at least 60% as compared to a control cell of a same type without the modification.

[00214] In some embodiments, the first gene or the second gene is selected from the group consisting of CD3 epsilon, CD3 gamma, CD3 delta, CD4, TRAC, TRBC1, TRBC2, PDCD1, CD30, CD33, CD7, CD52, B2M, CD70, CIITA, CD2, and CD5.

[00215] In some embodiments, the first gene or the second gene is selected from the group consisting of TRAC, CD2, CD5, CD7, and CD52.

[00216] In some embodiments, the second gene is TRAC.

[00217] In some embodiments, the immune cell further comprises modification in a single target nucleobase in two other gene sequences or regulatory elements thereof.

[00218] In some embodiments, the immune cell further comprises modification in a single target nucleobase in three other gene sequences or regulatory elements thereof.

[00219] In some embodiments, the immune cell further comprises modification in a single target nucleobase in four other gene sequences or regulatory elements thereof.

[00220] In some embodiments, the immune cell further comprises modification in a single target nucleobase in five other gene sequences or regulatory elements thereof.

[00221] In some embodiments, the immune cell further comprises modification in a single target nucleobase in six other gene sequences or regulatory elements thereof.

[00222] In some embodiments, the immune cell further comprises modification in a single target nucleobase in seven other gene sequences or regulatory elements thereof.

[00223] In some embodiments, the modification in a) is generated by a base editor comprising a deaminase and a nucleic acid programmable DNA binding protein (napDNAbp).

[00224] In some embodiments, the deaminase is a cytidine deaminase and the modification comprises conversion of a cytidine (C) to a thymine (T).

[00225] In some embodiments, the deaminase is an adenosine deaminase and wherein the modification comprises conversion of an adenine (A) to a guanine (G).

[00226] In some embodiments, the immune cell comprises less than 1% indels in the genome.

[00227] In some embodiments, the immune cell is a human cell.

[00228] In some embodiments, the immune cell is a cytotoxic T cell, a regulatory T cell, a T helper cell, a dendritic cell, a B cell, or a NK cell. 2026204466   11 Jun 2026

[00229] In some embodiments, the CAR specifically binds a marker associated with neoplasia.

[00230] In some embodiments, the CAR specifically binds CD7.

[00231] In some embodiments, the modification in b) is an insertion in exon 1 in the TRAC gene sequence. 5

[00232] In one aspect, provided herein is a population of modified immune cells, wherein a plurality of the population of immune cells comprises: a) a single target nucleobase modification in a first gene sequence or a regulatory element thereof in an immune cell; and b) a modification in a second gene sequence or a regulatory element thereof, wherein the modification is a Cas 12 polypeptide generated site-specific cleavage; wherein each of the first gene and the second gene 10 is a immunogenic gene, a checkpoint inhibitor gene, or an immune response regulation gene, and wherein the plurality of the population comprises an exogenous chimeric antigen receptor (CAR) or a functional fragment thereof.

[00233] In some embodiments, a polynucleotide encoding the CAR or the functional fragment thereof is inserted into the site specific cleavage generated by the Cas 12 polypeptide. 15

[00234] In one aspect, provided herein is a population of modified immune cells, wherein a plurality of the population of immune cells comprises: a) a single target nucleobase modification in a first gene sequence or a regulatory element thereof; and b) a modification in a second gene sequence or a regulatory sequence thereof, wherein the modification is an insertion of an exogenous chimeric antigen receptor (CAR) or a functional fragment thereof or an exogenous T 20 cell receptor or a functional fragment thereof; wherein each of the first gene and the second gene is a immunogenic gene, a checkpoint inhibitor gene, or immune response regulation gene, and wherein the plurality of cells with the modification in a) or b) exhibit reduced immunogenicity and / or increased anti-neoplasia activity. In some embodiments, the modification in b) is generated by a site-specific cleavage with a Cas 12b. In some embodiments, expression of the 25 first gene is reduced by at least 60% or wherein expression of the second gene is reduced by at least 60% in the plurality of cells with the modification in a) or b) as compared to plurality of control cells of a same type without the modification.

[00235] In some embodiments, the first gene or the second gene is selected from the group consisting of CD3 epsilon, CD3 gamma, CD3 delta, CD4, TRAC, TRBC1, TRBC2, PDCD1, 30   CD30, CD33, CD7, CD52, B2M, CD70, CIITA, CD2, and CD5. 2026204466   11 Jun 2026

[00236] In some embodiments, the first gene or the second gene is selected from the group consisting of TRAC, CIITA, CD2, CD5, , CD7, and CD52.

[00237] In some embodiments, the first gene is TRAC, CD7, or CD52.

[00238] In some embodiments, the second gene is TRAC. 5

[00239] In some embodiments, the plurality of cells with the modification in a) or b) further comprises a modification in a single target nucleobase in two other gene sequences or regulatory elements thereof.

[00240] In some embodiments, the plurality of cells with the modification in a) or b) further comprises a single target nucleobase in three, four, five, or six other gene sequences or 10 regulatory elements thereof.

[00241] In some embodiments, the modification in a) is generated by a base editor comprising a deaminase and a nucleic acid programmable DNA binding protein (napDNAbp) to form a base editor.

[00242] In some embodiments, the deaminase is a cytidine deaminase and wherein the 15 modification comprises conversion of a cytidine (C) to a thymine (T).

[00243] In some embodiments, the deaminase is an adenosine deaminase and wherein the modification comprises conversion of an adenine (A) to a guanine (G).

[00244] In some embodiments, the base editor further comprises a uracil glycosylase inhibitor.

[00245] In some embodiments, at least 60% of the population of immune cells are viable. 20

[00246] In some embodiments, at least 60% of the population of immune cells expand at least 80% of expansion rate of a population of control cells of a same type without the modification.

[00247] In some embodiments, the population of modified immune cells have increased yield of modified immune cells compared to a reference population of cells. In some embodiments, the reference population is a population of immune cells modified with a Cas9 nuclease. 25

[00248] In some embodiments, the immune cells are a human cells.

[00249] In some embodiments, the immune cells is are cytotoxic T cells, regulatory T cells, T helper cells, dendritic cells, B cells, or NK cells.

[00250] In some embodiments, the CAR specifically binds a marker associated with neoplasia.

[00251] In some embodiments, the CAR specifically binds CD7. 30

[00252] In some embodiments, the modification in b) is an insertion in exon 1 in the TRAC gene sequence. 2026204466   11 Jun 2026

[00253] In one aspect, provided herein is a method for producing a modified immune cell with increased anti-neoplasia activity, the method comprising: modifying a single target nucleobase in a Cbl Proto Oncogene B (CBLB) gene sequence or a regulatory element thereof in an immune cell, wherein the modification reduces an activation threshold of the immune cell compared with 5 an immune cell lacking the modification; thereby generating a modified immune cell with increased anti-neoplasia activity.

[00254] In one aspect, provided herein is a composition comprising a modified immune cell with increased anti-neoplasia activity, wherein the modified immune cell comprises: a modification in a single target nucleobase in a Cbl Proto-Oncogene B (CBLB) gene sequence or 10 a regulatory element thereof, wherein the modified immune cell exhibits a reduced activation threshold compared with a control immune cell of a same type without the modification.

[00255] In one aspect, provided herein is a population of immune cells, wherein a plurality of the population of immune cells comprises: a modification in a single target nucleobase in a CBLB gene sequence or a regulatory element thereof, wherein the plurality of the population of 15   the immune cells comprising the modification exhibit a reduced activation threshold compared with an control population of immune cells of a same type without the modification.

[00256] In one aspect, provided herein is a method for producing a population of modified immune cells with increased anti-neoplasia activity, the method comprising: modifying a single target nucleobase in a Cbl Proto Oncogene B (CBLB) gene sequence or a regulatory element 20 thereof in a population of immune cells, wherein at least 50% of the population of immune cells are modified to comprise the single target nucleobase modification.

[00257] In one aspect, provided herein is a composition comprising at least four different guide nucleic acid sequences for base editing. In some embodiments, the composition further comprising a polynucleotide encoding a base editor polypeptide, wherein the base editor 25 polypeptide comprises a nucleic acid programmable DNA binding protein (napDNAbp) and a deaminase. In some embodiments, the polynucleotide encoding the base editor is a mRNA sequence.

[00258] In some embodiments, the deaminase is a cytidine deaminase or an adenosine deaminase. 2026204466   11 Jun 2026

[00259] In some embodiments, the composition further comprises a base editor polypeptide, wherein the base editor polypeptide comprises a nucleic acid programmable DNA binding protein (napDNAbp) and a deaminase.

[00260] In some embodiments, the deaminase is a cytidine deaminase or an adenosine 5 deaminase.

[00261] In some embodiments, the composition further comprises a lipid nanoparticle.

[00262] In some embodiments, the at least four guide nucleic acid sequences each hybridize with a gene sequence selected from the group consisting of CD2, CD3 epsilon, CD3 gamma, CD3 delta, CD4, CD5, CD7, CD30, CD33, CD52, CD70, and CIITA. In some embodiments, the 10 at least 1, 2, 3, 4, 5, 6, 7, 8, or more genes or regulatory elements thereof are selected from CD2, CD3 epsilon, CD3 gamma, CD3 delta, CD4, CD5, CD7, CD30, CD33, CD52, CD70, and CIITA.

[00263] In some embodiments, the at least 1, 2, 3, 4, 5, 6, 7, 8, or more genes or regulatory elements thereof comprise one or more genes selected from CD2, CD3 epsilon, CD3 gamma, 15 CD3 delta, CD4, CD5, CD7, CD30, CD33, CD52, CD70, and CIITA. In some embodiments, the at least 1, 2, 3, 4, 5, 6, 7, 8, or more genes or regulatory elements thereof are selected from ACAT1, ACLY, ADORA2A, AXL, B2M , BATF, BCL2L11, BTLA, CAMK2D, cAMP, CASP8, Cblb, CCR5, CD2, CD3D, CD3E, CD3G, CD4, CD5, CD7, CD8A, CD33, CD38, CD52, CD70, CD82, CD86, CD96, CD123, CD160, CD244, CD276, CDK8, CDKN1B, Chi311, 20 CIITA, CISH, CSF2CSK, CTLA-4, CUL3, Cypl lai, DCK, DGKA, DGKZ, DHX37, ELOB(TCEB2), ENTPD1 (CD39), FADD, FAS, GATA3, IL6, IL6R, IL10, IL10RA, IRF4, IRF8, JUNB, Lag3,, LAIR-l (CD305), LDHA, LIF, LYN, MAP4K4, MAPK14, MCJ, MEF2D, MGAT5, NR4A1, NR4A2, NR4A3, NT5E (CD73), ODC1, OTULINL (FAM105A), PAG1, PDCD1, PDIA3, PHD1 (EGLN2), PHD2 (EGLN1), PHD3 (EGLN3), PIK3CD, PIKFYVE, 25 PPARa, PPARd, PRDMI1, PRKACA, PTEN, PTPN2, PTPN6, PTPN11, PVRIG (CD112R), RASA2, RFXANK, SELPG / PSGL1, SIGLEC15, SLA, SLAMF7, SOCS1, Spryl, Spry2, STK4, SUV39, H1TET2, TGFbRII, TIGIT, Tim-3, TMEM222, TNFAIP3, TNFRSF8 (CD30), TNFRSF10B, TOX, TOX2,, TRAC, TRBC1, TRBC2, UBASH3A, VHL, VISTA, In some embodiments, the at least four guide nucleic acid sequences each hybridize with a gene sequence 30 selected from the group consisting of CD3epsilon, CD3 delta, CD3 gamma, TRAC, TRBC1, and TRBC2, CD2, CD5, CD7, CD52, CD70, and CIITA. 2026204466   11 Jun 2026

[00264] In some embodiments, the at least four guide nucleic acid sequences comprise a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC, CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, 5 ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

[00265] In one aspect, provided herein is an immune cell comprising the composition of some of the embodiments described above, wherein the composition is introduced into the immune cell with electroporation. 10

[00266] In one aspect, provided herein is an immune cell comprising the composition of some of the embodiments described above, wherein the composition is introduced into the immune cell with electroporation, nucleofection, viral transduction, or a combination thereof.

[00267] Other features and advantages of the invention will be apparent from the detailed description, and from the claims. 15 Definitions

[00268] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used 20 in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise. 25

[00269] By “adenosine deaminase” is meant a polypeptide or fragment thereof capable of catalyzing the hydrolytic deamination of adenine or adenosine. In some embodiments, the deaminase or deaminase domain is an adenosine deaminase catalyzing the hydrolytic deamination of adenosine to inosine or deoxyadenosine to deoxyinosine. In some embodiments, the adenosine deaminase catalyzes the hydrolytic deamination of adenine or adenosine in 30 deoxyribonucleic acid (DNA). The adenosine deaminases (e.g., engineered adenosine deaminases, evolved adenosine deaminases) provided herein may be from any organism, such as 2026204466   11 Jun 2026 a bacterium. In some embodiments, the deaminase or deaminase domain is a variant of a naturally-occurring deaminase from an organism. In some embodiments, the deaminase or deaminase domain does not occur in nature. For example, in some embodiments, the deaminase or deaminase domain is at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at 5 least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to a naturally-occurring deaminase. In some embodiments, the adenosine deaminase is from a bacterium, such as, E. coli, S. aureus, S. typhi, S. putrefaciens, H. influenzae, or C. crescentus. In some embodiments, the adenosine deaminase is a TadA deaminase. In some embodiments, the TadA deaminase is an E. coli TadA 10 (ecTadA) deaminase or a fragment thereof.

[00270] For example, the truncated ecTadA may be missing one or more N-terminal amino acids relative to a full-length ecTadA. In some embodiments, the truncated ecTadA may be missing 1, 2, 3, 4, 5 ,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 6, 17, 18, 19, or 20 N-terminal amino acid residues relative to the full length ecTadA. In some embodiments, the truncated ecTadA may be 15 missing 1, 2, 3, 4, 5 ,6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 6, 17, 18, 19, or 20 C-terminal amino acid residues relative to the full length ecTadA. In some embodiments, the ecTadA deaminase does not comprise an N-terminal methionine. In some embodiments, the TadA deaminase is an N-terminal truncated TadA. In particular embodiments, the TadA is any one of the TadAs described in PCT / US2017 / 045381, which is incorporated herein by reference in its entirety. 20

[00271] In certain embodiments, the adenosine deaminase comprises the amino acid sequence: MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPT AHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKT GAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTD, which is termed “the TadA reference sequence.” 25

[00272] In some embodiments the TadA deaminase is a full-length E. coli TadA deaminase. For example, in certain embodiments, the adenosine deaminase comprises the amino acid sequence: MRRAFITGVFFLSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEG WNRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIG 30 RVVFGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEI KAQKKAQSSTD 2026204466   11 Jun 2026

[00273] It should be appreciated, however, that additional adenosine deaminases useful in the present application would be apparent to the skilled artisan and are within the scope of this disclosure. For example, the adenosine deaminase may be a homolog of adenosine deaminase acting on tRNA (AD AT). Exemplary AD AT homologs include, without limitation: 5

[00274] Staphylococcus aureus TadA: MGSHMTNDIYFMTLAIEEAKKAAQLGEVPIGAIITKDDEVIARAHNLRETLQQPTAH AEHIAIERAAKVLGSWRLEGCTLYVTLEPCVMCAGTIVMSRIPRVVYGADDPKGGCS GS LMNLLQQS NFNHRAIVDKG VLKE AC S TLLTTFFKNLRANKKS TN

[00275] Bacillus subtilis TadA: 10 MTQDELYMKEAIKEAKKAEEKGEVPIGAVLVINGEIIARAHNLRETEQRSIAHAEML VIDEACKALGTWRLEGATLYVTLEPCPMCAGAVVLSRVEKVVFGAFDPKGGC S GTLMN LLQEERFNHQAEVVSGVLEEECGGMLSAFFRELRKKKKAARKNLSE

[00276] Salmonella typhimurium (S. typhimurium) TadA: MPPAFITGVTSLSDVELDHEYWMRHALTLAKRAWDEREVPVGAVLVHNHRVIGEG 15 WNRPIGRHDPTAHAEIMALRQGGLVLQNYRLLDTTLYVTLEPCVMCAGAMVHSRIG RVVFGARDAKTGAAGSLIDVLHHPGMNHRVEIIEGVLRDECATLLSDFFRMRRQEIK ALKKADRAEGAGPAV

[00277] Shewanella putrefaciens (S. putrefaciens) TadA: MDE YWMQVAMQM AEKAEAAGE VPVGA VLVKDGQQIATGYNLS IS QHDPT AHAEI 20 LCLRSAGKKLENYRLLDATLYITLEPCAMCAGAMVHSRIARVVYGARDEKTGAAGT VVNLLQHPAFNHQVEVTSGVLAEACSAQLSRFFKRRRDEKKALKLAQRAQQGIE

[00278] Haemophilus influenzae F3031 (H. influenzae) TadA: MDAAKVRSEFDEKMMRYALELADKAEALGEIPVGAVLVDDARNIIGEGWNLSIVQS DPT AH AEIIALRNG AKNIQN YRLLNS TLY VTLEPCTMC AG AILHS RIKRLVFG AS D 25 YK TGAIGSRFHFFDDYKMNHTLEITSGVLAEECSQKLSTFFQKRREEKKIEKALLKSLSD K

[00279] Caulobacter crescentus (C. crescentus) TadA: 2026204466   11 Jun 2026 MRTDESEDQDHRMMRLALDAARAAAEAGETPVGAVILDPSTGEVIATAGNGPIAAH DPTAHAEIAAMRAAAAKLGNYRLTDLTLVVTLEPCAMCAGAISHARIGRVVFGADD PKGGAVVHGPKFFAQPTCHWRPEVTGGVLADESADLLRGFFRARRKAKI

[00280] Geobacter sulfurreducens (G. sulfurreducens) TadA: 5 MSSLKKTPIRDDAYWMGKAIREAAKAAARDEVPIGAVIVRDGAVIGRGHNLREGSN DPSAHAEMIAIRQAARRSANWRLTGATLYVTLEPCLMCMGAIILARLERVVFGCYDP KGGAAGSLYDLSADPRLNHQVRLSPGVCQEECGTMLSDFFRDLRRRKKAKATPALF IDERKVPPEP

[00281] TadA7.10 10  MSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEG WNRAIGLHDPTAH AEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAG SLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQSSTD

[00282] By “agent” is meant any small molecule chemical compound, antibody, nucleic acid molecule, or polypeptide, or fragments thereof. 15

[00283] By “alteration” is meant a change in the structure, expression levels or activity of a gene or polypeptide as detected by standard art known methods such as those described herein. As used herein, an alteration (e.g., increase or decrease) includes a 10% change in expression levels, a 25% change, a 40% change, and a 50% or greater change in expression levels.

[00284] "Allogeneic," as used herein, refers to cells of the same species that differ genetically 20 to the cell in comparison.

[00285] By “analog” is meant a molecule that is not identical, but has analogous functional or structural features. For example, a polypeptide analog retains the biological activity of a corresponding naturally-occurring polypeptide, while having certain sequence modifications that enhance the analog’s function relative to a naturally occurring polypeptide. Such modifications 25 could increase the analog’s protease resistance, membrane permeability, or half-life, without altering, for example, polynucleotide binding activity. In another example, a polynucleotide analog retains the biological activity of a corresponding naturally-occurring polynucleotide while having certain modifications that enhance the analog’s function relative to a naturally occurring polynucleotide. Such modifications could increase the polynucleotide’s affinity for DNA, half- 30 life, and / or nuclease resistance, an analog may include an unnatural nucleotide or amino acid. 2026204466   11 Jun 2026

[00286] By “anti-neoplasia activity” is meant preventing or inhibiting the maturation and / or proliferation of neoplasms.

[00287] "Autologous," as used herein, refers to cells from the same subject.

[00288] By “B cell maturation antigen, or tumor necrosis factor receptor superfamily member 5   17 polypeptide, (BCMA)” is meant a protein having at least about 85% amino acid sequence identify to NCBI Accession No. NP_001183 or a fragment thereof that is expressed on mature B lymphocytes. An exemplary BCMA polypeptide sequence is provided below.

[00289] >NP_001183.2 tumor necrosis factor receptor superfamily member 17 [Homo sapiens] MLQMAGQCSQNEYFDSLLHACIPCQLRCSSNTPPLTCQRYCNASVTNSVKGTNAILWTC 10 LGLSLIISLAVFVLMFLLRKINSEPLKDEFKNTGSGLLGMANIDLEKSRTGDEIILPRGLEY TVEECTCEDCIKSKPKVDSDHCFPLPAMEEGATILVTTKTNDYCKSLPAALSATEIEKSIS AR

[00290] This antigen can be targeted in relapsed or refractory multiple myeloma and other hematological neoplasia therapies. 15

[00291] By “B cell maturation antigen, or tumor necrosis factor receptor superfamily member 17, (BCMA) polynucleotide” is meant a nucleic acid molecule encoding a BCMA polypeptide. The BCMA gene encodes a cell surface receptor that recognizes B cell activating factor. An exemplary B2M polynucleotide sequence is provided below.

[00292] >NM_001192.2 Homo sapiens TNF receptor superfamily member 17 (TNFRSF17), 20 mRNA AAGACTCAAACTTAGAAACTTGAATTAGATGTGGTATTCAAATCCTTAGCTGCCGCG AAGACACAGACAGCCCCCGTAAGAACCCACGAAGCAGGCGAAGTTCATTGTTCTCA ACATTCTAGCTGCTCTTGCTGCATTTGCTCTGGAATTCTTGTAGAGATATTACTTGTC CTTCCAGGCTGTTCTTTCTGTAGCTCCCTTGTTTTCTTTTTGTGATCATGTTGCAGATG 25 GCTGGGCAGTGCTCCCAAAATGAATATTTTGACAGTTTGTTGCATGCTTGCATACCTT GTCAACTTCGATGTTCTTCTAATACTCCTCCTCTAACATGTCAGCGTTATTGTAATGC AAGTGTGACCAATTCAGTGAAAGGAACGAATGCGATTCTCTGGACCTGTTTGGGACT GAGCTTAATAATTTCTTTGGCAGTTTTCGTGCTAATGTTTTTGCTAAGGAAGATAAAC TCTGAACCATTAAAGGACGAGTTTAAAAACACAGGATCAGGTCTCCTGGGCATGGC 30 TAACATTGACCTGGAAAAGAGCAGGACTGGTGATGAAATTATTCTTCCGAGAGGCC TCGAGTACACGGTGGAAGAATGCACCTGTGAAGACTGCATCAAGAGCAAACCGAAG 2026204466   11 Jun 2026 GTCGACTCTGACCATTGCTTTCCACTCCCAGCTATGGAGGAAGGCGCAACCATTCTT GTCACCACGAAAACGAATGACTATTGCAAGAGCCTGCCAGCTGCTTTGAGTGCTACG GAGATAGAGAAATCAATTTCTGCTAGGTAATTAACCATTTCGACTCGAGCAGTGCCA CTTTAAAAATCTTTTGTCAGAATAGATGATGTGTCAGATCTCTTTAGGATGACTGTAT 5 TTTTCAGTTGCCGATACAGCTTTTTGTCCTCTAACTGTGGAAACTCTTTATGTTAGAT ATATTTCTCTAGGTTACTGTTGGGAGCTTAATGGTAGAAACTTCCTTGGTTTCATGAT TAAACTCTTTTTTTTCCTGA

[00293] By "base editor (BE)," or "nucleobase editor (NBE)" is meant an agent that binds a polynucleotide and has nucleobase modifying activity. In one embodiment, the agent binds the 10 polynucleotide at a specific sequence using a nucleic acid programmable DNA binding protein. In another embodiment, the base editor is an enzyme capable of modifying a cytidine base within a nucleic acid molecule (e.g., DNA). In some embodiments, the base editor is capable of deaminating a base within a nucleic acid molecule. In some embodiments, the base editor is capable of deaminating a base within a DNA molecule. In some embodiments, the base editor is 15 capable of deaminating a cytidine in DNA. In some embodiments, the base editor is a fusion protein comprising a cytidine deaminase or an adenosine deaminase. In some embodiments, the base editor is a Cas9 protein fused to a cytidine deaminase or an adenosine deaminase. In some embodiments, the base editor is a Cas9 nickase (nCas9) fused to a cytidine deaminase or an adenosine deaminase. In some embodiments, the base editor is fused to an inhibitor of base 20 excision repair, for example, a UGI domain. In some embodiments, the fusion protein comprises a Cas9 nickase fused to a deaminase and an inhibitor of base excision repair, such as a UGI domain. In some embodiments, the cytidine deaminase or an or an adenosine deaminase nucleobase editor polypeptide comprising the following domains A-B: NH2-[A-B]-COOH, 25

[00294] wherein A comprises a cytidine deaminase domain, an adenosine deaminase domain or an active fragment thereof, and wherein B comprises one or more domains having nucleic acid sequence specific binding activity. In one embodiment, the cytidine or adenosine deaminase Nucleobase Editor polypeptide of the previous aspect contains:

[00295] NH2-[An-Bo]-COOH, wherein A comprises: a cytidine deaminase domain, an 30 adenosine deaminase domain, or an active fragment thereof, wherein n is an integer: 1, 2, 3, 4, or 5; and wherein B comprises a domain having nucleic acid sequence specific binding activity; and 2026204466   11 Jun 2026 wherein o is an integer: 1,2,3, 4, or 5. In one embodiment, the polypeptide contains one or more nuclear localization sequences. In one embodiment, the polypeptide contains at least one of said nuclear localization sequences is at the N-terminus or C-terminus. In one embodiment, the polypeptide contains the nuclear localization signal is a bipartite nuclear localization signal. 5 In one embodiment, the polypeptide contains one or more domains linked by a linker.

[00296] In some embodiments, the base editor is a cytidine base editor (CBE). In some embodiments, the base editor is an adenosine base editor (ABE). In some embodiments, the base editor is an adenosine base editor (ABE) and a cytidine base editor (CBE). In some embodiments, the base editor is a nuclease-inactive Cas9 (dCas9) fused to an adenosine 10 deaminase. In some embodiments, the Cas9 is a circular permutant Cas9 (e.g., spCas9 or saCas9). Circular permutant Cas9s are known in the art and described, for example, in Oakes et al., Cell 176, 254-267, 2019. In some embodiments, the base editor is fused to an inhibitor of base excision repair, for example, a UGI domain, or a dISN domain. In some embodiments, the fusion protein comprises a Cas9 nickase fused to a deaminase and an inhibitor of base excision 15 repair, such as a UGI or dISN domain. In other embodiments the base editor is an abasic base editor.

[00297] In some embodiments, an adenosine deaminase is evolved from TadA. In some embodiments, the polynucleotide programmable DNA binding domain is a CRISPR associated (e.g., Cas or Cpfl) enzyme. In some embodiments, the base editor is a catalytically dead Cas9 20 (dCas9) fused to a deaminase domain. In some embodiments, the base editor is a Cas9 nickase (nCas9) fused to a deaminase domain. In some embodiments, the base editor is fused to an inhibitor of base excision repair (BER). In some embodiments, the inhibitor of base excision repair is a uracil DNA glycosylase inhibitor (UGI). In some embodiments, the inhibitor of base excision repair is an inosine base excision repair inhibitor. Details of base editors are described 25 in International PCT Application Nos. PCT / 2017 / 045381 (WO2018 / 027078) and PCT / US2016 / 058344 (WO2017 / 070632), each of which is incorporated herein by reference for its entirety. Also see Komor, A.C., et al., “Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage” Nature 533, 420-424 (2016); Gaudelli, N.M., et al., “Programmable base editing of ACT to G*C in genomic DNA without DNA cleavage” Nature 30   551,464-471 (2017); Komor, A.C., et al., “Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and 2026204466   11 Jun 2026 product purity” Science Advances 3:eaao4774 (2017), and Rees, H.A., et al., “Base editing: precision chemistry on the genome and transcriptome of living cells.” Nat Rev Genet. 2018 Dec;19(12):770-788. doi: 10.103 8 / s41576-018-0059-1, the entire contents of which are hereby incorporated by reference. 5

[00298] In some embodiments, base editors are generated by cloning an adenosine deaminase variant (e.g., TadA*7.10) into a scaffold that includes a circular permutant Cas9 (e.g., spCAS9) and a bipartite nuclear localization sequence. Circular permutant Cas9s are known in the art and described, for example, in Oakes et al., Cell 176, 254-267, 2019. Exemplary circular permutant sequences are set forth below, in which the bold sequence indicates sequence derived from Cas9, 10 the italics sequence denotes a linker sequence, and the underlined sequence denotes a bipartite nuclear localization sequence.

[00299] CP5 (with MSP “NGC=Pam Variant with mutations Regular Cas9 likes NGG” PID=Protein Interacting Domain and “D10A” nickase):

[00300] EIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGR 15 DFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGG FMQPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKE VKKDLIIKLPKYSLFELENGRKRMLASAKFLQKGNELALPSKYVNFLYLASHYEKL KGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPI REQAENIIHLFTLTNLGAPRAFKYFDTTIARKEYRSTKEVLDATLIHQSITGLYETRI 20 DLSQLGGDGG5GG5GG5GG5GG5GG5GGA7DKKYSIGLAIGTNSVGWAVITDEYKVPS KKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEI FSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKK LVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEE NPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNF 25 DL AEDAKLQL SKDTYDDDLDNLLAQIGDQYADLFLAAKNL SD AILL SDILRVNTEIT KAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQ EEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQ EDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVD KGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPA 30 FLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTY HDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQL 2026204466   11 Jun 2026 KRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKED IQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEM ARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQN GRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEE 5 VVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITK HVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEGADKRTADGSEFE SPKKKRKV*

[00301] The nucleobase components and the polynucleotide programmable nucleotide binding 10   component of a base editor system may be associated with each other covalently or non- covalently. For example, in some embodiments, the deaminase domain can be targeted to a target nucleotide sequence by a polynucleotide programmable nucleotide binding domain. In some embodiments, a polynucleotide programmable nucleotide binding domain can be fused or linked to a deaminase domain. In some embodiments, a polynucleotide programmable 15 nucleotide binding domain can target a deaminase domain to a target nucleotide sequence by non-covalently interacting with or associating with the deaminase domain. For example, in some embodiments, the nucleobase editing component, e.g., the deaminase component can comprise an additional heterologous portion or domain that is capable of interacting with, associating with, or capable of forming a complex with an additional heterologous portion or domain that is part of 20 a polynucleotide programmable nucleotide binding domain. In some embodiments, the additional heterologous portion may be capable of binding to, interacting with, associating with, or forming a complex with a polypeptide. In some embodiments, the additional heterologous portion may be capable of binding to, interacting with, associating with, or forming a complex with a polynucleotide. In some embodiments, the additional heterologous portion may be 25 capable of binding to a guide polynucleotide. In some embodiments, the additional heterologous portion may be capable of binding to a polypeptide linker. In some embodiments, the additional heterologous portion may be capable of binding to a polynucleotide linker. The additional heterologous portion may be a protein domain. In some embodiments, the additional heterologous portion may be a K Homology (KH) domain, a MS2 coat protein domain, a PP7 30 coat protein domain, a SfMu Com coat protein domain, a steril alpha motif, a telomerase Ku 2026204466   11 Jun 2026 binding motif and Ku protein, a telomerase Sm7 binding motif and Sm7 protein, or a RNA recognition motif.

[00302] A base editor system may further comprise a guide polynucleotide component. It should be appreciated that components of the base editor system may be associated with each 5 other via covalent bonds, noncovalent interactions, or any combination of associations and interactions thereof. In some embodiments, a deaminase domain can be targeted to a target nucleotide sequence by a guide polynucleotide. For example, in some embodiments, the nucleobase editing component of the base editor system, e.g., the deaminase component, can comprise an additional heterologous portion or domain (e.g., polynucleotide binding domain 10 such as an RNA or DNA binding protein) that is capable of interacting with, associating with, or capable of forming a complex with a portion or segment (e.g., a polynucleotide motif) of a guide polynucleotide. In some embodiments, the additional heterologous portion or domain (e.g., polynucleotide binding domain such as an RNA or DNA binding protein) can be fused or linked to the deaminase domain. In some embodiments, the additional heterologous portion may be 15 capable of binding to, interacting with, associating with, or forming a complex with a polypeptide. In some embodiments, the additional heterologous portion may be capable of binding to, interacting with, associating with, or forming a complex with a polynucleotide. In some embodiments, the additional heterologous portion may be capable of binding to a guide polynucleotide. In some embodiments, the additional heterologous portion may be capable of 20 binding to a polypeptide linker. In some embodiments, the additional heterologous portion may be capable of binding to a polynucleotide linker. The additional heterologous portion may be a protein domain. In some embodiments, the additional heterologous portion may be a K Homology (KH) domain, a MS2 coat protein domain, a PP7 coat protein domain, a SfMu Com coat protein domain, a sterile alpha motif, a telomerase Ku binding motif and Ku protein, a 25 telomerase Sm7 binding motif and Sm7 protein, or a RNA recognition motif.

[00303] In some embodiments, a base editor system can further comprise an inhibitor of base excision repair (BER) component. It should be appreciated that components of the base editor system may be associated with each other via covalent bonds, noncovalent interactions, or any combination of associations and interactions thereof. The inhibitor of BER component may 30 comprise a base excision repair inhibitor. In some embodiments, the inhibitor of base excision repair can be a uracil DNA glycosylase inhibitor (UGI). In some embodiments, the inhibitor of 2026204466   11 Jun 2026 base excision repair can be an inosine base excision repair inhibitor. In some embodiments, the inhibitor of base excision repair can be targeted to the target nucleotide sequence by the polynucleotide programmable nucleotide binding domain. In some embodiments, a polynucleotide programmable nucleotide binding domain can be fused or linked to an inhibitor 5 of base excision repair. In some embodiments, a polynucleotide programmable nucleotide binding domain can be fused or linked to a deaminase domain and an inhibitor of base excision repair. In some embodiments, a polynucleotide programmable nucleotide binding domain can target an inhibitor of base excision repair to a target nucleotide sequence by non-covalently interacting with or associating with the inhibitor of base excision repair. For example, in some 10 embodiments, the inhibitor of base excision repair component can comprise an additional heterologous portion or domain that is capable of interacting with, associating with, or capable of forming a complex with an additional heterologous portion or domain that is part of a polynucleotide programmable nucleotide binding domain. In some embodiments, the inhibitor of base excision repair can be targeted to the target nucleotide sequence by the guide 15 polynucleotide. For example, in some embodiments, the inhibitor of base excision repair can comprise an additional heterologous portion or domain (e.g., polynucleotide binding domain such as an RNA or DNA binding protein) that is capable of interacting with, associating with, or capable of forming a complex with a portion or segment (e.g., a polynucleotide motif) of a guide polynucleotide. In some embodiments, the additional heterologous portion or domain of the 20 guide polynucleotide (e.g., polynucleotide binding domain such as an RNA or DNA binding protein) can be fused or linked to the inhibitor of base excision repair. In some embodiments, the additional heterologous portion may be capable of binding to, interacting with, associating with, or forming a complex with a polynucleotide. In some embodiments, the additional heterologous portion may be capable of binding to a guide polynucleotide. In some 25 embodiments, the additional heterologous portion may be capable of binding to a polypeptide linker. In some embodiments, the additional heterologous portion may be capable of binding to a polynucleotide linker. The additional heterologous portion may be a protein domain. In some embodiments, the additional heterologous portion may be a K Homology (KH) domain, a MS2 coat protein domain, a PP7 coat protein domain, a SfMu Com coat protein domain, a sterile 30 alpha motif, a telomerase Ku binding motif and Ku protein, a telomerase Sm7 binding motif and Sm7 protein, or a RNA recognition motif. By “base editing activity” is meant acting to 2026204466   11 Jun 2026 chemically alter a base within a polynucleotide. In one embodiment, a first base is converted to a second base. In one embodiment, the base editing activity is cytidine deaminase activity, e.g., converting target OG to T»A. In another embodiment, the base editing activity is adenosine deaminase activity, e.g., converting A»T to G»C. 5

[00304] By “beta-2 microglobulin (B2M) polypeptide” is meant a protein having at least about 85% amino acid sequence identity to UniProt Accession No. P61769 or a fragment thereof and having immunomodulatory activity. An exemplary B2M polypeptide sequence is provided below. >sp|P61769|B2MG_HUMAN Beta-2-microglobulin OS=Homo sapiens OX=9606 GN=B2M 10 PE=1 SV=1 MSRSVALAVLALLSLSGLEAIQRTPKIQVYSRHPAENGKSNFLNCYVSGFHPSDIEVDLL KNGERIEKVEHSDLSFSKDWSFYLLYYTEFTPTEKDEYACRVNHVTLSQPKIVKWDRD M

[00305] By “beta-2-microglobulin (B2M) polynucleotide” is meant a nucleic acid molecule 15 encoding a B2M polypeptide. The beta-2-microglobulin gene encodes a serum protein associated with the major histocompatibility complex. B2M is involved in non-self recognition by host CD8+ T cells. An exemplary B2M polynucleotide sequence is provided below. >DQ217933.1 Homo sapiens beta-2-microglobin (B2M) gene, complete cds CATGTCATAAATGGTAAGTCCAAGAAAAATACAGGTATTCCCCCCCAAAGAAAACT 20 GTAAAATCGACTTTTTTCTATCTGTACTGTTTTTTATTGGTTTTTAAATTGGTTTTCCA AGTGAGTAAATCAGAATCTATCTGTAATGGATTTTAAATTTAGTGTTTCTCTGTGATG TAGTAAACAAGAAACTAGAGGCAAAAATAGCCCTGTCCCTTGCTAAACTTCTAAGG CACTTTTCTAGTACAACTCAACACTAACATTTCAGGCCTTTAGTGCCTTATATGAGTT TTTAAAAGGGGGAAAAGGGAGGGAGCAAGAGTGTCTTAACTCATACATTTAGGCAT 25 AACAATTATTCTCATATTTTAGTTATTGAGAGGGCTGGTAGAAAAACTAGGTAAATA ATATTAATAATTATAGCGCTTATTAAACACTACAGAACACTTACTATGTACCAGGCA TTGTGGGAGGCTCTCTCTTGTGCATTATCTCATTTCATTAGGTCCATGGAGAGTATTG CATTTTCTTAGTTTAGGCATGGCCTCCACAATAAAGATTATCAAAAGCCTAAAAATA TGTAAAAGAAACCTAGAAGTTATTTGTTGTGCTCCTTGGGGAAGCTAGGCAAATCCT 30 TTCAACTGAAAACCATGGTGACTTCCAAGATCTCTGCCCCTCCCCATCGCCATGGTC CACTTCCTCTTCTCACTGTTCCTCTTAGAAAAGATCTGTGGACTCCACCACCACGAA 2026204466   11 Jun 2026 ATGGCGGCACCTTATTTATGGTCACTTTAGAGGGTAGGTTTTCTTAATGGGTCTGCCT GTCATGTTTAACGTCCTTGGCTGGGTCCAAGGCAGATGCAGTCCAAACTCTCACTAA AATTGCCGAGCCCTTTGTCTTCCAGTGTCTAAAATATTAATGTCAATGGAATCAGGC CAGAGTTTGAATTCTAGTCTCTTAGCCTTTGTTTCCCCTGTCCATAAAATGAATGGGG 5 GTAATTCTTTCCTCCTACAGTTTATTTATATATTCACTAATTCATTCATTCATCCATCC ATTCGTTCATTCGGTTTACTGAGTACCTACTATGTGCCAGCCCCTGTTCTAGGGTGGA AACTAAGAGAATGATGTACCTAGAGGGCGCTGGAAGCTCTAAAGCCCTAGCAGTTA CTGCTTTTACTATTAGTGGTCGTTTTTTTCTCCCCCCCGCCCCCCGACAAATCAACAG AACAAAGAAAATTACCTAAACAGCAAGGACATAGGGAGGAACTTCTTGGCACAGAA 10 CTTTCCAAACACTTTTTCCTGAAGGGATACAAGAAGCAAGAAAGGTACTCTTTCACT AGGACCTTCTCTGAGCTGTCCTCAGGATGCTTTTGGGACTATTTTTCTTACCCAGAGA ATGGAGAAACCCTGCAGGGAATTCCCAAGCTGTAGTTATAAACAGAAGTTCTCCTTC TGCTAGGTAGCATTCAAAGATCTTAATCTTCTGGGTTTCCGTTTTCTCGAATGAAAAA TGCAGGTCCGAGCAGTTAACTGGCTGGGGCACCATTAGCAAGTCACTTAGCATCTCT 15 GGGGCCAGTCTGCAAAGCGAGGGGGCAGCCTTAATGTGCCTCCAGCCTGAAGTCCT AGAATGAGCGCCCGGTGTCCCAAGCTGGGGCGCGCACCCCAGATCGGAGGGCGCCG ATGTACAGACAGCAAACTCACCCAGTCTAGTGCATGCCTTCTTAAACATCACGAGAC TCTAAGAAAAGGAAACTGAAAACGGGAAAGTCCCTCTCTCTAACCTGGCACTGCGT CGCTGGCTTGGAGACAGGTGACGGTCCCTGCGGGCCTTGTCCTGATTGGCTGGGCAC 20 GCGTTTAATATAAGTGGAGGCGTCGCGCTGGCGGGCATTCCTGAAGCTGACAGCATT CGGGCCGAGATGTCTCGCTCCGTGGCCTTAGCTGTGCTCGCGCTACTCTCTCTTTCTG GCCTGGAGGCTATCCAGCGTGAGTCTCTCCTACCCTCCCGCTCTGGTCCTTCCTCTCC CGCTCTGCACCCTCTGTGGCCCTCGCTGTGCTCTCTCGCTCCGTGACTTCCCTTCTCC AAGTTCTCCTTGGTGGCCCGCCGTGGGGCTAGTCCAGGGCTGGATCTCGGGGAAGCG 25 GCGGGGTGGCCTGGGAGTGGGGAAGGGGGTGCGCACCCGGGACGCGCGCTACTTGC CCCTTTCGGCGGGGAGCAGGGGAGACCTTTGGCCTACGGCGACGGGAGGGTCGGGA CAAAGTTTAGGGCGTCGATAAGCGTCAGAGCGCCGAGGTTGGGGGAGGGTTTCTCT TCCGCTCTTTCGCGGGGCCTCTGGCTCCCCCAGCGCAGCTGGAGTGGGGGACGGGTA GGCTCGTCCCAAAGGCGCGGCGCTGAGGTTTGTGAACGCGTGGAGGGGCGCTTGGG 30 GTCTGGGGGAGGCGTCGCCCGGGTAAGCCTGTCTGCTGCGGCTCTGCTTCCCTTAGA CTGGAGAGCTGTGGACTTCGTCTAGGCGCCCGCTAAGTTCGCATGTCCTAGCACCTC 2026204466   11 Jun 2026 TGGGTCTATGTGGGGCCACACCGTGGGGAGGAAACAGCACGCGACGTTTGTAGAAT GCTTGGCTGTGATACAAAGCGGTTTCGAATAATTAACTTATTTGTTCCCATCACATGT CACTTTTAAAAAATTATAAGAACTACCCGTTATTGACATCTTTCTGTGTGCCAAGGA CTTTATGTGCTTTGCGTCATTTAATTTTGAAAACAGTTATCTTCCGCCATAGATAACT 5 ACTATGGTTATCTTCTGCCTCTCACAGATGAAGAAACTAAGGCACCGAGATTTTAAG AAACTTAATTACACAGGGGATAAATGGCAGCAATCGAGATTGAAGTCAAGCCTAAC CAGGGCTTTTGCGGGAGCGCATGCCTTTTGGCTGTAATTCGTGCATTTTTTTTTAAGA AAAACGCCTGCCTTCTGCGTGAGATTCTCCAGAGCAAACTGGGCGGCATGGGCCCT GTGGTCTTTTCGTACAGAGGGCTTCCTCTTTGGCTCTTTGCCTGGTTGTTTCCAAGAT 10 GTACTGTGCCTCTTACTTTCGGTTTTGAAAACATGAGGGGGTTGGGCGTGGTAGCTT ACGCCTGTAATCCCAGCACTTAGGGAGGCCGAGGCGGGAGGATGGCTTGAGGTCCG TAGTTGAGACCAGCCTGGCCAACATGGTGAAGCCTGGTCTCTACAAAAAATAATAA CAAAAATTAGCCGGGTGTGGTGGCTCGTGCCTGTGGTCCCAGCTGCTCCGGTGGCTG AGGCGGGAGGATCTCTTGAGCTTAGGCTTTTGAGCTATCATGGCGCCAGTGCACTCC 15 AGCGTGGGCAACAGAGCGAGACCCTGTCTCTCAAAAAAGAAAAAAAAAAAAAAAG AAAGAGAAAAGAAAAGAAAGAAAGAAGTGAAGGTTTGTCAGTCAGGGGAGCTGTA AAACCATTAATAAAGATAATCCAAGATGGTTACCAAGACTGTTGAGGACGCCAGAG ATCTTGAGCACTTTCTAAGTACCTGGCAATACACTAAGCGCGCTCACCTTTTCCTCTG GCAAAACATGATCGAAAGCAGAATGTTTTGATCATGAGAAAATTGCATTTAATTTGA 20 ATACAATTTATTTACAACATAAAGGATAATGTATATATCACCACCATTACTGGTATT TGCTGGTTATGTTAGATGTCATTTTAAAAAATAACAATCTGATATTTAAAAAAAAAT CTTATTTTGAAAATTTCCAAAGTAATACATGCCATGCATAGACCATTTCTGGAAGAT ACCACAAGAAACATGTAATGATGATTGCCTCTGAAGGTCTATTTTCCTCCTCTGACC TGTGTGTGGGTTTTGTTTTTGTTTTACTGTGGGCATAAATTAATTTTTCAGTTAAGTTT 25 TGGAAGCTTAAATAACTCTCCAAAAGTCATAAAGCCAGTAACTGGTTGAGCCCAAA TTCAAACCCAGCCTGTCTGATACTTGTCCTCTTCTTAGAAAAGATTACAGTGATGCTC TCACAAAATCTTGCCGCCTTCCCTCAAACAGAGAGTTCCAGGCAGGATGAATCTGTG CTCTGATCCCTGAGGCATTTAATATGTTCTTATTATTAGAAGCTCAGATGCAAAGAG CTCTCTTAGCTTTTAATGTTATGAAAAAAATCAGGTCTTCATTAGATTCCCCAATCCA 30 CCTCTTGATGGGGCTAGTAGCCTTTCCTTAATGATAGGGTGTTTCTAGAGAGATATA TCTGGTCAAGGTGGCCTGGTACTCCTCCTTCTCCCCACAGCCTCCCAGACAAGGAGG 2026204466   11 Jun 2026 AGTAGCTGCCTTTTAGTGATCATGTACCCTGAATATAAGTGTATTTAAAAGAATTTT ATACACATATATTTAGTGTCAATCTGTATATTTAGTAGCACTAACACTTCTCTTCATT TTCAATGAAAAATATAGAGTTTATAATATTTTCTTCCCACTTCCCCATGGATGGTCTA GTCATGCCTCTCATTTTGGAAAGTACTGTTTCTGAAACATTAGGCAATATATTCCCAA 5 CCTGGCTAGTTTACAGCAATCACCTGTGGATGCTAATTAAAACGCAAATCCCACTGT CACATGCATTACTCCATTTGATCATAATGGAAAGTATGTTCTGTCCCATTTGCCATAG TCCTCACCTATCCCTGTTGTATTTTATCGGGTCCAACTCAACCATTTAAGGTATTTGC CAGCTCTTGTATGCATTTAGGTTTTGTTTCTTTGTTTTTTAGCTCATGAAATTAGGTAC AAAGTCAGAGAGGGGTCTGGCATATAAAACCTCAGCAGAAATAAAGAGGTTTTGTT 10 GTTTGGTAAGAACATACCTTGGGTTGGTTGGGCACGGTGGCTCGTGCCTGTAATCCC AACACTTTGGGAGGCCAAGGCAGGCTGATCACTTGAAGTTGGGAGTTCAAGACCAG CCTGGCCAACATGGTGAAATCCCGTCTCTACTGAAAATACAAAAATTAACCAGGCAT GGTGGTGTGTGCCTGTAGTCCCAGGAATCACTTGAACCCAGGAGGCGGAGGTTGCA GTGAGCTGAGATCTCACCACTGCACACTGCACTCCAGCCTGGGCAATGGAATGAGA 15 TTCCATCCCAAAAAATAAAAAAATAAAAAAATAAAGAACATACCTTGGGTTGATCC ACTTAGGAACCTCAGATAATAACATCTGCCACGTATAGAGCAATTGCTATGTCCCAG GCACTCTACTAGACACTTCATACAGTTTAGAAAATCAGATGGGTGTAGATCAAGGCA GGAGCAGGAACCAAAAAGAAAGGCATAAACATAAGAAAAAAAATGGAAGGGGTGG AAACAGAGTACAATAACATGAGTAATTTGATGGGGGCTATTATGAACTGAGAAATG 20 AACTTTGAAAAGTATCTTGGGGCCAAATCATGTAGACTCTTGAGTGATGTGTTAAGG AATGCTATGAGTGCTGAGAGGGCATCAGAAGTCCTTGAGAGCCTCCAGAGAAAGGC TCTTAAAAATGCAGCGCAATCTCCAGTGACAGAAGATACTGCTAGAAATCTGCTAG AAAAAAAACAAAAAAGGCATGTATAGAGGAATTATGAGGGAAAGATACCAAGTCA CGGTTTATTCTTCAAAATGGAGGTGGCTTGTTGGGAAGGTGGAAGCTCATTTGGCCA 25 GAGTGGAAATGGAATTGGGAGAAATCGATGACCAAATGTAAACACTTGGTGCCTGA TATAGCTTGACACCAAGTTAGCCCCAAGTGAAATACCCTGGCAATATTAATGTGTCT TTTCCCGATATTCCTCAGGTACTCCAAAGATTCAGGTTTACTCACGTCATCCAGCAG AGAATGGAAAGTCAAATTTCCTGAATTGCTATGTGTCTGGGTTTCATCCATCCGACA TTGAAGTTGACTTACTGAAGAATGGAGAGAGAATTGAAAAAGTGGAGCATTCAGAC 30 TTGTCTTTCAGCAAGGACTGGTCTTTCTATCTCTTGTACTACACTGAATTCACCCCCA CTGAAAAAGATGAGTATGCCTGCCGTGTGAACCATGTGACTTTGTCACAGCCCAAGA 2026204466   11 Jun 2026 TAGTTAAGTGGGGTAAGTCTTACATTCTTTTGTAAGCTGCTGAAAGTTGTGTATGAG TAGTCATATCATAAAGCTGCTTTGATATAAAAAAGGTCTATGGCCATACTACCCTGA ATGAGTCCCATCCCATCTGATATAAACAATCTGCATATTGGGATTGTCAGGGAATGT TCTTAAAGATCAGATTAGTGGCACCTGCTGAGATACTGATGCACAGCATGGTTTCTG 5 AACCAGTAGTTTCCCTGCAGTTGAGCAGGGAGCAGCAGCAGCACTTGCACAAATAC ATATACACTCTTAACACTTCTTACCTACTGGCTTCCTCTAGCTTTTGTGGCAGCTTCA GGTATATTTAGCACTGAACGAACATCTCAAGAAGGTATAGGCCTTTGTTTGTAAGTC CTGCTGTCCTAGCATCCTATAATCCTGGACTTCTCCAGTACTTTCTGGCTGGATTGGT ATCTGAGGCTAGTAGGAAGGGCTTGTTCCTGCTGGGTAGCTCTAAACAATGTATTCA 10 TGGGTAGGAACAGCAGCCTATTCTGCCAGCCTTATTTCTAACCATTTTAGACATTTGT TAGTACATGGTATTTTAAAAGTAAAACTTAATGTCTTCCTTTTTTTTCTCCACTGTCTT TTTCATAGATCGAGACATGTAAGCAGCATCATGGAGGTAAGTTTTTGACCTTGAGAA AATGTTTTTGTTTCACTGTCCTGAGGACTATTTATAGACAGCTCTAACATGATAACCC TCACTATGTGGAGAACATTGACAGAGTAACATTTTAGCAGGGAAAGAAGAATCCTA 15 CAGGGTCATGTTCCCTTCTCCTGTGGAGTGGCATGAAGAAGGTGTATGGCCCCAGGT ATGGCCATATTACTGACCCTCTACAGAGAGGGCAAAGGAACTGCCAGTATGGTATT GCAGGATAAAGGCAGGTGGTTACCCACATTACCTGCAAGGCTTTGATCTTTCTTCTG CCATTTCCACATTGGACATCTCTGCTGAGGAGAGAAAATGAACCACTCTTTTCCTTT GTATAATGTTGTTTTATTCTTCAGACAGAAGAGAGGAGTTATACAGCTCTGCAGACA 20 TCCCATTCCTGTATGGGGACTGTGTTTGCCTCTTAGAGGTTCCCAGGCCACTAGAGG AGATAAAGGGAAACAGATTGTTATAACTTGATATAATGATACTATAATAGATGTAA CTACAAGGAGCTCCAGAAGCAAGAGAGAGGGAGGAACTTGGACTTCTCTGCATCTT TAGTTGGAGTCCAAAGGCTTTTCAATGAAATTCTACTGCCCAGGGTACATTGATGCT GAAACCCCATTCAAATCTCCTGTTATATTCTAGAACAGGGAATTGATTTGGGAGAGC 25 ATCAGGAAGGTGGATGATCTGCCCAGTCACACTGTTAGTAAATTGTAGAGCCAGGA CCTGAACTCTAATATAGTCATGTGTTACTTAATGACGGGGACATGTTCTGAGAAATG CTTACACAAACCTAGGTGTTGTAGCCTACTACACGCATAGGCTACATGGTATAGCCT ATTGCTCCTAGACTACAAACCTGTACAGCCTGTTACTGTACTGAATACTGTGGGCAG TTGTAACACAATGGTAAGTATTTGTGTATCTAAACATAGAAGTTGCAGTAAAAATAT 30 GCTATTTTAATCTTATGAGACCACTGTCATATATACAGTCCATCATTGACCAAAACA TCATATCAGCATTTTTTCTTCTAAGATTTTGGGAGCACCAAAGGGATACACTAACAG 2026204466   11 Jun 2026 GATATACTCTTTATAATGGGTTTGGAGAACTGTCTGCAGCTACTTCTTTTAAAAAGGT GATCTACACAGTAGAAATTAGACAAGTTTGGTAATGAGATCTGCAATCCAAATAAA ATAAATTCATTGCTAACCTTTTTCTTTTCTTTTCAGGTTTGAAGATGCCGCATTTGGA TTGGATGAATTCCAAATTCTGCTTGCTTGCTTTTTAATATTGATATGCTTATACACTT 5 ACACTTTATGCACAAAATGTAGGGTTATAATAATGTTAACATGGACATGATCTTCTT TATAATTCTACTTTGAGTGCTGTCTCCATGTTTGATGTATCTGAGCAGGTTGCTCCAC AGGTAGCTCTAGGAGGGCTGGCAACTTAGAGGTGGGGAGCAGAGAATTCTCTTATC CAACATCAACATCTTGGTCAGATTTGAACTCTTCAATCTCTTGCACTCAAAGCTTGTT AAGATAGTTAAGCGTGCATAAGTTAACTTCCAATTTACATACTCTGCTTAGAATTTG 10 GGGGAAAATTTAGAAATATAATTGACAGGATTATTGGAAATTTGTTATAATGAATGA AACATTTTGTCATATAAGATTCATATTTACTTCTTATACATTTGATAAAGTAAGGCAT GGTTGTGGTTAATCTGGTTTATTTTTGTTCCACAAGTTAAATAAATCATAAAACTTGA TGTGTTATCTCTTATATCTCACTCCCACTATTACCCCTTTATTTTCAAACAGGGAAAC AGTCTTCAAGTTCCACTTGGTAAAAAATGTGAACCCCTTGTATATAGAGTTTGGCTC 15 ACAGTGTAAAGGGCCTCAGTGATTCACATTTTCCAGATTAGGAATCTGATGCTCAAA GAAGTTAAATGGCATAGTTGGGGTGACACAGCTGTCTAGTGGGAGGCCAGCCTTCT ATATTTTAGCCAGCGTTCTTTCCTGCGGGCCAGGTCATGAGGAGTATGCAGACTCTA AGAGGGAGCAAAAGTATCTGAAGGATTTAATATTTTAGCAAGGAATAGATATACAA TCATCCCTTGGTCTCCCTGGGGGATTGGTTTCAGGACCCCTTCTTGGACACCAAATCT 20 ATGGATATTTAAGTCCCTTCTATAAAATGGTATAGTATTTGCATATAACCTATCCACA TCCTCCTGTATACTTTAAATCATTTCTAGATTACTTGTAATACCTAATACAATGTAAA TGCTATGCAAATAGTTGTTATTGTTTAAGGAATAATGACAAGAAAAAAAAGTCTGTA CATGCTCAGTAAAGACACAACCATCCCTTTTTTTCCCCAGTGTTTTTGATCCATGGTT TGCTGAATCCACAGATGTGGAGCCCCTGGATACGGAAGGCCCGCTGTACTTTGAATG 25 ACAAATAACAGATTTAAA

[00306] The term “Cas9” or “Cas9 domain” refers to an RNA-guided nuclease comprising a Cas9 protein, or a fragment thereof (e.g., a protein comprising an active, inactive, or partially active DNA cleavage domain of Cas9, and / or the gRNA binding domain of Cas9). A Cas9 nuclease is also referred to sometimes as a casnl nuclease or a CRISPR (“clustered regularly 30 interspaced short palindromic repeat”)-associated nuclease. CRISPR is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements 2026204466   11 Jun 2026 and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (me) 5 and a Cas9 protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently, Cas9 / crRNA / tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed 3 '-5' exonucleolytically. In nature, DNA-binding and cleavage typically requires protein and both RNAs. However, single guide RNAs (“sgRNA”, or 10 simply “gNRA”) can be engineered so as to incorporate aspects of both the crRNA and tracrRNA into a single RNA species. See, e.g., Jinek M., Chylinski K., Fonfara I., Hauer M., Doudna J.A., Charpentier E. Science 337:816-821(2012), the entire contents of which is hereby incorporated by reference. Cas9 recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus non-self. Cas9 nuclease 15 sequences and structures are well known to those of skill in the art (see, e.g., “Complete genome sequence of an Ml strain of Streptococcus pyogenes.” Ferretti et al., J. J., McShan W.M., Ajdic D.J., Savic D.J., Savic G., Lyon K., Primeaux C., Sezate S., Suvorov A.N., Kenton S., Lai H.S., Lin S.P., Qian Y., Jia H.G., Najar F.Z., Ren Q., Zhu H., Song L., White J., Yuan X., Clifton S.W., Roe B.A., McLaughlin R.E., Proc. Natl. Acad. Sci. U.S.A. 98:4658-4663(2001); “CRISPR 20 RNA maturation by trans-encoded small RNA and host factor RNase III.” Deltcheva E., Chylinski K., Sharma C.M., Gonzales K., Chao Y., Pirzada Z.A., Eckert M.R., Vogel J., Charpentier E., Nature 471:602-607(2011); and “A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity.” Jinek M., Chylinski K., Fonfara 1., Hauer M., Doudna J.A., Charpentier E. Science 337:816-821(2012), the entire contents of each of which are 25 incorporated herein by reference). Cas9 orthologs have been described in various species, including, but not limited to, S. pyogenes and S. thermophilus. Additional suitable Cas9 nucleases and sequences will be apparent to those of skill in the art based on this disclosure, and such Cas9 nucleases and sequences include Cas9 sequences from the organisms and loci disclosed in Chylinski, Rhun, and Charpentier, “The tracrRNA and Cas9 families of type II 30 CRISPR-Cas immunity systems” (2013) RNA Biology 10:5, 726-737; the entire contents of 2026204466   11 Jun 2026 which are incorporated herein by reference. In some embodiments, a Cas9 nuclease has an inactive (e.g., an inactivated) DNA cleavage domain, that is, the Cas9 is a nickase.

[00307] A nuclease-inactivated Cas9 protein may interchangeably be referred to as a “dCas9” protein (for nuclease-“dead” Cas9). Methods for generating a Cas9 protein (or a fragment 5 thereof) having an inactive DNA cleavage domain are known (See, e.g., Jinek et al., Science. 337:816-821(2012); Qi et al., “Repurposing CRISPR as an RNA-Guided Platform for SequenceSpecific Control of Gene Expression” (2013) Cell. 28; 152(5): 1173-83, the entire contents of each of which are incorporated herein by reference). For example, the DNA cleavage domain of Cas9 is known to include two subdomains, the HNH nuclease subdomain and the RuvC 1 10   subdomain. The HNH subdomain cleaves the strand complementary to the gRNA, whereas the RuvCl subdomain cleaves the non-complementary strand. Mutations within these subdomains can silence the nuclease activity of Cas9. For example, the mutations D10A and H840A completely inactivate the nuclease activity of S. pyogenes Cas9 (Jinek et al., Science. 337:816-821(2012); Qi et al., Cell. 28; 152(5): 1173-83 (2013)). In some embodiments, proteins 15 comprising fragments of Cas9 are provided. For example, in some embodiments, a protein comprises one of two Cas9 domains: (1) the gRNA binding domain of Cas9; or (2) the DNA cleavage domain of Cas9. In some embodiments, proteins comprising Cas9 or fragments thereof are referred to as “Cas9 variants.” A Cas9 variant shares homology to Cas9, or a fragment thereof. For example, a Cas9 variant is at least about 70% identical, at least about 80% identical, 20 at least about 90% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, at least about 99% identical, at least about 99.5% identical, or at least about 99.9% identical to wild type Cas9. In some embodiments, the Cas9 variant may have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 21, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 25   49, 50 or more amino acid changes compared to wild type Cas9. In some embodiments, the Cas9 variant comprises a fragment of Cas9 (e.g., a gRNA binding domain or a DNA-cleavage domain), such that the fragment is at least about 70% identical, at least about 80% identical, at least about 90% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, at least about 99% identical, at least about 99.5% 30 identical, or at least about 99.9% identical to the corresponding fragment of wild type Cas9. In some embodiments, the fragment is at least 30%, at least 35%, at least 40%, at least 45%, at least 2026204466   11 Jun 2026 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% identical, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of the amino acid length of a corresponding wild type Cas9.

[00308] In some embodiments, the fragment is at least 100 amino acids in length. In some 5 embodiments, the fragment is at least 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, or at least 1300 amino acids in length. In some embodiments, wild type Cas9 corresponds to Cas9 from Streptococcus pyogenes (NCBI Reference Sequence: NC_017053.1, nucleotide and amino acid sequences as follows). 10 ATGGATAAGAAATACTCAATAGGCTTAGATATCGGCACAAATAGCGTCGGATGGGC GGTGATCACTGATGATTATAAGGTTCCGTCTAAAAAGTTCAAGGTTCTGGGAAATAC AGACCGCCACAGTATCAAAAAAAATCTTATAGGGGCTCTTTTATTTGGCAGTGGAGA GACAGCGGAAGCGACTCGTCTCAAACGGACAGCTCGTAGAAGGTATACACGTCGGA 15 AGAATCGTATTTGTTATCTACAGGAGATTTTTTCAAATGAGATGGCGAAAGTAGATG ATAGTTTCTTTCATCGACTTGAAGAGTCTTTTTTGGTGGAAGAAGACAAGAAGCATG AACGTCATCCTATTTTTGGAAATATAGTAGATGAAGTTGCTTATCATGAGAAATATC CAACTATCTATCATCTGCGAAAAAAATTGGCAGATTCTACTGATAAAGCGGATTTGC GCTTAATCTATTTGGCCTTAGCGCATATGATTAAGTTTCGTGGTCATTTTTTGATTGA 20 GGGAGATTTAAATCCTGATAATAGTGATGTGGACAAACTATTTATCCAGTTGGTACA AATCTACAATCAATTATTTGAAGAAAACCCTATTAACGCAAGTAGAGTAGATGCTAA AGCGATTCTTTCTGCACGATTGAGTAAATCAAGACGATTAGAAAATCTCATTGCTCA GCTCCCCGGTGAGAAGAGAAATGGCTTGTTTGGGAATCTCATTGCTTTGTCATTGGG ATTGACCCCTAATTTTAAATCAAATTTTGATTTGGCAGAAGATGCTAAATTACAGCT 25 TTCAAAAGATACTTACGATGATGATTTAGATAATTTATTGGCGCAAATTGGAGATCA ATATGCTGATTTGTTTTTGGCAGCTAAGAATTTATCAGATGCTATTTTACTTTCAGAT ATCCTAAGAGTAAATAGTGAAATAACTAAGGCTCCCCTATCAGCTTCAATGATTAAG CGCTACGATGAACATCATCAAGACTTGACTCTTTTAAAAGCTTTAGTTCGACAACAA CTTCCAGAAAAGTATAAAGAAATCTTTTTTGATCAATCAAAAAACGGATATGCAGGT 30 TATATTGATGGGGGAGCTAGCCAAGAAGAATTTTATAAATTTATCAAACCAATTTTA GAAAAAATGGATGGTACTGAGGAATTATTGGTGAAACTAAATCGTGAAGATTTGCT 2026204466   11 Jun 2026 GCGCAAGCAACGGACCTTTGACAACGGCTCTATTCCCCATCAAATTCACTTGGGTGA GCTGCATGCTATTTTGAGAAGACAAGAAGACTTTTATCCATTTTTAAAAGACAATCG TGAGAAGATTGAAAAAATCTTGACTTTTCGAATTCCTTATTATGTTGGTCCATTGGCG CGTGGCAATAGTCGTTTTGCATGGATGACTCGGAAGTCTGAAGAAACAATTACCCCA 5 TGGAATTTTGAAGAAGTTGTCGATAAAGGTGCTTCAGCTCAATCATTTATTGAACGC ATGACAAACTTTGATAAAAATCTTCCAAATGAAAAAGTACTACCAAAACATAGTTTG CTTTATGAGTATTTTACGGTTTATAACGAATTGACAAAGGTCAAATATGTTACTGAG GGAATGCGAAAACCAGCATTTCTTTCAGGTGAACAGAAGAAAGCCATTGTTGATTTA CTCTTCAAAACAAATCGAAAAGTAACCGTTAAGCAATTAAAAGAAGATTATTTCAA 10 AAAAATAGAATGTTTTGATAGTGTTGAAATTTCAGGAGTTGAAGATAGATTTAATGC TTCATTAGGCGCCTACCATGATTTGCTAAAAATTATTAAAGATAAAGATTTTTTGGA TAATGAAGAAAATGAAGATATCTTAGAGGATATTGTTTTAACATTGACCTTATTTGA AGATAGGGGGATGATTGAGGAAAGACTTAAAACATATGCTCACCTCTTTGATGATA AGGTGATGAAACAGCTTAAACGTCGCCGTTATACTGGTTGGGGACGTTTGTCTCGAA 15 AATTGATTAATGGTATTAGGGATAAGCAATCTGGCAAAACAATATTAGATTTTTTGA AATCAGATGGTTTTGCCAATCGCAATTTTATGCAGCTGATCCATGATGATAGTTTGA CATTTAAAGAAGATATTCAAAAAGCACAGGTGTCTGGACAAGGCCATAGTTTACAT GAACAGATTGCTAACTTAGCTGGCAGTCCTGCTATTAAAAAAGGTATTTTACAGACT GTAAAAATTGTTGATGAACTGGTCAAAGTAATGGGGCATAAGCCAGAAAATATCGT 20 TATTGAAATGGCACGTGAAAATCAGACAACTCAAAAGGGCCAGAAAAATTCGCGAG AGCGTATGAAACGAATCGAAGAAGGTATCAAAGAATTAGGAAGTCAGATTCTTAAA GAGCATCCTGTTGAAAATACTCAATTGCAAAATGAAAAGCTCTATCTCTATTATCTA CAAAATGGAAGAGACATGTATGTGGACCAAGAATTAGATATTAATCGTTTAAGTGA TTATGATGTCGATCACATTGTTCCACAAAGTTTCATTAAAGACGATTCAATAGACAA 25 TAAGGTACTAACGCGTTCTGATAAAAATCGTGGTAAATCGGATAACGTTCCAAGTGA AGAAGTAGTCAAAAAGATGAAAAACTATTGGAGACAACTTCTAAACGCCAAGTTAA TCACTCAACGTAAGTTTGATAATTTAACGAAAGCTGAACGTGGAGGTTTGAGTGAAC TTGATAAAGCTGGTTTTATCAAACGCCAATTGGTTGAAACTCGCCAAATCACTAAGC ATGTGGCACAAATTTTGGATAGTCGCATGAATACTAAATACGATGAAAATGATAAA 30 CTTATTCGAGAGGTTAAAGTGATTACCTTAAAATCTAAATTAGTTTCTGACTTCCGA AAAGATTTCCAATTCTATAAAGTACGTGAGATTAACAATTACCATCATGCCCATGAT 2026204466   11 Jun 2026 GCGTATCTAAATGCCGTCGTTGGAACTGCTTTGATTAAGAAATATCCAAAACTTGAA TCGGAGTTTGTCTATGGTGATTATAAAGTTTATGATGTTCGTAAAATGATTGCTAAGT CTGAGCAAGAAATAGGCAAAGCAACCGCAAAATATTTCTTTTACTCTAATATCATGA ACTTCTTCAAAACAGAAATTACACTTGCAAATGGAGAGATTCGCAAACGCCCTCTAA 5 TCGAAACTAATGGGGAAACTGGAGAAATTGTCTGGGATAAAGGGCGAGATTTTGCC ACAGTGCGCAAAGTATTGTCCATGCCCCAAGTCAATATTGTCAAGAAAACAGAAGT ACAGACAGGCGGATTCTCCAAGGAGTCAATTTTACCAAAAAGAAATTCGGACAAGC TTATTGCTCGTAAAAAAGACTGGGATCCAAAAAAATATGGTGGTTTTGATAGTCCAA CGGTAGCTTATTCAGTCCTAGTGGTTGCTAAGGTGGAAAAAGGGAAATCGAAGAAG 10 TTAAAATCCGTTAAAGAGTTACTAGGGATCACAATTATGGAAAGAAGTTCCTTTGAA AAAAATCCGATTGACTTTTTAGAAGCTAAAGGATATAAGGAAGTTAAAAAAGACTT AATCATTAAACTACCTAAATATAGTCTTTTTGAGTTAGAAAACGGTCGTAAACGGAT GCTGGCTAGTGCCGGAGAATTACAAAAAGGAAATGAGCTGGCTCTGCCAAGCAAAT ATGTGAATTTTTTATATTTAGCTAGTCATTATGAAAAGTTGAAGGGTAGTCCAGAAG 15 ATAACGAACAAAAACAATTGTTTGTGGAGCAGCATAAGCATTATTTAGATGAGATT ATTGAGCAAATCAGTGAATTTTCTAAGCGTGTTATTTTAGCAGATGCCAATTTAGAT AAAGTTCTTAGTGCATATAACAAACATAGAGACAAACCAATACGTGAACAAGCAGA AAATATTATTCATTTATTTACGTTGACGAATCTTGGAGCTCCCGCTGCTTTTAAATAT TTTGATACAACAATTGATCGTAAACGATATACGTCTACAAAAGAAGTTTTAGATGCC 20 ACTCTTATCCATCAATCCATCACTGGTCTTTATGAAACACGCATTGATTTGAGTCAGC TAGGAGGTGACTGA

[00309] MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKKNLIGALLFGSGETA EATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF 25 GNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS DVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRLENLIAQLPGEKRNGLFGN LIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDA ILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYA GYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELH 30 AILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEV VDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAF 2026204466   11 Jun 2026 LSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGAYHDLL KIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEERLKTYAHLFDDKVMKQLKRRRYTG WGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQG HSLHEOIANLAGSPAIKKGILOTVKIVDELVKVMGHKPENIVIEMARENOTTOKGOKNS 5 RERMKRIEEGIKELGSOILKEHPVENTQLONEKLYLYYLONGRDMYVDQELDINRLSDY DVDHIVPOSFIKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWROLLNAKLITQ RKFDNLTKAERGGLSELDKAGFIKROLVETROITKHVAOILDSRMNTKYDENDKLIREV KVITLKSKLVSDFRKDFOFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGD YKVYDVRKMIAKSEOEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIV 10 WDKGRDFATVRKVLSMPOVNIVKKTEVOTGGFSKESIEPKRNSDKEIARKKDWDPKKY GGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEV KKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSP EDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD 15

[00310] (single underline: HNH domain; double underline: RuvC domain)

[00311] In some embodiments, wild type Cas9 corresponds to, or comprises the following nucleotide and / or amino acid sequences: 20 ATGGATAAAAAGTATTCTATTGGTTTAGACATCGGCACTAATTCCGTTGGATGGGCT GTCATAACCGATGAATACAAAGTACCTTCAAAGAAATTTAAGGTGTTGGGGAACAC AGACCGTCATTCGATTAAAAAGAATCTTATCGGTGCCCTCCTATTCGATAGTGGCGA AACGGCAGAGGCGACTCGCCTGAAACGAACCGCTCGGAGAAGGTATACACGTCGCA AGAACCGAATATGTTACTTACAAGAAATTTTTAGCAATGAGATGGCCAAAGTTGAC 25 GATTCTTTCTTTCACCGTTTGGAAGAGTCCTTCCTTGTCGAAGAGGACAAGAAACAT GAACGGCACCCCATCTTTGGAAACATAGTAGATGAGGTGGCATATCATGAAAAGTA CCCAACGATTTATCACCTCAGAAAAAAGCTAGTTGACTCAACTGATAAAGCGGACCT GAGGTTAATCTACTTGGCTCTTGCCCATATGATAAAGTTCCGTGGGCACTTTCTCATT GAGGGTGATCTAAATCCGGACAACTCGGATGTCGACAAACTGTTCATCCAGTTAGTA 30 CAAACCTATAATCAGTTGTTTGAAGAGAACCCTATAAATGCAAGTGGCGTGGATGC GAAGGCTATTCTTAGCGCCCGCCTCTCTAAATCCCGACGGCTAGAAAACCTGATCGC 2026204466   11 Jun 2026 ACAATTACCCGGAGAGAAGAAAAATGGGTTGTTCGGTAACCTTATAGCGCTCTCACT AGGCCTGACACCAAATTTTAAGTCGAACTTCGACTTAGCTGAAGATGCCAAATTGCA GCTTAGTAAGGACACGTACGATGACGATCTCGACAATCTACTGGCACAAATTGGAG ATCAGTATGCGGACTTATTTTTGGCTGCCAAAAACCTTAGCGATGCAATCCTCCTAT 5 CTGACATACTGAGAGTTAATACTGAGATTACCAAGGCGCCGTTATCCGCTTCAATGA TCAAAAGGTACGATGAACATCACCAAGACTTGACACTTCTCAAGGCCCTAGTCCGTC AGCAACTGCCTGAGAAATATAAGGAAATATTCTTTGATCAGTCGAAAAACGGGTAC GCAGGTTATATTGACGGCGGAGCGAGTCAAGAGGAATTCTACAAGTTTATCAAACC CATATTAGAGAAGATGGATGGGACGGAAGAGTTGCTTGTAAAACTCAATCGCGAAG 10 ATCTACTGCGAAAGCAGCGGACTTTCGACAACGGTAGCATTCCACATCAAATCCACT TAGGCGAATTGCATGCTATACTTAGAAGGCAGGAGGATTTTTATCCGTTCCTCAAAG ACAATCGTGAAAAGATTGAGAAAATCCTAACCTTTCGCATACCTTACTATGTGGGAC CCCTGGCCCGAGGGAACTCTCGGTTCGCATGGATGACAAGAAAGTCCGAAGAAACG ATTACTCCATGGAATTTTGAGGAAGTTGTCGATAAAGGTGCGTCAGCTCAATCGTTC 15 ATCGAGAGGATGACCAACTTTGACAAGAATTTACCGAACGAAAAAGTATTGCCTAA GCACAGTTTACTTTACGAGTATTTCACAGTGTACAATGAACTCACGAAAGTTAAGTA TGTCACTGAGGGCATGCGTAAACCCGCCTTTCTAAGCGGAGAACAGAAGAAAGCAA TAGTAGATCTGTTATTCAAGACCAACCGCAAAGTGACAGTTAAGCAATTGAAAGAG GACTACTTTAAGAAAATTGAATGCTTCGATTCTGTCGAGATCTCCGGGGTAGAAGAT 20 CGATTTAATGCGTCACTTGGTACGTATCATGACCTCCTAAAGATAATTAAAGATAAG GACTTCCTGGATAACGAAGAGAATGAAGATATCTTAGAAGATATAGTGTTGACTCTT ACCCTCTTTGAAGATCGGGAAATGATTGAGGAAAGACTAAAAACATACGCTCACCT GTTCGACGATAAGGTTATGAAACAGTTAAAGAGGCGTCGCTATACGGGCTGGGGAC GATTGTCGCGGAAACTTATCAACGGGATAAGAGACAAGCAAAGTGGTAAAACTATT 25 CTCGATTTTCTAAAGAGCGACGGCTTCGCCAATAGGAACTTTATGCAGCTGATCCAT GATGACTCTTTAACCTTCAAAGAGGATATACAAAAGGCACAGGTTTCCGGACAAGG GGACTCATTGCACGAACATATTGCGAATCTTGCTGGTTCGCCAGCCATCAAAAAGGG CATACTCCAGACAGTCAAAGTAGTGGATGAGCTAGTTAAGGTCATGGGACGTCACA AACCGGAAAACATTGTAATCGAGATGGCACGCGAAAATCAAACGACTCAGAAGGG 30 GCAAAAAAACAGTCGAGAGCGGATGAAGAGAATAGAAGAGGGTATTAAAGAACTG GGCAGCCAGATCTTAAAGGAGCATCCTGTGGAAAATACCCAATTGCAGAACGAGAA 2026204466   11 Jun 2026 ACTTTACCTCTATTACCTACAAAATGGAAGGGACATGTATGTTGATCAGGAACTGGA CATAAACCGTTTATCTGATTACGACGTCGATCACATTGTACCCCAATCCTTTTTGAAG GACGATTCAATCGACAATAAAGTGCTTACACGCTCGGATAAGAACCGAGGGAAAAG TGACAATGTTCCAAGCGAGGAAGTCGTAAAGAAAATGAAGAACTATTGGCGGCAGC 5 TCCTAAATGCGAAACTGATAACGCAAAGAAAGTTCGATAACTTAACTAAAGCTGAG AGGGGTGGCTTGTCTGAACTTGACAAGGCCGGATTTATTAAACGTCAGCTCGTGGAA ACCCGCCAAATCACAAAGCATGTTGCACAGATACTAGATTCCCGAATGAATACGAA ATACGACGAGAACGATAAGCTGATTCGGGAAGTCAAAGTAATCACTTTAAAGTCAA AATTGGTGTCGGACTTCAGAAAGGATTTTCAATTCTATAAAGTTAGGGAGATAAATA 10 ACTACCACCATGCGCACGACGCTTATCTTAATGCCGTCGTAGGGACCGCACTCATTA AGAAATACCCGAAGCTAGAAAGTGAGTTTGTGTATGGTGATTACAAAGTTTATGAC GTCCGTAAGATGATCGCGAAAAGCGAACAGGAGATAGGCAAGGCTACAGCCAAAT ACTTCTTTTATTCTAACATTATGAATTTCTTTAAGACGGAAATCACTCTGGCAAACGG AGAGATACGCAAACGACCTTTAATTGAAACCAATGGGGAGACAGGTGAAATCGTAT 15 GGGATAAGGGCCGGGACTTCGCGACGGTGAGAAAAGTTTTGTCCATGCCCCAAGTC AACATAGTAAAGAAAACTGAGGTGCAGACCGGAGGGTTTTCAAAGGAATCGATTCT TCCAAAAAGGAATAGTGATAAGCTCATCGCTCGTAAAAAGGACTGGGACCCGAAAA AGTACGGTGGCTTCGATAGCCCTACAGTTGCCTATTCTGTCCTAGTAGTGGCAAAAG TTGAGAAGGGAAAATCCAAGAAACTGAAGTCAGTCAAAGAATTATTGGGGATAACG 20 ATTATGGAGCGCTCGTCTTTTGAAAAGAACCCCATCGACTTCCTTGAGGCGAAAGGT TACAAGGAAGTAAAAAAGGATCTCATAATTAAACTACCAAAGTATAGTCTGTTTGA GTTAGAAAATGGCCGAAAACGGATGTTGGCTAGCGCCGGAGAGCTTCAAAAGGGGA ACGAACTCGCACTACCGTCTAAATACGTGAATTTCCTGTATTTAGCGTCCCATTACG AGAAGTTGAAAGGTTCACCTGAAGATAACGAACAGAAGCAACTTTTTGTTGAGCAG 25 CACAAACATTATCTCGACGAAATCATAGAGCAAATTTCGGAATTCAGTAAGAGAGT CATCCTAGCTGATGCCAATCTGGACAAAGTATTAAGCGCATACAACAAGCACAGGG ATAAACCCATACGTGAGCAGGCGGAAAATATTATCCATTTGTTTACTCTTACCAACC TCGGCGCTCCAGCCGCATTCAAGTATTTTGACACAACGATAGATCGCAAACGATACA CTTCTACCAAGGAGGTGCTAGACGCGACACTGATTCACCAATCCATCACGGGATTAT 30 ATGAAACTCGGATAGATTTGTCACAGCTTGGGGGTGACGGATCCCCCAAGAAGAAG 2026204466   11 Jun 2026 AGGAAAGTCTCGAGCGACTACAAAGACCATGACGGTGATTATAAAGATCATGACAT CGATTACAAGGATGACGATGACAAGGCTGCAGGA MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETA 5 EATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS DVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFG NLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSD AILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY 10 AGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGEL HAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEE VVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPA FLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLL KIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTG 15 WGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQG DSLHEHIANLAGSPAIKKGILOTVKVVDELVKVMGRHKPENIVIEMARENOTTOKGQKN SRERMKRIEEGIKELGSOILKEHPVENTOLONEKLYLYYLONGRDMYVDQELDINRLSD YDVDHIVPOSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLIT QRKFDNLTKAERGGLSELDKAGFIKROLVETROITKHVAOTLDSRMNTKYDENDKLIRE 20 VKVITLKSKLVSDFRKDFOFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYG DYKVYDVRKMIAKSEOEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEI VWDKGRDFATVRKVLSMPOVNIVKKTEVOTGGFSKESILPKRNSDKLIARKKDWDPKK YGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKE VKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGS 25 PEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENI IHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD

[00312] (single underline: HNH domain; double underline: RuvC domain)

[00313] In some embodiments, wild type Cas9 corresponds to Cas9 from Streptococcus 30 pyogenes (NCBI Reference Sequence: NC_002737.2 (nucleotide sequence as follows); and Uniprot Reference Sequence: Q99ZW2 (amino acid sequence as follows). 2026204466   11 Jun 2026 ATGGATAAGAAATACTCAATAGGCTTAGATATCGGCACAAATAGCGTCGGATGGGC GGTGATCACTGATGAATATAAGGTTCCGTCTAAAAAGTTCAAGGTTCTGGGAAATAC AGACCGCCACAGTATCAAAAAAAATCTTATAGGGGCTCTTTTATTTGACAGTGGAGA 5 GACAGCGGAAGCGACTCGTCTCAAACGGACAGCTCGTAGAAGGTATACACGTCGGA AGAATCGTATTTGTTATCTACAGGAGATTTTTTCAAATGAGATGGCGAAAGTAGATG ATAGTTTCTTTCATCGACTTGAAGAGTCTTTTTTGGTGGAAGAAGACAAGAAGCATG AACGTCATCCTATTTTTGGAAATATAGTAGATGAAGTTGCTTATCATGAGAAATATC CAACTATCTATCATCTGCGAAAAAAATTGGTAGATTCTACTGATAAAGCGGATTTGC 10 GCTTAATCTATTTGGCCTTAGCGCATATGATTAAGTTTCGTGGTCATTTTTTGATTGA GGGAGATTTAAATCCTGATAATAGTGATGTGGACAAACTATTTATCCAGTTGGTACA AACCTACAATCAATTATTTGAAGAAAACCCTATTAACGCAAGTGGAGTAGATGCTA AAGCGATTCTTTCTGCACGATTGAGTAAATCAAGACGATTAGAAAATCTCATTGCTC AGCTCCCCGGTGAGAAGAAAAATGGCTTATTTGGGAATCTCATTGCTTTGTCATTGG 15 GTTTGACCCCTAATTTTAAATCAAATTTTGATTTGGCAGAAGATGCTAAATTACAGC TTTCAAAAGATACTTACGATGATGATTTAGATAATTTATTGGCGCAAATTGGAGATC AATATGCTGATTTGTTTTTGGCAGCTAAGAATTTATCAGATGCTATTTTACTTTCAGA TATCCTAAGAGTAAATACTGAAATAACTAAGGCTCCCCTATCAGCTTCAATGATTAA ACGCTACGATGAACATCATCAAGACTTGACTCTTTTAAAAGCTTTAGTTCGACAACA 20 ACTTCCAGAAAAGTATAAAGAAATCTTTTTTGATCAATCAAAAAACGGATATGCAG GTTATATTGATGGGGGAGCTAGCCAAGAAGAATTTTATAAATTTATCAAACCAATTT TAGAAAAAATGGATGGTACTGAGGAATTATTGGTGAAACTAAATCGTGAAGATTTG CTGCGCAAGCAACGGACCTTTGACAACGGCTCTATTCCCCATCAAATTCACTTGGGT GAGCTGCATGCTATTTTGAGAAGACAAGAAGACTTTTATCCATTTTTAAAAGACAAT 25 CGTGAGAAGATTGAAAAAATCTTGACTTTTCGAATTCCTTATTATGTTGGTCCATTGG CGCGTGGCAATAGTCGTTTTGCATGGATGACTCGGAAGTCTGAAGAAACAATTACCC CATGGAATTTTGAAGAAGTTGTCGATAAAGGTGCTTCAGCTCAATCATTTATTGAAC GCATGACAAACTTTGATAAAAATCTTCCAAATGAAAAAGTACTACCAAAACATAGT TTGCTTTATGAGTATTTTACGGTTTATAACGAATTGACAAAGGTCAAATATGTTACTG 30 AAGGAATGCGAAAACCAGCATTTCTTTCAGGTGAACAGAAGAAAGCCATTGTTGAT TTACTCTTCAAAACAAATCGAAAAGTAACCGTTAAGCAATTAAAAGAAGATTATTTC 2026204466   11 Jun 2026 AAAAAAATAGAATGTTTTGATAGTGTTGAAATTTCAGGAGTTGAAGATAGATTTAAT GCTTCATTAGGTACCTACCATGATTTGCTAAAAATTATTAAAGATAAAGATTTTTTG GATAATGAAGAAAATGAAGATATCTTAGAGGATATTGTTTTAACATTGACCTTATTT GAAGATAGGGAGATGATTGAGGAAAGACTTAAAACATATGCTCACCTCTTTGATGA 5 TAAGGTGATGAAACAGCTTAAACGTCGCCGTTATACTGGTTGGGGACGTTTGTCTCG AAAATTGATTAATGGTATTAGGGATAAGCAATCTGGCAAAACAATATTAGATTTTTT GAAATCAGATGGTTTTGCCAATCGCAATTTTATGCAGCTGATCCATGATGATAGTTT GACATTTAAAGAAGACATTCAAAAAGCACAAGTGTCTGGACAAGGCGATAGTTTAC ATGAACATATTGCAAATTTAGCTGGTAGCCCTGCTATTAAAAAAGGTATTTTACAGA 10 CTGTAAAAGTTGTTGATGAATTGGTCAAAGTAATGGGGCGGCATAAGCCAGAAAAT ATCGTTATTGAAATGGCACGTGAAAATCAGACAACTCAAAAGGGCCAGAAAAATTC GCGAGAGCGTATGAAACGAATCGAAGAAGGTATCAAAGAATTAGGAAGTCAGATTC TTAAAGAGCATCCTGTTGAAAATACTCAATTGCAAAATGAAAAGCTCTATCTCTATT ATCTCCAAAATGGAAGAGACATGTATGTGGACCAAGAATTAGATATTAATCGTTTAA 15 GTGATTATGATGTCGATCACATTGTTCCACAAAGTTTCCTTAAAGACGATTCAATAG ACAATAAGGTCTTAACGCGTTCTGATAAAAATCGTGGTAAATCGGATAACGTTCCAA GTGAAGAAGTAGTCAAAAAGATGAAAAACTATTGGAGACAACTTCTAAACGCCAAG TTAATCACTCAACGTAAGTTTGATAATTTAACGAAAGCTGAACGTGGAGGTTTGAGT GAACTTGATAAAGCTGGTTTTATCAAACGCCAATTGGTTGAAACTCGCCAAATCACT 20 AAGCATGTGGCACAAATTTTGGATAGTCGCATGAATACTAAATACGATGAAAATGA TAAACTTATTCGAGAGGTTAAAGTGATTACCTTAAAATCTAAATTAGTTTCTGACTTC CGAAAAGATTTCCAATTCTATAAAGTACGTGAGATTAACAATTACCATCATGCCCAT GATGCGTATCTAAATGCCGTCGTTGGAACTGCTTTGATTAAGAAATATCCAAAACTT GAATCGGAGTTTGTCTATGGTGATTATAAAGTTTATGATGTTCGTAAAATGATTGCT 25 AAGTCTGAGCAAGAAATAGGCAAAGCAACCGCAAAATATTTCTTTTACTCTAATATC ATGAACTTCTTCAAAACAGAAATTACACTTGCAAATGGAGAGATTCGCAAACGCCCT CTAATCGAAACTAATGGGGAAACTGGAGAAATTGTCTGGGATAAAGGGCGAGATTT TGCCACAGTGCGCAAAGTATTGTCCATGCCCCAAGTCAATATTGTCAAGAAAACAG AAGTACAGACAGGCGGATTCTCCAAGGAGTCAATTTTACCAAAAAGAAATTCGGAC 30 AAGCTTATTGCTCGTAAAAAAGACTGGGATCCAAAAAAATATGGTGGTTTTGATAGT CCAACGGTAGCTTATTCAGTCCTAGTGGTTGCTAAGGTGGAAAAAGGGAAATCGAA 2026204466   11 Jun 2026 GAAGTTAAAATCCGTTAAAGAGTTACTAGGGATCACAATTATGGAAAGAAGTTCCTT TGAAAAAAATCCGATTGACTTTTTAGAAGCTAAAGGATATAAGGAAGTTAAAAAAG ACTTAATCATTAAACTACCTAAATATAGTCTTTTTGAGTTAGAAAACGGTCGTAAAC GGATGCTGGCTAGTGCCGGAGAATTACAAAAAGGAAATGAGCTGGCTCTGCCAAGC 5 AAATATGTGAATTTTTTATATTTAGCTAGTCATTATGAAAAGTTGAAGGGTAGTCCA GAAGATAACGAACAAAAACAATTGTTTGTGGAGCAGCATAAGCATTATTTAGATGA GATTATTGAGCAAATCAGTGAATTTTCTAAGCGTGTTATTTTAGCAGATGCCAATTT AGATAAAGTTCTTAGTGCATATAACAAACATAGAGACAAACCAATACGTGAACAAG CAGAAAATATTATTCATTTATTTACGTTGACGAATCTTGGAGCTCCCGCTGCTTTTAA 10 ATATTTTGATACAACAATTGATCGTAAACGATATACGTCTACAAAAGAAGTTTTAGA TGCCACTCTTATCCATCAATCCATCACTGGTCTTTATGAAACACGCATTGATTTGAGT CAGCTAGGAGGTGACTGA

[00314] MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLF 15 DSGETAEATRLKRTARRRYTRRKNRICYLOEIFSNEMAKVDDSFFHRLEESFLVEEDKK HERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEG DLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFL AAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFF 20 DQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETI TPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTE GMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASL GTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQL 25 KRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKA OVSGQGDSLHEHIANLAGSPAIKKGILOTVKVVDELVKVMGRHKPENIVIEMAR ENOTT OKGOKNSRERMKRIEEGIKELGSOILKEHPVENTOLONEKLYLYYLONGRDMYVDQEL DINRLSDYDVDHIVPOSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQL LNAKLITQRKFDNLTKAERGGLSELDKAGFIKROLVETROITKHVAOILDSRMNTKYDE 30 NDKLIREVKVITLKSKLVSDFRKDFOFYKVREINNYHHAHDAYLNAVVGTALIKKYPKL ESEFVYGDYKVYDVRKMIAKSEOEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIET 2026204466   11 Jun 2026 NGETGEIVWDKGRDFATVRKVLSMPOVNIVKKTEVOTGGFSKESIEPKRNSDKEIARKK DWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLE AKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHY EKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPI 5 REQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQ LGGD (single underline: HNH domain; double underline: RuvC domain)

[00315] In some embodiments, Cas9 refers to Cas9 from: Corynebacterium ulcerans (NCBI Refs: NC_015683.1, NC_017317.1); Corynebacterium diphtheria (NCBI Refs: NC_016782.1, 10 NC_016786.1); Spiroplasma syrphidicola (NCBI Ref: NC_021284.1); Prevotella intermedia (NCBI Ref: NC_017861.1); Spiroplasma taiwanense (NCBI Ref: NC_021846.1); Streptococcus iniae (NCBI Ref: NC_021314.1); Belliella baltica (NCBI Ref: NC_018010.1); Psychroflexus torquisl (NCBI Ref: NC_018721.1); Streptococcus thermophilus (NCBI Ref: YP_820832.1), Listeria innocua (NCBI Ref: NP_472073.1), Campylobacter jejuni (NCBI Ref: 15   YP_002344900.1) or Neisseria, meningitidis (NCBI Ref: YP_002342100.1) or to a Cas9 from any other organism.

[00316] In some embodiments, dCas9 corresponds to, or comprises in part or in whole, a Cas9 amino acid sequence having one or more mutations that inactivate the Cas9 nuclease activity. For example, in some embodiments, a dCas9 domain comprises D10A and an H840A 20 mutation or corresponding mutations in another Cas9. In some embodiments, the dCas9 comprises the amino acid sequence of dCas9 (D10A and H840A):

[00317] MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLF DSGETAEATREKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHREEFSFEVEEDKK 25 HERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEG DLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFL AAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFF DQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH 30 QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETI TPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTE 2026204466   11 Jun 2026 GMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASL GTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQL KRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKA OVSGOGDSLHEHIANLAGSPAIKKGILOTVKVVDELVKVMGRHKPENIVIEMARENOTT 5 OKGOKNSRERMKRIEEGIKELGSOILKEHPVENTOLONEKLYLYYLONGRDMYVDQEL DINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWROL LNAKLITQRKFDNLTKAERGGLSELDKAGFIKROLVETROITKHVAOILDSRMNTKYDE NDKLIREVKVITLKSKLVSDFRKDFOFYKVREINNYHHAHDAYLNAVVGTALIKKYPKL ESEFVYGDYKVYDVRKMIAKSEOEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIET 10 NGETGEIVWDKGRDFATVRKVLSMPOVNIVKKTEVOTGGFSKESIEPKRNSDKEIARKK DWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLE AKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHY EKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPI REQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQ 15 LGGD (single underline: HNH domain; double underline: RuvC domain).

[00318] In some embodiments, the Cas9 domain comprises a D10A mutation, while the residue at position 840 remains a histidine in the amino acid sequence provided above, or at corresponding positions in any of the amino acid sequences provided herein.

[00319] In other embodiments, dCas9 variants having mutations other than D10A and H840A 20 are provided, which, e.g., result in nuclease inactivated Cas9 (dCas9). Such mutations, by way of example, include other amino acid substitutions at D10 and H840, or other substitutions within the nuclease domains of Cas9 (e.g., substitutions in the HNH nuclease subdomain and / or the RuvCl subdomain).

[00320] In some embodiments, variants or homologues of dCas9 are provided which are at 25 least about 70% identical, at least about 80% identical, at least about 90% identical, at least about 95% identical, at least about 98% identical, at least about 99% identical, at least about 99.5% identical, or at least about 99.9% identical. In some embodiments, variants of dCas9 are provided having amino acid sequences which are shorter, or longer, by about 5 amino acids, by about 10 amino acids, by about 15 amino acids, by about 20 amino acids, by about 25 amino 30 acids, by about 30 amino acids, by about 40 amino acids, by about 50 amino acids, by about 75 amino acids, by about 100 amino acids or more. 2026204466   11 Jun 2026

[00321] In some embodiments, Cas9 fusion proteins as provided herein comprise the fulllength amino acid sequence of a Cas9 protein, e.g., one of the Cas9 sequences provided herein. In other embodiments, however, fusion proteins as provided herein do not comprise a full-length Cas9 sequence, but only a fragment thereof. For example, in some embodiments, a Cas9 fusion 5 protein provided herein comprises a Cas9 fragment, wherein the fragment binds crRNA and tracrRNA or sgRNA, but does not comprise a functional nuclease domain, e.g., in that it comprises only a truncated version of a nuclease domain or no nuclease domain at all.

[00322] Exemplary amino acid sequences of suitable Cas9 domains and Cas9 fragments are provided herein, and additional suitable sequences of Cas9 domains and fragments will be 10 apparent to those of skill in the art.

[00323] In some embodiments, Cas9 refers to Cas9 from: Corynebacterium ulcercms (NCBI Refs: NC_015683.1, NC_017317.1); Corynebacterium diphtheria (NCBI Refs: NC_016782.1, NC_016786.1); Spiroplasma syrphidicola (NCBI Ref: NC_021284.1); Prevotella intermedia (NCBI Ref: NC_017861.1); Spiroplasma taiwanense (NCBI Ref: NC_021846.1); Streptococcus 15 iniae (NCBI Ref: NC_021314.1); Belliella baltica (NCBI Ref: NC_018010.1); Psychroflexus torquisl (NCBI Ref: NC_018721.1); Streptococcus thermophilus (NCBI Ref: YP_820832.1); Listeria innocua (NCBI Ref: NP_472073.1); Campylobacter jejuni (NCBI Ref: YP_002344900.1); or Neisseria, meningitidis (NCBI Ref: YP_002342100.1).

[00324] It should be appreciated that additional Cas9 proteins (e.g., a nuclease dead Cas9 20 (dCas9), a Cas9 nickase (nCas9), or a nuclease active Cas9), including variants and homologs thereof, are within the scope of this disclosure. Exemplary Cas9 proteins include, without limitation, those provided below. In some embodiments, the Cas9 protein is a nuclease dead Cas9 (dCas9). In some embodiments, the Cas9 protein is a Cas9 nickase (nCas9). In some embodiments, the Cas9 protein is a nuclease active Cas9. 25

[00325] Exemplary catalytically inactive Cas9 (dCas9): DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEA TRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDV 30 DKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLI ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL 2026204466   11 Jun 2026 LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAG YIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAI LRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEW DKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLS 5 GEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKII KDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWG RLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRE RMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV 10 DAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRK FDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVI TLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYK VYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWD KGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGG 15 FDSPTVAYSVLWAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKK DLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD 20

[00326] Exemplary catalytically Cas9 nickase (nCas9): DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEA TRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDV DKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLI 25 ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAG YIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAI LRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEW DKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLS 30 GEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKII KDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWG 2026204466   11 Jun 2026 RLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRE RMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV DHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRK 5 FDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVI TLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYK VYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWD KGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGG FDSPTVAYSVLWAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKK 10 DLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD

[00327] Exemplary catalytically active Cas9: 15 DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEA TRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDV DKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLI ALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL 20 LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAG YIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAI LRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEW DKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLS GEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKII 25 KDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWG RLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSL HEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRE RMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV DHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRK 30 FDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVI TLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYK 2026204466   11 Jun 2026 VYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWD KGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGG FDSPTVAYSVLWAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKK DLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED 5 NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHL FTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD.

[00328] In some embodiments, Cas9 refers to a Cas9 from archaea (e.g. nanoarchaea), which constitute a domain and kingdom of single-celled prokaryotic microbes. In some embodiments, Cas9 refers to CasX or CasY, which have been described in, for example, Burstein et al., "New 10 CRISPR-Cas systems from uncultivated microbes." Cell Res. 2017 Feb 21. doi: 10.103 8 / cr.2017.21, the entire contents of which is hereby incorporated by reference. Using genome-resolved metagenomics, a number of CRISPR-Cas systems were identified, including the first reported Cas9 in the archaeal domain of life. This divergent Cas9 protein was found in little- studied nanoarchaea as part of an active CRISPR-Cas system. In bacteria, two previously 15 unknown systems were discovered, CRISPR-CasX and CRISPR-CasY, which are among the most compact systems yet discovered. In some embodiments, Cas9 refers to CasX, or a variant of CasX. In some embodiments, Cas9 refers to a CasY, or a variant of CasY. It should be appreciated that other RNA-guided DNA binding proteins may be used as a nucleic acid programmable DNA binding protein (napDNAbp), and are within the scope of this disclosure. 20

[00329] In some embodiments, the nucleic acid programmable DNA binding protein (napDNAbp) or any of the fusion proteins provided herein may be a CasX or CasY protein. In some embodiments, the napDNAbp is a CasX protein. In some embodiments, the napDNAbp is a CasY protein. In some embodiments, the napDNAbp comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at 25 least 96%, at least 97%, at least 98%, at least 99%, or at ease 99.5% identical to a naturally-occurring CasX or CasY protein. In some embodiments, the napDNAbp is a naturally-occurring CasX or CasY protein. In some embodiments, the napDNAbp comprises an amino acid sequence that is at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at ease 99.5% identical to any 30 CasX or CasY protein described herein. It should be appreciated that CasX and CasY from other bacterial species may also be used in accordance with the present disclosure. 2026204466   11 Jun 2026

[00330] CasX (uniprot.org / uniprot / F0NN87; uniprot.org / uniprot / F0NH53)

[00331] >tr|F0NN87|F0NN87_SULIH CRISPR-associated Casx protein OS = Sulfolobus islandicus (strain HVE10 / 4) GN = SiH_0402 PE=4 SV=1 5 MEVPLYNIFGDNYIIQVATEAENSTIYNNKVEIDDEELRNVLNLAYKIAKNNEDAAAERR GKAKKKKGEEGETTTSNIILPLSGNDKNPWTETLKCYNFPTTVALSEVFKNFSQVKECEE VSAPSFVKPEFYEFGRSPGMVERTRRVKLEVEPHYLIIAAAGWVLTRLGKAKVSEGDYV GVNVFTPTRGILYSLIQNVNGIVPGIKPETAFGLWIARKVVSSVTNPNVSWRIYTISDAV GQNPTTINGGFSIDLTKLLEKRYLLSERLEAIARNALSISSNMRERYIVLANYIYEYLTG 10 SKRLEDLLYFANRDLIMNLNSDDGKVRDLKLISAYVNGELIRGEG

[00332] >tr|F0NH53|F0NH53_SULIR CRISPR associated protein, Casx OS = Sulfolobus islandicus (strain REY15A) GN=SiRe_0771 PE=4 SV=1 MEVPLYNIFGDNYIIQVATEAENSTIYNNKVEIDDEELRNVLNLAYKIAKNNEDAAAERR 15 GKAKKKKGEEGETTTSNIILPLSGNDKNPWTETLKCYNFPTTVALSEVFKNFSQVKECEE VSAPSFVKPEFYKFGRSPGMVERTRRVKLEVEPHYLIMAAAGWVLTRLGKAKVSEGDY VGVNVFTPTRGILYSLIQNVNGIVPGIKPETAFGLWIARKVVSSVTNPNVSVVSIYTISDA VGQNPTTINGGFSIDLTKLLEKRDLLSERLEAIARNALSISSNMRERYIVLANYIYEYLTGS KRLEDLLYFANRDLIMNLNSDDGKVRDLKLISAYVNGELIRGEG 20

[00333] CasY (ncbi.nlm.nih.gov / protein / APG80656.1)

[00334] >APG80656.1 CRISPR-associated protein CasY [uncultured Parcubacteria group bacterium] MSKRHPRISGVKGYRLHAQRLEYTGKSGAMRTIKYPLYSSPSGGRTVPREIVSAINDDY 25 VGLYGLSNFDDLYNAEKRNEEKVYSVLDFWYDCVQYGAVFSYTAPGLLKNVAEVRGG SYELTKTLKGSHLYDELQIDKVIKFLNKKEISRANGSLDKLKKDIIDCFKAEYRERHKDQ CNKLADDIKNAKKDAGASLGERQKKLFRDFFGISEQSENDKPSFTNPLNLTCCLLPFDTV NNNRNRGEVLFNKLKEYAQKLDKNEGSLEMWEYIGIGNSGTAFSNFLGEGFLGRLREN KITELKKAMMDITDAWRGQEQEEELEKRLRILAALTIKLREPKFDNHWGGYRSDINGKL 30 SSWLQNYINQTVKIKEDLKGHKKDLKKAKEMINRFGESDTKEEAVVSSLLESIEKIVPDD SADDEKPDIPAIAIYRRFLSDGRLTLNRFVQREDVQEALIKERLEAEKKKKPKKRKKKSD 2026204466   11 Jun 2026 AEDEKETIDFKELFPHLAKPLKLVPNFYGDSKRELYKKYKNAAIYTDALWKAVEKIYKS AFSSSLKNSFFDTDFDKDFFIKRLQKIFSVYRRFNTDKWKPIVKNSFAPYCDIVSLAENEV LYKPKQSRSRKSAAIDKNRVRLPSTENIAKAGIALARELSVAGFDWKDLLKKEEHEEYID LIELHKTALALLLAVTETQLDISALDFVENGTVKDFMKTRDGNLVLEGRFLEMFSQSIVF 5 SELRGLAGLMSRKEFITRSAIQTMNGKQAELLYIPHEFQSAKITTPKEMSRAFLDLAPAEF ATSLEPESLSEKSLLKLKQMRYYPHYFGYELTRTGQGIDGGVAENALRLEKSPVKKREIK CKQYKTLGRGQNKIVLYVRSSYYQTQFLEWFLHRPKNVQTDVAVSGSFLIDEKKVKTR WNYDALTVALEPVSGSERVFVSQPFTIFPEKSAEEEGQRYLGIDIGEYGIAYTALEITGDS AKILDQNFISDPQLKTLREEVKGLKLDQRRGTFAMPSTKIARIRESLVHSLRNRIHHLALK 10 HKAKIVYELEVSRFEEGKQKIKKVYATLKKADVYSEIDADKNLQTTVWGKLAVASEISA SYTSQFCGACKKLWRAEMQVDETITTQELIGTVRVIKGGTLIDAIKDFMRPPIFDENDTPF PKYRDFCDKHHISKKMRGNSCLFICPFCRANADADIQASQTIALLRYVKEEKKVEDYFE RFRKLKN IKVLGQMKKI 15

[00335] The term “Cas 12b” or “Cas 12b domain” refers to an RNA-guided nuclease comprising a Casl2b / C2cl protein, or a fragment thereof (e.g., a protein comprising an active, inactive, or partially active DNA cleavage domain of Cas 12b, and / or the gRNA binding domain of Cas 12b). contents of each of which are incorporated herein by reference). Cas 12b orthologs have been described in various species, including, but not limited to, Alicyclobacillus 20 acidoterrestris, Alicyclobacillus acidophilus (Teng et al., Cell Discov. 2018 Nov 27;4:63), Bacillus hisashi, and Bacillus sp. V3-13. Additional suitable Cas 12b nucleases and sequences will be apparent to those of skill in the art based on this disclosure.

[00336] In some embodiments, proteins comprising Cas 12b or fragments thereof are referred to as “Casl2b variants.” A Casl2b variant shares homology to Casl2b, or a fragment thereof. 25 For example, a Casl2b variant is at least about 70% identical, at least about 80% identical, at least about 90% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, at least about 99% identical, at least about 99.5% identical, or at least about 99.9% identical to wild type Cas 12b. In some embodiments, the Cas 12b variant may have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 30   21, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or more amino acid changes compared to wild type Cas 12b. In some embodiments, the 2026204466   11 Jun 2026 Cas 12b variant comprises a fragment of Cas 12b (e.g., a gRNA binding domain or a DNA-cleavage domain), such that the fragment is at least about 70% identical, at least about 80% identical, at least about 90% identical, at least about 95% identical, at least about 96% identical, at least about 97% identical, at least about 98% identical, at least about 99% identical, at least 5 about 99.5% identical, or at least about 99.9% identical to the corresponding fragment of wild type Casl2b. In some embodiments, the fragment is at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% identical, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% of the amino acid length of a corresponding wild type 10 Casl2b. Exemplary Cas 12b polypeptides are listed below.

[00337] Casl2b / C2cl (uniprot.org / uniprot / T0D7A2#2)

[00338] sp|T0D7A2|C2Cl_ALIAG CRISPR-associated endo- nuclease C2cl OS = Alicyclobacillus acido- terrestris (strain ATCC 49025 / DSM 3922 / CIP 106132 / NCIMB 13137 / GD3B) GN=c2cl PE=1 SV=1 15 MAVKSIKVKLRLDDMPEIRAGLWKLHKEVNAGVRYYTEWLSLLRQENLYRRSPNGDG EQECDKTAEECKAELLERLRARQVENGHRGPAGSDDELLQLARQLYELLVPQAIGAKG DAQQIARKFLSPLADKDAVGGLGIAKAGNKPRWVRMREAGEPGWEEEKEKAETRKSA DRTADVLRALADFGLKPLMRVYTDSEMSSVEWKPLRKGQAVRTWDRDMFQQAIERM MSWESWNQRVGQEYAKLVEQKNRFEQKNFVGQEHLVHLVNQLQQDMKEASPGLESK 20 EQTAHYVTGRALRGSDKVFEKWGKLAPDAPFDLYDAEIKNVQRRNTRRFGSHDLFAKL AEPEYQALWREDASFLTRYAVYNSILRKLNHAKMFATFTLPDATAHPIWTRFDKLGGN LHQYTFLFNEFGERRHAIRFHKLLKVENGVAREVDDVTVPISMSEQLDNLLPRDPNEPIA LYFRDYGAEQHFTGEFGGAKIQCRRDQLAHMHRRRGARDVYLNVSVRVQSQSEARGE RRPPYAAVFRLVGDNHRAFVHFDKLSDYLAEHPDDGKLGSEGLLSGLRVMSVDLGLRT 25 SASISVFRVARKDELKPNSKGRVPFFFPIKGNDNLVAVHERSQLLKLPGETESKDLRAIRE ERQRTLRQLRTQLAYLRLLVRCGSEDVGRRERSWAKLIEQPVDAANHMTPDWREAFEN ELQKLKSLHGICSDKEWMDAVYESVRRVWRHMGKQVRDWRKDVRSGERPKIRGYAK DVVGGNSIEQIEYLERQYKFLKSWSFFGKVSGQVIRAEKGSRFAITLREHIDHAKEDRLK KLADRIIMEALGYVYALDERGKGKWVAKYPPCQLILLEELSEYQFNNDRPPSENNQLM 30 QWSHRGVFQELINQAQVHDLLVGTMYAAFSSRFDARTGAPGIRCRRVPARCTQEHNPE PFPWWLNKFVVEHTLDACPLRADDLIPTGEGEIFVSPFSAEEGDFHQIHADLNAAQNLQ 2026204466   11 Jun 2026 QRLWSDFDISQIRLRCDWGEVDGELVLIPRLTGKRTADSYSNKVFYTNTGVTYYERERG KKRRKVFAQEKLSEEEAELLVEADEAREKSVVLMRDPSGIINRGNWTRQKEFWSMV NQRIEGYLVKQIRSRVPLQDSACENTGDI

[00339] AacCasl2b (Alicyclobacillus acidiphilus) - WP_067623834 5 MAVKSMKVKLRLDNMPEIRAGLWKLHTEVNAGVRYYTEWLSLLRQENLYRRSPNGDG EQECYKTAEECKAELLERLRARQVENGHCGPAGSDDELLQLARQLYELLVPQAIGAKG DAQQIARKFLSPLADKDAVGGLGIAKAGNKPRWVRMREAGEPGWEEEKAKAEARKST DRTADVLRALADFGLKPLMRVYTDSDMSSVQWKPLRKGQAVRTWDRDMFQQAIERM MSWESWNQRVGEAYAKLVEQKSRFEQKNFVGQEHLVQLVNQLQQDMKEASHGLESK 10 EQTAHYLTGRALRGSDKVFEKWEKLDPDAPFDLYDTEIKNVQRRNTRRFGSHDLFAKL AEPKYQALWREDASFLTRYAVYNSIVRKLNHAKMFATFTLPDATAHPIWTRFDKLGGN LHQYTFLFNEFGEGRHAIRFQKLLTVEDGVAKEVDDVTVPISMSAQLDDLLPRDPHELV ALYFQDYGAEQHLAGEFGGAKIQYRRDQLNHLHARRGARDVYLNLSVRVQSQSEARG ERRPPYAAVFRLVGDNHRAFVHFDKLSDYLAEHPDDGKLGSEGLLSGLRVMSVDLGLR 15 TSASISVFRVARKDELKPNSEGRVPFCFPIEGNENLVAVHERSQLLKLPGETESKDLRAIR EERQRTLRQLRTQLAYLRLLVRCGSEDVGRRERSWAKLIEQPMDANQMTPDWREAFED ELQKLKSLYGICGDREWTEAVYESVRRVWRHMGKQVRDWRKDVRSGERPKIRGYQKD VVGGNSIEQIEYLERQYKFLKSWSFFGKVSGQVIRAEKGSRFAITLREHIDHAKEDRLKK LADRIIMEALGYVYALDDERGKGKWVAKYPPCQLILLEELSEYQFNNDRPPSENNQLM 20 QWSHRGVFQELLNQAQVHDLLVGTMYAAFSSRFDARTGAPGIRCRRVPARCAREQNPE PFPWWLNKFVAEHKLDGCPLRADDLIPTGEGEFFVSPFSAEEGDFHQIHADLNAAQNLQ RRLWSDFDISQIRLRCDWGEVDGEPVLIPRTTGKRTADSYGNKVFYTKTGVTYYERERG KKRRKVFAQEELSEEEAELLVEADEAREKSVVLMRDPSGIINRGDWTRQKEFWSMVNQ RIEGYLVKQIRSRVRLQESACENTGDI 25

[00340] BhCasl2b (Bacillus hisashii) NCBI Reference Sequence: WP 095142515 MAPKKKRKVGIHGVPAAATRSFILKIEPNEEVKKGLWKTHEVLNHGIAYYMNILKLIRQ EAIYEHHEQDPKNPKKVSKAEIQAELWDFVLKMQKCNSFTHEVDKDEVFNILRELYEEL VPSSVEKKGEANQLSNKFLYPLVDPNSQSGKGTASSGRKPRWYNLKIAGDPSWEEEKK 30 KWEEDKKKDPLAKILGKLAEYGLIPLFIPYTDSNEPIVKEIKWMEKSRNQSVRRLDKDM FIQALERFLSWESWNLKVKEEYEKVEKEYKTLEERIKEDIQALKALEQYEKERQEQLLR 2026204466   11 Jun 2026 DTLNTNEYRLSKRGLRGWREIIQKWLKMDENEPSEKYLEVFKDYQRKHPREAGDYSVY EFLSKKENHFIWRNHPEYPYLYATFCEIDKKKKDAKQQATFTLADPINHPLWVRFEERS GSNLNKYRILTEQLHTEKLKKKLTVQLDRLIYPTESGGWEEKGKVDIVLLPSRQFYNQIF LDIEEKGKHAFTYKDESIKFPLKGTLGGARVQFDRDHLRRYPHKVESGNVGRIYFNMTV 5 NIEPTESPVSKSLKIHRDDFPKVVNFKPKELTEWIKDSKGKKLKSGIESLEIGLRVMSIDL GQRQAAAASIFEVVDQKPDIEGKLFFPIKGTELYAVHRASFNIKLPGETLVKSREVLRKA REDNLKLMNQKLNFLRNVLHFQQFEDITEREKRVTKWISRQENSDVPLVYQDELIQIREL MYKPYKDWVAFLKQLHKRLEVEIGKEVKHWRKSLSDGRKGLYGISLKNIDEIDRTRKF LLRWSLRPTEPGEVRRLEPGQRFAIDQLNHLNALKEDRLKKMANTIIMHALGYCYDVR 10 KKKWQAKNPACQIILFEDLSNYNPYEERSRFENSKLMKWSRREIPRQVALQGEIYGLQV GEVGAQFSSRFHAKTGSPGIRCSVVTKEKLQDNRFFKNLQREGRLTLDKIAVLKEGDLY PDKGGEKFISLSKDRKCVTTHADINAAQNLQKRFWTRTHGFYKVYCKAYQVDGQTVYI PESKDQKQKIIEEFGEGYFILKDGVYE WVNAGKLKIKKGS SKQ S S SELVDSDILKD SFDL ASELKGEKLMLYRDPSGNVFPSDKWMAAGVFFGKLERILISKLTNQYSISTIEDDSSKQS 15 MKRPAATKKAGQAKKKK including the variant termed BvCasl2b V4 (S893R / K846R / E837G changes rel. to wt above)

[00341] BvCasl2b (Bacillus sp. V3-13) NCBI Reference Sequence: WP 101661451.1 MAIRSIKLKMKTNSGTDSIYLRKALWRTHQLINEGIAYYMNLLTLYRQEAIGDKTKEAY 20 QAELINIIRNQQRNNGSSEEHGSDQEILALLRQLYELIIPSSIGESGDANQLGNKFLYPLVD PNSQSGKGTSNAGRKPRWKRLKEEGNPDWELEKKKDEERKAKDPTVKIFDNLNKYGLL PLFPLFTNIQKDIEWLPLGKRQSVRKWDKDMFIQAIERLLSWESWNRRVADEYKQLKEK TESYYKEHLTGGEEWIEKIRKFEKERNMELEKNAFAPNDGYFITSRQIRGWDRVYEKWS KLPESASPEELWKVVAEQQNKMSEGFGDPKVFSFLANRENRDIWRGHSERIYHIAAYNG 25 LQKKLSRTKEQATFTLPDAIEHPLWIRYESPGGTNLNLFKLEEKQKKNYYVTLSKIIWPS EEKWIEKENIEIPLAPSIQFNRQIKLKQHVKGKQEISFSDYSSRISLDGVLGGSRIQFNRKYI KNHKELLGEGDIGPVFFNLVVDVAPLQETRNGRLQSPIGKALKVISSDFSKVIDYKPKEL MDWMNTGSASNSFGVASLLEGMRVMSIDMGQRTSASVSIFEVVKELPKDQEQKLFYSI NDTELFAIHKRSFLLNLPGEVVTKNNKQQRQERRKKRQFVRSQIRMLANVLRLETKKTP 30 DERKKAIHKLMEIVQSYDSWTASQKEVWEKELNLLTNMAAFNDEIWKESLVELHHRIE PYVGQIVSKWRKGLSEGRKNLAGISMWNIDELEDTRRLLISWSKRSRTPGEANRIETDEP 2026204466   11 Jun 2026 FGSSLLQHIQNVKDDRLKQMANLIIMTALGFKYDKEEKDRYKRWKETYPACQIILFENL NRYLFNLDRSRRENSRLMKWAHRSIPRTVSMQGEMFGLQVGDVRSEYSSRFHAKTGAP GIRCHALTEEDLKAGSNTLKRLIEDGFINESELAYLKKGDIIPSQGGELFVTLSKRYKKDS DNNELTVIHADINAAQNLQKRFWQQNSEVYRVPCQLARMGEDKLYIPKSQTETIKKYFG 5 KGSFVKNNTEQEVYKWEKSEKMKIKTDTTFDLQDLDGFEDISKTIELAQEQQKKYLTMF RDPSGYFFNNETWRPQKEYWSIVNNIIKSCLKKKILSNKVEL

[00342] By “Cbl proto-oncogene B (CBLB) polypeptide” is meant a protein having at least about 85% amino acid sequence identity to GenBank Accession No. ABC86700.1 or a fragment 10 thereof that is involved in the regulation of immune responses. An exemplary CBLB polypeptide sequence is provided below.

[00343] >ABC86700.1 CBL-B [Homo sapiens] MANSMNGRNPGGRGGNPRKGRILGIIDAIQDAVGPPKQAAADRRTVEKTWKLMDKVV RLCQNPKLQLKNSPPYILDILPDTYQHLRLILSKYDDNQKLAQLSENEYFKIYIDSLMKKS 15 KRAIRLFKEGKERMYEEQSQDRRNLTKLSLIFSHMLAEIKAIFPNGQFQGDNFRITKADA AEFWRKFFGDKTIVPWKVFRQCLHEVHQISSGLEAMALKSTIDLTCNDYISVFEFDIFTR LFQPWGSILRNWNFLAVTHPGYMAFLTYDEVKARLQKYSTKPGSYIFRLSCTRLGQWAI GYVTGDGNILQTIPHNKPLFQALIDGSREGFYLYPDGRSYNPDLTGLCEPTPHDHIKVTQ EQYELYCEMGSTFQLCKICAENDKDVKIEPCGHLMCTSCLTAWQESDGQGCPFCRCEIK 20 GTEPIIVDPFDPRDEGSRCCSIIDPFGMPMLDLDDDDDREESLMMNRLANVRKCTDRQN SPVTSPGSSPLAQRRKPQPDPLQIPHLSLPPVPPRLDLIQKGIVRSPCGSPTGSPKSSPCMV RKQDKPLPAPPPPLRDPPPPPPERPPPIPPDNRLSRHIHHVESVPSRDPPMPLEAWCPRDVF GTNQLVGCRLLGEGSPKPGITASSNVNGRHSRVGSDPVLMRKHRRHDLPLEGAKVFSN GHLGSEEYDVPPRLSPPPPVTTLLPSIKCTGPLANSLSEKTRDPVEEDDDEYKIPSSHPVSL 25 NSQPSHCHNVKPPVRSCDNGHCMLNGTHGPSSEKKSNIPDLSIYLKGDVFDSASDPVPLP PARPPTRDNPKHGSSLNRTPSDYDLLIPPLGEDAFDALPPSLPPPPPPARHSLIEHSKPPGSS SRPSSGQDLFLLPSDPFVDLASGQVPLPPARRLPGENVKTNRTSQDYDQLPSCSDGSQAP ARPPKPRPRRTAPEIHHRKPHGPEAALENVDAKIAKLMGEGYAFEEVKRALEIAQNNVE VARSILREFAFPPPVSPRLNL 30 By “Cbl proto-oncogene B (CBLB) polynucleotide” is meant a nucleic acid molecule encoding a CBLB polypeptide. The CBLB gene encodes an E3 ubiquitin ligase. An exemplary CBLB 2026204466   11 Jun 2026 nucleic acid sequence is provided below. Additional exemplary CBLB genomic sequences are indicated in NCBI Reference Sequence: NC_000003.12, or transcript reference NM_001321813.1. >DQ349203.1 Homo sapiens CBL-B mRNA, complete cds 5 ATGGCAAACTCAATGAATGGCAGAAACCCTGGTGGTCGAGGAGGAAATCCCCGAAA AGGTCGAATTTTGGGTATTATTGATGCTATTCAGGATGCAGTTGGACCCCCTAAGCA AGCTGCCGCAGATCGCAGGACCGTGGAGAAGACTTGGAAGCTCATGGACAAAGTGG TAAGACTGTGCCAAAATCCCAAACTTCAGTTGAAAAATAGCCCACCATATATACTTG ATATTTTGCCTGATACATATCAGCATTTACGACTTATATTGAGTAAATATGATGACA 10 ACCAGAAACTTGCCCAACTCAGTGAGAATGAGTACTTTAAAATCTACATTGATAGCC TTATGAAAAAGTCAAAACGGGCAATAAGACTCTTTAAAGAAGGCAAGGAGAGAATG TATGAAGAACAGTCACAGGACAGACGAAATCTCACAAAACTGTCCCTTATCTTCAGT CACATGCTGGCAGAAATCAAAGCAATCTTTCCCAATGGTCAATTCCAGGGAGATAA CTTTCGTATCACAAAAGCAGATGCTGCTGAATTCTGGAGAAAGTTTTTTGGAGACAA 15 AACTATCGTACCATGGAAAGTATTCAGACAGTGCCTTCATGAGGTCCACCAGATTAG CTCTGGCCTGGAAGCAATGGCTCTAAAATCAACAATTGATTTAACTTGCAATGATTA CATTTCAGTTTTTGAATTTGATATTTTTACCAGGCTGTTTCAGCCTTGGGGCTCTATTT TGCGGAATTGGAATTTCTTAGCTGTGACACATCCAGGTTACATGGCATTTCTCACAT ATGATGAAGTTAAAGCACGACTACAGAAATATAGCACCAAACCCGGAAGCTATATT 20 TTCCGGTTAAGTTGCACTCGATTGGGACAGTGGGCCATTGGCTATGTGACTGGGGAT GGGAATATCTTACAGACCATACCTCATAACAAGCCCTTATTTCAAGCCCTGATTGAT GGCAGCAGGGAAGGATTTTATCTTTATCCTGATGGGAGGAGTTATAATCCTGATTTA ACTGGATTATGTGAACCTACACCTCATGACCATATAAAAGTTACACAGGAACAATAT GAATTATATTGTGAAATGGGCTCCACTTTTCAGCTCTGTAAGATTTGTGCAGAGAAT 25 GACAAAGATGTCAAGATTGAGCCTTGTGGGCATTTGATGTGCACCTCTTGCCTTACG GCATGGCAGGAGTCGGATGGTCAGGGCTGCCCTTTCTGTCGTTGTGAAATAAAAGG AACTGAGCCCATAATCGTGGACCCCTTTGATCCAAGAGATGAAGGCTCCAGGTGTTG CAGCATCATTGACCCCTTTGGCATGCCGATGCTAGACTTGGACGACGATGATGATCG TGAGGAGTCCTTGATGATGAATCGGTTGGCAAACGTCCGAAAGTGCACTGACAGGC 30 AGAACTCACCAGTCACATCACCAGGATCCTCTCCCCTTGCCCAGAGAAGAAAGCCA CAGCCTGACCCACTCCAGATCCCACATCTAAGCCTGCCACCCGTGCCTCCTCGCCTG 2026204466   11 Jun 2026 GATCTAATTCAGAAAGGCATAGTTAGATCTCCCTGTGGCAGCCCAACGGGTTCACCA AAGTCTTCTCCTTGCATGGTGAGAAAACAAGATAAACCACTCCCAGCACCACCTCCT CCCTTAAGAGATCCTCCTCCACCGCCACCTGAAAGACCTCCACCAATCCCACCAGAC AATAGACTGAGTAGACACATCCATCATGTGGAAAGCGTGCCTTCCAGAGACCCGCC 5 AATGCCTCTTGAAGCATGGTGCCCTCGGGATGTGTTTGGGACTAATCAGCTTGTGGG ATGTCGACTCCTAGGGGAGGGCTCTCCAAAACCTGGAATCACAGCGAGTTCAAATG TCAATGGAAGGCACAGTAGAGTGGGCTCTGACCCAGTGCTTATGCGGAAACACAGA CGCCATGATTTGCCTTTAGAAGGAGCTAAGGTCTTTTCCAATGGTCACCTTGGAAGT GAAGAATATGATGTTCCTCCCCGGCTTTCTCCTCCTCCTCCAGTTACCACCCTCCTCC 10 CTAGCATAAAGTGTACTGGTCCGTTAGCAAATTCTCTTTCAGAGAAAACAAGAGACC CAGTAGAGGAAGATGATGATGAATACAAGATTCCTTCATCCCACCCTGTTTCCCTGA ATTCACAACCATCTCATTGTCATAATGTAAAACCTCCTGTTCGGTCTTGTGATAATGG TCACTGTATGCTGAATGGAACACATGGTCCATCTTCAGAGAAGAAATCAAACATCCC TGACTTAAGCATATATTTAAAGGGAGATGTTTTTGATTCAGCCTCTGATCCCGTGCC 15 ATTACCACCTGCCAGGCCTCCAACTCGGGACAATCCAAAGCATGGTTCTTCACTCAA CAGGACGCCCTCTGATTATGATCTTCTCATCCCTCCATTAGGTGAAGATGCTTTTGAT GCCCTCCCTCCATCTCTCCCACCTCCCCCACCTCCTGCAAGGCATAGTCTCATTGAAC ATTCAAAACCTCCTGGCTCCAGTAGCCGGCCATCCTCAGGACAGGATCTTTTTCTTCT TCCTTCAGATCCCTTTGTTGATCTAGCAAGTGGCCAAGTTCCTTTGCCTCCTGCTAGA 20 AGGTTACCAGGTGAAAATGTCAAAACTAACAGAACATCACAGGACTATGATCAGCT TCCTTCATGTTCAGATGGTTCACAGGCACCAGCCAGACCCCCTAAACCACGACCGCG CAGGACTGCACCAGAAATTCACCACAGAAAACCCCATGGGCCTGAGGCGGCATTGG AAAATGTCGATGCAAAAATTGCAAAACTCATGGGAGAGGGTTATGCCTTTGAAGAG GTGAAGAGAGCCTTAGAGATAGCCCAGAATAATGTCGAAGTTGCCCGGAGCATCCT 25 CCGAGAATTTGCCTTCCCTCCTCCAGTATCCCCACGTCTAAATCTATAG

[00344] By “chimeric antigen receptor” is meant a synthetic receptor comprising an extracellular antigen binding domain, a transmembrane domain, and an intracellular signaling domain that confers specificity for an antigen onto an immune cell.

[00345] In this disclosure, “comprises,” “comprising,” “containing” and “having” and the like 30 can have the meaning ascribed to them in U.S. Patent law and can mean “ includes,” “including,” and the like; “consisting essentially of’ or “consists essentially” likewise has the meaning 2026204466   11 Jun 2026 ascribed in U.S. Patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than that which is recited, but excludes prior art embodiments.

[00346] By “cluster of differentiation 2 (CD2)” is meant a protein having at least about 85% 5 amino acid sequence identity to NCBI Accession No. NP001315538.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_001315538.1 T-cell surface antigen CD2 isoform 1 precursor [Homo sapiens] MSFPCKFVASFLLIFNVSSKGAVSKEITNALETWGALGQDINLDIPSFQMSDDIDDIKWE KTSDKKKIAQFRKEKETFKEKDTYKLFKNGTLKIKHLKTDDQDIYKVSIYDTKGKNVLE 10 KIFDLKIQERVSKPKIS WTCINTTLTCEVMNGTDPELNLYQDGKHLKLSQRVITHKWTTS LSAKFKCTAGNKVSKESSVEPVSCPGGSILGQSNGLSAWTPPSHPTSLPFAEKGLDIYLII GICGGGSLLMVFVALLVFYITKRKKQRSRRNDEELETRAHRVATEERGRKPHQIPASTPQ NPATSQHPPPPPGHRSQAPSHRPPPPGHRVQHQPQKRPPAPSGTQVHQQKGPPLPRPRVQ PKPPHGAAENSLSPSSN 15

[00347] By “cluster of differentiation 2 (CD2)” is meant a nucleic acid encoding a CD2 polypeptide. An exemplary CD2 nucleic acid sequence is provided below. >NM_001328609.2 Homo sapiens CD2 molecule (CD2), transcript variant 1, mRNA AGTCTCACTTCAGTTCCTTTTGCATGAAGAGCTCAGAATCAAAAGAGGAAACCAACC CCTAAGATGAGCTTTCCATGTAAATTTGTAGCCAGCTTCCTTCTGATTTTCAATGTTT 20 CTTCCAAAGGTGCAGTCTCCAAAGAGATTACGAATGCCTTGGAAACCTGGGGTGCCT TGGGTCAGGACATCAACTTGGACATTCCTAGTTTTCAAATGAGTGATGATATTGACG ATATAAAATGGGAAAAAACTTCAGACAAGAAAAAGATTGCACAATTCAGAAAAGA GAAAGAGACTTTCAAGGAAAAAGATACATATAAGCTATTTAAAAATGGAACTCTGA AAATTAAGCATCTGAAGACCGATGATCAGGATATCTACAAGGTATCAATATATGAT 25 ACAAAAGGAAAAAATGTGTTGGAAAAAATATTTGATTTGAAGATTCAAGAGAGGGT CTCAAAACCAAAGATCTCCTGGACTTGTATCAACACAACCCTGACCTGTGAGGTAAT GAATGGAACTGACCCCGAATTAAACCTGTATCAAGATGGGAAACATCTAAAACTTT CTCAGAGGGTCATCACACACAAGTGGACCACCAGCCTGAGTGCAAAATTCAAGTGC ACAGCAGGGAACAAAGTCAGCAAGGAATCCAGTGTCGAGCCTGTCAGCTGTCCAGG 30 AGGCAGCATCCTTGGCCAGAGTAATGGGCTCTCTGCCTGGACCCCTCCCAGCCATCC CACTTCTCTTCCTTTTGCAGAGAAAGGTCTGGACATCTATCTCATCATTGGCATATGT 2026204466   11 Jun 2026 GGAGGAGGCAGCCTCTTGATGGTCTTTGTGGCACTGCTCGTTTTCTATATCACCAAA AGGAAAAAACAGAGGAGTCGGAGAAATGATGAGGAGCTGGAGACAAGAGCCCACA GAGTAGCTACTGAAGAAAGGGGCCGGAAGCCCCACCAAATTCCAGCTTCAACCCCT CAGAATCCAGCAACTTCCCAACATCCTCCTCCACCACCTGGTCATCGTTCCCAGGCA 5 CCTAGTCATCGTCCCCCGCCTCCTGGACACCGTGTTCAGCACCAGCCTCAGAAGAGG CCTCCTGCTCCGTCGGGCACACAAGTTCACCAGCAGAAAGGCCCGCCCCTCCCCAGA CCTCGAGTTCAGCCAAAACCTCCCCATGGGGCAGCAGAAAACTCATTGTCCCCTTCC TCTAATTAAAAAAGATAGAAACTGTCTTTTTCAATAAAAAGCACTGTGGATTTCTGC CCTCCTGATGTGCATATCCGTACTTCCATGAGGTGTTTTCTGTGTGCAGAACATTGTC 10 ACCTCCTGAGGCTGTGGGCCACAGCCACCTCTGCATCTTCGAACTCAGCCATGTGGT CAACATCTGGAGTTTTTGGTCTCCTCAGAGAGCTCCATCACACCAGTAAGGAGAAGC AATATAAGTGTGATTGCAAGAATGGTAGAGGACCGAGCACAGAAATCTTAGAGATT TCTTGTCCCCTCTCAGGTCATGTGTAGATGCGATAAATCAAGTGATTGGTGTGCCTG GGTCTCACTACAAGCAGCCTATCTGCTTAAGAGACTCTGGAGTTTCTTATGTGCCCT 15 GGTGGACACTTGCCCACCATCCTGTGAGTAAAAGTGAAATAAAAGCTTTGACTAGA

[00348] By “cluster of differentiation 3 epsilon (CD3e or CD3 epsilon)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_000724.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. 20   >NP_000724.1 T-cell surface glycoprotein CD3 epsilon chain precursor [Homo sapiens] MQSGTHWRVLGLCLLSVGVWGQDGNEEMGGITQTPYKVSISGTTVILTCPQYPGSEILW QHNDKNIGGDEDDKNIGSDEDHLSLKEFSELEQSGYYVCYPRGSKPEDANFYLYLRARV CENCMEMDVMSVATIVIVDICITGGLLLLVYYWSKNRKAKAKPVTRGAGAGGRQRGQ NKERPPPVPNPDYEPIRKGQRDLYSGLNQRRI 25

[00349] By “cluster of differentiation 3 epsilon (CD3e or CD3 epsilon)” is meant a nucleic acid encoding a CD3e polypeptide. An exemplary CD3e nucleic acid sequence is provided below. >NM_000733.4 Homo sapiens CD3e molecule (CD3E), mRNA AGAAACCCTCCTCCCCTCCCAGCCTCAGGTGCCTGCTTCAGAAAATGAAGTAGTAAG 30 TCTGCTGGCCTCCGCCATCTTAGTAAAGTAACAGTCCCATGAAACAAAGATGCAGTC GGGCACTCACTGGAGAGTTCTGGGCCTCTGCCTCTTATCAGTTGGCGTTTGGGGGCA 2026204466   11 Jun 2026 AGATGGTAATGAAGAAATGGGTGGTATTACACAGACACCATATAAAGTCTCCATCT CTGGAACCACAGTAATATTGACATGCCCTCAGTATCCTGGATCTGAAATACTATGGC AACACAATGATAAAAACATAGGCGGTGATGAGGATGATAAAAACATAGGCAGTGAT GAGGATCACCTGTCACTGAAGGAATTTTCAGAATTGGAGCAAAGTGGTTATTATGTC 5 TGCTACCCCAGAGGAAGCAAACCAGAAGATGCGAACTTTTATCTCTACCTGAGGGC AAGAGTGTGTGAGAACTGCATGGAGATGGATGTGATGTCGGTGGCCACAATTGTCA TAGTGGACATCTGCATCACTGGGGGCTTGCTGCTGCTGGTTTACTACTGGAGCAAGA ATAGAAAGGCCAAGGCCAAGCCTGTGACACGAGGAGCGGGTGCTGGCGGCAGGCA AAGGGGACAAAACAAGGAGAGGCCACCACCTGTTCCCAACCCAGACTATGAGCCCA 10 TCCGGAAAGGCCAGCGGGACCTGTATTCTGGCCTGAATCAGAGACGCATCTGACCC TCTGGAGAACACTGCCTCCCGCTGGCCCAGGTCTCCTCTCCAGTCCCCCTGCGACTC CCTGTTTCCTGGGCTAGTCTTGGACCCCACGAGAGAGAATCGTTCCTCAGCCTCATG GTGAACTCGCGCCCTCCAGCCTGATCCCCCGCTCCCTCCTCCCTGCCTTCTCTGCTGG TACCCAGTCCTAAAATATTGCTGCTTCCTCTTCCTTTGAAGCATCATCAGTAGTCACA 15 CCCTCACAGCTGGCCTGCCCTCTTGCCAGGATATTTATTTGTGCTATTCACTCCCTTC CCTTTGGATGTAACTTCTCCGTTCAGTTCCCTCCTTTTCTTGCATGTAAGTTGTCCCCC ATCCCAAAGTATTCCATCTACTTTTCTATCGCCGTCCCCTTTTGCAGCCCTCTCTGGG GATGGACTGGGTAAATGTTGACAGAGGCCCTGCCCCGTTCACAGATCCTGGCCCTGA GCCAGCCCTGTGCTCCTCCCTCCCCCAACACTCCCTACCAACCCCCTAATCCCCTACT 20 CCCTCCACCCCCCCTCCACTGTAGGCCACTGGATGGTCATTTGCATCTCCGTAAATGT GCTCTGCTCCTCAGCTGAGAGAGAAAAAAATAAACTGTATTTGGCTGCAA

[00350] By “cluster of differentiation 3 gamma (CD3g or CD3 gamma) is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_000064.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is 25 provided below. >NP_000064.1 T-cell surface glycoprotein CD3 gamma chain precursor [Homo sapiens] MEQGKGLAVLILAIILLQGTLAQSIKGNHLVKVYDYQEDGSVLLTCDAEAKNITWFKDG KMIGFLTEDKKKWNLGSNAKDPRGMYQCKGSQNKSKPLQVYYRMCQNCIELNAATIS GFLFAEIVSIFVLAVGVYFIAGQDGVRQSRASDKQTLLPNDQLYQPLKDREDDQYSHLQ 30 GNQLRRN 2026204466   11 Jun 2026

[00351] By “cluster of differentiation 3 gamma (CD3g or CD3 gamma)” is meant a nucleic acid encoding a CD3g polypeptide. An exemplary CD3g nucleic acid sequence is provided below. >NM_000073.3 Homo sapiens CD3g molecule (CD3G), mRNA 5 AGTCTAGCTGCTGCACAGGCTGGCTGGCTGGCTGGCTGCTAAGGGCTGCTCCACGCT TTTGCCGGAGGACAGAGACTGACATGGAACAGGGGAAGGGCCTGGCTGTCCTCATC CTGGCTATCATTCTTCTTCAAGGTACTTTGGCCCAGTCAATCAAAGGAAACCACTTG GTTAAGGTGTATGACTATCAAGAAGATGGTTCGGTACTTCTGACTTGTGATGCAGAA GCCAAAAATATCACATGGTTTAAAGATGGGAAGATGATCGGCTTCCTAACTGAAGA 10 TAAAAAAAAATGGAATCTGGGAAGTAATGCCAAGGACCCTCGAGGGATGTATCAGT GTAAAGGATCACAGAACAAGTCAAAACCACTCCAAGTGTATTACAGAATGTGTCAG AACTGCATTGAACTAAATGCAGCCACCATATCTGGCTTTCTCTTTGCTGAAATCGTC AGCATTTTCGTCCTTGCTGTTGGGGTCTACTTCATTGCTGGACAGGATGGAGTTCGCC AGTCGAGAGCTTCAGACAAGCAGACTCTGTTGCCCAATGACCAGCTCTACCAGCCCC 15 TCAAGGATCGAGAAGATGACCAGTACAGCCACCTTCAAGGAAACCAGTTGAGGAGG AATTGAACTCAGGACTCAGAGTAGTCCAGGTGTTCTCCTCCTATTCAGTTCCCAGAA TCAAAGCAATGCATTTTGGAAAGCTCCTAGCAGAGAGACTTTCAGCCCTAAATCTAG ACTCAAGGTTCCCAGAGATGACAAATGGAGAAGAAAGGCCATCAGAGCAAATTTGG GGGTTTCTCAAATAAAATAAAAATAAAAACAAATACTGTGTTTCAGAAGCGCCACC 20 TATTGGGGAAAATTGTAAAAGAAAAATGAAAAGATCAAATAACCCCCTGGATTTGA ATATAATTTTTTGTGTTGTAATTTTTATTTCGTTTTTGTATAGGTTATAATTCACATGG CTCAAATATTCAGTGAAAGCTCTCCCTCCACCGCCATCCCCTGCTACCCAGTGACCC TGTTGCCCTCTTCAGAGACAAATTAGTTTCTCTTTTTTTTTTTTTTTTTTTTTTTTTTGA GACAGTCTGGCTCTGTCACCCAGGCTGAAATGCAGTGGCACCATCTCGGCTCACTGC 25 AACCTCTGCCTCCTGGGTTCAAGCGATTCTCCTGCCTCAGCCTCCCGGGCAGCTGGG ATTACAGGCACACACTACCACACCTGGCTAATTTTTGTATTTTTAGTAGAGACAGGG TTTTGCTCTGTTGGCCAAGCTGGTCTCGAACTCCTGACCTCAAGTGATCCGCCCGCCT CAGCCTCCCAAAGTGCTGGGATTACAGGTGTGAGCCACCATGCCTGGTCTTAAAACC AGTTTCTTATATATCTCTCTGGAGGTATTCTAGGCATATATGAGCACATTCTCAAGTA 30 CATATTATCCTCCCTTCCCCTATCTTTTAGACAAATGATATCAAACTATACATCTTGT GAGATTATTGCATACCATTATATGAAGATACCATTATATCCTTTTTAATGCAACCATA 2026204466   11 Jun 2026 TTGTACAAATAGACTATGATTTATTTAACCTGTTATCTATCAGTGGATATTTAAGTTG GTAGTTGGTTCCAATCTTTTGCTCTTACAACAATTCTGCAATGACTAACATTGTATAA ATATCATTTTTAAAAATAATTGCATTGAAGCATAATGTACATGCCATAAAATCCACC CATCTTAAGTGATTTCACCTGTTCTCAGAAATTTTTAGTAAATTTAACTAATTGTACA 5 GCCATTACCATAATCCAGCTTTAGGACATTTTCTTTTTTTTCTTTTCTTTTCTTTTTTTT CTTTTTTTTTTTTTTTTGAAGTGGAATCTTGCTCTGTGGCCCAGGCTGGAGTGCAGTG GCGCGATCTCAGCTCACTGCAACCTCCACCTCCTGGGTTCAAGCGATTCTCTTGCCTT GGCCTCCCGAGTAGCTGAGACTACAGGCACATGCCACCACGCCCAGCTCATTTTTTG TGTATTTAGTATTTGTGTATCTAGTATTTGTGTACTTAGTAGAGACAGGGTTTCACCA 10 TGTTGGCCAGGCTGGTCTCCAATTCCTGACCTCAGGCGATCCACCCGCCTTGACCTC CCAAAGTGCTGGGATTACAGGTGTGAGCCACCGCGCCAGGCCCGTAACTGTATTTTA ATATAGCCATTCTATGGATTTAATATGGTATTTTATTATGGCCTTAATTTGCATTTCC CTAGATACTAACCATGCTGAGTGTCCTGTCTTGTGTTTATTAACCATTCATATATTTT TAGTGAAATGTGTATCAAATCTTTTGCCCATTTTTAAGTTGACTTATTTGTTTGTCTTC 15 TTACTATTGGGTTGCATATGTTTTTGATATAAGTCCTTTATCAGATATATGATTTGGA AATATTTTCTACCAATCTGTGGTTTGTTTTTCTTAATGGTGTCTTTTGAAGTGCAAAA GGTTTGAATTTTGAAGTACATTTTATTGATTTTTTCTTCTATATATTGTGCTTTTGGTA TCATGTCTAATAAATCTTTACCAAACCCACAGTTACAAAGATTTTCTCCTGTCTTCTT TTTATACTTTTTACAGCTTTATGGTTTTAGCTCTAACAATAAATGTGATTTTGAACAT 20 ACATAAGACTATTTGTAACAAACACAAATAAATTGAATTGTTGGGCA

[00352] By “cluster of differentiation 3 delta (CD3d or CD3 delta) is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_000723.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. 25   >NP_000723.1 T-cell surface glycoprotein CD3 delta chain isoform A precursor [Homo sapiens] MEHSTFLSGLVLATLLSQVSPFKIPIEELEDRVFVNCNTSITWVEGTVGTLLSDITRLDLG KRILDPRGIYRCNGTDIYKDKESTVQVHYRMCQSCVELDPATVAGIIVTDVIATLLLALG VFCFAGHETGRLSGAADTQALLRNDQVYQPLRDRDDAQYSHLGGNWARNK

[00353] By “cluster of differentiation 3 delta (CD3d or CD3 delta)” is meant a nucleic acid 30 encoding a CD3d polypeptide. An exemplary CD3d nucleic acid sequence is provided below. >NM_000732.4 Homo sapiens CD3d molecule (CD3D), transcript variant 1, mRNA 2026204466   11 Jun 2026 AGAGAAGCAGACATCTTCTAGTTCCTCCCCCACTCTCCTCTTTCCGGTACCTGTGAGT CAGCTAGGGGAGGGCAGCTCTCACCCAGGCTGATAGTTCGGTGACCTGGCTTTATCT ACTGGATGAGTTCCGCTGGGAGATGGAACATAGCACGTTTCTCTCTGGCCTGGTACT GGCTACCCTTCTCTCGCAAGTGAGCCCCTTCAAGATACCTATAGAGGAACTTGAGGA 5 CAGAGTGTTTGTGAATTGCAATACCAGCATCACATGGGTAGAGGGAACGGTGGGAA CACTGCTCTCAGACATTACAAGACTGGACCTGGGAAAACGCATCCTGGACCCACGA GGAATATATAGGTGTAATGGGACAGATATATACAAGGACAAAGAATCTACCGTGCA AGTTCATTATCGAATGTGCCAGAGCTGTGTGGAGCTGGATCCAGCCACCGTGGCTGG CATCATTGTCACTGATGTCATTGCCACTCTGCTCCTTGCTTTGGGAGTCTTCTGCTTT 10 GCTGGACATGAGACTGGAAGGCTGTCTGGGGCTGCCGACACACAAGCTCTGTTGAG GAATGACCAGGTCTATCAGCCCCTCCGAGATCGAGATGATGCTCAGTACAGCCACCT TGGAGGAAACTGGGCTCGGAACAAGTGAACCTGAGACTGGTGGCTTCTAGAAGCAG CCATTACCAACTGTACCTTCCCTTCTTGCTCAGCCAATAAATATATCCTCTTTCACTC AGAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA 15

[00354] By “cluster of differentiation 4 (CD4)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_000607.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_000607.1 T-cell surface glycoprotein CD4 isoform 1 precursor [Homo sapiens] MNRGVPFRHLLLVLQLALLPAATQGKKVVLGKKGDTVELTCTASQKKSIQFHWKNSNQ 20 IKILGNQGSFLTKGPSKLNDRADSRRSLWDQGNFPLIIKNLKIEDSDTYICEVEDQKEEVQ LLVFGLTANSDTHLLQGQSLTLTLESPPGSSPSVQCRSPRGKNIQGGKTLSVSQLELQDS GTWTCTVLQNQKKVEFKIDIVVLAFQKASSIVYKKEGEQVEFSFPLAFTVEKLTGSGEL WWQAERASSSKSWITFDLKNKEVSVKRVTQDPKLQMGKKLPLHLTLPQALPQYAGSG NLTLALEAKTGKLHQEVNLWMRATQLQKNLTCEVWGPTSPKLMLSLKLENKEAKVS 25 KREKAVWVLNPEAGMWQCLLSDSGQVLLESNIKVLPTWSTPVQPMALIVLGGVAGLLL FIGLGIFFCVRCRHRRRQAERMSQIKRLLSEKKTCQCPHRFQKTCSPI

[00355] By “cluster of differentiation 4 (CD4)” is meant a nucleic acid encoding a CD4 polypeptide. An exemplary CD4 nucleic acid sequence is provided below. >NM_000616.5 Homo sapiens CD4 molecule (CD4), transcript variant 1, mRNA 30 CTCTCTTCATTTAAGCACGACTCTGCAGAAGGAACAAAGCACCCTCCCCACTGGGCT CCTGGTTGCAGAGCTCCAAGTCCTCACACAGATACGCCTGTTTGAGAAGCAGCGGG 2026204466   11 Jun 2026 CAAGAAAGACGCAAGCCCAGAGGCCCTGCCATTTCTGTGGGCTCAGGTCCCTACTG GCTCAGGCCCCTGCCTCCCTCGGCAAGGCCACAATGAACCGGGGAGTCCCTTTTAGG CACTTGCTTCTGGTGCTGCAACTGGCGCTCCTCCCAGCAGCCACTCAGGGAAAGAAA GTGGTGCTGGGCAAAAAAGGGGATACAGTGGAACTGACCTGTACAGCTTCCCAGAA 5 GAAGAGCATACAATTCCACTGGAAAAACTCCAACCAGATAAAGATTCTGGGAAATC AGGGCTCCTTCTTAACTAAAGGTCCATCCAAGCTGAATGATCGCGCTGACTCAAGAA GAAGCCTTTGGGACCAAGGAAACTTTCCCCTGATCATCAAGAATCTTAAGATAGAA GACTCAGATACTTACATCTGTGAAGTGGAGGACCAGAAGGAGGAGGTGCAATTGCT AGTGTTCGGATTGACTGCCAACTCTGACACCCACCTGCTTCAGGGGCAGAGCCTGAC 10 CCTGACCTTGGAGAGCCCCCCTGGTAGTAGCCCCTCAGTGCAATGTAGGAGTCCAAG GGGTAAAAACATACAGGGGGGGAAGACCCTCTCCGTGTCTCAGCTGGAGCTCCAGG ATAGTGGCACCTGGACATGCACTGTCTTGCAGAACCAGAAGAAGGTGGAGTTCAAA ATAGACATCGTGGTGCTAGCTTTCCAGAAGGCCTCCAGCATAGTCTATAAGAAAGA GGGGGAACAGGTGGAGTTCTCCTTCCCACTCGCCTTTACAGTTGAAAAGCTGACGGG 15 CAGTGGCGAGCTGTGGTGGCAGGCGGAGAGGGCTTCCTCCTCCAAGTCTTGGATCAC CTTTGACCTGAAGAACAAGGAAGTGTCTGTAAAACGGGTTACCCAGGACCCTAAGC TCCAGATGGGCAAGAAGCTCCCGCTCCACCTCACCCTGCCCCAGGCCTTGCCTCAGT ATGCTGGCTCTGGAAACCTCACCCTGGCCCTTGAAGCGAAAACAGGAAAGTTGCAT CAGGAAGTGAACCTGGTGGTGATGAGAGCCACTCAGCTCCAGAAAAATTTGACCTG 20 TGAGGTGTGGGGACCCACCTCCCCTAAGCTGATGCTGAGTTTGAAACTGGAGAACA AGGAGGCAAAGGTCTCGAAGCGGGAGAAGGCGGTGTGGGTGCTGAACCCTGAGGC GGGGATGTGGCAGTGTCTGCTGAGTGACTCGGGACAGGTCCTGCTGGAATCCAACA TCAAGGTTCTGCCCACATGGTCCACCCCGGTGCAGCCAATGGCCCTGATTGTGCTGG GGGGCGTCGCCGGCCTCCTGCTTTTCATTGGGCTAGGCATCTTCTTCTGTGTCAGGTG 25 CCGGCACCGAAGGCGCCAAGCAGAGCGGATGTCTCAGATCAAGAGACTCCTCAGTG AGAAGAAGACCTGCCAGTGTCCTCACCGGTTTCAGAAGACATGTAGCCCCATTTGAG GCACGAGGCCAGGCAGATCCCACTTGCAGCCTCCCCAGGTGTCTGCCCCGCGTTTCC TGCCTGCGGACCAGATGAATGTAGCAGATCCCCAGCCTCTGGCCTCCTGTTCGCCTC CTCTACAATTTGCCATTGTTTCTCCTGGGTTAGGCCCCGGCTTCACTGGTTGAGTGTT 30 GCTCTCTAGTTTCCAGAGGCTTAATCACACCGTCCTCCACGCCATTTCCTTTTCCTTC AAGCCTAGCCCTTCTCTCATTATTTCTCTCTGACCCTCTCCCCACTGCTCATTTGGAT 2026204466   11 Jun 2026 CCCAGGGGAGTGTTCAGGGCCAGCCCTGGCTGGCATGGAGGGTGAGGCTGGGTGTC TGGAAGCATGGAGCATGGGACTGTTCTTTTACAAGACAGGACCCTGGGACCACAGA GGGCAGGAACTTGCACAAAATCACACAGCCAAGCCAGTCAAGGATGGATGCAGATC CAGAGGTTTCTGGCAGCCAGTACCTCCTGCCCCATGCTGCCCGCTTCTCACCCTATGT 5 GGGTGGGACCACAGACTCACATCCTGACCTTGCACAAACAGCCCCTCTGGACACAG CCCCATGTACACGGCCTCAAGGGATGTCTCACATCCTCTGTCTATTTGAGACTTAGA AAAATCCTACAAGGCTGGCAGTGACAGAACTAAGATGATCATCTCCAGTTTATAGA CCAGAACCAGAGCTCAGAGAGGCTAGATGATTGATTACCAAGTGCCGGACTAGCAA GTGCTGGAGTCGGGACTAACCCAGGTCCCTTGTCCCAAGTTCCACTGCTGCCTCTTG 10 AATGCAGGGACAAATGCCACACGGCTCTCACCAGTGGCTAGTGGTGGGTACTCAAT GTGTACTTTTGGGTTCACAGAAGCACAGCACCCATGGGAAGGGTCCATCTCAGAGA ATTTACGAGCAGGGATGAAGGCCTCCCTGTCTAAAATCCCTCCTTCATCCCCCGCTG GTGGCAGAATCTGTTACCAGAGGACAAAGCCTTTGGCTCTTCTAATCAGAGCGCAAG CTGGGAGCACAGGCACTGCAGGAGAGAATGCCCAGTGACCAGTCACTGACCCTGTG 15 CAGAACCTCCTGGAAGCGAGCTTTGCTGGGAGAGGGGGTAGCTAGCCTGAGAGGGA ACCCTCTAAGGGACCTCAAAGGTGATTGTGCCAGGCTCTGCGCCTGCCCCACACCCT CCCTTACCCTCCTCCAGACCATTCAGGACACAGGGAAATCAGGGTTACAAATCTTCT TGATCCACTTCTCTCAGGATCCCCTCTCTTCCTACCCTTCCTCACCACTTCCCTCAGTC CCAACTCCTTTTCCCTATTTCCTTCTCCTCCTGTCTTTAAAGCCTGCCTCTTCCAGGAA 20 GACCCCCCTATTGCTGCTGGGGCTCCCCATTTGCTTACTTTGCATTTGTGCCCACTCT CCACCCCTGCTCCCCTGAGCTGAAATAAAAATACAATAAACTTAC

[00356] By “cluster of differentiation 5 (CD5)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_001333385.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. 25   >NP_001333385.1 T-cell surface glycoprotein CD5 isoform 2 [Homo sapiens] MVCSQSWGRSSKQWEDPSQASKVCQRLNCGVPLSLGPFLVTYTPQSSIICYGQLGSFSN CSHSRNDMCHSLGLTCLEPQKTTPPTTRPPPTTTPEPTAPPRLQLVAQSGGQHCAGVVEF YSGSLGGTISYEAQDKTQDLENFLCNNLQCGSFLKHLPETEAGRAQDPGEPREHQPLPIQ WKIQNSSCTSLEHCFRKIKPQKSGRVLALLCSGFQPKVQSRLVGGSSICEGTVEVRQGAQ 30 WAALCDSSSARSSLRWEEVCREQQCGSVNSYRVLDAGDPTSRGLFCPHQKLSQCHELW ERNSYCKKVFVTCQDPNPAGLAAGTVASIILALVLLVVLLVVCGPLAYKKLVKKFRQK 2026204466   11 Jun 2026 KQRQWIGPTGMNQNMSFHRNHTATVRSHAENPTASHVDNEYSQPPRNSHLSAYPALEG ALHRS SMQPDNS SD SDYDLHGAQRL

[00357] By “cluster of differentiation 5 (CD5)” is meant a nucleic acid encoding a CD5 polypeptide. An exemplary CD5 nucleic acid sequence is provided below. >NM_001346456.1 5 Homo sapiens CD5 molecule (CD5), transcript variant 2, mRNA GAGTCTTGCTGATGCTCCCGGCTGAATAAACCCCTTCCTTCTTTAACTTGGTGTCTGA GGGGTTTTGTCTGTGGCTTGTCCTGCTACATTTCTTGGTTCCCTGACCAGGAAGCAAA GTGATTAACGGACAGTTGAGGCAGCCCCTTAGGCAGCTTAGGCCTGCCTTGTGGAGC ATCCCCGCGGGGAACTCTGGCCAGCTTGAGCGACACGGATCCTCAGAGCGCTCCCA 10 GGTAGGCAATTGCCCCAGTGGAATGCCTCGTCAGAGCAGTGCATGGCAGGCCCCTG TGGAGGATCAACGCAGTGGCTGAACACAGGGAAGGAACTGGCACTTGGAGTCCGGA CAACTGAAACTTGTCGCTTCCTGCCTCGGACGGCTCAGCTGGTATGACCCAGATTTC CAGGCAAGGCTCACCCGTTCCAACTCGAAGTGCCAGGGCCAGCTGGAGGTCTACCT CAAGGACGGATGGCACATGGTTTGCAGCCAGAGCTGGGGCCGGAGCTCCAAGCAGT 15 GGGAGGACCCCAGTCAAGCGTCAAAAGTCTGCCAGCGGCTGAACTGTGGGGTGCCC TTAAGCCTTGGCCCCTTCCTTGTCACCTACACACCTCAGAGCTCAATCATCTGCTACG GACAACTGGGCTCCTTCTCCAACTGCAGCCACAGCAGAAATGACATGTGTCACTCTC TGGGCCTGACCTGCTTAGAACCCCAGAAGACAACACCTCCAACGACAAGGCCCCCG CCCACCACAACTCCAGAGCCCACAGCTCCTCCCAGGCTGCAGCTGGTGGCACAGTCT 20 GGCGGCCAGCACTGTGCCGGCGTGGTGGAGTTCTACAGCGGCAGCCTGGGGGGTAC CATCAGCTATGAGGCCCAGGACAAGACCCAGGACCTGGAGAACTTCCTCTGCAACA ACCTCCAGTGTGGCTCCTTCTTGAAGCATCTGCCAGAGACTGAGGCAGGCAGAGCCC AAGACCCAGGGGAGCCACGGGAACACCAGCCCTTGCCAATCCAATGGAAGATCCAG AACTCAAGCTGTACCTCCCTGGAGCATTGCTTCAGGAAAATCAAGCCCCAGAAAAG 25 TGGCCGAGTTCTTGCCCTCCTTTGCTCAGGTTTCCAGCCCAAGGTGCAGAGCCGTCT GGTGGGGGGCAGCAGCATCTGTGAAGGCACCGTGGAGGTGCGCCAGGGGGCTCAGT GGGCAGCCCTGTGTGACAGCTCTTCAGCCAGGAGCTCGCTGCGGTGGGAGGAGGTG TGCCGGGAGCAGCAGTGTGGCAGCGTCAACTCCTATCGAGTGCTGGACGCTGGTGA CCCAACATCCCGGGGGCTCTTCTGTCCCCATCAGAAGCTGTCCCAGTGCCACGAACT 30 TTGGGAGAGAAATTCCTACTGCAAGAAGGTGTTTGTCACATGCCAGGATCCAAACCC CGCAGGCCTGGCCGCAGGCACGGTGGCAAGCATCATCCTGGCCCTGGTGCTCCTGGT 2026204466   11 Jun 2026 GGTGCTGCTGGTCGTGTGCGGCCCCCTTGCCTACAAGAAGCTAGTGAAGAAATTCCG CCAGAAGAAGCAGCGCCAGTGGATTGGCCCAACGGGAATGAACCAAAACATGTCTT TCCATCGCAACCACACGGCAACCGTCCGATCCCATGCTGAGAACCCCACAGCCTCCC ACGTGGATAACGAATACAGCCAACCTCCCAGGAACTCCCACCTGTCAGCTTATCCAG 5 CTCTGGAAGGGGCTCTGCATCGCTCCTCCATGCAGCCTGACAACTCCTCCGACAGTG ACTATGATCTGCATGGGGCTCAGAGGCTGTAAAGAACTGGGATCCATGAGCAAAAA GCCGAGAGCCAGACCTGTTTGTCCTGAGAAAACTGTCCGCTCTTCACTTGAAATCAT GTCCCTATTTCTACCCCGGCCAGAACATGGACAGAGGCCAGAAGCCTTCCGGACAG GCGCTGCTGCCCCGAGTGGCAGGCCAGCTCACACTCTGCTGCACAACAGCTCGGCC 10 GCCCCTCCACTTGTGGAAGCTGTGGTGGGCAGAGCCCCAAAACAAGCAGCCTTCCA ACTAGAGACTCGGGGGTGTCTGAAGGGGGCCCCCTTTCCCTGCCCGCTGGGGAGCG GCGTCTCAGTGAAATCGGCTTTCTCCTCAGACTCTGTCCCTGGTAAGGAGTGACAAG GAAGCTCACAGCTGGGCGAGTGCATTTTGAATAGTTTTTTGTAAGTAGTGCTTTTCCT CCTTCCTGACAAATCGAGCGCTTTGGCCTCTTCTGTGCAGCATCCACCCCTGCGGAT 15 CCCTCTGGGG AGGACAGGAAGGGGACTCCCGGAGACCTCTGCAGCCGTGGTGGTCA GAGGCTGCTCACCTGAGCACAAAGACAGCTCTGCACATTCACCGCAGCTGCCAGCC AGGGGTCTGGGTGGGCACCACCCTGACCCACAGCGTCACCCCACTCCCTCTGTCTTA TGACTCCCCTCCCCAACCCCCTCATCTAAAGACACCTTCCTTTCCACTGGCTGTCAAG CCCACAGGGCACCAGTGCCACCCAGGGCCCGGCACAAAGGGGCGCCTAGTAAACCT 20 TAACCAACTTGGTTTTTTGCTTCACCCAGCAATTAAAAGTCCCAAGCTGAGGTAGTT TCAGTCCATCACAGTTCATCTTCTAACCCAAGAGTCAGAGATGGGGCTGGTCATGTT CCTTTGGTTTGAATAACTCCCTTGACGAAAACAGACTCCTCTAGTACTTGGAGATCTT GGACGTACACCTAATCCCATGGGGCCTCGGCTTCCTTAACTGCAAGTGAGAAGAGG AGGTCTACCCAGGAGCCTCGGGTCTGATCAAGGGAGAGGCCAGGCGCAGCTCACTG 25 CGGCGGCTCCCTAAGAAGGTGAAGCAACATGGGAACACATCCTAAGACAGGTCCTT TCTCCACGCCATTTGATGCTGTATCTCCTGGGAGCACAGGCATCAATGGTCCAAGCC GCATAATAAGTCTGGAAGAGCAAAAGGGAGTTACTAGGATATGGGGTGGGCTGCTC CCAGAATCTGCTCAGCTTTCTGCCCCCACCAACACCCTCCAACCAGGCCTTGCCTTCT GAGAGCCCCCGTGGCCAAGCCCAGGTCACAGATCTTCCCCCGACCATGCTGGGAAT 30 CCAGAAACAGGGACCCCATTTGTCTTCCCATATCTGGTGGAGGTGAGGGGGCTCCTC AAAAGGGAACTGAGAGGCTGCTCTTAGGGAGGGCAAAGGTTCGGGGGCAGCCAGT 2026204466   11 Jun 2026 GTCTCCCATCAGTGCCTTTTTTAATAAAAGCTCTTTCATCTATAGTTTGGCCACCATA CAGTGGCCTCAAAGCAACCATGGCCTACTTAAAAACCAAACCAAAAATAAAGAGTT TAGTTGAGGAGAAAAAAAAAAAAAAAAAAAAAAAAA

[00358] By “cluster of differentiation 7 (CD7)” is meant a protein having at least about 85% 5 amino acid sequence identity to NCBI Accession No. NP_006128.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_006128.1 T-cell antigen CD7 precursor [Homo sapiens] MAGPPRLLLLPLLLALARGLPGALAAQEVQQSPHCTTVPVGASVNITCSTSGGLRGIYLR QLGPQPQDIIYYEDGVVPTTDRRFRGRIDFSGSQDNLTITMHRLQLSDTGTYTCQAITEV 10 NVYGSGTLVLVTEEQSQGWHRCSDAPPRASALPAPPTGSALPDPQTASALPDPPAASAL PAALAVISFLLGLGLGVACVLARTQIKKLCSWRDKNSAACVVYEDMSHSRCNTLSSPNQ YQ

[00359] By “cluster of differentiation 7 (CD7)” is meant a nucleic acid encoding a CD7 polypeptide. An exemplary CD7 nucleic acid sequence is provided below. 15   >NM_006137.7 Homo sapiens CD7 molecule (CD7), mRNA CTCTCTGAGCTCTGAGCGCCTGCGGTCTCCTGTGTGCTGCTCTCTGTGGGGTCCTGTA GACCCAGAGAGGCTCAGCTGCACTCGCCCGGCTGGGAGAGCTGGGTGTGGGGAACA TGGCCGGGCCTCCGAGGCTCCTGCTGCTGCCCCTGCTTCTGGCGCTGGCTCGCGGCC TGCCTGGGGCCCTGGCTGCCCAAGAGGTGCAGCAGTCTCCCCACTGCACGACTGTCC 20 CCGTGGGAGCCTCCGTCAACATCACCTGCTCCACCAGCGGGGGCCTGCGTGGGATCT ACCTGAGGCAGCTCGGGCCACAGCCCCAAGACATCATTTACTACGAGGACGGGGTG GTGCCCACTACGGACAGACGGTTCCGGGGCCGCATCGACTTCTCAGGGTCCCAGGA CAACCTGACTATCACCATGCACCGCCTGCAGCTGTCGGACACTGGCACCTACACCTG CCAGGCCATCACGGAGGTCAATGTCTACGGCTCCGGCACCCTGGTCCTGGTGACAG 25 AGGAACAGTCCCAAGGATGGCACAGATGCTCGGACGCCCCACCAAGGGCCTCTGCC CTCCCTGCCCCACCGACAGGCTCCGCCCTCCCTGACCCGCAGACAGCCTCTGCCCTC CCTGACCCGCCAGCAGCCTCTGCCCTCCCTGCGGCCCTGGCGGTGATCTCCTTCCTCC TCGGGCTGGGCCTGGGGGTGGCGTGTGTGCTGGCGAGGACACAGATAAAGAAACTG TGCTCGTGGCGGGATAAGAATTCGGCGGCATGTGTGGTGTACGAGGACATGTCGCA 30 CAGCCGCTGCAACACGCTGTCCTCCCCCAACCAGTACCAGTGACCCAGTGGGCCCCT GCACGTCCCGCCTGTGGTCCCCCCAGCACCTTCCCTGCCCCACCATGCCCCCCACCC 2026204466   11 Jun 2026 TGCCACACCCCTCACCCTGCTGTCCTCCCACGGCTGCAGCAGAGTTTGAAGGGCCCA GCCGTGCCCAGCTCCAAGCAGACACACAGGCAGTGGCCAGGCCCCACGGTGCTTCT CAGTGGACAATGATGCCTCCTCCGGGAAGCCTTCCCTGCCCAGCCCACGCCGCCACC GGGAGGAAGCCTGACTGTCCTTTGGCTGCATCTCCCGACCATGGCCAAGGAGGGCTT 5 TTCTGTGGGATGGGCCTGGGCACGCGGCCCTCTCCTGTCAGTGCCGGCCCACCCACC AGCAGGCCCCCAACCCCCAGGCAGCCCGGCAGAGGACGGGAGGAGACCAGTCCCC CACCCAGCCGTACCAGAAATAAAGGCTTCTGTGCTTCC

[00360] By “cluster of differentiation 30 (CD30)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_001234.3 or fragment thereof and 10 having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_001234.3 tumor necrosis factor receptor superfamily member 8 isoform 1 precursor [Homo sapiens] MRVLLAALGLLFLGALRAFPQDRPFEDTCHGNPSHYYDKAVRRCCYRCPMGLFPTQQC PQRPTDCRKQCEPDYYLDEADRCTACVTCSRDDLVEKTPCAWNSSRVCECRPGMFCST 15 SAVNSCARCFFHSVCPAGMIVKFPGTAQKNTVCEPASPGVSPACASPENCKEPSSGTIPQ AKPTPVSPATSSASTMPVRGGTRLAQEAASKLTRAPDSPSSVGRPSSDPGLSPTQPCPEGS GDCRKQCEPDYYLDEAGRCTACVSCSRDDLVEKTPCAWNSSRTCECRPGMICATSATN SCARCVPYPICAAETVTKPQDMAEKDTTFEAPPLGTQPDCNPTPENGEAPASTSPTQSLL VDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSSAFLLCHRRACRKR 20 IRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSV GAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPE GRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK

[00361] By “cluster of differentiation 30 (CD30)” is meant a nucleic acid encoding a CD30 polypeptide. An exemplary CD30 nucleic acid sequence is provided below. >NM_001243.5 25 Homo sapiens TNF receptor superfamily member 8 (TNFRSF8), transcript variant 1, mRNA CTGAGTCATCTCTGCACGTGTTTGCCCCCTTTTTTCTTCGCTGCTTGTAGCTAAGTGTT CCTGGAACCAATTTGATACGGGAGAACTAAGGCTGAAACCTCGGAGGAACAACCAC TTTTGAAGTGACTTCGCGGCGTGCGTTGGGTGCGGACTAGGTGGCCGCGGCGGGAGT GTGCTGGAGCCTGAAGTCCACGCGCGCGGCTGAGAACCGCCGGGACCGCACGTGGG 30 CGCCGCGCGCTTCCCCCGCTTCCCAGGTGGGCGCCGGCCGCCAGGCCACCTCACGTC CGGCCCCGGGGATGCGCGTCCTCCTCGCCGCGCTGGGACTGCTGTTCCTGGGGGCGC 2026204466   11 Jun 2026 TACGAGCCTTCCCACAGGATCGACCCTTCGAGGACACCTGTCATGGAAACCCCAGCC ACTACTATGACAAGGCTGTCAGGAGGTGCTGTTACCGCTGCCCCATGGGGCTGTTCC CGACACAGCAGTGCCCACAGAGGCCTACTGACTGCAGGAAGCAGTGTGAGCCTGAC TACTACCTGGATGAGGCCGACCGCTGTACAGCCTGCGTGACTTGTTCTCGAGACGAC 5 CTCGTGGAGAAGACGCCGTGTGCATGGAACTCCTCCCGTGTCTGCGAATGTCGACCC GGCATGTTCTGTTCCACGTCTGCCGTCAACTCCTGTGCCCGCTGCTTCTTCCATTCTG TCTGTCCGGCAGGGATGATTGTCAAGTTCCCAGGCACGGCGCAGAAGAACACGGTC TGTGAGCCGGCTTCCCCAGGGGTCAGCCCTGCCTGTGCCAGCCCAGAGAACTGCAA GGAACCCTCCAGTGGCACCATCCCCCAGGCCAAGCCCACCCCGGTGTCCCCAGCAA 10 CCTCCAGTGCCAGCACCATGCCTGTAAGAGGGGGCACCCGCCTCGCCCAGGAAGCT GCTTCTAAACTGACGAGGGCTCCCGACTCTCCCTCCTCTGTGGGAAGGCCTAGTTCA GATCCAGGTCTGTCCCCAACACAGCCATGCCCAGAGGGGTCTGGTGATTGCAGAAA GCAGTGTGAGCCCGACTACTACCTGGACGAGGCCGGCCGCTGCACGGCCTGCGTGA GCTGTTCTCGAGATGACCTTGTGGAGAAGACGCCATGTGCATGGAACTCCTCCCGCA 15 CCTGCGAATGTCGACCTGGCATGATCTGTGCCACATCAGCCACCAACTCCTGTGCCC GCTGTGTCCCCTACCCAATCTGTGCAGCAGAGACGGTCACCAAGCCCCAGGATATG GCTGAGAAGGACACCACCTTTGAGGCGCCACCCCTGGGGACCCAGCCGGACTGCAA CCCCACCCCAGAGAATGGCGAGGCGCCTGCCAGCACCAGCCCCACTCAGAGCTTGC TGGTGGACTCCCAGGCCAGTAAGACGCTGCCCATCCCAACCAGCGCTCCCGTCGCTC 20 TCTCCTCCACGGGGAAGCCCGTTCTGGATGCAGGGCCAGTGCTCTTCTGGGTGATCC TGGTGTTGGTTGTGGTGGTCGGCTCCAGCGCCTTCCTCCTGTGCCACCGGAGGGCCT GCAGGAAGCGAATTCGGCAGAAGCTCCACCTGTGCTACCCGGTCCAGACCTCCCAG CCCAAGCTAGAGCTTGTGGATTCCAGACCCAGGAGGAGCTCAACGCAGCTGAGGAG TGGTGCGTCGGTGACAGAACCCGTCGCGGAAGAGCGAGGGTTAATGAGCCAGCCAC 25 TGATGGAGACCTGCCACAGCGTGGGGGCAGCCTACCTGGAGAGCCTGCCGCTGCAG GATGCCAGCCCGGCCGGGGGCCCCTCGTCCCCCAGGGACCTTCCTGAGCCCCGGGT GTCCACGGAGCACACCAATAACAAGATTGAGAAAATCTACATCATGAAGGCTGACA CCGTGATCGTGGGGACCGTGAAGGCTGAGCTGCCGGAGGGCCGGGGCCTGGCGGGG CCAGCAGAGCCCGAGTTGGAGGAGGAGCTGGAGGCGGACCATACCCCCCACTACCC 30 CGAGCAGGAGACAGAACCGCCTCTGGGCAGCTGCAGCGATGTCATGCTCTCAGTGG AAGAGGAAGGGAAAGAAGACCCCTTGCCCACAGCTGCCTCTGGAAAGTGAGGCCTG 2026204466   11 Jun 2026 GGCTGGGCTGGGGCTAGGAGGGCAGCAGGGTGGCCTCTGGGAGGCCAGGATGGCAC TGTTGGCACCGAGGTTGGGGGCAGAGGCCCATCTGGCCTGAACTGAGGCTCCAGCA TCTAGTGGTGGACCGGCCGGTCACTGCAGGGGTCTGGTGGTCTCTGCTTGCATCCCC AACTTAGCTGTCCCCTGACCCAGAGCCTAGGGGATCCGGGGCTTGTACAGAAGAGA 5 CAGTCCAAGGGGACTGGATCCCAGCAGTGATGTTGGTTGAGGCAGCAAACAGATGG CAGGATGGGCACTGCCGAGAACAGCATTGGTCCCAGAGCCCTGGGCATCAGACCTT AACCACCAGGCCCACAGCCCAGCGAGGGAGAGGTCGTGAGGCCAGCTCCCGGGGCC CCTGTAACCCTACTCTCCTCTCTCCCTGGACCTCAGAGGTGACACCCATTGGGCCCTT CCGGCATGCCCCCAGTTACTGTAAATGTGGCCCCCAGTGGGCATGGAGCCAGTGCCT 10 GTGGTTGTTTCTCCAGAGTCAAAAGGGAAGTCGAGGGATGGGGCGTCGTCAGCTGG CACTGTCTCTGCTGCAGCGGCCACACTGTACTCTGCACTGGTGTGAGGGCCCCTGCC TGGACTGTGGGACCCTCCTGGTGCTGCCCACCTTCCCTGTCCTGTAGCCCCCTCGGTG GGCCCAGGGCCTAGGGCCCAGGATCAAGTCACTCATCTCAGAATGTCCCCACCAAT CCCCGCCACAGCAGGCGCCTCGGGTCCCAGATGTCTGCAGCCCTCAGCAGCTGCAG 15 ACCGCCCCTCACCAACCCAGAGAACCTGCTTTACTTTGCCCAGGGACTTCCTCCCCA TGTGAACATGGGGAACTTCGGGCCCTGCCTGGAGTCCTTGACCGCTCTCTGTGGGCC CCACCCACTCTGTCCTGGGAAATGAAGAAGCATCTTCCTTAGGTCTGCCCTGCTTGC AAATCCACTAGCACCGACCCCACCACCTGGTTCCGGCTCTGCACGCTTTGGGGTGTG GATGTCGAGAGGCACCACGGCCTCACCCAGGCATCTGCTTTACTCTGGACCATAGGA 20 AACAAGACCGTTTGGAGGTTTCATCAGGATTTTGGGTTTTTCACATTTCACGCTAAG GAGTAGTGGCCCTGACTTCCGGTCGGCTGGCCAGCTGACTCCCTAGGGCCTTCAGAC GTGTATGCAAATGAGTGATGGATAAGGATGAGTCTTGGAGTTGCGGGCAGCCTGGA GACTCGTGGACTTACCGCCTGGAGGCAGGCCCGGGAAGGCTGCTGTTTACTCATCGG GCAGCCACGTGCTCTCTGGAGGAAGTGATAGTTTCTGAAACCGCTCAGATGTTTTGG 25 GGAAAGTTGGAGAAGCCGTGGCCTTGCGAGAGGTGGTTACACCAGAACCTGGACAT TGGCCAGAAGAAGCTTAAGTGGGCAGACACTGTTTGCCCAGTGTTTGTGCAAGGAT GGAGTGGGTGTCTCTGCATCACCCACAGCCGCAGCTGTAAGGCACGCTGGAAGGCA CACGCCTGCCAGGCAGGGCAGTCTGGCGCCCATGATGGGAGGGATTGACATGTTTC AACAAAATAATGCACTTCCTTACCTAGTGGCCCTTCACACAACTTTTGAATCTCTAA 30 AAATCCATAAAATCCTTAAAGAACTGTAA 2026204466   11 Jun 2026

[00362] By “cluster of differentiation 33 (CD33)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_001763.3 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_001763.3 myeloid cell surface antigen CD33 isoform 1 precursor [Homo sapiens] 5 MPLLLLLPLLWAGALAMDPNFWLQVQESVTVQEGLCVLVPCTFFHPIPYYDKNSPVHG YWFREGAIISRDSPVATNKLDQEVQEETQGRFRLLGDPSRNNCSLSIVDARRRDNGSYFF RMERGSTKYSYKSPQLSVHVTDLTHRPKILIPGTLEPGHSKNLTCSVSWACEQGTPPIFS WLSAAPTSLGPRTTHSSVLIITPRPQDHGTNLTCQVKFAGAGVTTERTIQLNVTYVPQNP TTGIFPGDGSGKQETRAGVVHGAIGGAGVTALLALCLCLIFFIVKTHRRKAARTAVGRN 10 DTHPTTGSASPKHQKKSKLHGPTETSSCSGAAPTVEMDEELHYASLNFHGMNPSKDTST EYSEVRTQ

[00363] By “cluster of differentiation 33 (CD33)” is meant a nucleic acid encoding a CD33 polypeptide. An exemplary CD33 nucleic acid sequence is provided below. >NM_001772.4 Homo sapiens CD33 molecule (CD33), transcript variant 1, mRNA 15 CTGCTCACACAGGAAGCCCTGGAAGCTGCTTCCTCAGACATGCCGCTGCTGCTACTG CTGCCCCTGCTGTGGGCAGGGGCCCTGGCTATGGATCCAAATTTCTGGCTGCAAGTG CAGGAGTCAGTGACGGTACAGGAGGGTTTGTGCGTCCTCGTGCCCTGCACTTTCTTC CATCCCATACCCTACTACGACAAGAACTCCCCAGTTCATGGTTACTGGTTCCGGGAA GGAGCCATTATATCCAGGGACTCTCCAGTGGCCACAAACAAGCTAGATCAAGAAGT 20 ACAGGAGGAGACTCAGGGCAGATTCCGCCTCCTTGGGGATCCCAGTAGGAACAACT GCTCCCTGAGCATCGTAGACGCCAGGAGGAGGGATAATGGTTCATACTTCTTTCGGA TGGAGAGAGGAAGTACCAAATACAGTTACAAATCTCCCCAGCTCTCTGTGCATGTGA CAGACTTGACCCACAGGCCCAAAATCCTCATCCCTGGCACTCTAGAACCCGGCCACT CCAAAAACCTGACCTGCTCTGTGTCCTGGGCCTGTGAGCAGGGAACACCCCCGATCT 25 TCTCCTGGTTGTCAGCTGCCCCCACCTCCCTGGGCCCCAGGACTACTCACTCCTCGGT GCTCATAATCACCCCACGGCCCCAGGACCACGGCACCAACCTGACCTGTCAGGTGA AGTTCGCTGGAGCTGGTGTGACTACGGAGAGAACCATCCAGCTCAACGTCACCTATG TTCCACAGAACCCAACAACTGGTATCTTTCCAGGAGATGGCTCAGGGAAACAAGAG ACCAGAGCAGGAGTGGTTCATGGGGCCATTGGAGGAGCTGGTGTTACAGCCCTGCT 30 CGCTCTTTGTCTCTGCCTCATCTTCTTCATAGTGAAGACCCACAGGAGGAAAGCAGC CAGGACAGCAGTGGGCAGGAATGACACCCACCCTACCACAGGGTCAGCCTCCCCGA 2026204466   11 Jun 2026 AACACCAGAAGAAGTCCAAGTTACATGGCCCCACTGAAACCTCAAGCTGTTCAGGT GCCGCCCCTACTGTGGAGATGGATGAGGAGCTGCATTATGCTTCCCTCAACTTTCAT GGGATGAATCCTTCCAAGGACACCTCCACCGAATACTCAGAGGTCAGGACCCAGTG AGGAACCCACAAGAGCATCAGGCTCAGCTAGAAGATCCACATCCTCTACAGGTCGG 5 GGACCAAAGGCTGATTCTTGGAGATTTAACACCCCACAGGCAATGGGTTTATAGAC ATTATGTGAGTTTCCTGCTATATTAACATCATCTTAGACTTTGCAAGCAGAGAGTCGT GGAATCAAATCTGTGCTCTTTCATTTGCTAAGTGTATGATGTCACACAAGCTCCTTAA CCTTCCATGTCTCCATTTTCTTCTCTGTGAAGTAGGTATAAGAAGTCCTATCTCATAG GGATGCTGTGAGCATTAAATAAAGGTACACATGGAAAACACCA 10

[00364] By “cluster of differentiation 52 (CD52)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_001794.2 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_001794.2 CAMPATH-1 antigen precursor [Homo sapiens]

[00365] MKRFLFLLLTISLLVMVQIQTGLSGQNDTSQTSSPSASSNISGGIFLFFVANAIIH 15 LFCFS

[00366] By “cluster of differentiation 52 (CD52)” is meant a nucleic acid encoding a CD52 polypeptide. An exemplary CD52 nucleic acid sequence is provided below. >NM_001803.3 Homo sapiens CD52 molecule (CD52), mRNA AGACAGCCCTGAGATCACCTAAAAAGCTGCTACCAAGACAGCCACGAAGATCCTAC 20 CAAAATGAAGCGCTTCCTCTTCCTCCTACTCACCATCAGCCTCCTGGTTATGGTACA GATACAAACTGGACTCTCAGGACAAAACGACACCAGCCAAACCAGCAGCCCCTCAG CATCCAGCAACATAAGCGGAGGCATTTTCCTTTTCTTCGTGGCCAATGCCATAATCC ACCTCTTCTGCTTCAGTTGAGGTGACACGTCTCAGCCTTAGCCCTGTGCCCCCTGAA ACAGCTGCCACCATCACTCGCAAGAGAATCCCCTCCATCTTTGGGAGGGGTTGATGC 25 CAGACATCACCAGGTTGTAGAAGTTGACAGGCAGTGCCATGGGGGCAACAGCCAAA ATAGGGGGGTAATGATGTAGGGGCCAAGCAGTGCCCAGCTGGGGGTCAATAAAGTT ACCCTTGTACTTGCA

[00367] By “cluster of differentiation 70 (CD70)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP_001243.1 or fragment thereof and 30 having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_001243.1 CD70 antigen isoform 1 [Homo sapiens] 2026204466   11 Jun 2026 MPEEGSGCSVRRRPYGCVLRAALVPLVAGLVICLVVCIQRFAQAQQQLPLESLGWDVA ELQLNHTGPQQDPRLYWQGGPALGRSFLHGPELDKGQLRIHRDGIYMVHIQVTLAICSS TTASRHHPTTLAVGICSPASRSISLLRLSFHQGCTIASQRLTPLARGDTLCTNLTGTLLPSR NTDETFFGVQWVRP 5

[00368] By “cluster of differentiation 70 (CD70)” is meant a nucleic acid encoding a CD70 polypeptide. An exemplary CD70 nucleic acid sequence is provided below. >NM_001252.5 Homo sapiens CD70 molecule (CD70), transcript variant 1, mRNA AGAGAGGGGCAGGCTGGTCCCCTGACAGGTTGAAGCAAGTAGACGCCCAGGAGCCC CGGGAGGGGGCTGCAGTTTCCTTCCTTCCTTCTCGGCAGCGCTCCGCGCCCCCATCG 10 CCCCTCCTGCGCTAGCGGAGGTGATCGCCGCGGCGATGCCGGAGGAGGGTTCGGGC TGCTCGGTGCGGCGCAGGCCCTATGGGTGCGTCCTGCGGGCTGCTTTGGTCCCATTG GTCGCGGGCTTGGTGATCTGCCTCGTGGTGTGCATCCAGCGCTTCGCACAGGCTCAG CAGCAGCTGCCGCTCGAGTCACTTGGGTGGGACGTAGCTGAGCTGCAGCTGAATCA CACAGGACCTCAGCAGGACCCCAGGCTATACTGGCAGGGGGGCCCAGCACTGGGCC 15 GCTCCTTCCTGCATGGACCAGAGCTGGACAAGGGGCAGCTACGTATCCATCGTGATG GCATCTACATGGTACACATCCAGGTGACGCTGGCCATCTGCTCCTCCACGACGGCCT CCAGGCACCACCCCACCACCCTGGCCGTGGGAATCTGCTCTCCCGCCTCCCGTAGCA TCAGCCTGCTGCGTCTCAGCTTCCACCAAGGTTGTACCATTGCCTCCCAGCGCCTGA CGCCCCTGGCCCGAGGGGACACACTCTGCACCAACCTCACTGGGACACTTTTGCCTT 20 CCCGAAACACTGATGAGACCTTCTTTGGAGTGCAGTGGGTGCGCCCCTGACCACTGC TGCTGATTAGGGTTTTTTAAATTTTATTTTATTTTATTTAAGTTCAAGAGAAAAAGTG TACACACAGGGGCCACCCGGGGTTGGGGTGGGAGTGTGGTGGGGGGTAGTGGTGGC AGGACAAGAGAAGGCATTGAGCTTTTTCTTTCATTTTCCTATTAAAAAATACAAAAA TCA 25

[00369] By “class II, major histocompatibility complex, transactivator (CIITA)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NP 001273331.1 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. >NP_001273331.1 MHC class II transactivator isoform 1 [Homo sapiens] 30 MRCLAPRPAGSYLSEPQGSSQCATMELGPLEGGYLELLNSDADPLCLYHFYDQMDLAG EEEIELYSEPDTDTINCDQFSRLLCDMEGDEETREAYANIAELDQYVFQDSQLEGLSKDIF 2026204466   11 Jun 2026 IEHIGPDEVIGESMEMPAEVGQKSQKRPFPEELPADLKHWKPAEPPTVVTGSLLVGPVSD CSTLPCLPLPALFNQEPASGQMRLEKTDQIPMPFSSSSLSCLNLPEGPIQFVPTISTLPHGL WQISEAGTGVSSIFIYHGEVPQASQVPPPSGFTVHGLPTSPDRPGSTSPFAPSATDLPSMPE PALTSRANMTEHKTSPTQCPAAGEVSNKLPKWPEPVEQFYRSLQDTYGAEPAGPDGILV 5 EVDLVQARLERSSSKSLERELATPDWAERQLAQGGLAEVLLAAKEHRRPRETRVIAVLG KAGQGKSYWAGAVSRAWACGRLPQYDFVFSVPCHCLNRPGDAYGLQDLLFSLGPQPL VAADEVFSHILKRPDRVLLILDGFEELEAQDGFLHSTCGPAPAEPCSLRGLLAGLFQKKL LRGCTLLLTARPRGRLVQSLSKADALFELSGFSMEQAQAYVMRYFESSGMTEHQDRAL TLLRDRPLLLSHSHSPTLCRAVCQLSEALLELGEDAKLPSTLTGLYVGLLGRAALDSPPG 10 ALAELAKLAWELGRRHQSTLQEDQFPSADVRTWAMAKGLVQHPPRAAESELAFPSFLL QCFLGALWLALSGEIKDKELPQYLALTPRKKRPYDNWLEGVPRFLAGLIFQPPARCLGA LLGPSAAASVDRKQKVLARYLKRLQPGTLRARQLLELLHCAHEAEEAGIWQHVVQELP GRLSFLGTRLTPPDAHVLGKALEAAGQDFSLDLRSTGICPSGLGSLVGLSCVTRFRAALS DTVALWESLQQHGETKLLQAAEEKFTIEPFKAKSLKDVEDLGKLVQTQRTRSSSEDTAG 15 ELPAVRDLKKLEFALGPVSGPQAFPKLVRILTAFSSLQHLDLDALSENKIGDEGVSQLSA TFPQLKSLETLNLSQNNITDLGAYKLAEALPSLAASLLRLSLYNNCICDVGAESLARVLP DMVSLRVMDVQYNKFTAAGAQQLAASLRRCPHVETLAMWTPTIPFSVQEHLQQQDSRI SLR

[00370] By “class II, major histocompatibility complex, transactivator (CIITA)” is meant a 20 nucleic acid encoding a CIITA polypeptide. An exemplary CIITA nucleic acid sequence is provided below. >NM_001286402.1 Homo sapiens class II major histocompatibility complex transactivator (CIITA), transcript variant 1, mRNA GGTTAGTGATGAGGCTAGTGATGAGGCTGTGTGCTTCTGAGCTGGGCATCCGAAGGC 25 ATCCTTGGGGAAGCTGAGGGCACGAGGAGGGGCTGCCAGACTCCGGGAGCTGCTGC CTGGCTGGGATTCCTACACAATGCGTTGCCTGGCTCCACGCCCTGCTGGGTCCTACC TGTCAGAGCCCCAAGGCAGCTCACAGTGTGCCACCATGGAGTTGGGGCCCCTAGAA GGTGGCTACCTGGAGCTTCTTAACAGCGATGCTGACCCCCTGTGCCTCTACCACTTC TATGACCAGATGGACCTGGCTGGAGAAGAAGAGATTGAGCTCTACTCAGAACCCGA 30 CACAGACACCATCAACTGCGACCAGTTCAGCAGGCTGTTGTGTGACATGGAAGGTG ATGAAGAGACCAGGGAGGCTTATGCCAATATCGCGGAACTGGACCAGTATGTCTTC 2026204466   11 Jun 2026 CAGGACTCCCAGCTGGAGGGCCTGAGCAAGGACATTTTCATAGAGCACATAGGACC AGATGAAGTGATCGGTGAGAGTATGGAGATGCCAGCAGAAGTTGGGCAGAAAAGTC AGAAAAGACCCTTCCCAGAGGAGCTTCCGGCAGACCTGAAGCACTGGAAGCCAGCT GAGCCCCCCACTGTGGTGACTGGCAGTCTCCTAGTGGGACCAGTGAGCGACTGCTCC 5 ACCCTGCCCTGCCTGCCACTGCCTGCGCTGTTCAACCAGGAGCCAGCCTCCGGCCAG ATGCGCCTGGAGAAAACCGACCAGATTCCCATGCCTTTCTCCAGTTCCTCGTTGAGC TGCCTGAATCTCCCTGAGGGACCCATCCAGTTTGTCCCCACCATCTCCACTCTGCCCC ATGGGCTCTGGCAAATCTCTGAGGCTGGAACAGGGGTCTCCAGTATATTCATCTACC ATGGTGAGGTGCCCCAGGCCAGCCAAGTACCCCCTCCCAGTGGATTCACTGTCCACG 10 GCCTCCCAACATCTCCAGACCGGCCAGGCTCCACCAGCCCCTTCGCTCCATCAGCCA CTGACCTGCCCAGCATGCCTGAACCTGCCCTGACCTCCCGAGCAAACATGACAGAG CACAAGACGTCCCCCACCCAATGCCCGGCAGCTGGAGAGGTCTCCAACAAGCTTCC AAAATGGCCTGAGCCGGTGGAGCAGTTCTACCGCTCACTGCAGGACACGTATGGTG CCGAGCCCGCAGGCCCGGATGGCATCCTAGTGGAGGTGGATCTGGTGCAGGCCAGG 15 CTGGAGAGGAGCAGCAGCAAGAGCCTGGAGCGGGAACTGGCCACCCCGGACTGGG CAGAACGGCAGCTGGCCCAAGGAGGCCTGGCTGAGGTGCTGTTGGCTGCCAAGGAG CACCGGCGGCCGCGTGAGACACGAGTGATTGCTGTGCTGGGCAAAGCTGGTCAGGG CAAGAGCTATTGGGCTGGGGCAGTGAGCCGGGCCTGGGCTTGTGGCCGGCTTCCCC AGTACGACTTTGTCTTCTCTGTCCCCTGCCATTGCTTGAACCGTCCGGGGGATGCCTA 20 TGGCCTGCAGGATCTGCTCTTCTCCCTGGGCCCACAGCCACTCGTGGCGGCCGATGA GGTTTTCAGCCACATCTTGAAGAGACCTGACCGCGTTCTGCTCATCCTAGACGGCTT CGAGGAGCTGGAAGCGCAAGATGGCTTCCTGCACAGCACGTGCGGACCGGCACCGG CGGAGCCCTGCTCCCTCCGGGGGCTGCTGGCCGGCCTTTTCCAGAAGAAGCTGCTCC GAGGTTGCACCCTCCTCCTCACAGCCCGGCCCCGGGGCCGCCTGGTCCAGAGCCTGA 25 GCAAGGCCGACGCCCTATTTGAGCTGTCCGGCTTCTCCATGGAGCAGGCCCAGGCAT ACGTGATGCGCTACTTTGAGAGCTCAGGGATGACAGAGCACCAAGACAGAGCCCTG ACGCTCCTCCGGGACCGGCCACTTCTTCTCAGTCACAGCCACAGCCCTACTTTGTGC CGGGCAGTGTGCCAGCTCTCAGAGGCCCTGCTGGAGCTTGGGGAGGACGCCAAGCT GCCCTCCACGCTCACGGGACTCTATGTCGGCCTGCTGGGCCGTGCAGCCCTCGACAG 30 CCCCCCCGGGGCCCTGGCAGAGCTGGCCAAGCTGGCCTGGGAGCTGGGCCGCAGAC ATCAAAGTACCCTACAGGAGGACCAGTTCCCATCCGCAGACGTGAGGACCTGGGCG 2026204466   11 Jun 2026 ATGGCCAAAGGCTTAGTCCAACACCCACCGCGGGCCGCAGAGTCCGAGCTGGCCTT CCCCAGCTTCCTCCTGCAATGCTTCCTGGGGGCCCTGTGGCTGGCTCTGAGTGGCGA AATCAAGGACAAGGAGCTCCCGCAGTACCTAGCATTGACCCCAAGGAAGAAGAGGC CCTATGACAACTGGCTGGAGGGCGTGCCACGCTTTCTGGCTGGGCTGATCTTCCAGC 5 CTCCCGCCCGCTGCCTGGGAGCCCTACTCGGGCCATCGGCGGCTGCCTCGGTGGACA GGAAGCAGAAGGTGCTTGCGAGGTACCTGAAGCGGCTGCAGCCGGGGACACTGCGG GCGCGGCAGCTGCTGGAGCTGCTGCACTGCGCCCACGAGGCCGAGGAGGCTGGAAT TTGGCAGCACGTGGTACAGGAGCTCCCCGGCCGCCTCTCTTTTCTGGGCACCCGCCT CACGCCTCCTGATGCACATGTACTGGGCAAGGCCTTGGAGGCGGCGGGCCAAGACT 10 TCTCCCTGGACCTCCGCAGCACTGGCATTTGCCCCTCTGGATTGGGGAGCCTCGTGG GACTCAGCTGTGTCACCCGTTTCAGGGCTGCCTTGAGCGACACGGTGGCGCTGTGGG AGTCCCTGCAGCAGCATGGGGAGACCAAGCTACTTCAGGCAGCAGAGGAGAAGTTC ACCATCGAGCCTTTCAAAGCCAAGTCCCTGAAGGATGTGGAAGACCTGGGAAAGCT TGTGCAGACTCAGAGGACGAGAAGTTCCTCGGAAGACACAGCTGGGGAGCTCCCTG 15 CTGTTCGGGACCTAAAGAAACTGGAGTTTGCGCTGGGCCCTGTCTCAGGCCCCCAGG CTTTCCCCAAACTGGTGCGGATCCTCACGGCCTTTTCCTCCCTGCAGCATCTGGACCT GGATGCGCTGAGTGAGAACAAGATCGGGGACGAGGGTGTCTCGCAGCTCTCAGCCA CCTTCCCCCAGCTGAAGTCCTTGGAAACCCTCAATCTGTCCCAGAACAACATCACTG ACCTGGGTGCCTACAAACTCGCCGAGGCCCTGCCTTCGCTCGCTGCATCCCTGCTCA 20 GGCTAAGCTTGTACAATAACTGCATCTGCGACGTGGGAGCCGAGAGCTTGGCTCGTG TGCTTCCGGACATGGTGTCCCTCCGGGTGATGGACGTCCAGTACAACAAGTTCACGG CTGCCGGGGCCCAGCAGCTCGCTGCCAGCCTTCGGAGGTGTCCTCATGTGGAGACGC TGGCGATGTGGACGCCCACCATCCCATTCAGTGTCCAGGAACACCTGCAACAACAG GATTCACGGATCAGCCTGAGATGATCCCAGCTGTGCTCTGGACAGGCATGTTCTCTG 25 AGGACACTAACCACGCTGGACCTTGAACTGGGTACTTGTGGACACAGCTCTTCTCCA GGCTGTATCCCATGAGCCTCAGCATCCTGGCACCCGGCCCCTGCTGGTTCAGGGTTG GCCCCTGCCCGGCTGCGGAATGAACCACATCTTGCTCTGCTGACAGACACAGGCCCG GCTCCAGGCTCCTTTAGCGCCCAGTTGGGTGGATGCCTGGTGGCAGCTGCGGTCCAC CCAGGAGCCCCGAGGCCTTCTCTGAAGGACATTGCGGACAGCCACGGCCAGGCCAG 30 AGGGAGTGACAGAGGCAGCCCCATTCTGCCTGCCCAGGCCCCTGCCACCCTGGGGA GAAAGTACTTCTTTTTTTTTATTTTTAGACAGAGTCTCACTGTTGCCCAGGCTGGCGT 2026204466   11 Jun 2026 GCAGTGGTGCGATCTGGGTTCACTGCAACCTCCGCCTCTTGGGTTCAAGCGATTCTT CTGCTTCAGCCTCCCGAGTAGCTGGGACTACAGGCACCCACCATCATGTCTGGCTAA TTTTTCATTTTTAGTAGAGACAGGGTTTTGCCATGTTGGCCAGGCTGGTCTCAAACTC TTGACCTCAGGTGATCCACCCACCTCAGCCTCCCAAAGTGCTGGGATTACAAGCGTG 5 AGCCACTGCACCGGGCCACAGAGAAAGTACTTCTCCACCCTGCTCTCCGACCAGAC ACCTTGACAGGGCACACCGGGCACTCAGAAGACACTGATGGGCAACCCCCAGCCTG CTAATTCCCCAGATTGCAACAGGCTGGGCTTCAGTGGCAGCTGCTTTTGTCTATGGG ACTCAATGCACTGACATTGTTGGCCAAAGCCAAAGCTAGGCCTGGCCAGATGCACC AGCCCTTAGCAGGGAAACAGCTAATGGGACACTAATGGGGCGGTGAGAGGGGAAC 10 AGACTGGAAGCACAGCTTCATTTCCTGTGTCTTTTTTCACTACATTATAAATGTCTCT TTAATGTCACAGGCAGGTCCAGGGTTTGAGTTCATACCCTGTTACCATTTTGGGGTA CCCACTGCTCTGGTTATCTAATATGTAACAAGCCACCCCAAATCATAGTGGCTTAAA ACAACACTCACATTTA

[00371] By “cytotoxic T-lymphocyte associated protein 4 (CTLA-4) polypeptide” is meant a 15 protein having at least about 85% sequence identity to NCBI Accession No. EAW70354.1 or a fragment thereof. An exemplary amino acid sequence is provided below: >EAW70354.1 cytotoxic T-lymphocyte-associated protein 4 [Homo sapiens] MACLGFQRHKAQLNLATRTWPCTLLFFLLFIPVFCKAMHVAQPAVVLASSRGIASFVCE YASPGKATEVRVTVLRQADSQVTEVCAATYMMGNELTFLDDSICTGTSSGNQVNLTIQ 20 GLRAMDTGLYICKVELMYPPPYYLGIGNGTQIYVIDPEPCPDSDFLLWILAAVSSGLFFY SFLLTAVSLSKMLKKRSPLTTGVYVKMPPTEPECEKQFQPYFIPIN

[00372] By “cytotoxic T-lymphocyte associated protein 4 (CTLA-4) polynucleotide” is meant a nucleic acid molecule encoding a CTLA-4 polypeptide. The CTLA-4 gene encodes an immunoglobulin superfamily and encodes a protein which transmits an inhibitory signal to T 25 cells. An exemplary CTLA-4 nucleic acid sequence is provided below. >BC074842.2 Homo sapiens cytotoxic T-lymphocyte-associated protein 4, mRNA (cDNA clone MGC:104099 IMAGE:30915552), complete cds GACCTGAACACCGCTCCCATAAAGCCATGGCTTGCCTTGGATTTCAGCGGCACAAGG CTCAGCTGAACCTGGCTACCAGGACCTGGCCCTGCACTCTCCTGTTTTTTCTTCTCTT 30 CATCCCTGTCTTCTGCAAAGCAATGCACGTGGCCCAGCCTGCTGTGGTACTGGCCAG CAGCCGAGGCATCGCCAGCTTTGTGTGTGAGTATGCATCTCCAGGCAAAGCCACTGA 2026204466   11 Jun 2026 GGTCCGGGTGACAGTGCTTCGGCAGGCTGACAGCCAGGTGACTGAAGTCTGTGCGG CAACCTACATGATGGGGAATGAGTTGACCTTCCTAGATGATTCCATCTGCACGGGCA CCTCCAGTGGAAATCAAGTGAACCTCACTATCCAAGGACTGAGGGCCATGGACACG GGACTCTACATCTGCAAGGTGGAGCTCATGTACCCACCGCCATACTACCTGGGCATA 5 GGCAACGGAACCCAGATTTATGTAATTGATCCAGAACCGTGCCCAGATTCTGACTTC CTCCTCTGGATCCTTGCAGCAGTTAGTTCGGGGTTGTTTTTTTATAGCTTTCTCCTCAC AGCTGTTTCTTTGAGCAAAATGCTAAAGAAAAGAAGCCCTCTTACAACAGGGGTCTA TGTGAAAATGCCCCCAACAGAGCCAGAATGTGAAAAGCAATTTCAGCCTTATTTTAT TCCCATCAATTGAGAAACCATTATGAAGAAGAGAGTCCATATTTCAATTTCCAAGAG 10 CTGAGG

[00373] By “cytidine deaminase” is meant a polypeptide or fragment thereof capable of catalyzing a deamination reaction that converts an amino group to a carbonyl group. In one embodiment, the cytidine deaminase converts cytosine to uracil or 5-methylcytosine to thymine. PmCDAl derived from Petromyzon marinus (Petromyzon marinus cytosine deaminase 1), or 15 AID (Activation-induced cytidine deaminase; AICDA) derived from mammal (e.g., human, swine, bovine, horse, monkey etc.), and APOBEC are exemplary cytidine deaminases.

[00374] The base sequence and amino acid sequence of PmCDAl and the base sequence and amino acid sequence of human AID are shown below. >tr|A5H718|A5H718_PETMA Cytosine deaminase OS=Petromyzon marinus OX=7757 PE=2 20 SV=1 MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQ SGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHT LKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENR WLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV 25 >EF094822.1 Petromyzon marinus isolate PmCDA.21 cytosine deaminase mRNA, complete cds TGACACGACACAGCCGTGTATATGAGGAAGGGTAGCTGGATGGGGGGGGGGGGAA TACGTTCAGAGAGGACATTAGCGAGCGTCTTGTTGGTGGCCTTGAGTCTAGACACCT GCAGACATGACCGACGCTGAGTACGTGAGAATCCATGAGAAGTTGGACATCTACAC 30 GTTTAAGAAACAGTTTTTCAACAACAAAAAATCCGTGTCGCATAGATGCTACGTTCT CTTTGAATTAAAACGACGGGGTGAACGTAGAGCGTGTTTTTGGGGCTATGCTGTGAA 2026204466   11 Jun 2026 TAAACCACAGAGCGGGACAGAACGTGGAATTCACGCCGAAATCTTTAGCATTAGAA AAGTCGAAGAATACCTGCGCGACAACCCCGGACAATTCACGATAAATTGGTACTCA TCCTGGAGTCCTTGTGCAGATTGCGCTGAAAAGATCTTAGAATGGTATAACCAGGAG CTGCGGGGGAACGGCCACACTTTGAAAATCTGGGCTTGCAAACTCTATTACGAGAA 5 AAATGCGAGGAATCAAATTGGGCTGTGGAACCTCAGAGATAACGGGGTTGGGTTGA ATGTAATGGTAAGTGAACACTACCAATGTTGCAGGAAAATATTCATCCAATCGTCGC ACAATCAATTGAATGAGAATAGATGGCTTGAGAAGACTTTGAAGCGAGCTGAAAAA CGACGGAGCGAGTTGTCCATTATGATTCAGGTAAAAATACTCCACACCACTAAGAGT CCTGCTGTTTAAGAGGCTATGCGGATGGTTTTC 10 >tr|Q6QJ80|Q6QJ80_HUMAN Activation-induced cytidine deaminase OS=Homo sapiens OX=9606 GN=AICDA PE=2 SV=1 MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHV ELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTARLYFC 15 EDRKAEPEGLRRLHRAGVQIAIMTFKAPV >NG_011588.1:5001-15681 Homo sapiens activation induced cytidine deaminase (AICDA), RefSeqGene (LRG17) on chromosome 12 AGAGAACCATCATTAATTGAAGTGAGATTTTTCTGGCCTGAGACTTGCAGGGAGGCA 20 AGAAGACACTCTGGACACCACTATGGACAGGTAAAGAGGCAGTCTTCTCGTGGGTG ATTGCACTGGCCTTCCTCTCAGAGCAAATCTGAGTAATGAGACTGGTAGCTATCCCT TTCTCTCATGTAACTGTCTGACTGATAAGATCAGCTTGATCAATATGCATATATATTT TTTGATCTGTCTCCTTTTCTTCTATTCAGATCTTATACGCTGTCAGCCCAATTCTTTCT GTTTCAGACTTCTCTTGATTTCCCTCTTTTTCATGTGGCAAAAGAAGTAGTGCGTACA 25 ATGTACTGATTCGTCCTGAGATTTGTACCATGGTTGAAACTAATTTATGGTAATAAT ATTAACATAGCAAATCTTTAGAGACTCAAATCATGAAAAGGTAATAGCAGTACTGT ACTAAAAACGGTAGTGCTAATTTTCGTAATAATTTTGTAAATATTCAACAGTAAAAC AACTTGAAGACACACTTTCCTAGGGAGGCGTTACTGAAATAATTTAGCTATAGTAAG AAAATTTGTAATTTTAGAAATGCCAAGCATTCTAAATTAATTGCTTGAAAGTCACTA 30 TGATTGTGTCCATTATAAGGAGACAAATTCATTCAAGCAAGTTATTTAATGTTAAAG GCCCAATTGTTAGGCAGTTAATGGCACTTTTACTATTAACTAATCTTTCCATTTGTTC 2026204466   11 Jun 2026 AGACGTAGCTTAACTTACCTCTTAGGTGTGAATTTGGTTAAGGTCCTCATAATGTCTT TATGTGCAGTTTTTGATAGGTTATTGTCATAGAACTTATTCTATTCCTACATTTATGA TTACTATGGATGTATGAGAATAACACCTAATCCTTATACTTTACCTCAATTTAACTCC TTTATAAAGAACTTACATTACAGAATAAAGATTTTTTAAAAATATATTTTTTTGTAGA 5 GACAGGGTCTTAGCCCAGCCGAGGCTGGTCTCTAAGTCCTGGCCCAAGCGATCCTCC TGCCTGGGCCTCCTAAAGTGCTGGAATTATAGACATGAGCCATCACATCCAATATAC AGAATAAAGATTTTTAATGGAGGATTTAATGTTCTTCAGAAAATTTTCTTGAGGTCA GACAATGTCAAATGTCTCCTCAGTTTACACTGAGATTTTGAAAACAAGTCTGAGCTA TAGGTCCTTGTGAAGGGTCCATTGGAAATACTTGTTCAAAGTAAAATGGAAAGCAA 10 AGGTAAAATCAGCAGTTGAAATTCAGAGAAAGACAGAAAAGGAGAAAAGATGAAA TTCAACAGGACAGAAGGGAAATATATTATCATTAAGGAGGACAGTATCTGTAGAGC TCATTAGTGATGGCAAAATGACTTGGTCAGGATTATTTTTAACCCGCTTGTTTCTGGT TTGCACGGCTGGGGATGCAGCTAGGGTTCTGCCTCAGGGAGCACAGCTGTCCAGAG CAGCTGTCAGCCTGCAAGCCTGAAACACTCCCTCGGTAAAGTCCTTCCTACTCAGGA 15 CAGAAATGACGAGAACAGGGAGCTGGAAACAGGCCCCTAACCAGAGAAGGGAAGT AATGGATCAACAAAGTTAACTAGCAGGTCAGGATCACGCAATTCATTTCACTCTGAC TGGTAACATGTGACAGAAACAGTGTAGGCTTATTGTATTTTCATGTAGAGTAGGACC CAAAAATCCACCCAAAGTCCTTTATCTATGCCACATCCTTCTTATCTATACTTCCAGG ACACTTTTTCTTCCTTATGATAAGGCTCTCTCTCTCTCCACACACACACACACACACA 20 CACACACACACACACACACACACACAAACACACACCCCGCCAACCAAGGTGCATGT AAAAAGATGTAGATTCCTCTGCCTTTCTCATCTACACAGCCCAGGAGGGTAAGTTAA TATAAGAGGGATTTATTGGTAAGAGATGATGCTTAATCTGTTTAACACTGGGCCTCA AAGAGAGAATTTCTTTTCTTCTGTACTTATTAAGCACCTATTATGTGTTGAGCTTATA TATACAAAGGGTTATTATATGCTAATATAGTAATAGTAATGGTGGTTGGTACTATGG 25 TAATTACCATAAAAATTATTATCCTTTTAAAATAAAGCTAATTATTATTGGATCTTTT TTAGTATTCATTTTATGTTTTTTATGTTTTTGATTTTTTAAAAGACAATCTCACCCTGT TACCCAGGCTGGAGTGCAGTGGTGCAATCATAGCTTTCTGCAGTCTTGAACTCCTGG GCTCAAGCAATCCTCCTGCCTTGGCCTCCCAAAGTGTTGGGATACAGTCATGAGCCA CTGCATCTGGCCTAGGATCCATTTAGATTAAAATATGCATTTTAAATTTTAAAATAAT 30 ATGGCTAATTTTTACCTTATGTAATGTGTATACTGGCAATAAATCTAGTTTGCTGCCT AAAGTTTAAAGTGCTTTCCAGTAAGCTTCATGTACGTGAGGGGAGACATTTAAAGTG 2026204466   11 Jun 2026 AAACAGACAGCCAGGTGTGGTGGCTCACGCCTGTAATCCCAGCACTCTGGGAGGCT GAGGTGGGTGGATCGCTTGAGCCCTGGAGTTCAAGACCAGCCTGAGCAACATGGCA AAACGCTGTTTCTATAACAAAAATTAGCCGGGCATGGTGGCATGTGCCTGTGGTCCC AGCTACTAGGGGGCTGAGGCAGGAGAATCGTTGGAGCCCAGGAGGTCAAGGCTGCA 5 CTGAGCAGTGCTTGCGCCACTGCACTCCAGCCTGGGTGACAGGACCAGACCTTGCCT CAAAAAAATAAGAAGAAAAATTAAAAATAAATGGAAACAACTACAAAGAGCTGTT GTCCTAGATGAGCTACTTAGTTAGGCTGATATTTTGGTATTTAACTTTTAAAGTCAGG GTCTGTCACCTGCACTACATTATTAAAATATCAATTCTCAATGTATATCCACACAAA GACTGGTACGTGAATGTTCATAGTACCTTTATTCACAAAACCCCAAAGTAGAGACTA 10 TCCAAATATCCATCAACAAGTGAACAAATAAACAAAATGTGCTATATCCATGCAAT GGAATACCACCCTGCAGTACAAAGAAGCTACTTGGGGATGAATCCCAAAGTCATGA CGCTAAATGAAAGAGTCAGACATGAAGGAGGAGATAATGTATGCCATACGAAATTC TAGAAAATGAAAGTAACTTATAGTTACAGAAAGCAAATCAGGGCAGGCATAGAGGC TCACACCTGTAATCCCAGCACTTTGAGAGGCCACGTGGGAAGATTGCTAGAACTCAG 15 GAGTTCAAGACCAGCCTGGGCAACACAGTGAAACTCCATTCTCCACAAAAATGGGA AAAAAAGAAAGCAAATCAGTGGTTGTCCTGTGGGGAGGGGAAGGACTGCAAAGAG GGAAGAAGCTCTGGTGGGGTGAGGGTGGTGATTCAGGTTCTGTATCCTGACTGTGGT AGCAGTTTGGGGTGTTTACATCCAAAAATATTCGTAGAATTATGCATCTTAAATGGG TGGAGTTTACTGTATGTAAATTATACCTCAATGTAAGAAAAAATAATGTGTAAGAAA 20 ACTTTCAATTCTCTTGCCAGCAAACGTTATTCAAATTCCTGAGCCCTTTACTTCGCAA ATTCTCTGCACTTCTGCCCCGTACCATTAGGTGACAGCACTAGCTCCACAAATTGGA TAAATGCATTTCTGGAAAAGACTAGGGACAAAATCCAGGCATCACTTGTGCTTTCAT ATCAACCATGCTGTACAGCTTGTGTTGCTGTCTGCAGCTGCAATGGGGACTCTTGAT TTCTTTAAGGAAACTTGGGTTACCAGAGTATTTCCACAAATGCTATTCAAATTAGTG 25 CTTATGATATGCAAGACACTGTGCTAGGAGCCAGAAAACAAAGAGGAGGAGAAATC AGTCATTATGTGGGAACAACATAGCAAGATATTTAGATCATTTTGACTAGTTAAAAA AGCAGCAGAGTACAAAATCACACATGCAATCAGTATAATCCAAATCATGTAAATAT GTGCCTGTAGAAAGACTAGAGGAATAAACACAAGAATCTTAACAGTCATTGTCATT AGACACTAAGTCTAATTATTATTATTAGACACTATGATATTTGAGATTTAAAAAATC 3 0 TTTAAT ATTTT AAAATTTAGAGCTCTTCTATTTTTCCATAGTATTCAAGTTTGACAAT GATCAAGTATTACTCTTTCTTTTTTTTTTTTTTTTTTTTTTTTTGAGATGGAGTTTTGGT 2026204466   11 Jun 2026 CTTGTTGCCCATGCTGGAGTGGAATGGCATGACCATAGCTCACTGCAACCTCCACCT CCTGGGTTCAAGCAAAGCTGTCGCCTCAGCCTCCCGGGTAGATGGGATTACAGGCG CCCACCACCACACTCGGCTAATGTTTGTATTTTTAGTAGAGATGGGGTTTCACCATGT TGGCCAGGCTGGTCTCAAACTCCTGACCTCAGAGGATCCACCTGCCTCAGCCTCCCA 5 AAGTGCTGGGATTACAGATGTAGGCCACTGCGCCCGGCCAAGTATTGCTCTTATACA TTAAAAAACAGGTGTGAGCCACTGCGCCCAGCCAGGTATTGCTCTTATACATTAAAA AATAGGCCGGTGCAGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAAGCCAAGGC GGGCAGAACACCCGAGGTCAGGAGTCCAAGGCCAGCCTGGCCAAGATGGTGAAAC CCCGTCTCTATTAAAAATACAAACATTACCTGGGCATGATGGTGGGCGCCTGTAATC 10 CCAGCTACTCAGGAGGCTGAGGCAGGAGGATCCGCGGAGCCTGGCAGATCTGCCTG AGCCTGGGAGGTTGAGGCTACAGTAAGCCAAGATCATGCCAGTATACTTCAGCCTG GGCGACAAAGTGAGACCGTAACAAAAAAAAAAAAATTTAAAAAAAGAAATTTAGA TCAAGATCCAACTGTAAAAAGTGGCCTAAACACCACATTAAAGAGTTTGGAGTTTAT TCTGCAGGCAGAAGAGAACCATCAGGGGGTCTTCAGCATGGGAATGGCATGGTGCA 15 CCTGGTTTTTGTGAGATCATGGTGGTGACAGTGTGGGGAATGTTATTTTGGAGGGAC TGGAGGCAGACAGACCGGTTAAAAGGCCAGCACAACAGATAAGGAGGAAGAAGAT GAGGGCTTGGACCGAAGCAGAGAAGAGCAAACAGGGAAGGTACAAATTCAAGAAA TATTGGGGGGTTTGAATCAACACATTTAGATGATTAATTAAATATGAGGACTGAGGA ATAAGAAATGAGTCAAGGATGGTTCCAGGCTGCTAGGCTGCTTACCTGAGGTGGCA 20 AAGTCGGGAGGAGTGGCAGTTTAGGACAGGGGGCAGTTGAGGAATATTGTTTTGAT CATTTTGAGTTTGAGGTACAAGTTGGACACTTAGGTAAAGACTGGAGGGGAAATCT GAATATACAATTATGGGACTGAGGAACAAGTTTATTTTATTTTTTGTTTCGTTTTCTT GTTGAAGAACAAATTTAATTGTAATCCCAAGTCATCAGCATCTAGAAGACAGTGGC AGGAGGTGACTGTCTTGTGGGTAAGGGTTTGGGGTCCTTGATGAGTATCTCTCAATT 25 GGCCTTAAATATAAGCAGGAAAAGGAGTTTATGATGGATTCCAGGCTCAGCAGGGC TCAGGAGGGCTCAGGCAGCCAGCAGAGGAAGTCAGAGCATCTTCTTTGGTTTAGCC CAAGTAATGACTTCCTTAAAAAGCTGAAGGAAAATCCAGAGTGACCAGATTATAAA CTGTACTCTTGCATTTTCTCTCCCTCCTCTCACCCACAGCCTCTTGATGAACCGGAGG AAGTTTCTTTACCAATTCAAAAATGTCCGCTGGGCTAAGGGTCGGCGTGAGACCTAC 30 CTGTGCTACGTAGTGAAGAGGCGTGACAGTGCTACATCCTTTTCACTGGACTTTGGT TATCTTCGCAATAAGGTATCAATTAAAGTCGGCTTTGCAAGCAGTTTAATGGTCAAC 2026204466   11 Jun 2026 TGTGAGTGCTTTTAGAGCCACCTGCTGATGGTATTACTTCCATCCTTTTTTGGCATTT GTGTCTCTATCACATTCCTCAAATCCTTTTTTTTATTTCTTTTTCCATGTCCATGCACC CATATTAGACATGGCCCAAAATATGTGATTTAATTCCTCCCCAGTAATGCTGGGCAC CCTAATACCACTCCTTCCTTCAGTGCCAAGAACAACTGCTCCCAAACTGTTTACCAG 5 CTTTCCTCAGCATCTGAATTGCCTTTGAGATTAATTAAGCTAAAAGCATTTTTATATG GGAGAATATTATCAGCTTGTCCAAGCAAAAATTTTAAATGTGAAAAACAAATTGTGT CTTAAGCATTTTTGAAAATTAAGGAAGAAGAATTTGGGAAAAAATTAACGGTGGCT CAATTCTGTCTTCCAAATGATTTCTTTTCCCTCCTACTCACATGGGTCGTAGGCCAGT GAATACATTCAACATGGTGATCCCCAGAAAACTCAGAGAAGCCTCGGCTGATGATT 10 AATTAAATTGATCTTTCGGCTACCCGAGAGAATTACATTTCCAAGAGACTTCTTCAC CAAAATCCAGATGGGTTTACATAAACTTCTGCCCACGGGTATCTCCTCTCTCCTAAC ACGCTGTGACGTCTGGGCTTGGTGGAATCTCAGGGAAGCATCCGTGGGGTGGAAGG TCATCGTCTGGCTCGTTGTTTGATGGTTATATTACCATGCAATTTTCTTTGCCTACATT TGTATTGAATACATCCCAATCTCCTTCCTATTCGGTGACATGACACATTCTATTTCAG 15 AAGGCTTTGATTTTATCAAGCACTTTCATTTACTTCTCATGGCAGTGCCTATTACTTC TCTTACAATACCCATCTGTCTGCTTTACCAAAATCTATTTCCCCTTTTCAGATCCTCCC AAATGGTCCTCATAAACTGTCCTGCCTCCACCTAGTGGTCCAGGTATATTTCCACAA TGTTACATCAACAGGCACTTCTAGCCATTTTCCTTCTCAAAAGGTGCAAAAAGCAAC TTCATAAACACAAATTAAATCTTCGGTGAGGTAGTGTGATGCTGCTTCCTCCCAACT 20 CAGCGCACTTCGTCTTCCTCATTCCACAAAAACCCATAGCCTTCCTTCACTCTGCAGG ACTAGTGCTGCCAAGGGTTCAGCTCTACCTACTGGTGTGCTCTTTTGAGCAAGTTGCT TAGCCTCTCTGTAACACAAGGACAATAGCTGCAAGCATCCCCAAAGATCATTGCAG GAGACAATGACTAAGGCTACCAGAGCCGCAATAAAAGTCAGTGAATTTTAGCGTGG TCCTCTCTGTCTCTCCAGAACGGCTGCCACGTGGAATTGCTCTTCCTCCGCTACATCT 25 CGGACTGGGACCTAGACCCTGGCCGCTGCTACCGCGTCACCTGGTTCACCTCCTGGA GCCCCTGCTACGACTGTGCCCGACATGTGGCCGACTTTCTGCGAGGGAACCCCAACC TCAGTCTGAGGATCTTCACCGCGCGCCTCTACTTCTGTGAGGACCGCAAGGCTGAGC CCGAGGGGCTGCGGCGGCTGCACCGCGCCGGGGTGCAAATAGCCATCATGACCTTC AAAGGTGCGAAAGGGCCTTCCGCGCAGGCGCAGTGCAGCAGCCCGCATTCGGGATT 30 GCGATGCGGAATGAATGAGTTAGTGGGGAAGCTCGAGGGGAAGAAGTGGGCGGGG ATTCTGGTTCACCTCTGGAGCCGAAATTAAAGATTAGAAGCAGAGAAAAGAGTGAA 2026204466   11 Jun 2026 TGGCTCAGAGACAAGGCCCCGAGGAAATGAGAAAATGGGGCCAGGGTTGCTTCTTT CCCCTCGATTTGGAACCTGAACTGTCTTCTACCCCCATATCCCCGCCTTTTTTTCCTTT TTTTTTTTTTGAAGATTATTTTTACTGCTGGAATACTTTTGTAGAAAACCACGAAAGA ACTTTCAAAGCCTGGGAAGGGCTGCATGAAAATTCAGTTCGTCTCTCCAGACAGCTT 5 CGGCGCATCCTTTTGGTAAGGGGCTTCCTCGCTTTTTAAATTTTCTTTCTTTCTCTACA GTCTTTTTTGGAGTTTCGTATATTTCTTATATTTTCTTATTGTTCAATCACTCTCAGTT TTCATCTGATGAAAACTTTATTTCTCCTCCACATCAGCTTTTTCTTCTGCTGTTTCACC ATTCAGAGCCCTCTGCTAAGGTTCCTTTTCCCTCCCTTTTCTTTCTTTTGTTGTTTCAC ATCTTTAAATTTCTGTCTCTCCCCAGGGTTGCGTTTCCTTCCTGGTCAGAATTCTTTTC 10 TCCTTTTTTTTTTTTTTTTTTTTTTTTTTTAAACAAACAAACAAAAAACCCAAAAAAAC TCTTTCCCAATTTACTTTCTTCCAACATGTTACAAAGCCATCCACTCAGTTTAGAAGA CTCTCCGGCCCCACCGACCCCCAACCTCGTTTTGAAGCCATTCACTCAATTTGCTTCT CTCTTTCTCTACAGCCCCTGTATGAGGTTGATGACTTACGAGACGCATTTCGTACTTT GGGACTTTGATAGCAACTTCCAGGAATGTCACACACGATGAAATATCTCTGCTGAAG 15 ACAGTGGATAAAAAACAGTCCTTCAAGTCTTCTCTGTTTTTATTCTTCAACTCTCACT TTCTTAGAGTTTACAGAAAAAATATTTATATACGACTCTTTAAAAAGATCTATGTCTT GAAAATAGAGAAGGAACACAGGTCTGGCCAGGGACGTGCTGCAATTGGTGCAGTTT TGAATGCAACATTGTCCCCTACTGGGAATAACAGAACTGCAGGACCTGGGAGCATC CTAAAGTGTCAACGTTTTTCTATGACTTTTAGGTAGGATGAGAGCAGAAGGTAGATC 20 CTAAAAAGCATGGTGAGAGGATCAAATGTTTTTATATCAACATCCTTTATTATTTGA TTCATTTGAGTTAACAGTGGTGTTAGTGATAGATTTTTCTATTCTTTTCCCTTGACGTT TACTTTCAAGTAACACAAACTCTTCCATCAGGCCATGATCTATAGGACCTCCTAATG AGAGTATCTGGGTGATTGTGACCCCAAACCATCTCTCCAAAGCATTAATATCCAATC ATGCGCTGTATGTTTTAATCAGCAGAAGCATGTTTTTATGTTTGTACAAAAGAAGAT 25 TGTTATGGGTGGGGATGGAGGTATAGACCATGCATGGTCACCTTCAAGCTACTTTAA TAAAGGATCTTAAAATGGGCAGGAGGACTGTGAACAAGACACCCTAATAATGGGTT GATGTCTGAAGTAGCAAATCTTCTGGAAACGCAAACTCTTTTAAGGAAGTCCCTAAT TTAGAAACACCCACAAACTTCACATATCATAATTAGCAAACAATTGGAAGGAAGTT GCTTGAATGTTGGGGAGAGGAAAATCTATTGGCTCTCGTGGGTCTCTTCATCTCAGA 30 AATGCCAATCAGGTCAAGGTTTGCTACATTTTGTATGTGTGTGATGCTTCTCCCAAA GGTATATTAACTATATAAGAGAGTTGTGACAAAACAGAATGATAAAGCTGCGAACC 2026204466   11 Jun 2026 GTGGCACACGCTCATAGTTCTAGCTGCTTGGGAGGTTGAGGAGGGAGGATGGCTTG AACACAGGTGTTCAAGGCCAGCCTGGGCAACATAACAAGATCCTGTCTCTCAAAAA AAAAAAAAAAAAAAAGAAAGAGAGAGGGCCGGGCGTGGTGGCTCACGCCTGTAAT CCCAGCACTTTGGGAGGCCGAGCCGGGCGGATCACCTGTGGTCAGGAGTTTGAGAC 5 CAGCCTGGCCAACATGGCAAAACCCCGTCTGTACTCAAAATGCAAAAATTAGCCAG GCGTGGTAGCAGGCACCTGTAATCCCAGCTACTTGGGAGGCTGAGGCAGGAGAATC GCTTGAACCCAGGAGGTGGAGGTTGCAGTAAGCTGAGATCGTGCCGTTGCACTCCA GCCTGGGCGACAAGAGCAAGACTCTGTCTCAGAAAAAAAAAAAAAAAAGAGAGAG AGAGAGAAAGAGAACAATATTTGGGAGAGAAGGATGGGGAAGCATTGCAAGGAAA 10 TTGTGCTTTATCCAACAAAATGTAAGGAGCCAATAAGGGATCCCTATTTGTCTCTTTT GGTGTCTATTTGTCCCTAACAACTGTCTTTGACAGTGAGAAAAATATTCAGAATAAC CATATCCCTGTGCCGTTATTACCTAGCAACCCTTGCAATGAAGATGAGCAGATCCAC AGGAAAACTTGAATGCACAACTGTCTTATTTTAATCTTATTGTACATAAGTTTGTAA AAGAGTTAAAAATTGTTACTTCATGTATTCATTTATATTTTATATTATTTTGCGTCTA 15 ATGATTTTTTATTAACATGATTTCCTTTTCTGATATATTGAAATGGAGTCTCAAAGCT TCATAAATTTATAACTTTAGAAATGATTCTAATAACAACGTATGTAATTGTAACATT GCAGTAATGGTGCTACGAAGCCATTTCTCTTGATTTTTAGTAAACTTTTATGACAGCA AATTTGCTTCTGGCTCACTTTCAATCAGTTAAATAAATGATAAATAATTTTGGAAGCT GTGAAGATAAAATACCAAATAAAATAATATAAAAGTGATTTATATGAAGTTAAAAT 20 AAAAAATCAGTATGATGGAATAAACTTG

[00375] Apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like (APOBEC) is a family of evolutionarily conserved cytidine deaminases. Members of this family are C-to-U editing enzymes. The N-terminal domain of APOBEC like proteins is the catalytic domain, while the C-terminal domain is a pseudocatalytic domain. More specifically, the catalytic domain is a 25 zinc dependent cytidine deaminase domain and is important for cytidine deamination. APOBEC family members include APOBEC1, APOBEC2, APOBEC3A, APOBEC3B, APOBEC3C, APOBEC3D ("APOBEC3E" now refers to this), APOBEC3F, APOBEC3G, APOBEC3H, APOBEC4, and Activation-induced (cytidine) deaminase. Many modified cytidine deaminases are commercially available, including but not limited to SaBE3, SaKKH-BE3, VQR-BE3, EQR- 30 BE3, VRER-BE3, YE1-BE3, EE-BE3, YE2-BE3, and YEE-BE3, which are available from Addgene (plasmids 85169, 85170, 85171, 85172, 85173, 85174, 85175, 85176, 85177). 2026204466   11 Jun 2026

[00376] Other exemplary deaminases that can be fused to Cas9 according to aspects of this disclosure are provided below. It should be understood that, in some embodiments, the active domain of the respective sequence can be used, e.g., the domain without a localizing signal (nuclear localization sequence, without nuclear export signal, cytoplasmic localizing signal). 5

[00377] Human AID: MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHV ELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTARLYFC EDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWEGLHENSVRLSR OLRRILLPLYEVDDLRDAFRTLGL (underline: nuclear localization sequence; double 10 underline: nuclear export signal)

[00378] Mouse AID: MDSLLMKOKKFLYHFKNVRWAKGRHETYLCYVVKRRDSATSCSLDFGHLRNKSGCHV ELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVAEFLRWNPNLSLRIFTARLYFC EDRKAEPEGLRRLHRAGVQIGIMTFKDYFYCWNTFVENRERTFKAWEGLHENSVRLTR 15 OLRRILLPLYEVDDLRDAFRMLGF (underline: nuclear localization sequence; double underline: nuclear export signal)

[00379] Canine AID: MDSLLMKQRKFLYHFKNVRWAKGRHETYLCYVVKRRDSATSFSLDFGHLRNKSGCHV ELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGYPNLSLRIFAARLYFC 20 EDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENREKTFKAWEGLHENSVRLSR OLRRILLPLYEVDDLRDAFRTLGL (underline: nuclear localization sequence; double underline: nuclear export signal)

[00380] Bovine AID: MDSLLKKOROFLYOFKNVRWAKGRHETYLCYVVKRRDSPTSFSLDFGHLRNKAGCHV 25 ELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGYPNLSLRIFTARLYFC DKERKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWEGLHENSVRLS ROLRRILLPLYEVDDLRDAFRTLGL (underline: nuclear localization sequence; double underline: nuclear export signal)

[00381] Rat AID 30 MAVGSKPKAALVGPHWERERIWCFLCSTGLGTOOTGOTSRWLRPAATQDPVSPPRSLL MKQRKFLYHFKNVRWAKGRHETYLCYVVKRRDSATSFSLDFGYLRNKSGCHVELLFL 2026204466   11 Jun 2026 RYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTARLTGWGALP AGT.MSPARPSDYFYCWNTFVENHERTFKAWEGLHENSVRLSRRLRRIELPLYEVDDLR DAFRTLGL (underline: nuclear localization sequence; double underline: nuclear export signal) 5

[00382] Mouse APOBEC-3 MGPFCLGCSHRKCYSPIRNLISQETFKFHFKNLGYAKGRKDTFLCYEVTRKDCDSPVSLH HGVFKNKDNIHzl£ZCFLFlEF77DKFLKmPRE£FKZriPyWR3PCF£CAEQIVRFLATHHN LSLDIFSSRLYNVQDPETQQNLCRLVQEGAQVAAMDLYEFKKCWKKFVDNGGRRFRP WKRLLTNFRYQDSKLQEILRPCYIPVPSSSSSTLSNICLTKGLPETRFCVEGRRMDPLSEE 10 EF YSQF YNQRVKH LC Y YH R M KP Y LC YQLEQFNGQ A PLKGC LLS EKG KQH4 EILFLDKIR W£LSgmrCyL7WPCPACAWQLAAFKRDRPDLILHIYTSRLYFHWKRPFQKGLCSLW QSGILVDVMDLPQFTDCWTNFVNPKRPFWPWKGLEIISRRTQRRLRRIKESWGLQDLVN DFGNLQLGPPMS (italic: nucleic acid editing domain)

[00383] Rat APOBEC-3 : 15 MGPFCLGCSHRKCYSPIRNLISQETFKFHFKNRLRYAIDRKDTFLCYEVTRKDCDSPVSL HHGVFKNKDNIHzl£ZCFLnm7DKFLKmPR££FKZriPyWRYPCF£CAEQVLRFLATHH NLSLDIFSSRLYNIRDPENQQNLCRLVQEGAQVAAMDLYEFKKCWKKFVDNGGRRFRP WKKLLTNFRYQDSKLQEILRPCYIPVPSSSSSTLSNICLTKGLPETRFCVERRRVHLLSEEE FYSQFYNQRVKHLCYYHGVKPYLCYQLEQFNGQAPLKGCLLSEKGKQEM^ZLFLDKZR.S' 20 dl£LSgFZZFCFL7WPCPACAWQLAAFKRDRPDLILHIYTSRLYFHWKRPFQKGLCSLWQ SGILVDVMDLPQFTDCWTNFVNPKRPFWPWKGLEIISRRTQRRLHRIKESWGLQDLVND FGNLQLGPPMS (italic: nucleic acid editing domain) 2026204466   11 Jun 2026

[00384] Rhesus macaque APOBEC-3 G: MVEPMDPRTFVSNFNNRPILSGLNTVWLCCEVKTKDPSGPPLDAKIFQGKVYSKAKYHP EMRFLRWFHKWRQLHHDQEYKVTWYVSWSPCTRCANSVATFLAKDPKVTLTIFVARL YYFWKPDYQQALRILCQKRGGPHATMKIMNYNEFQDCWNKFVDGRGKPFKPRNNLPK 5 HYTLLQATLGELLRHLMDPGTFTSNFNNKPWVSGQHETYLCYKVERLHNDTWVPLNQ HRGFLRNQAPNIHGFPKGRHAELCFLDLIPFWKLDGQQYRVTCFTSWSPCFSCAQEMAK FISNNEHVSLCIFAARIYDDQGRYQEGLRALHRDGAKIAMMNYSEFEYCWDTFVDRQG RPFQPWDGLDEHSQALSGRLRAI (italic: nucleic acid editing domain; underline: cytoplasmic localization signal) 10

[00385] Chimpanzee APOBEC-3 G: MKPHFRNPVERMYQDTFSDNFYNRPILSHRNTVWLCYEVKTKGPSRPPLDAKIFRGOVY SKEKAHPEMRFFHWFSKWRKLHRDQEYEVTWYISWSPCTKCYPF>VAYFEAEDPKVYEY1F VARLYYFWDPDYQEALRSLCQKRDGPRATMKIMNYDEFQHCWSKFVYSQRELFEPWN NLPKYYILLHIMLGEILRHSMDPPTFTSNFNNELWVRGRHETYLCYEVERLHNDTWVLL 15 NQRRGFLCNQ APHKHGFLEGRK4EL CFLD VIPFWKLDLHQD YR VTCFTSWSPCFSCAQEM AKFISNNKHVSLCIFAARIYDDQGRCQEGLRTLAKAGAKISIMTYSEFKHCWDTFVDHQ GCPFQP WDGLEEHS QALS GRLRAILQNQGN

[00386] (italic: nucleic acid editing domain; underline: cytoplasmic localization signal)

[00387] Green monkey APOBEC-3 G: 20 MNPQIRNMVEOMEPDIFVYYFNNRPILSGRNTVWLCYEVKTKDPSGPPLDANIFQGKLY ]^A¥J5HPEMKFLHWFRKWRQLHRDQEYEVTWYVSWSPCTRCANSN ATFEAEFYPYAJTETIF VARLYYFWKPDYQQALRILCQERGGPHATMKIMNYNEFQHCWNEFVDGQGKPFKPRK NLPKHYTLLHATLGELLRHVMDPGTFTSNFNNKPWVSGQRETYLCYKVERSHNDTWV LLNQHRGFLRNQAPDRHGFPKGRH / lELCFLDL / PFlFAYDD^m^ 25 MAKFISNNKHVSLCIFAARIYDDQGRCQEGLRTLHRDGAKIAVMNYSEFEYCWDTFVD RQGRPFQPWDGLDEHSQALSGRLRAI

[00388] (italic: nucleic acid editing domain; underline: cytoplasmic localization signal)

[00389] Human APOBEC-3G: MKPHFRNTVERMYRDTFSYNFYNRPILSRRNTVWLCYEVKTKGPSRPPLDAKIFRGOVY 30 SELKAHPEMRFFHWFSKWRKLHRDQEYEVTWYISWSPCTKCTRDMAYFLAEDPKAYLY1F VARLYYFWDPDYQEALRSLCQKRDGPRATMKIMNYDEFQHCWSKFVYSQRELFEPWN 2026204466   11 Jun 2026 NLPKYYILLHIMLGEILRHSMDPPTFTFNFNNEPWVRGRHETYLCYEVERMHNDTWVLL NQRRGFLCNQAPHKHGFLEGRKdELCFLDFZPFOZDLDgDFRFrCFTYRYPCFECAQEM AKFISKNKHVSLCIFTARIYDDQGRCQEGLRTLAEAGAKISIMTYSEFKHCWDTFVDHQ GCPFQPWDGLDEHSQDLSGRLRAILQNQEN 5 (italic: nucleic acid editing domain; underline: cytoplasmic localization signal)

[00390] Human APOBEC-3F: MKPHFRNTVERMYRDTFSYNFYNRPILSRRNTVWLCYEVKTKGPSRPRLDAKIFRGQV YSQPEHH / lE / WCF£5>FCGAX)£ / YirAX.’F2 / ^^^^ AARLYYYWERDYRRALCRLSQAGARVKIMDDEEFAYCWENFVYSEGQPFMPWYKFD 10 DNYAFLHRTLKEILRNPMEAMYPHIFYFHFKNLRKAYGRNES WLCFTMEVVKHHSPVS WKRGVFRNQVDPEYHCHAERCFLSWFCDDILSPNTNYEVTWYTSWSPCPECAGEVAEFEA RHSNVNLTIFTARLYYFWDTDYQEGLRSLSQEGASVEIMGYKDFKYCWENFVYNDDEP FKPWKGLKYNFLFLDSKLQEILE (italic: nucleic acid editing domain) 15

[00391] Human APOBEC-3B: MNPQIRNPMERMYRDTFYDNFENEPILYGRSYTWLCYEVKIKRGRSNLLWDTGVFRGQ VYFKPQYTEdEAfCFLERyCGAGLP^F^CFgZFRyFERTPCPDCVAKLAEFLSEHPNVTLTI SAARLYYYWERDYRRALCRLSQAGARVTIMDYEEFAYCWENFVYNEGQQFMPWYKF DENYAFLHRTLKEILRYLMDPDTFTFNFNNDPLVLRRRQTYLCYEVERLDNGTWVLMD 20 QHMGFLCNEAKNLLCGFYGR7E4ELRFZZ>L VPSLQLDPAQIYR VTWFISWSPCFSWGCAGE VRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFEYCWDTFVY RQGCPFQPWDGLEEHSQALSGRLRAILQNQGN (italic: nucleic acid editing domain) 2026204466   11 Jun 2026

[00392] Rat APOBEC-3B: MQPQGLGPNAGMGPVCLGCSHRRPYSPIRNPLKKLYQQTFYFHFKNVRYAWGRKNNF LCYEVNGMDCALPVPLRQGVFRKQGHIHAELCFIYWFHDKVLRVLSPMEEFKVTWYM SWSPCSKCAEQVARFLAAHRNLSLAIFSSRLYYYLRNPNYQQKLCRLIQEGVHVAAMD 5 LPEFKKCWNKFVDNDGQPFRPWMRLRINFSFYDCKLQEIFSRMNLLREDVFYLQFNNSH RVKPVQNRYYRRKSYLCYQLERANGQEPLKGYLLYKKGEQHVEILFLEKMRSMELSQV RITCYLTWSPCPNCARQLAAFKKDHPDLILRIYTSRLYFWRKKFQKGLCTLWRSGIHVD VMDLPQFADCWTNFVNPQRPFRPWNELEKNSWRIQRRLRRIKESWGL

[00393] Bovine APOBEC-3B: 10 DGWEVAFRSGTVLKAGVLGVSMTEGWAGSGHPGQGACVWTPGTRNTMNLLREVLFK QQFGNQPRVPAPYYRRKTYLCYQLKQRNDLTLDRGCFRNKKQRHAERFIDKINSLDLNP SQSYKIICYITWSPCPNCANELVNFITRNNHLKLEIFASRLYFHWIKSFKMGLQDLQNAGI SVAVMTHTEFEDCWEQFVDNQSRPFQPWDKLEQYSASIRRRLQRILTAPI

[00394] Chimpanzee APOBEC-3 B: 15 MNPQIRNPMEWMYQRTFYYNFENEPILYGRS YTWLCYEVKIRRGHSNLLWDTGVFRGQ MYSQPEHHAEMCFLSWFCGNQLSAYKCFQITWFVSWTPCPDCVAKLAKFLAEHPNVTL TISAARLYYYWERDYRRALCRLSQAGARVKIMDDEEFAYCWENFVYNEGQPFMPWYK FDDNYAFLHRTLKEIIRHLMDPDTFTFNFNNDPLVLRRHQTYLCYEVERLDNGTWVLM DQHMGFLCNEAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGC 20 AGQVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFEYCWDT FVYRQGCPFQPWDGLEEHSQALSGRLRAILQVRASSLCMVPHRPPPPPQSPGPCLPLCSE PPLGSLLPTGRPAPSLPFLLTASFSFPPPASLPPLPSLSLSPGHLPVPSFHSLTSCSIQPPCSSR IRETEGWASVSKEGRDLG

[00395] Human APOBEC-3C: 25 MNPQIRNPMKAMYPGTFYFQFKNLWEANDRNETWLCFTVEGIKRRSVVSWKTGVFRN QVDSETHC7i4JERCFZ5RyCZ)Z)ZL5PA7XFgFrRT7YRYPCPZ)CAGEVAEFLARHSNVNLTI FTARLYYFQYPCYQEGLRSLSQEGVAVEIMDYEDFKYCWENFVYNDNEPFKPWKGLKT NFRLLKRRLRESLQ (italic: nucleic acid editing domain) 30

[00396] Gorilla APOBEC3C 2026204466   11 Jun 2026 MNPQIRNPMKAMYPGTFYFQFKNLWEANDRNETWLCFTVEGIKRRSVVSWKTGVFRN QVDSETHCT^ERCFESlFECDDZESPArAFemPmRYPCF’ECAGEVAEFLARHSNVNLTI FTARLYYFQDTDYQEGLRSLSQEGVAVKIMDYKDFKYCWENFVYNDDEPFKPWKGLK YNFRFLKRRLQEILE 5 (italic: nucleic acid editing domain)

[00397] Human APOBEC-3A: MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFLHNQ A KN LLCG FYG RHA ELR FLDL VPSLQLDPA QIYR VTWFISWSPCFSWGCAGENRAFLQENYH VRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFVDHQGCPFQPWD 10 GLDEHSQALSGRLRAILQNQGN (italic: nucleic acid editing domain)

[00398] Rhesus macaque APOBEC-3A: MDGSPASRPRHLMDPNTFTFNFNNDLSVRGRHQTYLCYEVERLDNGTWVPMDERRGF ECNKAKNVPCGDAGCHVELRFLCEVPSWQLDPAQTYRVTWFISWSPCFRRGCAGQVRVFE 15 QENKHVRLRIFAARIYDYDPLYQEALRTLRDAGAQVSIMTYEEFKHCWDTFVDRQGRP FQPWDGLDEHSQALSGRLRAILQNQGN (italic: nucleic acid editing domain)

[00399] Bovine APOBEC-3A: MDEYTFTENFNNQGWPSKTYLCYEMERLDGDATIPLDEYKGFVRNKGLDQPEKPC7E4E 20 L YFLGKIHSWNLDRNQHYRLTCFISWSPCADCAQKEYYFEKENHH1SEH1EASRTYYWRFG CHQSGLCELQAAGARITIMTFEDFKHCWETFVDHKGKPFQPWEGLNVKSQALCTELQAI LKTQQN (italic: nucleic acid editing domain)

[00400] Human APOBEC-3H: 25 MALLTAETFRLQFNNKRRLRRPYYPRKALLCYQLTPQNGSTPTRGYFENKKKC7E4EZCE7 AEZKWGEDETgCFeFTCFEERNPCSYCAWELVDFIKAHDHLNLGIFASRLYYHWCKPQQ KGLRLLCGSQVPVEVMGFPKFADCWENFVDHEKPLSFNPYKMLEELDKNSRAIKRRLE RIKIPGVRAQGRYMDILCDAEV (italic: nucleic acid editing domain) 30

[00401] Rhesus macaque APOBEC-3H: 2026204466   11 Jun 2026 MALLTAKTFSLQFNNKRRVNKPYYPRKALLCYQLTPQNGSTPTRGHLKNKKKDHAEIR FINKIKSMGLDETQCYQVTCYLTWSPCPSCAGELVDFIKAHRHLNLRIFASRLYYHWRP NYQEGLLLLCGSQVPVEVMGLPEFTDCWENFVDHKEPPSFNPSEKLEELDKNSQAIKRR LERIKSRSVDVLENGLRSLQLGPVTPSSSIRNSR 5

[00402] Human APOBEC-3D: MNPQIRNPMERMYRDTFYDNFENEPILYGRSYTWLCYEVKIKRGRSNLLWDTGVFRGP VLPKRQSNHRQEVYFRFENT^SHCFLSlFFCGARCP^ARRFgZriPFFSWPCLPCVVKVT KFLAEHPNVTLTISAARLYYYRDRDWRWVLLRLHKAGARVKIMDYEDFAYCWENFVC NEGQPFMPWYKFDDNYASLHRTLKEILRNPMEAMYPHIFYFHFKNLLKACGRNESWLC 10 FTMEVTKHHS ANFRKRGVFRNQVDPETHCHAERCFLSWFCDDILSPNTNYEVTWYTSWSP CP£CAGEVAEFLARHSNVNLTIFTARLCYFWDTDYQEGLCSLSQEGASVKIMGYKDFVS CWKNFVYSDDEPFKPWKGLQTNFRLLKRRLREILQ (italic: nucleic acid editing domain)

[00403] Human APOBEC-1 : 15 MTSEKGPSTGDPTLRRRIEPWEFDVFYDPRELRKEACLLYEIKWGMSRKIWRSSGKNTT NHVEVNFIKKFTSERDFHPSMSCSITWFLSWSPCWECSQAIREFLSRHPGVTLVIYVARLF WHMDQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGDEAHWPQYPPLWMM LYALELHCIILSLPPCLKISRRWQNHLTFFRLHLQNCHYQTIPPHILLATGLIHPSVAWR

[00404] Mouse APOBEC-1 : 20 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSVWRHTSQNTSN HVEVNFLEKFTTERYFRPNTRCSITWFLSWSPCGECSRAITEFLSRHPYVTLFIYIARLYH HTDQRNRQGLRDLISSGVTIQIMTEQEYCYCWRNFVNYPPSNEAYWPRYPHLWVKLYV LELYCIILGLPPCLKILRRKQPQLTFFTITLQTCHYQRIPPHLLWATGLK

[00405] Rat APOBEC-1 : 25 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNK HVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHH ADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVL ELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK

[00406] Human APOBEC-2: 30 MAQKEEAAVATEAASQNGEDLENLDDPEKLKELIELPPFEIVTGERLPANFFKFQFRNVE YSSGRNKTFLCYVVEAQGKGGQVQASRGYLEDEHAAAHAEEAFFNTILPAFDPALRYN 2026204466   11 Jun 2026 VTWYVS S SPCAACADRIIKTLSKTKNLRLLILVGRLFMWEEPEIQAALKKLKEAGCKLRI MKPQDFEYVWQNFVEQEEGESKAFQPWEDIQENFLYYEEKLADILK

[00407] Mouse APOBEC-2: MAQKEEAAEAAAPASQNGDDLENLEDPEKLKELIDLPPFEIVTGVRLPVNFFKFQFRNV 5 EYS S GRNKTFLCYVVE VQSKGGQAQ ATQG YLEDEHAGAHAEEAFFNTILPAFDP ALKY NVTWYVSSSPCAACADRILKTLSKTKNLRLLILVSRLFMWEEPEVQAALKKLKEAGCKL RIMKPQDFEYIWQNFVEQEEGESKAFEPWEDIQENFLYYEEKLADILK

[00408] Rat APOBEC-2: MAQKEEAAEAAAPASQNGDDLENLEDPEKLKELIDLPPFEIVTGVRLPVNFFKFQFRNV 10 EYS S GRNKTFLCYVVE AQSKGGQVQ ATQG YLEDEHAGAHAEEAFFNTILPAFDP ALKY NVTWYVSSSPCAACADRILKTLSKTKNLRLLILVSRLFMWEEPEVQAALKKLKEAGCKL RIMKPQDFEYLWQNFVEQEEGESKAFEPWEDIQENFLYYEEKLADILK

[00409] Bovine APOBEC-2: MAQKEEAAAAAEPASQNGEEVENLEDPEKLKELIELPPFEIVTGERLPAHYFKFQFRNVE 15 YSSGRNKTFLCYVVEAQSKGGQVQASRGYLEDEHATNHAEEAFFNSIMPTFDPALRYM VTWYVS SSPCAACADRIVKTLNKTKNLRLLILVGRLFMWEEPEIQAALRKLKEAGCRLR IMKPQDFEYIWQNFVEQEEGESKAFEPWEDIQENFLYYEEKLADILK

[00410] Petromyzon marinus CDA1 (pmCDAl) MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQ 20 SGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHT LKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENR WLEKTLKRAEKRRSELSFMIQVKILHTTKSPAV

[00411] Human APOBEC3G D316R D317R MKPHFRNTVERMYRDTFSYNFYNRPILSRRNTVWLCYEVKTKGPSRPPLDAKIFRGQVY 25 SELKYHPEMRFFHWFSKWRKLHRDQEYEVTWYISWSPCTKCTRDMATFLAEDPKVTLT IFVARLYYFWDPDYQEALRSLCQKRDGPRATMKFNYDEFQHCWSKFVYSQRELFEPWN NLPKYYILLHFMLGEILRHSMDPPTFTFNFNNEPWVRGRHETYLCYEVERMHNDTWVL LNQRRGFLCNQAPHKHGFLEGRHAELCFLDVIPFWKLDLDQDYRVTC FTSWSPCFSCAQEMAKFISKKHVSLCIFTARIYRRQGRCQEGLRTLAEAGAKISFTYSEFK 3 0 HC WDTFVDHQGCPFQP WDGLDEHS QDLS GRLRAILQNQEN

[00412] Human APOBEC3G chain A 2026204466   11 Jun 2026 MDPPTFTFNFNNEPWWGRHETYLCYEVERMHNDTWVLLNQRRGFLCNQAPHKHGFLE GRHAELCFLDVIPFWKLDLDQDYRVTCFTSWSPCFSCAQEMAKFISKNKHVSLCIFTARI YDDQGRCQEGLRTLAEAGAKISFTYSEFKHCWDTFVDHQGCPFQPWDGLD EHSQDLSGRLRAILQ 5

[00413] Human APOBEC3G chain A D120RD121R MDPPTFTFNFNNEPWVRGRHETYLCYEVERMHNDTWVLLNQRRGFLCNQAPHKHGFL EGRHAELCFLDVIPFWKLDLDQDYRVTCFTSWSPCFSCAQEMAKFISKNKHVSLCIFTAR IYRRQGRCQEGLRTLAEAGAKISFMTYSEFKHCWDTFVDHQGCPFQPWDGLDEHSQDL SGRLRAILQ 10

[00414] The term "deaminase" or "deaminase domain" refers to a protein or fragment thereof that catalyzes a deamination reaction. In some embodiments, the deaminase or deaminase domain is a variant of a naturally-occurring deaminase from an organism. In some embodiments, the deaminase or deaminase domain does not occur in nature. For example, in some embodiments, the deaminase or deaminase domain is at least 50%, at least 55%, at least 60%, at 15 least 65%, at least 70%, at least 75% at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or at least 99.5% identical to a naturally-occurring deaminase. In some embodiments, the deaminase is a cytosine deaminase or an adenosine deaminase.

[00415] “Detect” refers to identifying the presence, absence or amount of the analyte to be 20 detected.

[00416] By"detectable label" is meant a composition that when linked to a molecule of interest renders the latter detectable, via spectroscopic, photochemical, biochemical, immunochemical, or chemical means. For example, useful labels include radioactive isotopes, magnetic beads, metallic beads, colloidal particles, fluorescent dyes, electron-dense reagents, enzymes (for 25 example, as commonly used in an ELISA), biotin, digoxigenin, or haptens.

[00417] By “disease” is meant any condition or disorder that damages or interferes with the normal function of a cell, tissue, or organ. In one embodiment, the disease is a neoplasia or cancer (e.g., multiple myeloma).

[00418] The term “effective amount,” as used herein, refers to an amount of a biologically 30 active agent that is sufficient to elicit a desired biological response. In some embodiments, an effective amount of a fusion protein provided herein, e.g., of a cytidine deaminase or an 2026204466   11 Jun 2026 adenosine deaminase nucleobase editor comprising a nCas9 domain and one or more deaminase domains (e.g., cytidine deaminase, adenosine deaminase) may refer to the amount of the fusion protein that is sufficient to induce editing of a target site specifically bound and edited by the cytidine deaminase or adenosine deaminase nucleobase editors. As will be appreciated by the 5 skilled artisan, the effective amount of an agent, e.g., a fusion protein, may vary depending on various factors as, for example, on the desired biological response, e.g., on the specific allele, genome, or target site to be edited, on the cell or tissue being targeted, and on the agent being used. In the context of a CAR-T cell, “an effective amount refers” to the quantity of cells necessary to administer to a patient to achieve a therapeutic response. 10

[00419] In some embodiments, an effective amount of a fusion protein provided herein, e.g., of a fusion protein comprising a nCas9 domain and a cytidine deaminase or adenosine deaminase may refer to the amount of the fusion protein that is sufficient to induce editing of a target site specifically bound and edited by the fusion protein. As will be appreciated by the skilled artisan, the effective amount of an agent, e.g., a fusion protein, a nuclease, a cytidine deaminase or 15 adenosine deaminase, a hybrid protein, a protein dimer, a complex of a protein (or protein dimer) and a polynucleotide, or a polynucleotide, may vary depending on various factors as, for example, on the desired biological response, e.g., on the specific allele, genome, or target site to be edited, on the cell or tissue being targeted, and on the agent being used.

[00420] “Epitope,” as used herein, means an antigenic determinant. An epitope is the part of 20 an antigen molecule that by its structure determines the specific antibody molecule that will recognize and bind it.

[00421] By "fragment" is meant a portion of a polypeptide or nucleic acid molecule. This portion contains, at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90% of the entire length of the reference nucleic acid molecule or polypeptide. A fragment may contain 10, 25   20, 30, 40, 50, 60, 70, 80, 90, or 100, 200, 300, 400, 500, 600, 700, 800, 900, or 1000 nucleotides or amino acids.

[00422] “Graft versus host disease” (GVHD) refers to a pathological condition where transplanted cells of a donor generate an immune response against cells of the host.

[00423] “Host versus graft disease” (HVGD) refers to a pathological condition where the 30 immune system of a host generates an immune response against transplanted cells of a donor. 2026204466   11 Jun 2026

[00424] "Hybridization" means hydrogen bonding, which may be Watson-Crick, Hoogsteen or reversed Hoogsteen hydrogen bonding, between complementary nucleobases. For example, adenine and thymine are complementary nucleobases that pair through the formation of hydrogen bonds. 5

[00425] By “immune cell” is meant a cell of the immune system capable of generating an immune response.

[00426] By “immune effector cell” is meant a lymphocyte, once activated, capable of effecting an immune response upon a target cell. A T cell is an exemplary immune effector cell.

[00427] By “immune response regulation gene” or “immune response regulator” is meant a 10 gene that encodes a polypeptide that is involved in regulation of a immune response. An immune response regulation gene may regulate immune response in multiple mechanisms or on different levels. For example, an immune response regulation gene may inhibit or facilitate the activation of an immune cell, e.g. a T cell. An immune response regulation gene may increase or decrease the activation threshold of a immune cell. In some embodiments, the immune response 15 regulation gene positively regulates an immune cell signal transduction pathway. In some embodiments, the immune response regulation gene negatively regulates an immune cell signal transduction pathway. In some embodiments, the immune response regulation gene encodes an antigen, an antibody, a cytokine, or a neuroendocrine. In some embodiments, the immune response regulation gene encodes a Cblb protein. 20

[00428] By “immunogenic gene” is meant a gene that encodes a polypeptide that is able to elicit an immune response. For example, an immunogenic gene may encode an immunogen that elicits an immune response. In some embodiments, an immunogenic gene encodes a cell surface protein. In some embodiments, an immunogenic gene encodes a cell surface antigen or a cell surface marker. In some embodiments, the cell surface marker is a T cell marker or a B cell 25 marker. In some embodiments, an immunogenic gene encodes a CD2, CD3e, CD3 delta, CD3 gamma, TRAC, TRBC1, TRBC2, CD4, CD5, CD7, CD8, CD19, CD23, CD27, CD28, CD30, CD33, CD52, CD70, CD127, CD122, CD130, CD132, CD38, CD69, CD1 la, CD58, CD99, CD 103, CCR4, CCR5, CCR6, CCR9, CCR10, CXCR3, CXCR4, CLA, CD 161, B2M, or CIITA polypeptide. 30

[00429] The term "inhibitor of base repair" or "IBR" refers to a protein that is capable in inhibiting the activity of a nucleic acid repair enzyme, for example a base excision repair 2026204466   11 Jun 2026 enzyme. In some embodiments, the IBR is an inhibitor of inosine base excision repair. Exemplary inhibitors of base repair include inhibitors of APE1, Endo III, Endo IV, Endo V, Endo VIII, Fpg, hOGGl, hNEILl, T7 Endol, T4PDG, UDG, hSMUGl, and hAAG. In some embodiments, the IBR is an inhibitor of Endo V or hAAG. In some embodiments, the IBR is a 5 catalytically inactive EndoV or a catalytically inactive hAAG.

[00430] The terms "isolated," "purified," or "biologically pure" refer to material that is free to varying degrees from components which normally accompany it as found in its native state. "Isolate" denotes a degree of separation from original source or surroundings. "Purify" denotes a degree of separation that is higher than isolation. A "purified" or "biologically pure" protein is 10    sufficiently free of other materials such that any impurities do not materially affect the biological properties of the protein or cause other adverse consequences. That is, a nucleic acid or peptide of this invention is purified if it is substantially free of cellular material, viral material, or culture medium when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. Purity and homogeneity are typically determined using 15 analytical chemistry techniques, for example, polyacrylamide gel electrophoresis or high-performance liquid chromatography. The term "purified" can denote that a nucleic acid or protein gives rise to essentially one band in an electrophoretic gel. For a protein that can be subjected to modifications, for example, phosphorylation or glycosylation, different modifications may give rise to different isolated proteins, which can be separately purified. 20

[00431] By "isolated polynucleotide" is meant a nucleic acid (e.g., a DNA) that is free of the genes which, in the naturally-occurring genome of the organism from which the nucleic acid molecule of the invention is derived, flank the gene. The term therefore includes, for example, a recombinant DNA that is incorporated into a vector; into an autonomously replicating plasmid or virus; or into the genomic DNA of a prokaryote or eukaryote; or that exists as a separate 25 molecule (for example, a cDNA or a genomic or cDNA fragment produced by PCR or restriction endonuclease digestion) independent of other sequences. In addition, the term includes an RNA molecule that is transcribed from a DNA molecule, as well as a recombinant DNA that is part of a hybrid gene encoding additional polypeptide sequence.

[00432] By an "isolated polypeptide" is meant a polypeptide of the invention that has been 30 separated from components that naturally accompany it. Typically, the polypeptide is isolated when it is at least 60%, by weight, free from the proteins and naturally-occurring organic 2026204466   11 Jun 2026 molecules with which it is naturally associated. Preferably, the preparation is at least 75%, more preferably at least 90%, and most preferably at least 99%, by weight, a polypeptide of the invention. An isolated polypeptide of the invention may be obtained, for example, by extraction from a natural source, by expression of a recombinant nucleic acid encoding such a polypeptide; 5 or by chemically synthesizing the protein. Purity can be measured by any appropriate method, for example, column chromatography, polyacrylamide gel electrophoresis, or by HPLC analysis.

[00433] The term "linker," as used herein, refers to a bond (e.g., covalent bond), chemical group, or a molecule linking two molecules or moieties, e.g., two domains of a fusion protein, such as, for example, a nuclease-inactive Cas9 domain and a nucleic acid-editing domain (e.g., a 10   cytidine deaminase, adenosine deaminase) or in the context of a chimeric antigen receptor, a linker linking a variable heavy (VH) region to a constant heavy (CH) region. In some embodiments, the linker joins two domains of a fusion protein, such as, for example, a nucleaseinactive Cas9 domain and a nucleic acid-editing domain (e.g., a cytidine deaminase, adenosine deaminase). In some embodiments, a linker joins a gRNA binding domain of an RNA- 15 programmable nuclease, including a Cas9 nuclease domain, and the catalytic domain of a nucleic-acid editing protein. In some embodiments, a linker joins a dCas9 and a nucleic-acid editing protein. Typically, the linker is positioned between, or flanked by, two groups, molecules, or other moieties and connected to each one via a covalent bond, thus connecting the two. In some embodiments, the linker is an amino acid or a plurality of amino acids (e.g., a 20 peptide or protein). In some embodiments, the linker is an organic molecule, group, polymer, or chemical moiety. In some embodiments, the linker is 5-100 amino acids in length, for example, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 35, 45, 50, 55, 60, 60, 65, 70, 70, 75, 80, 85, 90, 90, 95, 100, 101, 102, 103, 104, 105, 110, 120, 130, 140, 150, 160, 175, 180, 190, or 200 amino acids in length. Longer or shorter linkers are also contemplated. In some 25 embodiments, a linker comprises the amino acid sequence SGSETPGTSESATPES, which may also be referred to as the XTEN linker. In some embodiments, a linker comprises the amino acid sequence SGGS. In some embodiments, a linker comprises (SGGS)n, (GGGS)n, (GGGGS) n, (G)n, (EAAAK)n, (GGS)n, SGSETPGTSESATPES, or (XP)n motif, or a combination of any of these, wherein n is independently an integer between 1 and 30, and wherein X is any amino acid. 30 In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. 2026204466   11 Jun 2026

[00434] In some embodiments, the chimeric antigen receptor comprises at least one linker. The at least one linker joins, or links, a variable heavy (VH) region to a constant heavy (CH) region of the extracellular binding domain of the chimeric antigen receptor. Linkers can also link a variable light (VL) region to a variable constant (VC) region of the extracellular binding 5 domain.

[00435] In some embodiments, the domains of the cytidine deaminase or adenosine deaminase nucleobase editor are fused via a linker that comprises the amino acid sequence of SGGSSGSETPGTSESATPESSGGS, SGGSSGGSSGSETPGTSESATPESSGGSSGGS, or GGSGGSPGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTE 10 PSEGSAPGTSTEPSEGSAPGTSESATPESGPGSEPATSGGSGGS. In some embodiments, domains of the cytidine deaminase or adenosine deaminase nucleobase editor are fused via a linker comprising the amino acid sequence SGSETPGTSESATPES, which may also be referred to as the XTEN linker. In some embodiments, the linker is 24 amino acids in length. In some embodiments, the linker comprises the amino acid sequence 15 SGGSSGGSSGSETPGTSESATPES. In some embodiments, the linker is 40 amino acids in length. In some embodiments, the linker comprises the amino acid sequence SGGSSGGSSGSETPGTSESATPESSGGSSGGSSGGSSGGS. In some embodiments, the linker is 64 amino acids in length. In some embodiments, the linker comprises the amino acid sequence SGGSSGGSSGSETPGTSESATPESSGGSSGGSSGGSSGGSSGSETPGTSESATPESSGGS 20 SGGS. In some embodiments, the linker is 92 amino acids in length. In some embodiments, the linker comprises the amino acid sequence PGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAP GTSTEPSEGSAPGTSESATPESGPGSEPATS.

[00436] By “marker” is meant any protein or polynucleotide having an alteration in expression 25 level or activity that is associated with a disease or disorder.

[00437] The term “mutation,” as used herein, refers to a substitution of a residue within a sequence, e.g., a nucleic acid or amino acid sequence, with another residue, or a deletion or insertion of one or more residues within a sequence. Mutations are typically described herein by identifying the original residue followed by the position of the residue within the sequence and 30   by the identity of the newly substituted residue. Various methods for making the amino acid substitutions (mutations) provided herein are well known in the art, and are provided by, for 2026204466   11 Jun 2026 example, Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (2012)).

[00438] “Neoplasia” refers to cells or tissues exhibiting abnormal growth or proliferation. The term neoplasia encompasses cancer and solid tumors. 5

[00439] By “nuclear factor of activated T cells 1 (NFATcl) polypeptide” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. NM_172390.2 or a fragment thereof and is a component of the activated T cell DNA-binding transcription complex. An exemplary amino acid sequence is provided below.

[00440] >NP_765978.1 nuclear factor of activated T-cells, cytoplasmic 1 isoform A [Homo 10 sapiens] MPSTSFPVPSKFPLGPAAAVFGRGETLGPAPRAGGTMKSAEEEHYGYASSNVSPALPLPT AHSTLPAPCHNLQTSTPGIIPPADHPSGYGAALDGGPAGYFLSSGHTRPDGAPALESPRIE ITSCLGLYHNNNQFFHDVEVEDVLPSSKRSPSTATLSLPSLEAYRDPSCLSPASSLSSRSC NSEASSYESNYSYPYASPQTSPWQSPCVSPKTTDPEEGFPRGLGACTLLGSPRHSPSTSPR 15 ASVTEESWLGARSSRPASPCNKRKYSLNGRQPPYSPHHSPTPSPHGSPRVSVTDDSWLG NTTQYTSSAIVAAINALTTDSSLDLGDGVPVKSRKTTLEQPPSVALKVEPVGEDLGSPPPP ADFAPEDYSSFQHIRKGGFCDQYLAVPQHPYQWAKPKPLSPTSYMSPTLPALDWQLPSH SGPYELRIEVQPKSHHRAHYETEGSRGAVKASAGGHPIVQLHGYLENEPLMLQLFIGTA DDRLLRPHAFYQVHRITGKTVSTTSHEAILSNTKVLEIPLLPENSMRAVIDCAGILKLRNS 20 DIELRKGETDIGRKNTRVRLVFRVHVPQPSGRTLSLQVASNPIECSQRSAQELPLVEKQST DSYPVVGGKKMVLSGHNFLQDSKVIFVEKAPDGHHVWEMEAKTDRDLCKPNSLVVEIP PFRNQRITSPVHVSFYVCNGKRKRSQYQRFTYLPANGNAIFLTVSREHERVGCFF

[00441] By “nuclear factor of activated T cells 1 (NFATcl) polynucleotide” is meant a nucleic acid molecule encoding a NFATcl polypeptide. The NFATcl gene encodes a protein that is 25 involved in in the inducible expression of cytokine genes, especially IL-2 and IL-4, in T-cells. An exemplary nucleic acid sequenced is provided below.

[00442] >NM_172390.2 Homo sapiens nuclear factor of activated T cells 1 (NFATC1), transcript variant 1, mRNA GGCGGGCGCTCGGCGACTCGTCCCCGGGGCCCCGCGCGGGCCCGGGCAGCAGGGGCGTGAT 30 GTCACGGCA GGGAGGGGGCGCGGGAGCCGCCGGGCCGGCGGGGAGGCGGGGGAGGTGTTTTCCAGCTTTA AAAAGGCAG 2026204466   11 Jun 2026 GAGGCAGAGCGCGGCCCTGCGTCAGAGCGAGACTCAGAGGCTCCGAACTCGCCGGCGGAGT CGCCGCGCC AGATCCCAGCAGCAGGGCGCGGGCACCGGGGCGCGGGCAGGGCTCGGAGCCACCGCGCAG GTCCTAGGGC 5 CGCGGCCGGGCCCCGCCACGCGCGCACACGCCCCTCGATGACTTTCCTCCGGGGCGCGCGGC GCTGAGCC CGGGGCGAGGGCTGTCTTCCCGGAGACCCGACCCCGGCAGCGCGGGGCGGCCGCTTCTCCT GTGCCTCCG CCCGCCGCTCCACTCCCCGCCGCCGCCGCGCGGATGCCAAGCACCAGCTTTCCAGTCCCTTC 10 CAAGTTTC CACTTGGCCCTGCGGCTGCGGTCTTCGGGAGAGGAGAAACTTTGGGGCCCGCGCCGCGCGCC GGCGGCAC CATGAAGTCAGCGGAGGAAGAACACTATGGCTATGCATCCTCCAACGTCAGCCCCGCCCTGC CGCTCCCC 15 ACGGCGCACTCCACCCTGCCGGCCCCGTGCCACAACCTTCAGACCTCCACACCGGGCATCAT CCCGCCGG CGGATCACCCCTCGGGGTACGGAGCAGCTTTGGACGGTGGGCCCGCGGGCTACTTCCTCTCC TCCGGCCA CACCAGGCCTGATGGGGCCCCTGCCCTGGAGAGTCCTCGCATCGAGATAACCTCGTGCTTGG 20 GCCTGTAC CACAACAATAACCAGTTTTTCCACGATGTGGAGGTGGAAGACGTCCTCCCTAGCTCCAAACG GTCCCCCT CCACGGCCACGCTGAGTCTGCCCAGCCTGGAGGCCTACAGAGACCCCTCGTGCCTGAGCCCG GCCAGCAG 25 CCTGTCCTCCCGGAGCTGCAACTCAGAGGCCTCCTCCTACGAGTCCAACTACTCGTACCCGT ACGCGTCC CCCCAGACGTCGCCATGGCAGTCTCCCTGCGTGTCTCCCAAGACCACGGACCCCGAGGAGGG CTTTCCCC GCGGGCTGGGGGCCTGCACACTGCTGGGTTCCCCGCGGCACTCCCCCTCCACCTCGCCCCGC 30 GCCAGCGT CACTGAGGAGAGCTGGCTGGGTGCCCGCTCCTCCAGACCCGCGTCCCCTTGCAACAAGAGG AAGTACAGC CTCAACGGCCGGCAGCCGCCCTACTCACCCCACCACTCGCCCACGCCGTCCCCGCACGGCTC CCCGCGGG 2026204466   11 Jun 2026 TCAGCGTGACCGACGACTCGTGGTTGGGCAACACCACCCAGTACACCAGCTCGGCCATCGTG GCCGCCAT CAACGCGCTGACCACCGACAGCAGCCTGGACCTGGGAGATGGCGTCCCTGTCAAGTCCCGC AAGACCACC 5 CTGGAGCAGCCGCCCTCAGTGGCGCTCAAGGTGGAGCCCGTCGGGGAGGACCTGGGCAGCC CCCCGCCCC CGGCCGACTTCGCGCCCGAAGACTACTCCTCTTTCCAGCACATCAGGAAGGGCGGCTTCTGC GACCAGTA CCTGGCGGTGCCGCAGCACCCCTACCAGTGGGCGAAGCCCAAGCCCCTGTCCCCTACGTCCT 10 ACATGAGC CCGACCCTGCCCGCCCTGGACTGGCAGCTGCCGTCCCACTCAGGCCCGTATGAGCTTCGGAT TGAGGTGC AGCCCAAGTCCCACCACCGAGCCCACTACGAGACGGAGGGCAGCCGGGGGGCCGTGAAGGC GTCGGCCGG 15 AGGACACCCCATCGTGCAGCTGCATGGCTACTTGGAGAATGAGCCGCTGATGCTGCAGCTTT TCATTGGG ACGGCGGACGACCGCCTGCTGCGCCCGCACGCCTTCTACCAGGTGCACCGCATCACAGGGA AGACCGTGT CCACCACCAGCCACGAGGCCATCCTCTCCAACACCAAAGTCCTGGAGATCCCACTCCTGCCG 20 GAGAACAG CATGCGAGCCGTCATTGACTGTGCCGGAATCCTGAAACTCAGAAACTCCGACATTGAACTTC GGAAAGGA GAGACGGACATCGGGAGGAAGAACACACGGGTACGGCTGGTGTTCCGCGTTCACGTCCCGC AACCCAGCG 25 GCCGCACGCTGTCCCTGCAGGTGGCCTCCAACCCCATCGAATGCTCCCAGCGCTCAGCTCAG GAGCTGCC TCTGGTGGAGAAGCAGAGCACGGACAGCTATCCGGTCGTGGGCGGGAAGAAGATGGTCCTG TCTGGCCAC AACTTCCTGCAGGACTCCAAGGTCATTTTCGTGGAGAAAGCCCCAGATGGCCACCATGTCTG 30 GGAGATGG AAGCGAAAACTGACCGGGACCTGTGCAAGCCGAATTCTCTGGTGGTTGAGATCCCGCCATTT CGGAATCA GAGGATAACCAGCCCCGTTCACGTCAGTTTCTACGTCTGCAACGGGAAGAGAAAGCGAAGC CAGTACCAG 2026204466   11 Jun 2026 CGTTTCACCTACCTTCCCGCCAACGGTAACGCCATCTTTCTAACCGTAAGCCGTGAACATGA GCGCGTGG GGTGCTTTTTCTAAAGACGCAGAAACGACGTCGCCGTAAAGCAGCGTGGCGTGTTGCACATT TAACTGTG 5 TGATGTCCCGTTAGTGAGACCGAGCCATCGATGCCCTGAAAAGGAAAGGAAAAGGGAAGCT TCGGATGCA TTTTCCTTGATCCCTGTTGGGGGTGGGGGGCGGGGGTTGCATACTCAGATAGTCACGGTTAT TTTGCTTC TTGCGAATGTATAACAGCCAAGGGGAAAACATGGCTCTTCTGCTCCAAAAAACTGAGGGGG 10 TCCTGGTGT GCATTTGCACCCTAAAGCTGCTTACGGTGAAAAGGCAAATAGGTATAGCTATTTTGCAGGCA CCTTTAGGAATAAACTTTGCTTTTAAGCCTGTAAAAAAAAAAAAAA

[00443] The term “nuclear localization sequence,” “nuclear localization signal,” or “NLS” refers to an amino acid sequence that promotes import of a protein into the cell nucleus. Nuclear 15 localization sequences are known in the art and described, for example, in Plank et al., International PCT application, PCT / EP2000 / 011690, filed November 23, 2000, published as WO / 2001 / 038547 on May 31, 2001, the contents of which are incorporated herein by reference for their disclosure of exemplary nuclear localization sequences. In other embodiments, the NLS is an optimized NLS described, for example, by Koblan et aL, Nature Biotech. 2018 20   doi:10.1038 / nbt.4172. Optimized sequences useful in the methods of the invention are shown at FIGS. 8A-8E and 9. In some embodiments, an NLS comprises the amino acid sequence PKKKRKVEGADKRTADGSEFES PKKKRKV, KRTADGSEFESPKKKRKV, KRPAATKKAGQAKKKK, KKTELQTTNAENKTKKL, KRGINDRNFWRGENGRKTR, RKSGKIAAIVVKRPRK, PKKKRKV, or MDSLLMNRRKFLYQFKNVRWAKGRRETYLC. 25

[00444] The terms “nucleic acid” and “nucleic acid molecule,” as used herein, refer to a compound comprising a nucleobase and an acidic moiety, e.g., a nucleoside, a nucleotide, or a polymer of nucleotides. Typically, polymeric nucleic acids, e.g., nucleic acid molecules comprising three or more nucleotides are linear molecules, in which adjacent nucleotides are linked to each other via a phosphodiester linkage. In some embodiments, “nucleic acid” refers to 30 individual nucleic acid residues (e.g. nucleotides and / or nucleosides). In some embodiments, “nucleic acid” refers to an oligonucleotide chain comprising three or more individual nucleotide residues. As used herein, the terms “oligonucleotide” and “polynucleotide” can be used 2026204466   11 Jun 2026 interchangeably to refer to a polymer of nucleotides (e.g., a string of at least three nucleotides). In some embodiments, “nucleic acid” encompasses RNA as well as single and / or doublestranded DNA. Nucleic acids may be naturally occurring, for example, in the context of a genome, a transcript, an mRNA, tRNA, rRNA, siRNA, snRNA, a plasmid, cosmid, chromosome, 5 chromatid, or other naturally occurring nucleic acid molecule. On the other hand, a nucleic acid molecule may be a non-naturally occurring molecule, e.g., a recombinant DNA or RNA, an artificial chromosome, an engineered genome, or fragment thereof, or a synthetic DNA, RNA, DNA / RNA hybrid, or including non-naturally occurring nucleotides or nucleosides. Furthermore, the terms “nucleic acid,” “DNA,” “RNA,” and / or similar terms include nucleic acid 10 analogs, e.g., analogs having other than a phosphodiester backbone. Nucleic acids can be purified from natural sources, produced using recombinant expression systems and optionally purified, chemically synthesized, etc. Where appropriate, e.g., in the case of chemically synthesized molecules, nucleic acids can comprise nucleoside analogs such as analogs having chemically modified bases or sugars, and backbone modifications. A nucleic acid sequence is 15 presented in the 5' to 3' direction unless otherwise indicated. In some embodiments, a nucleic acid is or comprises natural nucleosides (e.g. adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine); nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5- 20   propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7- deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); intercalated bases; modified sugars (., 2'-e.g.,fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose); and / or modified phosphate groups (e.g., phosphorothioates and 5'-A-phosphoramidite 25 linkages).

[00445] The term "nucleic acid programmable DNA binding protein" or "napDNAbp" refers to a protein that associates with a nucleic acid (e.g., DNA or RNA), such as a guide nucleic acid, that guides the napDNAbp to a specific nucleic acid sequence. For example, a Cas9 protein can associate with a guide RNA that guides the Cas9 protein to a specific DNA sequence that has 30 complementary to the guide RNA. In some embodiments, the napDNAbp, the napDNAbp is a Cas9 domain, for example a nuclease active Cas9, a Cas9 nickase (nCas9), or a nuclease inactive 2026204466   11 Jun 2026 Cas9 (dCas9). Examples of nucleic acid programmable DNA binding proteins include, without limitation, Cas9 (e.g., dCas9 and nCas9), CasX, CasY, Cpfl, Casl2b / C2cl, and Casl2c / C2c3. Other nucleic acid programmable DNA binding proteins are also within the scope of this disclosure, though they may not be specifically listed in this disclosure. 5

[00446] As used herein, “obtaining” as in “obtaining an agent” includes synthesizing, purchasing, or otherwise acquiring the agent.

[00447] By “Programmed cell death 1 (PDCD1 or PD-1) polypeptide” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. AJS10360.1 or a fragment thereof. The PD-1 protein is thought to be involved in T cell function regulation during 10   immune reactions and in tolerance conditions. An exemplary B2M polypeptide sequence is provided below.

[00448] >AJS 10360.1 programmed cell death 1 protein [Homo sapiens] MQIPQAPWPVVWAVLQLGWRPGWFLDSPDRPWNPPTFSPALLVVTEGDNATFTCSFSN TSESFVLNWYRMSPSNQTDKLAAFPEDRSQPGQDCRFRVTQLPNGRDFHMSVVRARRN 15 DSGTYLCGAISLAPKAQIKESLRAELRVTERRAEVPTAHPSPSPRPAGQFQTLVVGVVGG LLGSLVLLVWVLAVICSRAARGTIGARRTGQPLKEDPSAVPVFSVDYGELDFQWREKTP EPPVPCVPEQTEYATIVFPSGMGTSSPARRGSADGPRSAQPLRPEDGHCSWPL

[00449] By “Programmed cell death 1 (PDCD1 or PD-1) polynucleotide” is meant a nucleic acid molecule encoding a PD-1 polypeptide. The PDCD 1 gene encodes an inhibitory cell 20 surface receptor that inhibits T-cell effector functions in an antigen-specific manner. An exemplary PDCD1 nucleic acid sequence is provided below.

[00450] AY238517.1 Homo sapiens programmed cell death 1 (PDCD1) mRNA, complete cds ATGCAGATCCCACAGGCGCCCTGGCCAGTCGTCTGGGCGGTGCTACAACTGGGCTG GCGGCCAGGATGGTTCTTAGACTCCCCAGACAGGCCCTGGAACCCCCCCACCTTCTC 25 CCCAGCCCTGCTCGTGGTGACCGAAGGGGACAACGCCACCTTCACCTGCAGCTTCTC CAACACATCGGAGAGCTTCGTGCTAAACTGGTACCGCATGAGCCCCAGCAACCAGA CGGACAAGCTGGCCGCCTTCCCCGAGGACCGCAGCCAGCCCGGCCAGGACTGCCGC TTCCGTGTCACACAACTGCCCAACGGGCGTGACTTCCACATGAGCGTGGTCAGGGCC CGGCGCAATGACAGCGGCACCTACCTCTGTGGGGCCATCTCCCTGGCCCCCAAGGC 30 GCAGATCAAAGAGAGCCTGCGGGCAGAGCTCAGGGTGACAGAGAGAAGGGCAGAA GTGCCCACAGCCCACCCCAGCCCCTCACCCAGGCCAGCCGGCCAGTTCCAAACCCTG 2026204466   11 Jun 2026 GTGGTTGGTGTCGTGGGCGGCCTGCTGGGCAGCCTGGTGCTGCTAGTCTGGGTCCTG GCCGTCATCTGCTCCCGGGCCGCACGAGGGACAATAGGAGCCAGGCGCACCGGCCA GCCCCTGAAGGAGGACCCCTCAGCCGTGCCTGTGTTCTCTGTGGACTATGGGGAGCT GGATTTCCAGTGGCGAGAGAAGACCCCGGAGCCCCCCGTGCCCTGTGTCCCTGAGC 5 AGACGGAGTATGCCACCATTGTCTTTCCTAGCGGAATGGGCACCTCATCCCCCGCCC GCAGGGGCTCAGCTGACGGCCCTCGGAGTGCCCAGCCACTGAGGCCTGAGGATGGA CACTGCTCTTGGCCCCTCTGA

[00451] The term “recombinant" as used herein in the context of proteins or nucleic acids refers to proteins or nucleic acids that do not occur in nature, but are the product of human 10 engineering. For example, in some embodiments, a recombinant protein or nucleic acid molecule comprises an amino acid or nucleotide sequence that comprises at least one, at least two, at least three, at least four, at least five, at least six, or at least seven mutations as compared to any naturally occurring sequence.

[00452] By “reduces” or “increases” is meant a negative or positive alteration, respectively, of 15 at least 10%, 25%, 50%, 75%, or 100%.

[00453] By “reference” is meant a standard or control condition.

[00454] A "reference sequence" is a defined sequence used as a basis for sequence comparison. A reference sequence may be a subset of or the entirety of a specified sequence; for example, a segment of a full-length cDNA or gene sequence, or the complete cDNA or gene sequence. For 20 polypeptides, the length of the reference polypeptide sequence will generally be at least about 16 amino acids, at least about 20 amino acids, more at least about 25 amino acids, and even more preferably about 35 amino acids, about 50 amino acids, or about 100 amino acids. For nucleic acids, the length of the reference nucleic acid sequence will generally be at least about 50 nucleotides, at least about 60 nucleotides, at least about 75 nucleotides, and about 100 25 nucleotides or about 300 nucleotides or any integer thereabout or therebetween.

[00455] The term "RNA-programmable nuclease," and "RNA-guided nuclease" are used with (e.g., binds or associates with) one or more RNA(s) that is not a target for cleavage. In some embodiments, an RNA-programmable nuclease, when in a complex with an RNA, may be referred to as a nuclease:RNA complex. Typically, the bound RNA(s) is referred to as a guide 30 RNA (gRNA). gRNAs can exist as a complex of two or more RNAs, or as a single RNA molecule. gRNAs that exist as a single RNA molecule may be referred to as single-guide RNAs 2026204466   11 Jun 2026 (sgRNAs), though "gRNA" is used interchangeably to refer to guide RNAs that exist as either single molecules or as a complex of two or more molecules. Typically, gRNAs that exist as single RNA species comprise two domains: (1) a domain that shares homology to a target nucleic acid (e.g., and directs binding of a Cas9 complex to the target); and (2) a domain that 5 binds a Cas9 protein. In some embodiments, domain (2) corresponds to a sequence known as a tracrRNA, and comprises a stem-loop structure. For example, in some embodiments, domain (2) is identical or homologous to a tracrRNA as provided in Jinek et ah, Science 337:816-821(2012), the entire contents of which is incorporated herein by reference. Other examples of gRNAs (e.g., those including domain 2) can be found in U.S. Provisional Patent Application No. 61 / 874,682, 10 filed September 6, 2013, entitled "Switchable Cas9 Nucleases and Uses Thereof," and U.S. Provisional Patent Application, No. 61 / 874,746, filed September 6, 2013, entitled "Delivery System For Functional Nucleases," the entire contents of each are hereby incorporated by reference in their entirety. In some embodiments, a gRNA comprises two or more of domains (1) and (2), and may be referred to as an "extended gRNA." For example, an extended gRNA will, 15   e.g., bind two or more Cas9 proteins and bind a target nucleic acid at two or more distinct regions, as described herein. The gRNA comprises a nucleotide sequence that complements a target site, which mediates binding of the nuclease / RNA complex to said target site, providing the sequence specificity of the nuclease:RNA complex. In some embodiments, the RNA-programmable nuclease is the (CRIS PR-associated system) Cas9 endonuclease, for example, 20 Cas9 (Csnl) from Streptococcus pyogenes (see, e.g., "Complete genome sequence of an Ml strain of Streptococcus pyogenes." Ferretti J.J., McShan W.M., Ajdic D.J., Savic D.J., Savic G., Lyon K., Primeaux C, Sezate S., Suvorov A.N., Kenton S., Lai H.S., Lin S.P., Qian Y., Jia H.G., Najar F.Z., Ren Q., Zhu H., Song L., White J., Yuan X., Clifton S.W., Roe B.A., McLaughlin R.E., Proc. Natl. Acad. Sci. U.S.A. 98:4658-4663(2001); "CRISPR RNA maturation by trans-encoded 25 small RNA and host factor RNase III." Deltcheva E., Chylinski K., Sharma CM., Gonzales K., Chao Y., Pirzada Z.A., Eckert M.R., Vogel J., Charpentier E., Nature 471:602-607(2011).

[00456] By "specifically binds" is meant a nucleic acid molecule, polypeptide, or complex thereof (e.g., a nucleic acid programmable DNA binding protein, a guide nucleic acid, and a chimeric antigen receptor), but which does not substantially recognize and bind other molecules 30 in a sample, for example, a biological sample. For example, a chimeric antigen receptor 2026204466   11 Jun 2026 specifically binds to a particular marker expressed on the surface of a cell, but does not bind to other polypeptides, carbohydrates, lipids, or any other compound on the surface of the cell.

[00457] Nucleic acid molecules useful in the methods of the invention include any nucleic acid molecule that encodes a polypeptide of the invention or a fragment thereof. Such nucleic acid 5 molecules need not be 100% identical with an endogenous nucleic acid sequence, but will typically exhibit substantial identity. Polynucleotides having “substantial identity” to an endogenous sequence are typically capable of hybridizing with at least one strand of a doublestranded nucleic acid molecule. Nucleic acid molecules useful in the methods of the invention include any nucleic acid molecule that encodes a polypeptide of the invention or a fragment 10 thereof. Such nucleic acid molecules need not be 100% identical with an endogenous nucleic acid sequence, but will typically exhibit substantial identity. Polynucleotides having “substantial identity” to an endogenous sequence are typically capable of hybridizing with at least one strand of a double-stranded nucleic acid molecule. By "hybridize" is meant pair to form a doublestranded molecule between complementary polynucleotide sequences (e.g., a gene described 15 herein), or portions thereof, under various conditions of stringency. (See, e.g., Wahl, G. M. and S. L. Berger (1987) Methods Enzymol. 152:399; Kimmel, A. R. (1987) Methods Enzymol. 152:507).

[00458] For example, stringent salt concentration will ordinarily be less than about 750 mM NaCl and 75 mM trisodium citrate, preferably less than about 500 mM NaCl and 50 mM 20 trisodium citrate, and more preferably less than about 250 mM NaCl and 25 mM trisodium citrate. Low stringency hybridization can be obtained in the absence of organic solvent, e.g., formamide, while high stringency hybridization can be obtained in the presence of at least about 35% formamide, and more preferably at least about 50% formamide. Stringent temperature conditions will ordinarily include temperatures of at least about 30° C, more preferably of at least 25 about 37° C, and most preferably of at least about 42° C. Varying additional parameters, such as hybridization time, the concentration of detergent, e.g., sodium dodecyl sulfate (SDS), and the inclusion or exclusion of carrier DNA, are well known to those skilled in the art. Various levels of stringency are accomplished by combining these various conditions as needed. In a one: embodiment, hybridization will occur at 30° C in 750 mM NaCl, 75 mM trisodium citrate, and 30   1% SDS. In another embodiment, hybridization will occur at 37° C in 500 mM NaCl, 50 mM trisodium citrate, 1% SDS, 35% formamide, and 100 pgml denatured salmon sperm DNA 2026204466   11 Jun 2026 (ssDNA). In another embodiment, hybridization will occur at 42° C in 250 mM NaCl, 25 mM trisodium citrate, 1% SDS, 50% formamide, and 200 pg / ml ssDNA. Useful variations on these conditions will be apparent to those skilled in the art.

[00459] For most applications, washing steps that follow hybridization will also vary in 5 stringency. Wash stringency conditions can be defined by salt concentration and by temperature. As above, wash stringency can be increased by decreasing salt concentration or by increasing temperature. For example, stringent salt concentration for the wash steps will preferably be less than about 30 mM NaCl and 3 mM trisodium citrate, and most preferably less than about 15 mM NaCl and 1.5 mM trisodium citrate. Stringent temperature conditions for the wash steps will 10 ordinarily include a temperature of at least about 25° C, more preferably of at least about 42° C, and even more preferably of at least about 68° C. In an embodiment, wash steps will occur at 25° C in 30 mM NaCl, 3 mM trisodium citrate, and 0.1% SDS. In a more preferred embodiment, wash steps will occur at 42 C in 15 mM NaCl, 1.5 mM trisodium citrate, and 0.1% SDS. In a more preferred embodiment, wash steps will occur at 68° C in 15 mM NaCl, 1.5 mM trisodium 15 citrate, and 0.1% SDS. Additional variations on these conditions will be apparent to those skilled in the art. Hybridization techniques are well known to those skilled in the art and are described, for example, in Benton and Davis (Science 196:180, 1977); Grunstein and Hogness (Proc. Natl. Acad. Sci., USA 72:3961, 1975); Ausubel et al. (Current Protocols in Molecular Biology, Wiley Interscience, New York, 2001); Berger and Kimmel (Guide to Molecular 20 Cloning Techniques, 1987, Academic Press, New York); and Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, New York.

[00460] By "subject" is meant a mammal, including, but not limited to, a human or non-human mammal, such as a bovine, equine, canine, ovine, or feline. Subjects include livestock, domesticated animals raised to produce labor and to provide commodities, such as food, 25 including without limitation, cattle, goats, chickens, horses, pigs, rabbits, and sheep.

[00461] By "substantially identical" is meant a polypeptide or nucleic acid molecule exhibiting at least 50% identity to a reference amino acid sequence (for example, any one of the amino acid sequences described herein) or nucleic acid sequence (for example, any one of the nucleic acid sequences described herein). In one embodiment, such a sequence is at least 60%, 80% or 85%, 30   90%, 95% or even 99% identical at the amino acid level or nucleic acid to the sequence used for comparison. 2026204466   11 Jun 2026

[00462] Sequence identity is typically measured using sequence analysis software (for example, Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705, BLAST, BESTFIT, GAP, or PILEUP / PRETTYBOX programs). Such software matches identical or 5 similar sequences by assigning degrees of homology to various substitutions, deletions, and / or other modifications. Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine. In an exemplary approach to determining the degree of identity, a BLAST program may be used, with 10 a probability score between e-3 and e-100 indicating a closely related sequence.

[00463] Because RNA-programmable nucleases (e.g., Cas9) use RNA:DNA hybridization to target DNA cleavage sites, these proteins can be targeted, in principle, to any sequence specified by the guide RNA. Methods of using RNA-programmable nucleases, such as Cas9, for sitespecific cleavage (e.g., to modify a genome) are known in the art (see e.g., Cong, L. et ah, 15 Multiplex genome engineering using CRISPR / Cas systems. Science 339, 819-823 (2013); Mali, P. et ah, RNA-guided human genome engineering via Cas9. Science 339, 823-826 (2013); Hwang, W.Y. et ah, Efficient genome editing in zebrafish using a CRISPR-Cas system. Nature biotechnology 31, 227-229 (2013); Jinek, M. et ah, RNA-programmed genome editing in human cells. eLife 2, e00471 (2013); Dicarlo, J.E. et ah, Genome engineering in Saccharomyces 20 cerevisiae using CRISPR-Cas systems. Nucleic acids research (2013); Jiang, W. et ah RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Nature biotechnology 31, 233239 (2013); the entire contents of each of which are incorporated herein by reference).

[00464] By “tet methylcytosine dioxygenase 2 (TET2) polypeptide” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. FM992369.1 or 25 a fragment thereof and having catalytic activity to convert methyl cytosine to 5- hydroxymethylcytosine. Defects in the gene have been associated with myeloproliferative disorders, and the enzyme’s ability to methylate cytosine contributes to transcriptional regulation. An exemplary TET2 amino acid sequence is provided below. >CAX30492.1 tet oncogene family member 2 [Homo sapiens] 30 MEQDRTNHVEGNRLSPFLIPSPPICQTEPLATKLQNGSPLPERAHPEVNGDTKWHSFKSY YGIPCMKGSQNSRVSPDFTQESRGYSKCLQNGGIKRTVSEPSLSGLLQIKKLKQDQKAN 2026204466   11 Jun 2026 GERRNFGVSQERNPGESSQPNVSDLSDKKESVSSVAQENAVKDFTSFSTHNCSGPENPEL QILNEQEGKSANYHDKNIVLLKNKAVLMPNGATVSASSVEHTHGELLEKTLSQYYPDC VSIAVQKTTSHINAINSQATNELSCEITHPSHTSGQINSAQTSNSELPPKPAAVVSEACDA DDADNASKLAAMLNTCSFQKPEQLQQQKSVFEICPSPAENNIQGTTKLASGEEFCSGSSS 5 NLQAPGGSSERYLKQNEMNGAYFKQSSVFTKDSFSATTTPPPPSQLLLSPPPPLPQVPQLP SEGKSTLNGGVLEEHHHYPNQSNTTLLREVKIEGKPEAPPSQSPNPSTHVCSPSPMLSERP QNNCVNRNDIQTAGTMTVPLCSEKTRPMSEHLKHNPPIFGSSGELQDNCQQLMRNKEQ EILKGRDKEQTRDLVPPTQHYLKPGWIELKAPRFHQAESHLKRNEASLPSILQYQPNLSN QMTSKQYTGNSNMPGGLPRQAYTQKTTQLEHKSQMYQVEMNQGQSQGTVDQHLQFQ 10 KPSHQVHFSKTDHLPKAHVQSLCGTRFHFQQRADSQTEKLMSPVLKQHLNQQASETEPF SNSHLLQHKPHKQAAQTQPSQSSHLPQNQQQQQKLQIKNKEEILQTFPHPQSNNDQQRE GSFFGQTKVEECFHGENQYSKSSEFETHNVQMGLEEVQNINRRNSPYSQTMKSSACKIQ VSCSNNTHLVSENKEQTTHPELFAGNKTQNLHHMQYFPNNVIPKQDLLHRCFQEQEQK SQQASVLQGYKNRNQDMSGQQAAQLAQQRYLIHNHANVFPVPDQGGSHTQTPPQKDT 15 QKHAALRWHLLQKQEQQQTQQPQTESCHSQMHRPIKVEPGCKPHACMHTAPPENKTW KKVTKQENPPASCDNVQQKSIIETMEQHLKQFHAKSLFDHKALTLKSQKQVKVEMSGP VTVLTRQTTAAELDSHTPALEQQTTSSEKTPTKRTAASVLNNFIESPSKLLDTPIKNLLDT PVKTQYDFPSCRCVEQIIEKDEGPFYTHLGAGPNVAAIREIMEERFGQKGKAIRIERVIYT GKEGKSSQGCPIAKWVVRRSSSEEKLLCLVRERAGHTCEAAVIVILILVWEGIPLSLADK 20 LYSELTETLRKYGTLTNRRCALNEERTCACQGLDPETCGASFSFGCSWSMYYNGCKFA RSKIPRKFKLLGDDPKEEEKLESHLQNLSTLMAPTYKKLAPDAYNNQIEYEHRAPECRL GLKEGRPFSGVTACLDFCAHAHRDLHNMQNGSTLVCTLTREDNREFGGKPEDEQLHVL PLYKVSDVDEFGSVEAQEEKKRSGAIQVLSSFRRKVRMLAEPVKTCRQRKLEAKKAAA EKLSSLENSSNKNEKEKSAPSRTKQTENASQAKQLAELLRLSGPVMQQSQQPQPLQKQP 25 PQPQQQQRPQQQQPHHPQTESVNSYSASGSTNPYMRRPNPVSPYPNSSHTSDIYGSTSPM NFYSTSSQAAGSYLNSSNPMNPYPGLLNQNTQYPSYQCNGNLSVDNCSPYLGSYSPQSQ PMDLYRYPSQDPLSKLSLPPIHTLYQPRFGNSQSFTSKYLGYGNQNMQGDGFSSCTIRPN VHHVGKLPPYPTHEMDGHFMGATSRLPPNLSNPNMDYKNGEHHSPSHIIHNYSAAPGM FNSSLHALHLQNKENDMLSHTANGLSKMLPALNHDRTACVQGGLHKLSDANGQEKQP 30 LALVQGVASGAEDNDEVWSDSEQSFLDPDIGGVAVAPTHGSILIECAKRELHATTPLKN 2026204466   11 Jun 2026 PNRNHPTRISLVFYQHKSMNEPKHGLALWEAKMAEKAREKEEECEKYGPDYVPQKSH GKKVKREPAEPHETSEPTYLRFIKSLAERTMSVTTDSTVTTSPYAFTRVTGPYNRYI

[00465] By “tet methylcytosine dioxygenase 2 (TET2) polynucleotide” is meant a nucleic acid molecule encoding a TET2 polypeptide. The TETs polypeptide encodes a methylcytosine 5 dioxygenase and has transcription regulatory activity. An exemplary TET2 nucleic acid is presented below. >FM992369.1 Homo sapiens mRNA for tet oncogene family member 2 (TET2 gene) CCGTGCCATCCCAACCTCCCACCTCGCCCCCAACCTTCGCGCTTGCTCTGCTTCTTCT CCCAGGGGTGGAGACCCGCCGAGGTCCCCGGGGTTCCCGAGGGCTGCACCCTTCCC 10 CGCGCTCGCCAGCCCTGGCCCCTACTCCGCGCTGGTCCGGGCGCACCACTCCCCCCG CGCCACTGCACGGCGTGAGGGCAGCCCAGGTCTCCACTGCGCGCCCCGCTGTACGG CCCCAGGTGCCGCCGGCCTTTGTGCTGGACGCCCGGTGCGGGGGGCTAATTCCCTGG GAGCCGGGGCTGAGGGCCCCAGGGCGGCGGCGCAGGCCGGGGCGGAGCGGGAGGA GGCCGGGGCGGAGCAGGAGGAGGCCCGGGCGGAGGAGGAGAGCCGGCGGTAGCGG 15 CAGTGGCAGCGGCGAGAGCTTGGGCGGCCGCCGCCGCCTCCTCGCGAGCGCCGCGC GCCCGGGTCCCGCTCGCATGCAAGTCACGTCCGCCCCCTCGGCGCGGCCGCCCCGAG ACGCCGGCCCCGCTGAGTGATGAGAACAGACGTCAAACTGCCTTATGAATATTGAT GCGGAGGCTAGGCTGCTTTCGTAGAGAAGCAGAAGGAAGCAAGATGGCTGCCCTTT AGGATTTGTTAGAAAGGAGACCCGACTGCAACTGCTGGATTGCTGCAAGGCTGAGG 20 GACGAGAACGAGGCTGGCAAACATTCAGCAGCACACCCTCTCAAGATTGTTTACTTG CCTTTGCTCCTGTTGAGTTACAACGCTTGGAAGCAGGAGATGGGCTCAGCAGCAGCC AATAGGACATGATCCAGGAAGAGCAAATTCAACTAGAGGGCAGCCTTGTGGATGGC CCCGAAGCAAGCCTGATGGAACAGGATAGAACCAACCATGTTGAGGGCAACAGACT AAGTCCATTCCTGATACCATCACCTCCCATTTGCCAGACAGAACCTCTGGCTACAAA 25 GCTCCAGAATGGAAGCCCACTGCCTGAGAGAGCTCATCCAGAAGTAAATGGAGACA CCAAGTGGCACTCTTTCAAAAGTTATTATGGAATACCCTGTATGAAGGGAAGCCAGA ATAGTCGTGTGAGTCCTGACTTTACACAAGAAAGTAGAGGGTATTCCAAGTGTTTGC AAAATGGAGGAATAAAACGCACAGTTAGTGAACCTTCTCTCTCTGGGCTCCTTCAGA TCAAGAAATTGAAACAAGACCAAAAGGCTAATGGAGAAAGACGTAACTTCGGGGTA 30 AGCCAAGAAAGAAATCCAGGTGAAAGCAGTCAACCAAATGTCTCCGATTTGAGTGA TAAGAAAGAATCTGTGAGTTCTGTAGCCCAAGAAAATGCAGTTAAAGATTTCACCA 2026204466   11 Jun 2026 GTTTTTCAACACATAACTGCAGTGGGCCTGAAAATCCAGAGCTTCAGATTCTGAATG AGCAGGAGGGGAAAAGTGCTAATTACCATGACAAGAACATTGTATTACTTAAAAAC AAGGCAGTGCTAATGCCTAATGGTGCTACAGTTTCTGCCTCTTCCGTGGAACACACA CATGGTGAACTCCTGGAAAAAACACTGTCTCAATATTATCCAGATTGTGTTTCCATT 5 GCGGTGCAGAAAACCACATCTCACATAAATGCCATTAACAGTCAGGCTACTAATGA GTTGTCCTGTGAGATCACTCACCCATCGCATACCTCAGGGCAGATCAATTCCGCACA GACCTCTAACTCTGAGCTGCCTCCAAAGCCAGCTGCAGTGGTGAGTGAGGCCTGTGA TGCTGATGATGCTGATAATGCCAGTAAACTAGCTGCAATGCTAAATACCTGTTCCTT TCAGAAACCAGAACAACTACAACAACAAAAATCAGTTTTTGAGATATGCCCATCTCC 10 TGCAGAAAATAACATCCAGGGAACCACAAAGCTAGCGTCTGGTGAAGAATTCTGTT CAGGTTCCAGCAGCAATTTGCAAGCTCCTGGTGGCAGCTCTGAACGGTATTTAAAAC AAAATGAAATGAATGGTGCTTACTTCAAGCAAAGCTCAGTGTTCACTAAGGATTCCT TTTCTGCCACTACCACACCACCACCACCATCACAATTGCTTCTTTCTCCCCCTCCTCC TCTTCCACAGGTTCCTCAGCTTCCTTCAGAAGGAAAAAGCACTCTGAATGGTGGAGT 15 TTTAGAAGAACACCACCACTACCCCAACCAAAGTAACACAACACTTTTAAGGGAAG TGAAAATAGAGGGTAAACCTGAGGCACCACCTTCCCAGAGTCCTAATCCATCTACA CATGTATGCAGCCCTTCTCCGATGCTTTCTGAAAGGCCTCAGAATAATTGTGTGAAC AGGAATGACATACAGACTGCAGGGACAATGACTGTTCCATTGTGTTCTGAGAAAAC AAGACCAATGTCAGAACACCTCAAGCATAACCCACCAATTTTTGGTAGCAGTGGAG 20 AGCTACAGGACAACTGCCAGCAGTTGATGAGAAACAAAGAGCAAGAGATTCTGAAG GGTCGAGACAAGGAGCAAACACGAGATCTTGTGCCCCCAACACAGCACTATCTGAA ACCAGGATGGATTGAATTGAAGGCCCCTCGTTTTCACCAAGCGGAATCCCATCTAAA ACGTAATGAGGCATCACTGCCATCAATTCTTCAGTATCAACCCAATCTCTCCAATCA AATGACCTCCAAACAATACACTGGAAATTCCAACATGCCTGGGGGGCTCCCAAGGC 25 AAGCTTACACCCAGAAAACAACACAGCTGGAGCACAAGTCACAAATGTACCAAGTT GAAATGAATCAAGGGCAGTCCCAAGGTACAGTGGACCAACATCTCCAGTTCCAAAA ACCCTCACACCAGGTGCACTTCTCCAAAACAGACCATTTACCAAAAGCTCATGTGCA GTCACTGTGTGGCACTAGATTTCATTTTCAACAAAGAGCAGATTCCCAAACTGAAAA ACTTATGTCCCCAGTGTTGAAACAGCACTTGAATCAACAGGCTTCAGAGACTGAGCC 30 ATTTTCAAACTCACACCTTTTGCAACATAAGCCTCATAAACAGGCAGCACAAACACA ACCATCCCAGAGTTCACATCTCCCTCAAAACCAGCAACAGCAGCAAAAATTACAAA 2026204466   11 Jun 2026 TAAAGAATAAAGAGGAAATACTCCAGACTTTTCCTCACCCCCAAAGCAACAATGAT CAGCAAAGAGAAGGATCATTCTTTGGCCAGACTAAAGTGGAAGAATGTTTTCATGG TGAAAATCAGTATTCAAAATCAAGCGAGTTCGAGACTCATAATGTCCAAATGGGAC TGGAGGAAGTACAGAATATAAATCGTAGAAATTCCCCTTATAGTCAGACCATGAAA 5 TCAAGTGCATGCAAAATACAGGTTTCTTGTTCAAACAATACACACCTAGTTTCAGAG AATAAAGAACAGACTACACATCCTGAACTTTTTGCAGGAAACAAGACCCAAAACTT GCATCACATGCAATATTTTCCAAATAATGTGATCCCAAAGCAAGATCTTCTTCACAG GTGCTTTCAAGAACAGGAGCAGAAGTCACAACAAGCTTCAGTTCTACAGGGATATA AAAATAGAAACCAAGATATGTCTGGTCAACAAGCTGCGCAACTTGCTCAGCAAAGG 10 TACTTGATACATAACCATGCAAATGTTTTTCCTGTGCCTGACCAGGGAGGAAGTCAC ACTCAGACCCCTCCCCAGAAGGACACTCAAAAGCATGCTGCTCTAAGGTGGCATCTC TTACAGAAGCAAGAACAGCAGCAAACACAGCAACCCCAAACTGAGTCTTGCCATAG TCAGATGCACAGGCCAATTAAGGTGGAACCTGGATGCAAGCCACATGCCTGTATGC ACACAGCACCACCAGAAAACAAAACATGGAAAAAGGTAACTAAGCAAGAGAATCC 15 ACCTGCAAGCTGTGATAATGTGCAGCAAAAGAGCATCATTGAGACCATGGAGCAGC ATCTGAAGCAGTTTCACGCCAAGTCGTTATTTGACCATAAGGCTCTTACTCTCAAAT CACAGAAGCAAGTAAAAGTTGAAATGTCAGGGCCAGTCACAGTTTTGACTAGACAA ACCACTGCTGCAGAACTTGATAGCCACACCCCAGCTTTAGAGCAGCAAACAACTTCT TCAGAAAAGACACCAACCAAAAGAACAGCTGCTTCTGTTCTCAATAATTTTATAGAG 20 TCACCTTCCAAATTACTAGATACTCCTATAAAAAATTTATTGGATACACCTGTCAAG ACTCAATATGATTTCCCATCTTGCAGATGTGTAGAGCAAATTATTGAAAAAGATGAA GGTCCTTTTTATACCCATCTAGGAGCAGGTCCTAATGTGGCAGCTATTAGAGAAATC ATGGAAGAAAGGTTTGGACAGAAGGGTAAAGCTATTAGGATTGAAAGAGTCATCTA TACTGGTAAAGAAGGCAAAAGTTCTCAGGGATGTCCTATTGCTAAGTGGGTGGTTCG 25 CAGAAGCAGCAGTGAAGAGAAGCTACTGTGTTTGGTGCGGGAGCGAGCTGGCCACA CCTGTGAGGCTGCAGTGATTGTGATTCTCATCCTGGTGTGGGAAGGAATCCCGCTGT CTCTGGCTGACAAACTCTACTCGGAGCTTACCGAGACGCTGAGGAAATACGGCACG CTCACCAATCGCCGGTGTGCCTTGAATGAAGAGAGAACTTGCGCCTGTCAGGGGCTG GATCCAGAAACCTGTGGTGCCTCCTTCTCTTTTGGTTGTTCATGGAGCATGTACTACA 30 ATGGATGTAAGTTTGCCAGAAGCAAGATCCCAAGGAAGTTTAAGCTGCTTGGGGAT GACCCAAAAGAGGAAGAGAAACTGGAGTCTCATTTGCAAAACCTGTCCACTCTTAT 2026204466   11 Jun 2026 GGCACCAACATATAAGAAACTTGCACCTGATGCATATAATAATCAGATTGAATATG AACACAGAGCACCAGAGTGCCGTCTGGGTCTGAAGGAAGGCCGTCCATTCTCAGGG GTCACTGCATGTTTGGACTTCTGTGCTCATGCCCACAGAGACTTGCACAACATGCAG AATGGCAGCACATTGGTATGCACTCTCACTAGAGAAGACAATCGAGAATTTGGAGG 5 AAAACCTGAGGATGAGCAGCTTCACGTTCTGCCTTTATACAAAGTCTCTGACGTGGA TGAGTTTGGGAGTGTGGAAGCTCAGGAGGAGAAAAAACGGAGTGGTGCCATTCAGG TACTGAGTTCTTTTCGGCGAAAAGTCAGGATGTTAGCAGAGCCAGTCAAGACTTGCC GACAAAGGAAACTAGAAGCCAAGAAAGCTGCAGCTGAAAAGCTTTCCTCCCTGGAG AACAGCTCAAATAAAAATGAAAAGGAAAAGTCAGCCCCATCACGTACAAAACAAA 10 CTGAAAACGCAAGCCAGGCTAAACAGTTGGCAGAACTTTTGCGACTTTCAGGACCA GTCATGCAGCAGTCCCAGCAGCCCCAGCCTCTACAGAAGCAGCCACCACAGCCCCA GCAGCAGCAGAGACCCCAGCAGCAGCAGCCACATCACCCTCAGACAGAGTCTGTCA ACTCTTATTCTGCTTCTGGATCCACCAATCCATACATGAGACGGCCCAATCCAGTTA GTCCTTATCCAAACTCTTCACACACTTCAGATATCTATGGAAGCACCAGCCCTATGA 15 ACTTCTATTCCACCTCATCTCAAGCTGCAGGTTCATATTTGAATTCTTCTAATCCCAT GAACCCTTACCCTGGGCTTTTGAATCAGAATACCCAATATCCATCATATCAATGCAA TGGAAACCTATCAGTGGACAACTGCTCCCCATATCTGGGTTCCTATTCTCCCCAGTCT CAGCCGATGGATCTGTATAGGTATCCAAGCCAAGACCCTCTGTCTAAGCTCAGTCTA CCACCCATCCATACACTTTACCAGCCAAGGTTTGGAAATAGCCAGAGTTTTACATCT 20 AAATACTTAGGTTATGGAAACCAAAATATGCAGGGAGATGGTTTCAGCAGTTGTAC CATTAGACCAAATGTACATCATGTAGGGAAATTGCCTCCTTATCCCACTCATGAGAT GGATGGCCACTTCATGGGAGCCACCTCTAGATTACCACCCAATCTGAGCAATCCAAA CATGGACTATAAAAATGGTGAACATCATTCACCTTCTCACATAATCCATAACTACAG TGCAGCTCCGGGCATGTTCAACAGCTCTCTTCATGCCCTGCATCTCCAAAACAAGGA 25 GAATGACATGCTTTCCCACACAGCTAATGGGTTATCAAAGATGCTTCCAGCTCTTAA CCATGATAGAACTGCTTGTGTCCAAGGAGGCTTACACAAATTAAGTGATGCTAATGG TCAGGAAAAGCAGCCATTGGCACTAGTCCAGGGTGTGGCTTCTGGTGCAGAGGACA ACGATGAGGTCTGGTCAGACAGCGAGCAGAGCTTTCTGGATCCTGACATTGGGGGA GTGGCCGTGGCTCCAACTCATGGGTCAATTCTCATTGAGTGTGCAAAGCGTGAGCTG 30 CATGCCACAACCCCTTTAAAGAATCCCAATAGGAATCACCCCACCAGGATCTCCCTC GTCTTTTACCAGCATAAGAGCATGAATGAGCCAAAACATGGCTTGGCTCTTTGGGAA 2026204466   11 Jun 2026 GCCAAAATGGCTGAAAAAGCCCGTGAGAAAGAGGAAGAGTGTGAAAAGTATGGCC CAGACTATGTGCCTCAGAAATCCCATGGCAAAAAAGTGAAACGGGAGCCTGCTGAG CCACATGAAACTTCAGAGCCCACTTACCTGCGTTTCATCAAGTCTCTTGCCGAAAGG ACCATGTCCGTGACCACAGACTCCACAGTAACTACATCTCCATATGCCTTCACTCGG 5 GTCACAGGGCCTTACAACAGATATATATGAAGATATATATGATATCACCCCCTTTTG TTGGTTACCTCACTTGAAAAGACCACAACCAACCTGTCAGTAGTATAGTTCTCATGA CGTGGGCAGTGGGGAAAGGTCACAGTATTCATGACAAATGTGGTGGGAAAAACCTC AGCTCACCAGCAACAAAAGAGGTTATCTTACCATAGCACTTAATTTTCACTGGCTCC CAAGTGGTCACAGATGGCATCTAGGAAAAGACCAAAGCATTCTATGCAAAAAGAAG 10 GTGGGGAAGAAAGTGTTCCGCAATTTACATTTTTAAACACTGGTTCTATTATTGGAC GAGATGATATGTAAATGTGATCCCCCCCCCCCGCTTACAACTCTACACATCTGTGAC CACTTTTAATAATATCAAGTTTGCATAGTCATGGAACACAAATCAAACAAGTACTGT AGTATTACAGTGACAGGAATCTTAAAATACCATCTGGTGCTGAATATATGATGTACT GAAATACTGGAATTATGGCTTTTTGAAATGCAGTTTTTACTGTAATCTTAACTTTTAT 15 TTATCAAAATAGCTACAGGAAACATGAATAGCAGGAAAACACTGAATTTGTTTGGA TGTTCTAAGAAATGGTGCTAAGAAAATGGTGTCTTTAATAGCTAAAAATTTAATGCC TTTATATCATCAAGATGCTATCAGTGTACTCCAGTGCCCTTGAATAATAGGGGTACC TTTTCATTCAAGTTTTTATCATAATTACCTATTCTTACACAAGCTTAGTTTTTAAAATG TGGACATTTTAAAGGCCTCTGGATTTTGCTCATCCAGTGAAGTCCTTGTAGGACAAT 20 AAACGTATATATGTACATATATACACAAACATGTATATGTGCACACACATGTATATG TATAAATATTTTAAATGGTGTTTTAGAAGCACTTTGTCTACCTAAGCTTTGACAACTT GAACAATGCTAAGGTACTGAGATGTTTAAAAAACAAGTTTACTTTCATTTTAGAATG CAAAGTTGATTTTTTTAAGGAAACAAAGAAAGCTTTTAAAATATTTTTGCTTTTAGCC ATGCATCTGCTGATGAGCAATTGTGTCCATTTTTAACACAGCCAGTTAAATCCACCA 25 TGGGGCTTACTGGATTCAAGGGAATACGTTAGTCCACAAAACATGTTTTCTGGTGCT CATCTCACATGCTATACTGTAAAACAGTTTTATACAAAATTGTATGACAAGTTCATT GCTCAAAAATGTACAGTTTTAAGAATTTTCTATTAACTGCAGGTAATAATTAGCTGC ATGCTGCAGACTCAACAAAGCTAGTTCACTGAAGCCTATGCTATTTTATGGATCATA GGCTCTTCAGAGAACTGAATGGCAGTCTGCCTTTGTGTTGATAATTATGTACATTGT 30 GACGTTGTCATTTCTTAGCTTAAGTGTCCTCTTTAACAAGAGGATTGAGCAGACTGA TGCCTGCATAAGATGAATAAACAGGGTTAGTTCCATGTGAATCTGTCAGTTAAAAAG 2026204466   11 Jun 2026 AAACAAAAACAGGCAGCTGGTTTGCTGTGGTGGTTTTAAATCATTAATTTGTATAAA GAAGTGAAAGAGTTGTATAGTAAATTAAATTGTAAACAAAACTTTTTTAATGCAATG CTTTAGTATTTTAGTACTGTAAAAAAATTAAATATATACATATATATATATATATATA TATATATATATATGAGTTTGAAGCAGAATTCACATCATGATGGTGCTACTCAGCCTG 5 CTACAAATATATCATAATGTGAGCTAAGAATTCATTAAATGTTTGAGTGATGTTCCT ACTTGTCATATACCTCAACACTAGTTTGGCAATAGGATATTGAACTGAGAGTGAAAG CATTGTGTACCATCATTTTTTTCCAAGTCCTTTTTTTTATTGTTAAAAAAAAAAGCAT ACCTTTTTTCAATACTTGATTTCTTAGCAAGTATAACTTGAACTTCAACCTTTTTGTTC TAAAAATTCAGGGATATTTCAGCTCATGCTCTCCCTATGCCAACATGTCACCTGTGTT 10 TATGTAAAATTGTTGTAGGTTAATAAATATATTCTTTGTCAGGGATTTAACCCTTTTA TTTTGAATCCCTTCTATTTTACTTGTACATGTGCTGATGTAACTAAAACTAATTTTGT AAATCTGTTGGCTCTTTTTATTGTAAAGAAAAGCATTTTAAAAGTTTGAGGAATCTTT TGACTGTTTCAAGCAGGAAAAAAAAATTACATGAAAATAGAATGCACTGAGTTGAT AAAGGGAAAAATTGTAAGGCAGGAGTTTGGCAAGTGGCTGTTGGCCAGAGACTTAC 15 TTGTAACTCTCTAAATGAAGTTTTTTTGATCCTGTAATCACTGAAGGTACATACTCCA TGTGGACTTCCCTTAAACAGGCAAACACCTACAGGTATGGTGTGCAACAGATTGTAC AATTACATTTTGGCCTAAATACATTTTTGCTTACTAGTATTTAAAATAAATTCTTAAT CAGAGGAGGCCTTTGGGTTTTATTGGTCAAATCTTTGTAAGCTGGCTTTTGTCTTTTT AAAAAATTTCTTGAATTTGTGGTTGTGTCCAATTTGCAAACATTTCCAAAAATGTTTG 20 CTTTGCTTACAAACCACATGATTTTAATGTTTTTTGTATACCATAATATCTAGCCCCA AACATTTGATTACTACATGTGCATTGGTGATTTTGATCATCCATTCTTAATATTTGAT TTCTGTGTCACCTACTGTCATTTGTTAAACTGCTGGCCAACAAGAACAGGAAGTATA GTTTGGGGGGTTGGGGAGAGTTTACATAAGGAAGAGAAGAAATTGAGTGGCATATT GTAAATATCAGATCTATAATTGTAAATATAAAACCTGCCTCAGTTAGAATGAATGGA 25 AAGCAGATCTACAATTTGCTAATATAGGAATATCAGGTTGACTATATAGCCATACTT GAAAATGCTTCTGAGTGGTGTCAACTTTACTTGAATGAATTTTTCATCTTGATTGACG CACAGTGATGTACAGTTCACTTCTGAAGCTAGTGGTTAACTTGTGTAGGAAACTTTT GCAGTTTGACACTAAGATAACTTCTGTGTGCATTTTTCTATGCTTTTTTAAAAACTAG TTTCATTTCATTTTCATGAGATGTTTGGTTTATAAGATCTGAGGATGGTTATAAATAC 30 TGTAAGTATTGTAATGTTATGAATGCAGGTTATTTGAAAGCTGTTTATTATTATATCA 2026204466   11 Jun 2026 TTCCTGATAATGCTATGTGAGTGTTTTTAATAAAATTTATATTTATTTAATGCACTCT AAGTGTTGTCTTCCT

[00466] By “transforming growth factor receptor 2 (TGFBRII) polypeptide” is meant a protein having at least about 85% sequence identity to NCBI Accession No. ABG65632.1 or a fragment 5 thereof and having immunosuppressive activity. An exemplary amino acid sequence is provided below. >ABG65632.1 transforming growth factor beta receptor II [Homo sapiens] MGRGLLRGLWPLHIVLWTRIASTIPPHVQKSVNNDMIVTDNNGAVKFPQLCKFCDVRFS TCDNQKSCMSNCSITSICEKPQEVCVAVWRKNDENITLETVCHDPKLPYHDFILEDAASP 10 KCIMKEKKKPGETFFMCSCSSDECNDNIIFSEEYNTSNPDLLLVIFQVTGISLLPPLGVAIS VIIIFYCYRVNRQQKLSSTWETGKTRKLMEFSEHCAIILEDDRSDISSTCANNINHNTELLP IELDTLVGKGRFAEVYKAKLKQNTSEQFETVAVKIFPYEEYASWKTEKDIFSDINLKHEN ILQFLTAEERKTELGKQYWLITAFHAKGNLQEYLTRHVISWEDLRKLGSSLARGIAHLHS DHTPCGRPKMPIVHRDLKSSNILVKNDLTCCLCDFGLSLRLDPTLSVDDLANSGQVGTA 15 RYMAPEVLESRMNLENVESFKQTDVYSMALVLWEMTSRCNAVGEVKDYEPPFGSKVR EHPCVESMKDNVLRDRGRPEIPSFWLNHQGIQMVCETLTECWDHDPEARLTAQCVAER FSELEHLDRLSGRSCSEEKIPEDGSLNTTK

[00467] By “transforming growth factor receptor 2 (TGFBRII) polynucleotide” is meant a nucleic acid that encodes a TGFBRII polypeptide. The TGFBRII gene encodes a transmembrane 20 protein having serine / threonine kinase activity. An exemplary TGFBRII nucleic acid is provided below. >M85079.1 Human TGF-beta type II receptor mRNA, complete cds GTTGGCGAGGAGTTTCCTGTTTCCCCCGCAGCGCTGAGTTGAAGTTGAGTGAGTCAC TCGCGCGCACGGAGCGACGACACCCCCGCGCGTGCACCCGCTCGGGACAGGAGCCG 25 GACTCCTGTGCAGCTTCCCTCGGCCGCCGGGGGCCTCCCCGCGCCTCGCCGGCCTCC AGGCCCCTCCTGGCTGGCGAGCGGGCGCCACATCTGGCCCGCACATCTGCGCTGCCG GCCCGGCGCGGGGTCCGGAGAGGGCGCGGCGCGGAGCGCAGCCAGGGGTCCGGGA AGGCGCCGTCCGTGCGCTGGGGGCTCGGTCTATGACGAGCAGCGGGGTCTGCCATG GGTCGGGGGCTGCTCAGGGGCCTGTGGCCGCTGCACATCGTCCTGTGGACGCGTATC 30 GCCAGCACGATCCCACCGCACGTTCAGAAGTCGGTTAATAACGACATGATAGTCAC TGACAACAACGGTGCAGTCAAGTTTCCACAACTGTGTAAATTTTGTGATGTGAGATT 2026204466   11 Jun 2026 TTCCACCTGTGACAACCAGAAATCCTGCATGAGCAACTGCAGCATCACCTCCATCTG TGAGAAGCCACAGGAAGTCTGTGTGGCTGTATGGAGAAAGAATGACGAGAACATAA CACTAGAGACAGTTTGCCATGACCCCAAGCTCCCCTACCATGACTTTATTCTGGAAG ATGCTGCTTCTCCAAAGTGCATTATGAAGGAAAAAAAAAAGCCTGGTGAGACTTTCT 5 TCATGTGTTCCTGTAGCTCTGATGAGTGCAATGACAACATCATCTTCTCAGAAGAAT ATAACACCAGCAATCCTGACTTGTTGCTAGTCATATTTCAAGTGACAGGCATCAGCC TCCTGCCACCACTGGGAGTTGCCATATCTGTCATCATCATCTTCTACTGCTACCGCGT TAACCGGCAGCAGAAGCTGAGTTCAACCTGGGAAACCGGCAAGACGCGGAAGCTCA TGGAGTTCAGCGAGCACTGTGCCATCATCCTGGAAGATGACCGCTCTGACATCAGCT 10 CCACGTGTGCCAACAACATCAACCACAACACAGAGCTGCTGCCCATTGAGCTGGAC ACCCTGGTGGGGAAAGGTCGCTTTGCTGAGGTCTATAAGGCCAAGCTGAAGCAGAA CACTTCAGAGCAGTTTGAGACAGTGGCAGTCAAGATCTTTCCCTATGAGGAGTATGC CTCTTGGAAGACAGAGAAGGACATCTTCTCAGACATCAATCTGAAGCATGAGAACA TACTCCAGTTCCTGACGGCTGAGGAGCGGAAGACGGAGTTGGGGAAACAATACTGG 15 CTGATCACCGCCTTCCACGCCAAGGGCAACCTACAGGAGTACCTGACGCGGCATGT CATCAGCTGGGAGGACCTGCGCAAGCTGGGCAGCTCCCTCGCCCGGGGGATTGCTC ACCTCCACAGTGATCACACTCCATGTGGGAGGCCCAAGATGCCCATCGTGCACAGG GACCTCAAGAGCTCCAATATCCTCGTGAAGAACGACCTAACCTGCTGCCTGTGTGAC TTTGGGCTTTCCCTGCGTCTGGACCCTACTCTGTCTGTGGATGACCTGGCTAACAGTG 20 GGCAGGTGGGAACTGCAAGATACATGGCTCCAGAAGTCCTAGAATCCAGGATGAAT TTGGAGAATGCTGAGTCCTTCAAGCAGACCGATGTCTACTCCATGGCTCTGGTGCTC TGGGAAATGACATCTCGCTGTAATGCAGTGGGAGAAGTAAAAGATTATGAGCCTCC ATTTGGTTCCAAGGTGCGGGAGCACCCCTGTGTCGAAAGCATGAAGGACAACGTGT TGAGAGATCGAGGGCGACCAGAAATTCCCAGCTTCTGGCTCAACCACCAGGGCATC 25 CAGATGGTGTGTGAGACGTTGACTGAGTGCTGGGACCACGACCCAGAGGCCCGTCT CACAGCCCAGTGTGTGGCAGAACGCTTCAGTGAGCTGGAGCATCTGGACAGGCTCT CGGGGAGGAGCTGCTCGGAGGAGAAGATTCCTGAAGACGGCTCCCTAAACACTACC AAATAGCTCTTATGGGGCAGGCTGGGCATGTCCAAAGAGGCTGCCCCTCTCACCAA A 30

[00468] By “T Cell Immunoreceptor with Ig and ITIM Domains (TIGIT) polypeptide” is meant a protein having at least about 85% sequence identity to NCBI Accession No. 2026204466   11 Jun 2026 ACD74757.1 or a fragment thereof and having immunomodulatory activity. An exemplary TIGIT amino acid sequence is provided below. >ACD74757.1 T cell immunoreceptor with Ig and ITIM domains [Homo sapiens] MRWCLLLIWAQGLRQAPLASGMMTGTIETTGNISAEKGGSIILQCHLSSTTAQVTQVNW 5 EQQDQLLAICNADLGWHISPSFKDRVAPGPGLGLTLQSLTVNDTGEYFCIYHTYPDGTY TGRIFLEVLESSVAEHGARFQIPLLGAMAATLVVICTAVIVVVALTRKKKALRIHSVEGD LRRKSAGQEEWSPSAPSPPGSCVQAEAAPAGLCGEQRGEDCAELHDYFNVLSYRSLGN CSFFTETG

[00469] By “T Cell Immunoreceptor With Ig And ITIM Domains (TIGIT) 10 polynucleotide” is meant a nucleic acid encoding a TIGIT polypeptide. The TIGIT gene encodes an inhibitory immune receptor that is associated with neoplasia and T cell exhaustion. An exemplary nucleic acid sequence is provided below. >EU675310.1 Homo sapiens T cell immunoreceptor with Ig and ITIM domains (TIGIT) mRNA, complete cds 15 CGTCCTATCTGCAGTCGGCTACTTTCAGTGGCAGAAGAGGCCACATCTGCTTCCTGT AGGCCCTCTGGGCAGAAGCATGCGCTGGTGTCTCCTCCTGATCTGGGCCCAGGGGCT GAGGCAGGCTCCCCTCGCCTCAGGAATGATGACAGGCACAATAGAAACAACGGGGA ACATTTCTGCAGAGAAAGGTGGCTCTATCATCTTACAATGTCACCTCTCCTCCACCA CGGCACAAGTGACCCAGGTCAACTGGGAGCAGCAGGACCAGCTTCTGGCCATTTGT 20 AATGCTGACTTGGGGTGGCACATCTCCCCATCCTTCAAGGATCGAGTGGCCCCAGGT CCCGGCCTGGGCCTCACCCTCCAGTCGCTGACCGTGAACGATACAGGGGAGTACTTC TGCATCTATCACACCTACCCTGATGGGACGTACACTGGGAGAATCTTCCTGGAGGTC CTAGAAAGCTCAGTGGCTGAGCACGGTGCCAGGTTCCAGATTCCATTGCTTGGAGCC ATGGCCGCGACGCTGGTGGTCATCTGCACAGCAGTCATCGTGGTGGTCGCGTTGACT 25 AGAAAGAAGAAAGCCCTCAGAATCCATTCTGTGGAAGGTGACCTCAGGAGAAAATC AGCTGGACAGGAGGAATGGAGCCCCAGTGCTCCCTCACCCCCAGGAAGCTGTGTCC AGGCAGAAGCTGCACCTGCTGGGCTCTGTGGAGAGCAGCGGGGAGAGGACTGTGCC GAGCTGCATGACTACTTCAATGTCCTGAGTTACAGAAGCCTGGGTAACTGCAGCTTC TTCACAGAGACTGGTTAGCAACCAGAGGCATCTTCTGG 30

[00470] By “T Cell Receptor Alpha Constant (TRAC) polypeptide” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. P01848.2 or fragment 2026204466   11 Jun 2026 thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. >sp|P01848.2|TRAC_HUMAN RecName: Full=T cell receptor alpha constant IQNPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSN 5 SAVAWSNKSDFACANAFNNSIIPEDTFFPSPESSCDVKLVEKSFETDTNLNFQNLSVIGFR ILLLKVAGFNLLMTLRLWSS

[00471] By “T Cell Receptor Alpha Constant (TRAC) polynucleotide” is meant a nucleic acid encoding a TRAC polypeptide. Exemplary TRAC nucleic acid sequences are provided below. 10 UCSC human genome database, Gene ENSG00000277734.8 Human T-cell receptor alpha chain (TCR-alpha) catgctaatcctccggcaaacctctgtttcctcctcaaaaggcaggaggtcggaaagaataaacaatgagagtcacattaaaaacacaaaat cctacggaaatactgaagaatgagtctcagcactaaggaaaagcctccagcagctcctgctttctgagggtgaaggatagacgctgtggct ctgcatgactcactagcactctatcacggccatattctggcagggtcagtggctccaactaacatttgtttggtactttacagtttattaaatagat 15 gtttatatggagaagctctcatttctttctcagaagagcctggctaggaaggtggatgaggcaccatattcattttgcaggtgaaattcctgaga tgtaaggagctgctgtgacttgctcaaggccttatatcgagtaaacggtagtgctggggcttagacgcaggtgttctgatttatagttcaaaac ctctatcaatgagagagcaatctcctggtaatgtgatagatttcccaacttaatgccaacataccataaacctcccattctgctaatgcccagcc taagttggggagaccactccagattccaagatgtacagtttgctttgctgggcctttttcccatgcctgcctttactctgccagagttatattgctg gggttttgaagaagatcctattaaataaaagaataagcagtattattaagtagccctgcatttcaggtttccttgagtggcaggccaggcctgg 20 ccgtgaacgttcactgaaatcatggcctcttggccaagattgatagcttgtgcctgtccctgagtcccagtccatcacgagcagctggtttcta agatgctatttcccgtataaagcatgagaccgtgacttgccagccccacagagccccgcccttgtccatcactggcatctggactccagcct gggttggggcaaagagggaaatgagatcatgtcctaaccctgatcctcttgtcccacagATATCCAGAACCCTGACCCT GCCGTGTACCAGCTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACC GATTTTGATTCTCAAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACA 25 GACAAAACTGTGCTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGC CTGGAGCAACAAATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCC AGAAGACACCTTCTTCCCCAGCCCAGgtaagggcagctttggtgccttcgcaggctgtttccttgcttcaggaatgg ccaggttctgcccagagctctggtcaatgatgtctaaaactcctctgattggtggtctcggccttatccattgccaccaaaaccctctttttacta agaaacagtgagccttgttctggcagtccagagaatgacacgggaaaaaagcagatgaagagaaggtggcaggagagggcacgtggc 30 ccagcctcagtctctccaactgagttcctgcctgcctgcctttgctcagactgtttgccccttactgctcttctaggcctcattctaagccccttct ccaagttgcctctccttatttctccctgtctgccaaaaaatctttcccagctcactaagtcagtctcacgcagtcactcattaacccaccaatcact 2026204466   11 Jun 2026 gattgtgccggcacatgaatgcaccaggtgttgaagtggaggaattaaaaagtcagatgaggggtgtgcccagaggaagcaccattctagt tgggggagcccatctgtcagctgggaaaagtccaaataacttcagattggaatgtgttttaactcagggttgagaaaacagctaccttcagga caaaagtcagggaagggctctctgaagaaatgctacttgaagataccagccctaccaagggcagggagaggaccctatagaggcctggg acaggagctcaatgagaaaggagaagagcagcaggcatgagttgaatgaaggaggcagggccgggtcacagggccttctaggccatg 5 agagggtagacagtattctaaggacgccagaaagctgttgatcggcttcaagcaggggagggacacctaatttgcttttcttttttttttttttttttt tttttttttttgagatggagttttgctcttgttgcccaggctggagtgcaatggtgcatcttggctcactgcaacctccgcctcccaggttcaagtg attctcctgcctcagcctcccgagtagctgagattacaggcacccgccaccatgcctggctaattttttgtatttttagtagagacagggtttcac tatgttggccaggctggtctcgaactcctgacctcaggtgatccacccgcttcagcctcccaaagtgctgggattacaggcgtgagccacca cacccggcctgcttttcttaaagatcaatctgagtgctgtacggagagtgggttgtaagccaagagtagaagcagaaagggagcagttgca 10 gcagagagatgatggaggcctgggcagggtggtggcagggaggtaaccaacaccattcaggtttcaaaggtagaaccatgcagggatg agaaagcaaagaggggatcaaggaaggcagctggattttggcctgagcagctgagtcaatgatagtgccgtttactaagaagaaaccaag gaaaaaatttggggtgcagggatcaaaactttttggaacatatgaaagtacgtgtttatactctttatggcccttgtcactatgtatgcctcgctgc ctccattggactctagaatgaagccaggcaagagcagggtctatgtgtgatggcacatgtggccagggtcatgcaacatgtactttgtacaa acagtgtatattgagtaaatagaaatggtgtccaggagccgaggtatcggtcctgccagggccaggggctctccctagcaggtgctcatatg 15 ctgtaagttccctccagatctctccacaaggaggcatggaaaggctgtagttgttcacctgcccaagaactaggaggtctggggtgggaga gtcagcctgctctggatgctgaaagaatgtctgtttttccttttagAAAGTTCCTGTGATGTCAAGCTGGTCGAGA AAAGCTTTGAAACAGgtaagacaggggtctagcctgggtttgcacaggattgcggaagtgatgaacccgcaataaccctgc ctggatgagggagtgggaagaaattagtagatgtgggaatgaatgatgaggaatggaaacagcggttcaagacctgcccagagctgggt ggggtctctcctgaatccctctcaccatctctgactttccattctaagcactttgaggatgagtttctagcttcaatagaccaaggactctctccta 20 ggcctctgtattcctttcaacagctccactgtcaagagagccagagagagcttctgggtggcccagctgtgaaatttctgagtcccttagggat agccctaaacgaaccagatcatcctgaggacagccaagaggttttgccttctttcaagacaagcaacagtactcacataggctgtgggcaat ggtcctgtctctcaagaatcccctgccactcctcacacccaccctgggcccatattcatttccatttgagttgttcttattgagtcatccttcctgtg gtagcggaactcactaaggggcccatctggacccgaggtattgtgatgataaattctgagcacctaccccatccccagaagggctcagaaa taaaataagagccaagtctagtcggtgtttcctgtcttgaaacacaatactgttggccctggaagaatgcacagaatctgtttgtaaggggatat 25 gcacagaagctgcaagggacaggaggtgcaggagctgcaggcctcccccacccagcctgctctgccttggggaaaaccgtgggtgtgt cctgcaggccatgcaggcctgggacatgcaagcccataaccgctgtggcctcttggttttacagATACGAACCTAAACTTT CAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCTGAAAGTGGCCGGGTTTAAT CTGCTCATGACGCTGCGGCTGTGGTCCAGCTGAGgtgaggggccttgaagctgggagtggggtttaggga cgcgggtctctgggtgcatcctaagctctgagagcaaacctccctgcagggtcttgcttttaagtccaaagcctgagcccaccaaactctcct 30 acttcttcctgttacaaattcctcttgtgcaataataatggcctgaaacgctgtaaaatatcctcatttcagccgcctcagttgcacttctcccctat gaggtaggaagaacagttgtttagaaacgaagaaactgaggccccacagctaatgagtggaggaagagagacacttgtgtacaccacatg 2026204466   11 Jun 2026 ccttgtgttgtacttctctcaccgtgtaacctcctcatgtcctctctccccagtacggctctcttagctcagtagaaagaagacattacactcatatt acaccccaatcctggctagagtctccgcaccctcctcccccagggtccccagtcgtcttgctgacaactgcatcctgttccatcaccatcaaa aaaaaactccaggctgggtgcgggggctcacacctgtaatcccagcactttgggaggcagaggcaggaggagcacaggagctggaga ccagcctgggcaacacagggagaccccgcctctacaaaaagtgaaaaaattaaccaggtgtggtgctgcacacctgtagtcccagctactt 5 aagaggctgagatgggaggatcgcttgagccctggaatgttgaggctacaatgagctgtgattgcgtcactgcactccagcctggaagaca aagcaagatcctgtctcaaataataaaaaaaataagaactccagggtacatttgctcctagaactctaccacatagccccaaacagagccatc accatcacatccctaacagtcctgggtcttcctcagtgtccagcctgacttctgttcttcctcattccagATCTGCAAGATTGTAA GACAGCCTGTGCTCCCTCGCTCCTTCCTCTGCATTGCCCCTCTTCTCCCTCTCCAAAC AGAGGGAACTCTCCTACCCCCAAGGAGGTGAAAGCTGCTACCACCTCTGTGCCCCCC 10 CGGCAATGCCACCAACTGGATCCTACCCGAATTTATGATTAAGATTGCTGAAGAGCT GCCAAACACTGCTGCCACCCCCTCTGTTCCCTTATTGCTGCTTGTCACTGCCTGACAT TCACGGCAGAGGCAAGGCTGCTGCAGCCTCCCCTGGCTGTGCACATTCCCTCCTGCT CCCCAGAGACTGCCTCCGCCATCCCACAGATGATGGATCTTCAGTGGGTTCTCTTGG GCTCTAGGTCCTGCAGAATGTTGTGAGGGGTTTATTTTTTTTTAATAGTGTTCATAAA 15 GAAATACATAGTATTCTTCTTCTCAAGACGTGGGGGGAAATTATCTCATTATCGAGG CCCTGCTATGCTGTGTATCTGGGCGTGTTGTATGTCCTGCTGCCGATGCCTTCATTAA AATGATTTGGAAGAGCAGA

[00472] Nucleotides in lower cases above are untranslated regions or introns, and nucleotides in upper cases are exons. 20

[00473] >X02592.1 Human mRNA for T-cell receptor alpha chain (TCR-alpha) TTTTGAAACCCTTCAAAGGCAGAGACTTGTCCAGCCTAACCTGCCTGCTGCTCCTAG CTCCTGAGGCTCAGGGCCCTTGGCTTCTGTCCGCTCTGCTCAGGGCCCTCCAGCGTG GCCACTGCTCAGCCATGCTCCTGCTGCTCGTCCCAGTGCTCGAGGTGATTTTTACCCT GGGAGGAACCAGAGCCCAGTCGGTGACCCAGCTTGGCAGCCACGTCTCTGTCTCTG 25 AAGGAGCCCTGGTTCTGCTGAGGTGCAACTACTCATCGTCTGTTCCACCATATCTCTT CTGGTATGTGCAATACCCCAACCAAGGACTCCAGCTTCTCCTGAAGTACACATCAGC GGCCACCCTGGTTAAAGGCATCAACGGTTTTGAGGCTGAATTTAAGAAGAGTGAAA CCTCCTTCCACCTGACGAAACCCTCAGCCCATATGAGCGACGCGGCTGAGTACTTCT GTGCTGTGAGTGATCTCGAACCGAACAGCAGTGCTTCCAAGATAATCTTTGGATCAG 30 GGACCAGACTCAGCATCCGGCCAAATATCCAGAACCCTGACCCTGCCGTGTACCAG CTGAGAGACTCTAAATCCAGTGACAAGTCTGTCTGCCTATTCACCGATTTTGATTCTC 2026204466   11 Jun 2026 AAACAAATGTGTCACAAAGTAAGGATTCTGATGTGTATATCACAGACAAAACTGTG CTAGACATGAGGTCTATGGACTTCAAGAGCAACAGTGCTGTGGCCTGGAGCAACAA ATCTGACTTTGCATGTGCAAACGCCTTCAACAACAGCATTATTCCAGAAGACACCTT CTTCCCCAGCCCAGAAAGTTCCTGTGATGTCAAGCTGGTCGAGAAAAGCTTTGAAAC 5 AGATACGAACCTAAACTTTCAAAACCTGTCAGTGATTGGGTTCCGAATCCTCCTCCT GAAAGTGGCCGGGTTTAATCTGCTCATGACGCTGCGGCTGTGGTCCAGCTGAGATCT GCAAGATTGTAAGACAGCCTGTGCTCCCTCGCTCCTTCCTCTGCATTGCCCCTCTTCT CCCTCTCCAAACAGAGGGAACTCTCCTACCCCCAAGGAGGTGAAAGCTGCTACCAC CTCTGTGCCCCCCCGGTAATGCCACCAACTGGATCCTACCCGAATTTATGATTAAGA 10 TTGCTGAAGAGCTGCCAAACACTGCTGCCACCCCCTCTGTTCCCTTATTGCTGCTTGT CACTGCCTGACATTCACGGCAGAGGCAAGGCTGCTGCAGCCTCCCCTGGCTGTGCAC ATTCCCTCCTGCTCCCCAGAGACTGCCTCCGCCATCCCACAGATGATGGATCTTCAG TGGGTTCTCTTGGGCTCTAGGTCCTGGAGAATGTTGTGAGGGGTTTATTTTTTTTTAA TAGTGTTCATAAAGAAATACATAGTATTCTTCTTCTCAAGACGTGGGGGGAAATTAT 15 CTCATTATCGAGGCCCTGCTATGCTGTGTGTCTGGGCGTGTTGTATGTCCTGCTGCCG ATGCCTTCATTAAAATGATTTGGAA

[00474] By “T cell receptor beta constant 1 polypeptide (TRBC1)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. P01850 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided 20 below.

[00475] ,>sp|P01850|TRBCl_HUMAN T cell receptor beta constant 1 OS=Homo sapiens OX=9606 GN=TRBC1 PE=1 SV=4DLNKVFPPEVAVFEPSEAEISHTQKATLVCLATGFFPDHVELSWWVNGKEVHSGV STDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRA 25 KPVTQIVSAEAWGRADCGFTSVSYQQGVLSATILYEILLGKATLYAVLVSALVLMAMV KRKDF

[00476] By “T cell receptor beta constant 1 polynucleotide (TRBC1)” is meant a nucleic acid encoding a TRBC1 polypeptide. An exemplary TRBC1 nucleic acid sequence is provided below. > 30 X00437.1CTGGTCTAGAATATTCCACATCTGCTCTCACTCTGCCATGGACTCCTGGACC TTCTGCTGTGTGTCCCTTTGCATCCTGGTAGCGAAGCATACAGATGCTGGAGTTATCC 2026204466   11 Jun 2026 AGTCACCCCGCCATGAGGTGACAGAGATGGGACAAGAAGTGACTCTGAGATGTAAA CCAATTTCAGGCCACAACTCCCTTTTCTGGTACAGACAGACCATGATGCGGGGACTG GAGTTGCTCATTTACTTTAACAACAACGTTCCGATAGATGATTCAGGGATGCCCGAG GATCGATTCTCAGCTAAGATGCCTAATGCATCATTCTCCACTCTGAAGATCCAGCCC 5 TCAGAACCCAGGGACTCAGCTGTGTACTTCTGTGCCAGCAGTTTCTCGACCTGTTCG GCTAACTATGGCTACACCTTCGGTTCGGGGACCAGGTTAACCGTTGTAGAGGACCTG AACAAGGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCAGAGATCTCC CACACCCAAAAGGCCACACTGGTGTGCCTGGCCACAGGCTTCTTCCCCGACCACGTG GAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAGCACAGACCC 10 GCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCCTGAGCAGCC GCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCCGCTGTCAAG TCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGGGCCAAACCC GTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGACTGTGGCTTTACCTC GGTGTCCTACCAGCAAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCCTGCTAGG 15 GAAGGCCACCCTGTATGCTGTGCTGGTCAGCGCCCTTGTGTTGATGGCCATGGTCAA GAGAAAGGATTTCTGAAGGCAGCCCTGGAAGTGGAGTTAGGAGCTTCTAACCCGTC ATGGTTCAATACACATTCTTCTTTTGCCAGCGCTTCTGAAGAGCTGCTCTCACCTCTC TGCATCCCAATAGATATCCCCCTATGTGCATGCACACCTGCACACTCACGGCTGAAA TCTCCCTAACCCAGGGGGAC 20

[00477] By “T cell receptor beta constant 2 polypeptide (TRBC2)” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. A0A5B9 or fragment thereof and having immunomodulatory activity. An exemplary amino acid sequence is provided below. ,>sp|A0A5B9|TRBC2_HUMAN T cell receptor beta constant 2 OS=Homo sapiens OX=9606 25 GN=TRBC2 PE=1 SV=2DLKNVFPPKVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGV STDPQPLKEQPALNDSRYCLSSRLRVSATFWQNPRNHFRCQVQFYGLSENDEWTQDRA KPVTQIVSAEAWGRADCGFTSESYQQGVLSATILYEILLGKATLYAVLVSALVLMAMV KRKDSRG 30

[00478] By “T cell receptor beta constant 2 polynucleotide (TRBC2)” is meant a nucleic acid encoding a TRAC polypeptide. An exemplary TRBC2 nucleic acid sequence is provided below. 2026204466   11 Jun 2026

[00479] >NG_001333.2:655095-656583 Homo sapiens T cell receptor beta locus (TRB) on chromosome? AGGACCTGAAAAACGTGTTCCCACCCGAGGTCGCTGTGTTTGAGCCATCAGAAGCA GAGATCTCCCACACCCAAAAGGCCACACTGGTATGCCTGGCCACAGGCTTCTACCCC 5 GACCACGTGGAGCTGAGCTGGTGGGTGAATGGGAAGGAGGTGCACAGTGGGGTCAG CACAGACCCGCAGCCCCTCAAGGAGCAGCCCGCCCTCAATGACTCCAGATACTGCC TGAGCAGCCGCCTGAGGGTCTCGGCCACCTTCTGGCAGAACCCCCGCAACCACTTCC GCTGTCAAGTCCAGTTCTACGGGCTCTCGGAGAATGACGAGTGGACCCAGGATAGG GCCAAACCCGTCACCCAGATCGTCAGCGCCGAGGCCTGGGGTAGAGCAGGTGAGTG 10 GGGCCTGGGGAGATGCCTGGAGGAGATTAGGTGAGACCAGCTACCAGGGAAAATG GAAAGATCCAGGTAGCGGACAAGACTAGATCCAGAAGAAAGCCAGAGTGGACAAG GTGGGATGATCAAGGTTCACAGGGTCAGCAAAGCACGGTGTGCACTTCCCCCACCA AGAAGCATAGAGGCTGAATGGAGCACCTCAAGCTCATTCTTCCTTCAGATCCTGACA CCTTAGAGCTAAGCTTTCAAGTCTCCCTGAGGACCAGCCATACAGCTCAGCATCTGA 15 GTGGTGTGCATCCCATTCTCTTCTGGGGTCCTGGTTTCCTAAGATCATAGTGACCACT TCGCTGGCACTGGAGCAGCATGAGGGAGACAGAACCAGGGCTATCAAAGGAGGCTG ACTTTGTACTATCTGATATGCATGTGTTTGTGGCCTGTGAGTCTGTGATGTAAGGCTC AATGTCCTTACAAAGCAGCATTCTCTCATCCATTTTTCTTCCCCTGTTTTCTTTCAGAC TGTGGCTTCACCTCCGGTAAGTGAGTCTCTCCTTTTTCTCTCTATCTTTCGCCGTCTCT 20 GCTCTCGAACCAGGGCATGGAGAATCCACGGACACAGGGGCGTGAGGGAGGCCAG AGCCACCTGTGCACAGGTGCCTACATGCTCTGTTCTTGTCAACAGAGTCTTACCAGC AAGGGGTCCTGTCTGCCACCATCCTCTATGAGATCTTGCTAGGGAAGGCCACCTTGT ATGCCGTGCTGGTCAGTGCCCTCGTGCTGATGGCCATGGTAAGGAGGAGGGTGGGA TAGGGCAGATGATGGGGGCAGGGGATGGAACATCACACATGGGCATAAAGGAATCT 25 CAGAGCCAGAGCACAGCCTAATATATCCTATCACCTCAATGAAACCATAATGAAGC CAGACTGGGGAGAAAATGCAGGGAATATCACAGAATGCATCATGGGAGGATGGAG ACAACCAGCGAGCCCTACTCAAATTAGGCCTCAGAGCCCGCCTCCCCTGCCCTACTC CTGCTGTGCCATAGCCCCTGAAACCCTGAAAATGTTCTCTCTTCCACAGGTCAAGAG AAAGGATTCCAGAGGCTAG 30

[00480] As used herein “transduction” means to transfer a gene or genetic material to a cell via a viral vector. 2026204466   11 Jun 2026

[00481] “Transformation,” as used herein refers to the process of introducing a genetic change in a cell produced by the introduction of exogenous nucleic acid.

[00482] “Transfection” refers to the transfer of a gene or genetical material to a cell via a chemical or physical means. 5

[00483] By “translocation” is meant the rearrangement of nucleic acid segments between non- homologous chromosomes.

[00484] As used herein, the terms “treat,” treating,” “treatment,” and the like refer to reducing or ameliorating a disorder and / or a symptom associated therewith. It will be appreciated that, although not precluded, treating a disorder or condition does not require that the disorder, 10   condition or symptoms associated therewith be eliminated.

[00485] The term "uracil glycosylase inhibitor" or "UGI," as used herein, refers to a protein that is capable of inhibiting a uracil-DNA glycosylase base-excision repair enzyme. In some embodiments, the polypeptide further contains one or more (e.g., 1,2, 3, 4, 5) Uracil glycosylase inhibitors. In some embodiments, a UGI domain comprises a wild-type UGI or a modified 15 version thereof. In some embodiments, the UGI proteins provided herein include fragments of UGI and proteins homologous to a UGI or a UGI fragment. For example, in some embodiments, a UGI domain comprises a fragment of the amino acid sequence set forth herein below. In some embodiments, a UGI fragment comprises an amino acid sequence that comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at 20 least 96%, at least 97%, at least 98%, at least 99%, or 100% of an exemplary UGI sequence provided herein. In some embodiments, a UGI comprises an amino acid sequence homologous to the amino acid sequence set forth herein below, or an amino acid sequence homologous to a fragment of the amino acid sequence set forth herein below. In some embodiments, proteins comprising UGI or fragments of UGI or homologs of UGI or UGI fragments are referred to as 25 "UGI variants." A UGI variant shares homology to UGI, or a fragment thereof. For example, a UGI variant is at least 70% identical, at least 75% identical, at least 80% identical, at least 85% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, at least 99% identical, at least 99.5% identical, or at least 99.9% identical to a wild type UGI or a UGI as set forth herein. In some embodiments, the UGI variant 30 comprises a fragment of UGI, such that the fragment is at least 70% identical, at least 80% identical, at least 90% identical, at least 95% identical, at least 96% identical, at least 97% 2026204466   11 Jun 2026 identical, at least 98% identical, at least 99% identical, at least 99.5% identical, or at least 99.9% to the corresponding fragment of wild-type UGI or a UGI as set forth below. In some embodiments, the UGI comprises the following amino acid sequence:

[00486] >splP14739IUNGI_BPPB2 Uracil-DNA glycosylase inhibitor 5

[00487] MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENV MLLT S D APE YKPW ALVIQDS NGENKIKML

[00488] The term “vector” refers to a means of introducing a nucleic acid sequence into a cell, resulting in a transformed cell. Vectors include plasmids, transposons, phages, viruses, liposomes, and episome. “Expression vectors” are nucleic acid sequences comprising the 10 nucleotide sequence to be expressed in the recipient cell. Expression vectors may include additional nucleic acid sequences to promote and / or facilitate the expression of the of the introduced sequence such as start, stop, enhancer, promoter, and secretion sequences.

[00489] By “zeta chain of T cell receptor associated protein kinase 70 (ZAP70) polypeptide” is meant a protein having at least about 85% amino acid sequence identity to NCBI Accession No. 15   AAH53878.1 and having kinase activity. An exemplary amino acid sequence is provided below.

[00490] >AAH53878.1 Zeta-chain (TCR) associated protein kinase 70kDa [Homo sapiens] MPDPAAHLPFFYGSISRAEAEEHLKLAGMADGLFLLRQCLRSLGGYVLSLVHDVRFHHF PIERQLNGTYAIAGGKAHCGPAELCEFYSRDPDGLPCNLRKPCNRPSGLEPQPGVFDCLR DAMVRDYVRQTWKLEGEALEQAIISQAPQVEKLIATTAHERMPWYHSSLTREEAERKL 20 YSGAQTDGKFLLRPRKEQGTYALSLIYGKTVYHYLISQDKAGKYCIPEGTKFDTLWQLV EYLKLKADGLIYCLKEACPNSSASNASGAAAPTLPAHPSTLTHPQRRIDTLNSDGYTPEP ARITSPDKPRPMPMDTSVYESPYSDPEELKDKKLFLKRDNLLIADIELGCGNFGSVRQGV YRMRKKQIDVAIKVLKQGTEKADTEEMMREAQIMHQLDNPYIVRLIGVCQAEALMLV MEMAGGGPLHKFLVGKREEIPVSNVAELLHQVSMGMKYLEEKNFVHRDLAARNVLLV 25 NRHYAKISDFGLSKALGADDSYYTARSAGKWPLKWYAPECINFRKFSSRSDVWSYGVT MWEALSYGQKPYKKMKGPEVMAFIEQGKRMECPPECPPELYALMSDCWIYKWEDRPD FLTVEQRMRACYYSLASKVEGPPGSTQKAEAACA

[00491] By “zeta chain of T cell receptor associated protein kinase 70 (ZAP70) polynucleotide” is meant a nucleic acid encoding a ZAP70 polypeptide. The ZAP70 gene 30 encodes a tyrosine kinase that is involved in T cell development and lymphocyte activation. 2026204466   11 Jun 2026 Absence of functional ZAP 10 can lead to a severe combined immunodeficiency characterized by the lack of CD8+ T cells. An exemplary ZAP70 nucleic acid sequence is provided below.

[00492] >BC053878.1 Homo sapiens zeta-chain (TCR) associated protein kinase 70kDa, mRNA (cDNA clone MGC:61743 IMAGE:5757161), complete cds 5 GCTTGCCGGAGCTCAGCAGACACCAGGCCTTCCGGGCAGGCCTGGCCCACCGTGGG CCTCAGAGCTGCTGCTGGGGCATTCAGAACCGGCTCTCCATTGGCATTGGGACCAGA GACCCCGCAAGTGGCCTGTTTGCCTGGACATCCACCTGTACGTCCCCAGGTTTCGGG AGGCCCAGGGGCGATGCCAGACCCCGCGGCGCACCTGCCCTTCTTCTACGGCAGCA TCTCGCGTGCCGAGGCCGAGGAGCACCTGAAGCTGGCGGGCATGGCGGACGGGCTC 10 TTCCTGCTGCGCCAGTGCCTGCGCTCGCTGGGCGGCTATGTGCTGTCGCTCGTGCAC GATGTGCGCTTCCACCACTTTCCCATCGAGCGCCAGCTCAACGGCACCTACGCCATT GCCGGCGGCAAAGCGCACTGTGGACCGGCAGAGCTCTGCGAGTTCTACTCGCGCGA CCCCGACGGGCTGCCCTGCAACCTGCGCAAGCCGTGCAACCGGCCGTCGGGCCTCG AGCCGCAGCCGGGGGTCTTCGACTGCCTGCGAGACGCCATGGTGCGTGACTACGTG 15 CGCCAGACGTGGAAGCTGGAGGGCGAGGCCCTGGAGCAGGCCATCATCAGCCAGGC CCCGCAGGTGGAGAAGCTCATTGCTACGACGGCCCACGAGCGGATGCCCTGGTACC ACAGCAGCCTGACGCGTGAGGAGGCCGAGCGCAAACTTTACTCTGGGGCGCAGACC GACGGCAAGTTCCTGCTGAGGCCGCGGAAGGAGCAGGGCACATACGCCCTGTCCCT CATCTATGGGAAGACGGTGTACCACTACCTCATCAGCCAAGACAAGGCGGGCAAGT 20 ACTGCATTCCCGAGGGCACCAAGTTTGACACGCTCTGGCAGCTGGTGGAGTATCTGA AGCTGAAGGCGGACGGGCTCATCTACTGCCTGAAGGAGGCCTGCCCCAACAGCAGT GCCAGCAACGCCTCAGGGGCTGCTGCTCCCACACTCCCAGCCCACCCATCCACGTTG ACTCATCCTCAGAGACGAATCGACACCCTCAACTCAGATGGATACACCCCTGAGCC AGCACGCATAACGTCCCCAGACAAACCGCGGCCGATGCCCATGGACACGAGCGTGT 25 ATGAGAGCCCCTACAGCGACCCAGAGGAGCTCAAGGACAAGAAGCTCTTCCTGAAG CGCGATAACCTCCTCATAGCTGACATTGAACTTGGCTGCGGCAACTTTGGCTCAGTG CGCCAGGGCGTGTACCGCATGCGCAAGAAGCAGATCGACGTGGCCATCAAGGTGCT GAAGCAGGGCACGGAGAAGGCAGACACGGAAGAGATGATGCGCGAGGCGCAGATC ATGCACCAGCTGGACAACCCCTACATCGTGCGGCTCATTGGCGTCTGCCAGGCCGAG 30 GCCCTCATGCTGGTCATGGAGATGGCTGGGGGCGGGCCGCTGCACAAGTTCCTGGTC GGCAAGAGGGAGGAGATCCCTGTGAGCAATGTGGCCGAGCTGCTGCACCAGGTGTC 2026204466   11 Jun 2026 CATGGGGATGAAGTACCTGGAGGAGAAGAACTTTGTGCACCGTGACCTGGCGGCCC GCAACGTCCTGCTGGTTAACCGGCACTACGCCAAGATCAGCGACTTTGGCCTCTCCA AAGCACTGGGTGCCGACGACAGCTACTACACTGCCCGCTCAGCAGGGAAGTGGCCG CTCAAGTGGTACGCACCCGAATGCATCAACTTCCGCAAGTTCTCCAGCCGCAGCGAT 5 GTCTGGAGCTATGGGGTCACCATGTGGGAGGCCTTGTCCTACGGCCAGAAGCCCTAC AAGAAGATGAAAGGGCCGGAGGTCATGGCCTTCATCGAGCAGGGCAAGCGGATGG AATGCCCACCAGAGTGTCCACCCGAACTGTACGCACTCATGAGTGACTGCTGGATCT ACAAGTGGGAGGATCGCCCCGACTTCCTGACCGTGGAGCAGCGCATGCGAGCCTGT TACTACAGCCTGGCCAGCAAGGTGGAAGGGCCCCCAGGCAGCACACAGAAGGCTGA 10 GGCTGCCTGTGCCTGAGCTCCCGCTGCCCAGGGGAGCCCTCCACACCGGCTCTTCCC CACCCTCAGCCCCACCCCAGGTCCTGCAGTCTGGCTGAGCCCTGCTTGGTTGTCTCC ACACACAGCTGGGCTGTGGTAGGGGGTGTCTCAGGCCACACCGGCCTTGCATTGCCT GCCTGGCCCCCTGTCCTCTCTGGCTGGGGAGCAGGGAGGTCCGGGAGGGTGCGGCT GTGCAGCCTGTCCTGGGCTGGTGGCTCCCGGAGGGCCCTGAGCTGAGGGCATTGCTT 15 ACACGGATGCCTTCCCCTGGGCCCTGACATTGGAGCCTGGGCATCCTCAGGTGGTCA GGCGTAGATCACCAGAATAAACCCAGCTTCCCTCTTGAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA

[00493] Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, 20 the terms "a", "an", and "the" are understood to be singular or plural.

[00494] Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from 25 context, all numerical values provided herein are modified by the term about.

[00495] Ranges provided herein are understood to be shorthand for all the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 30   42, 43, 44, 45, 46, 47, 48, 49, or 50. 2026204466   11 Jun 2026

[00496] The recitation of a listing of chemical groups in any definition of a variable herein includes definitions of that variable as any single group or combination of listed groups. The recitation of an embodiment for a variable or aspect herein includes that embodiment as any single embodiment or in combination with any other embodiments or portions thereof. 5

[00497] Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein. BRIEF DESCRIPTION OF THE DRAWINGS

[00498] FIGs. 1A-1B are illustrations of three proteins that impact T cell function. FIG. 10 1A is an illustratration of the TRAC protein, which is a key component in graft versus host disease. FIG. IB is an illustratration of the B2M protein, a component of the MHC class 1 antigen presenting complex present on nucleated cells that can be recognized by a host’s CD8+ T cells. FIG. IC is an illustratration of T cell signaling that leads to expression of the PDCD1 gene, and the resulting PD-1 protein acts to inhibit the T cell signaling. 15

[00499] FIG. 2 is a graph of the percentage of cells with knocked down expression of target genes after base editing. “EP” denotes electroporation.

[00500] FIG. 3 is a graph of the percentages of the observed types of genetic modification in untransduced cells or in cells transduced with a BE4 base editing system or a Cas9 nuclease.

[00501] FIG. 4 is a graph depicting target nucleotide modification percentage as measured 20 by percentage of cells that are negative for target protein expression as determined by flow cytometry (FC) in cells transduced with BE4 and sgRNAs directing BE4 to splice site acceptors (SA) or donors (SD) or that generate a STOP codon. Control cells were mock electroporated (EP).

[00502] FIG. 5 is a diagram of the BE4 system disrupting splice site acceptors (SA), splice 25 donors (SD), or generate STOP codons.

[00503] FIG. 6 is a chart summarizing off-target binding sites of sgRNAs employed to disrupt target genes.

[00504] FIG. 7 is a graph summarizing flow cytometry (FC) data of the percentage of cells edited with BE4 or Cas9 that exhibit reduced protein expression. Cells were either gated to B2M 30 or CD3, the latter being a proxy for TRAC expression. 2026204466   11 Jun 2026

[00505] FIG. 8A is a scatter plot of FACS data of unedited control cells. FIG. 8B is a scatter plot of FACS data of cells that have been edited at the B2M, TRAC, and PD1 loci.

[00506] FIG. 9 is a graph illustratrating the effectiveness of the base editing techniques described herein to modify specific genes that can negatively impact CAR-T immunotherapy. 5

[00507] FIG. 10 is a diagram depicting a droplet digital PCR (ddPCR) protocol to detect and quantify gene modifications and translocations.

[00508] FIG. 11 presents two graphs showing the data generated from next generation sequencing (NGS) analysis or ddPCR of cells edited using either the BE4 system or the Cas9 system. 10

[00509] FIG. 12 is a schematic diagram that illustrates the role Cbl-b plays in suppressing T cell activation.

[00510] FIG. 13 is a graph depicting the efficiency of Cbl-b knockdown by disruption of splice sites. SA = Splice Acceptor; SD = Splice Donor; STOP - generated STOP codon; 2° Only = secondary antibody only; C373 refers to a loss of function variant (C373R); RL1-A::APC-A = 15    laser; ICS = intracellular staining.

[00511] FIG. 14 is a graph illustrating the rate of Cas 12b-mediated indels in the GRIN2B and DNMT1 genes in T cells. EP denotes electroporation.

[00512] FIG. 15 is a graph summarizing fluorescence assisted cell sorting (FACS) data of ...

Claims

1. A method for producing a modified immune cell with reduced immunogenicity and / or5         increased anti-neoplasia activity by multiplexed editing, the method comprising:modifying at least four gene sequences or regulatory elements thereof, at a single target nucleobase in each thereof in an immune cell, thereby generating the modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity.

2. A method for producing a population of modified immune cells with reduced10         immunogenicity and / or increased anti-neoplasia activity by multiplexed editing, themethod comprising: modifying at least four gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in a population of immune cells, thereby generating the population of modified immune cells with reduced immunogenicity and / or increased anti-neoplasia activity.15   3. The method of claim 1 or 2, wherein at least one of the at least four gene sequences is acheckpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.

4. The method of any one of claims 1-3, wherein the modifying reduces expression of at least one of the at least four gene sequences.20   5. The method of any one of claim 1, wherein expression of at least one of the at least fourgenes is reduced by at least 80% as compared to a control cell without the modification.

6. The method of claim 5, wherein expression of each one of the at least four genes is reduced by at least 80% as compared to a control cell without the modification.

7. The method of claim 2, wherein expression of at least one of the at least four genes is25 reduced in at least 50% of the population of immune cells.

8. The method of 7, wherein expression of each one of the at least four genes is reduced in at least 50% of the population of immune cells.

9. The method of any one of the preceding claims, wherein the at least four gene sequences comprise a TCR complex gene sequence.30   10. The method of claim 9, wherein the at least four gene sequences comprise a TRACgene sequence.

11. The method of any one of claims 1-8, wherein the at least four gene sequences comprise a check point inhibitor gene sequence.2026204466   11 Jun 202612. The method of claim 10, wherein the at least four gene sequences comprise a PDCD1 gene sequence.

13. The method of any one of claims 1-8, wherein the at least four gene sequences comprise a T cell marker gene sequence.5   14. The method of claim 13, wherein the at least four gene sequences comprise a CD52gene sequence.

15. The method of claim 13, wherein the at least four gene sequences comprises a CD7 gene sequence.

16. The method of any one of claims 1-15, wherein the at least four gene sequences10 comprise a TRAC gene sequence, a PDCD1 gene sequence, a CD52 gene sequence, or a CD7 gene sequence.

17. The method of any one of claims 1-16, wherein the at least four sequences comprises a TCR complex gene sequence, a CD7 gene sequence, a CD52 gene sequence ,and a gene sequence selected from the group consisting of a CD2 gene sequence, a CD4 gene15 sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence18. The method of claim any one of claims 1-17, wherein the at least four gene sequences comprise a gene sequence selected from the group consisting of a CD2 gene sequence, a20 TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence.25   19. The method of any one of claims 1, 5, and 6, comprising modifying five gene sequencesor regulatory elements thereof at a single target nucleobase in each thereof in the immune cell.

20. The method of any one of claims 1, 5, and 6, comprising modifying six gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the30 immune cell.

21. The method of any one of claims 1, 5, and 6, comprising modifying seven gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the immune cell.2026204466   11 Jun 202622. The method of any one of claims 1, 5, and 6, comprising modifying eight gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the immune cell.

23. The method of any one of claims 2, 7, and 8, comprising modifying five gene sequences 5 or regulatory elements thereof at a single target nucleobase in each thereof in the population of immune cells.

24. The method of any one of claims 2, 7, and 8, comprising modifying six gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in the population of immune cells.10   25. The method of any one of claims 2, 7, and 8, comprising modifying seven genesequences or regulatory elements thereof at a single target nucleobase in each thereof in the population of immune cells.

26. The method of any one of claims 2, 7, and 8, comprising modifying eight gene sequences or regulatory elements thereof at a single target nucleobase in each thereof in 15         the population of immune cells.

27. The method of any one of claims 19-26, wherein the five, six, seven, or eight gene sequences or regulatory elements thereof are selected from the group consisting of a CD2 gene sequence, a TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC220 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence.

28. The method of any one of claims 19-27, wherein the five, six, seven, or eight gene sequences or regulatory elements thereof at comprises a CD3 gene sequence, a CD725 gene sequence, a a CD2 gene sequence, a CD5 gene sequence, and a CD52 gene sequence.

29. The method of any one of the preceding claims, wherein the modifying comprises deaminating the single target nucleobase.

30. The method of claim 29, wherein the deaminating is performed by a polypeptide30 comprising a deaminase.

31. The method of claim 30, wherein the deaminase is associated with a nucleic acid programmable DNA binding protein (napDNAbp) to form a base editor.

32. The method of claim 31, wherein the deaminase is fused to the nucleic acid programmable DNA binding protein (napDNAbp).2026204466   11 Jun 202633. The method of claim 32, wherein the napDNAbp comprises a Cas9 polypeptide or a portion thereof.

34. The method of claim 33, wherein the napDNAbp comprises a Cas9 nickase or nuclease dead Cas9.5   35. The method of any one of claims 30-34, wherein the deaminase is a cytidine deaminase.

36. The method of claim 35, wherein the single target nucleobase is a cytosine (C) and wherein the modification comprises conversion of the C to a thymine (T).

37. The method of claim 36, wherein the base editor further comprises a uracil glycosylase inhibitor.10   38. The method of any one of claims 30-34, wherein the deaminase is an adenosinedeaminase.

39. The method of claim 38, wherein the single target nucleobase is a adenosine (A) and wherein the modification comprises conversion of the A to a guanine (G).

40. The method of any one of claims 30-39, wherein the modifying comprises contacting 15 the immune cell with a guide nucleic acid sequences.

41. The method of claim 40, wherein the modifying comprises contacting the immune cell with at least four guide nucleic acid sequences, wherein each guide nucleic acid sequence targets the napDNAbp to one of the at least four gene sequences or regulatory elements thereof.20   42. The method of claim 40, wherein the guide nucleic acid sequence comprises a sequenceselected from guide RNA sequences of table 8A, table 8B, or table 8C.

43. The method of claim 40, wherein the guide nucleic acid sequence comprises a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC,25 CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

44. The method of any one of claims 1-28, wherein the modifying comprises replacing the single target nucleobase with a different nucleobase by target-primed reverse30 transcription with a reverse transcriptase and an extended guide nucleic acid sequence.

45. The method of claim 44, wherein the extended guide nucleic acid sequence comprises a reverse transcription template sequence, a reverse transcription primer binding site, or a combination thereof.2026204466   11 Jun 202646. The method of any one of claims 1-45, wherein the single target nucleobase is in an exon.

47. The method of claim 46, wherein the modifying generates a premature stop codon in the exon.5       48. The method of claim 46 or 47, wherein the single target nucleobase is within an exon1, an exon 2, or an exon 3 of the TRAC gene sequence.

49. The method of claim 46 or 47, wherein the single target nucleobase is within an exon 1, an exon 2, or an exon 5 of the PCDC1 gene sequence.

50. The method of claim 46 or 47, wherein the single target nucleobase is within an exon 10          1 or an exon 2 of the CD52 gene sequence.

51. The method of claim 46 or 47, wherein the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of the CD7 gene sequence.

52. The method of claim 46 or 47, wherein the single target nucleobase is within an exon 1 or an exon 2 of the B2M gene sequence.15       53. The method of claim 46 or 47, wherein the single target nucleobase is within an exon2, an exon 3, an exon 4, an exon 5, an exon 6, an exon 7, or an exon 8 of the CD5 gene sequence.

54. The method of claim 46 or 47, wherein the single target nucleobase is within an exon 2, an exon 3, an exon 4, or an exon 5 of the CD2 gene sequence.20      55. The method of claim 46 or 47, wherein the single target nucleobase is within an exon1, an exon 2, an exon 4, an exon 7, an exon 8, an exon 9, an exon 10, an exon 11, an exon 12, an exon 14, an exon 15, an exon 18, or an exon 19 of the CIITA gene sequence.

56. The method of any one of claims 1-45, wherein the single target nucleobase is in a25           splice donor site or a splice acceptor site.

57. The method of claim 50, wherein the single target nucleobase is in a exon 1 splice acceptor site, a exon 1 splice donor site, or a exon 3 splice acceptor site of the TRAC gene sequence.

58. The method of claim 50, wherein the single target nucleobase is in a exon 1 splice30           acceptor site, a exon 1 splice donor site, an exon 2 splice acceptor site, an exon 3splice donor site, an exon 4 splice acceptor site, an exon 4 splice donor site, or a exon 5 splice acceptor site of the PDCD1 gene sequence.

59. The method of claim 50, wherein the single target nucleobase is in a exon 1 splice donor site, or an exon 2 splice acceptor site of the CD52 gene sequence.2026204466   11 Jun 202660. The method of claim 50, wherein the single target nucleobase is in a exon 1 splice donor site, a exon 2 splice donor site, an exon 2 splice acceptor site, or an exon 3 splice acceptor site of the CD7 gene sequence.

61. The method of claim 50, wherein the single target nucleobase is in a exon 1 splice5           donor site, a exon 2 splice donor site, an exon 2 splice acceptor site, or an exon 3splice acceptor site of the B2M gene sequence.

62. The method of claim 50, wherein the single target nucleobase is in an exon 3 splice donor site of the CD2 gene sequence.

63. The method of claim 50, wherein the single target nucleobase is in an exon 1 splice10           donor site, an exon 1 splice acceptor site, an exon 3 splice acceptor site, an exon 3splice donor site, an exon 4 splice acceptor site, an exon 5 splice donor site, an exon 6 splice acceptor site, an exon 9 splice donor site, an exon 10 splice acceptor site of the CD5 gene sequence.

64. The method of claim 50, wherein the single target nucleobase is in an exon 1 splice15           donor site, an exon 7 splice donor site, an exon 8 splice acceptor site, an exon 9 slicedonor site, an exon 10 splice acceptor site, an exon 11 splice acceptor site, an exon 14 splice acceptor site, an exon 14 splice donor site, an exon 15 splice donor site, an exon 16 splice acceptor site, an exon 16 splice donor site, an exon 17 splice acceptor site, an exon 17 splice donor site, or an exon 19 splice acceptor site of the CIITA gene20          sequence.

65. The method of any one of claims 4-64, wherein the immune cell is a human cell.

66. The method of claim 65, wherein the immune cell is a cytotoxic T cell, a regulatory T cell, a T helper cell, a dendritic cell, a B cell, or a NK cell.

67. The method of any one of claims 7-66, wherein the population of immune cells are 25          human cells.

68. The method of claim 67, wherein the population of immune cells are cytotoxic T cells, regulatory T cells, T helper cells, dendritic cells, B cells, or NK cells.

69. The method of any one of claims 1-68, wherein the modifying is ex vivo.

70. The method of any one of claims 1-69, wherein the immune cell or the population of 30          immune cells are derived from a single human donor.

71. The method of any one of claims 1-70, further comprising contacting the immune cell or the population of immune cells with a polynucleotide that encodes an exogenous functional chimeric antigen receptor (CAR) or a functional fragment thereof.2026204466   11 Jun 202672. The method of claim 71, comprising contacting the immune cell or the population of immune cells with a lentivirus comprising the polynucleotide that encodes the CAR.

73. The method of claim 71, comprising contacting the immune cell or the population of immune cells with a napDNAbp and a donor DNA sequence comprising the5          polynucleotide that encodes the CAR.

74. The method of claim 73, wherein the napDNAbp is a Casl2b.

75. The method of any one of claims 71-74, wherein the CAR specifically binds a marker associated with neoplasia.

76. The method of claim 75, wherein the neoplasia is a T cell cancer, a B cell cancer, a10          lymphoma, a leukemia, or a multiple myeloma.

77. The method of claim 76, wherein the CAR specifically binds CD7.

78. The method of claim 76, wherein the CAR specifically binds BCMA.

79. The method of any one of claims 2-78, wherein the immune cell or the population of immune cells comprises no detectable translocation.15       80. The method of claim 79, wherein at least 50% of the population of immune cellsexpress the CAR.

81. The method of claim 79, wherein at least 50% of the population of immune cells are viable.

82. The method of claim 79,wherein at least 50% of the population of immune cells20          expand at least 80% of expansion rate of a population of control cells of a same typewithout the modification.

83. The method of any one of claims 4-79, wherein the modifying generates less than 1% of indels in the immune cell.

84. The method of any one of claims 4-79, wherein the modifying generates less than 5% 25           of non-target edits in the immune cell.

85. The method of any one of claims 4-79, wherein the modifying generates less than 5% of off-target edits in the immune cell.

86. A modified immune cell produced according to the method of any one of claims 4-85.

87. A population of modified immune cells produced according to the method of any one 30          of claims 7-85.

88. A modified immune cell with reduced immunogenicity or increased anti-neoplasia activity, wherein the modified immune cell comprises a single target nucleobase modification in each one of at least four gene sequences or regulatory elements thereof.2026204466   11 Jun 202689. The modified immune cell of claim 88, wherein each one of the at least four gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence,or an immunogenic gene sequence.

90. The modified immune cell of claim 88 or 89, wherein the at least four gene sequences 5          comprise a TCR complex gene sequence.

91. The modified immune cell of claim 90, wherein the at least four gene sequences comprise a TRAC gene sequence.

92. The modified immune cell of claim 88 or 89, wherein the at least four gene sequences comprise a check point inhibitor gene sequence.10      93. The modified immune cell of claim 92, wherein the at least four gene sequencescomprise a PDCD1 gene sequence.

94. The modified immune cell of claim 88 or 89, wherein the at least four gene sequences comprise a T cell marker gene sequence.

95. The modified immune cell of claim 94, wherein the at least four gene sequences15          comprise CD52 gene sequence.

96. The modified immune cell of claim 94, wherein the at least four gene sequences comprises a CD7 gene sequence.

97. The modified immune cell of any one of claims 88-96, wherein expression of one of the at least four genes is reduced by at least 80% as compared to a control cell without 20          the modification.

98. The modified immune cell of claim 97, wherein expression of each one of the at least four genes is reduced by at least 90% as compared to a control cell without the modification.

99. The modified immune cell of any one of claims 88-98, wherein the immune cell 25           comprises a modification at a single target nucleobase in each one of five genesequences or regulatory elements thereof, wherein each one of the five gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.

100. The modified immune cell of any one of claims 88-98, wherein the immune 30          cell comprises a modification at a single target nucleobase in each one of six genesequences or regulatory elements thereof, wherein each one of the six gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.2026204466   11 Jun 2026101.      The modified immune cell of any one of claims 88-98, wherein the immunecell comprises a modification at a single target nucleobase in each one of seven gene sequences or regulatory elements thereof, wherein each one of the seven gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation5         gene sequence or an immunogenic gene sequence.

102. The modified immune cell of any one of claims 88-98, wherein the immune cell comprises a modification at a single target nucleobase in each one of eight gene sequences or regulatory elements thereof, wherein each one of the eight gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation 10         gene sequence, or an immunogenic gene sequence103. The modified immune cell of any one of claims 99-102, wherein expression of at least one of the five, six, seven or eight genes is reduced by at least 90% as compared to a control cell without the modification.

104. The modified immune cell of any one of claims 99-102, wherein expression of 15           each one of the five, six, seven, or eight genes is reduced by at least 90% as comparedto a control cell without the modification.

105. The modified immune cell of any one of claims 99-104, wherein the five, six, seven, or eight gene sequences or regulatory elements thereof comprise a sequence selected from the group consisting of a CD2 gene sequence, a TRAC gene sequence, a20         CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta genesequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence.25       106. A modified immune cell comprising a single target nucleobase modification ineach one of a CD3 gene sequence, a CD5 gene sequence, a CD52 gene sequence, and a CD7 gene sequence, wherein the modified immune cell exhibits reduced immunogenicity or increased anti-neoplasia activity as compared to a control cell of a same type without the modification.30       107. The modified immune cell of claim 106, where in the immune cell furthercomprises a single target nucleobase modification in a CD2 gene sequence,CIITA or a regulatory element of each thereof.

108. The modified immune cell of claim 106, wherein the immune cell comprises a single target nucleobase modification in a TRAC gene sequence, a CD3 epsilon gene2026204466   11 Jun 2026sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, or a TRBC2 gene sequence further comprises a single target nucleobase modification in a gene sequence a CD4 gene sequence, a CD30 gene sequence, a CD33 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA5           gene sequence or a regulatory element of each thereof.

109. The modified immune cell of claim 107, wherein the immune cell comprises a single nucleobase modification in each one of a TRAC gene sequence, a PDCD1 gene sequence, a CD52 gene sequence, a CD7 gene sequence, a CD2 gene sequence, a CD5 gene sequence, a CIITA gene sequence, and a B2M gene sequence.10      110. The modified immune cell of any one of claims 88-109, wherein the immunecell comprises no detectable translocation.

111. The modified immune cell of any one of claims 88-110, wherein the immune cell comprises less than 1% of indels.

112. The modified immune cell of any one of claims 88-110, wherein the immune 15           cell comprises less than 5% of non-target edits.

113. The modified immune cell of any one of claims 88-110, wherein the immune cell comprises less than 5% of off-target edits.

114. The modified immune cell of any one of claims 88-110, wherein the immune cell is a mammalian cell.20      115. The modified immune cell of any one of claims 88-110, wherein the immunecell is a human cell.

116. The modified immune cell of any one of claims 88-115, wherein the immune cell is a cytotoxic T cell, a regulatory T cell, a T helper cell, a dendritic cell, a B cell, or a NK cell.25      117. The modified immune cell of any one of claims 88-116, wherein the immunecell is in an ex vivo culture.

118. The modified immune cell of any one of claims 88-117, wherein the immune cell is derived from a single human donor.

119. The modified immune cell of any one of claims 88-118, wherein the immune 30          cell further comprises a polynucleotide that encodes an exogenous functional chimericantigen receptor (CAR) or a functional fragment thereof.

120. The modified immune cell of claim 119, wherein the polynucleotide that encodes the CAR is integrated in the genome of the immune cell.2026204466   11 Jun 2026121. The modified immune cell of claim 119 or 120, wherein the CAR specifically binds a marker associated with neoplasia.

122. The modified immune cell of claim 119 or 120, wherein the neoplasia is a T cell cancer, a B cell cancer, a lymphoma, a leukemia, or a multiple myeloma.5      123. The modified immune cell of any one of claims 119-122, wherein the CARspecifically binds CD7.

124. The modified immune cell of any one of claims 119-122, wherein the CAR specifically binds BCMA.

125. The modified immune cell of any one of claims 88-124, wherein the single10           target nucleobase is in an exon.

126. The modified immune cell of claim 125, wherein the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of the TRAC gene sequence.

127. The modified immune cell of claim 125, wherein the single target nucleobase is within an exon 1, an exon 2, or an exon 5 of the PCDC1 gene sequence.15       128. The modified immune cell of claim 125, wherein the single target nucleobaseis within an exon 1 or an exon 2 of the CD52 gene sequence.

129. The modified immune cell of claim 125, wherein the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of a CD7 gene sequence.

130. The modified immune cell of any one of claims 88-124, wherein the single 20           target nucleobase is in a splice donor site or a splice acceptor site.

131. The modified immune cell of claim 130, wherein the single target nucleobase is in a exon 1 splice acceptor site, a exon 1 splice donor site, or a exon 3 splice acceptor site of the TRAC gene sequence.

132. The modified immune cell of claim 130, wherein the single target nucleobase 25           is in a exon 1 splice acceptor site, a exon 1 splice donor site, an exon 2 splice acceptorsite, an exon 3 splice donor site, an exon 4 splice acceptor site, an exon 4 splice donor site, or a exon 5 splice acceptor site of the PDCD1 gene sequence.

133. The modified immune cell of claim 130, wherein the single target nucleobase is in a exon 1 splice donor site, or an exon 2 splice acceptor site of the CD52 gene 30          sequence.

134. The modified immune cell of claim 130, wherein the single target nucleobase is in a exon 1 splice donor site, a exon 2 splice donor site, an exon 2 splice acceptor site, or an exon 3 splice acceptor site of the CD7 gene sequence.2026204466   11 Jun 2026135. A population of modified immune cells, wherein a plurality of the population of cells comprise a single target nucleobase modification in each one of at least four gene sequences or regulatory elements thereof, and wherein the plurality of the population of cells having the modification exhibit reduced immunogenicity or5           increased anti-neoplasia activity as compared to a plurality of control cells of a sametype without the modification.

136. The population of modified immune cells of claim 135, wherein the plurality of cells comprises at least 50% of the population.

137. The population of modified immune cells of claim 135 or 136, wherein each 10          one of the at least four gene sequences is a checkpoint inhibitor gene sequence, animmune response regulation gene sequence, or an immunogenic gene sequence138. The population of modified immune cells of claim 137, wherein the at least four gene sequences comprise a TCR component gene sequence, a check point inhibitor gene sequence, or a T cell marker gene sequence.15       139. The population of modified immune cells of claim 137, wherein the at leastfour gene sequences comprise a TRAC gene sequence.

140. The population of modified immune cells of claim 137, wherein the at least four gene sequences comprise a PDCD1 gene sequence.

141. The population of modified immune cells of claim 137, wherein the at least20         four gene sequences comprise CD52 gene sequence.

142. The population of modified immune cells of claim 137, wherein the at least four gene sequences comprises a CD7 gene sequence.

143. The population of modified immune cells of any one of claims 135-142, wherein expression of at least one of the at least four genes is reduced by at least 80% 25           in the plurality of cells having the modification as compared to a control cell withoutthe modification144. The population of modified immune cells of claim 143, wherein expression of each one of the at least four genes is reduced by at least 80% in the plurality of cells having the modification as compared to a control cell without the modification.30       145. The population of modified immune cells of any one of claims 135-144,wherein the plurality of the population comprises a modification at a single target nucleobase in each one of five gene sequences or regulatory elements thereof, wherein each one of the five gene sequences is a checkpoint inhibitor gene sequence, an immune response regulation gene sequence, or an immunogenic gene sequence.2026204466   11 Jun 2026146. The population of modified immune cells of any one of claims 135-144, wherein the plurality of the population comprises a modification at a single target nucleobase in each one of six gene sequences or regulatory elements thereof, wherein each one of the six sequences is a checkpoint inhibitor gene sequence, an immune5          response regulation gene sequence, or an immunogenic gene sequence.

147. The population of modified immune cells of any one of claims 135-144, wherein the plurality of the population comprises a modification at a single target nucleobase in each one of seven gene sequences or regulatory elements thereof, wherein each one of the seven gene sequences is a checkpoint inhibitor gene10          sequence, an immune response regulation gene sequence, or an immunogenic genesequence.

148. The population of modified immune cells of any one of claims 135-144, wherein the plurality of the population comprises a modification at a single target nucleobase in each one of eight gene sequences or regulatory elements thereof,15          wherein each one of the eight gene sequences is a checkpoint inhibitor gene sequence,an immune response regulation gene sequence, or an immunogenic gene sequence.

149. The population of modified immune cells of any one of claims 145-148, wherein expression of at least one of the five, six, seven, or eight genes is reduced by at least 90% in the plurality of cells having the modification as compared to a control 20          cell without the modification150. The population of modified immune cells of any one of claims 145-148, wherein expression of each one of the five, six, seven, or eight genes is reduced by at least 90% in the plurality of cells having the modification as compared to a control cell without the modification25       151. The population of modified immune cells of any one of claims 145-148,wherein the five, six, seven, or eight gene sequences or regulatory elements thereof are selected from the group consisting of a CD2 gene sequence, a TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene30         sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, aCD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M gene sequence, and a CIITA gene sequence.

152. A population of modified immune cells, wherein a plurality of the populationcomprise a single target nucleobase modification in each one of a TRAC gene2026204466   11 Jun 2026sequence, a PDCD1 gene sequence, a CD52 gene sequence, and a CD7 gene sequence, and wherein the plurality of the population having the modification exhibit reduced immunogenicity or increased anti-neoplasia activity as compared to a plurality of control cells of a same type without the modification.5       153.      The population of modified immune cells of claim 152, wherein the pluralityof the population further comprises a single target nucleobase modification in a CD2 gene sequence, a CD5 gene sequence, a CIITA gene sequence, a B2M gene sequence, or a regulatory element of each thereof.

154. The population of modified immune cells of claim 152, wherein the plurality10          of the population further comprises a single target nucleobase modification in a genesequence of a gene selected from the group consisting of a CD2 gene sequence, a TRAC gene sequence, a CD3 epsilon gene sequence, a CD3 gamma gene sequence, a CD3 delta gene sequence, a TRBC1 gene sequence, a TRBC2 gene sequence, a CD4 gene sequence, a CD5 gene sequence, a CD7 gene sequence, a CD30 gene sequence, 15          a CD33 gene sequence, a CD52 gene sequence, a CD70 gene sequence, a B2M genesequence, and a CIITA gene sequence or a regulatory element of each thereof.

155. The population of modified immune cells of claim 153, wherein the pluralityof the population comprises a single nucleobase modification in each one of a TRAC gene sequence, a PDCD1 gene sequence, a CD52 gene sequence, a CD7 gene20         sequence, a CD2 gene sequence, a CD5 gene sequence, a CIITA gene sequence, and aB2M gene sequence.

156. The population of modified immune cells of any one of claims 135-155,wherein the plurality of the population comprises no detectable translocation.

157. The population of modified immune cells of any one of claims 135-156,25          wherein at least 60% of the population of immune cells are viable.

158. The population of modified immune cells of any one of claims 135-156,wherein at least 60% of the population of immune cells expand at least 80% of expansion rate of a population of control cells of a same type without the modification.30       159.      The population of modified immune cells of any one of claims 135-158,wherein population of immune cells are human cells.

160. The population of modified immune cells of any one of claims 135-159,wherein the population of immune cells are cytotoxic T cells, regulatory T cells, T helper cells, dendritic cells, B cells, or NK cells.2026204466   11 Jun 2026161. The population of modified immune cells of any one of claims 135-160, wherein the population of immune cells are derived from a single human donor.

162. The population of modified immune cells of any one of claims 135-161, wherein the plurality of cells having the modification further comprises a5          polynucleotide that encodes an exogenous functional chimeric antigen receptor(CAR) or a functional fragment thereof.

163. The population of modified immune cells of claim 162, wherein at least 50% of the population of immune cells express the CAR.

164. The population of modified immune cells of claim 162 or 163, wherein the10          CAR specifically binds a marker associated with neoplasia.

165. The population of modified immune cells of claim 164, wherein the neoplasia is a T cell cancer, a B cell cancer, a lymphoma, a leukemia, or a multiple myeloma.

166. The population of modified immune cells of claim 165, wherein the CAR specifically binds CD7.15      167. The population of modified immune cells of claim 165, wherein the CARspecifically binds BCMA.

168. The population of modified immune cells of claim of any one of claims 135167, wherein the single target nucleobase is in an exon.

169. The population of modified immune cells of claim 168, wherein the single20          target nucleobase is within an exon 1, an exon 2, or an exon 3 of the TRAC genesequence.

170. The population of modified immune cells of claim 168, wherein the single target nucleobase is within an exon 1, an exon 2, or an exon 5 of the PCDC1 gene sequence.25       171. The population of modified immune cells of claim 168, wherein the singletarget nucleobase is within an exon 1 or an exon 2 of the CD52 gene sequence.

172. The population of modified immune cells of claim 168, wherein the single target nucleobase is within an exon 1, an exon 2, or an exon 3 of a CD7 gene sequence.30       173. The population of modified immune cells of any one of claims 135-167,wherein the single target nucleobase is in a splice donor site or a splice acceptor site.

174. The population of modified immune cells of claim 173, wherein the single target nucleobase is in a exon 1 splice acceptor site, a exon 1 splice donor site, or a exon 3 splice acceptor site of the TRAC gene sequence.2026204466   11 Jun 2026175.      The population of modified immune cells of claim 173, wherein the singletarget nucleobase is in a exon 1 splice acceptor site, a exon 1 splice donor site, an exon 2 splice acceptor site, an exon 3 splice donor site, an exon 4 splice acceptor site, an exon 4 splice donor site, or a exon 5 splice acceptor site of the PDCD1 gene5           sequence.

176. The population of modified immune cells of claim 173, wherein the singletarget nucleobase is in a exon 1 splice donor site, or an exon 2 splice acceptor site of the CD52 gene sequence.

177. The population of modified immune cells of claim 173, wherein the single10           target nucleobase is in a exon 1 splice donor site, a exon 2 splice donor site, an exon 2splice acceptor site, or an exon 3 splice acceptor site of the CD7 gene sequence.

178. A composition comprising deaminase and a nucleic acid sequence, wherein the guide nucleic acid sequence comprises a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, 15       CUCUUACCUGUACCAUAACC, CACCUACCUAAGAACCAUCC,ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

179. The composition of claim 178, wherein the deaminase is associated with a nucleic acid programmable DNA binding protein (napDNAbp) to form a base editor.20      180. The composition of claim 179, wherein the napDNAbp comprises a Cas9nickase or nuclease dead Cas9 and wherein the deaminase is a cytidine deaminase.

181. The composition of claim 180, wherein the base editor further comprises auracil glycosylase inhibitor.

182. The composition of claim 166, wherein the napDNAbp comprises a Cas9 25          nickase or nuclease dead Cas9 and wherein the deaminase is a adenosine deaminase.

183. A composition comprising a polymerase and a guide nucleic acid sequence, wherein the guide nucleic acid sequence comprises a sequence selected from the group consisting of the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC,30      CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC,ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

184. The composition of claim 170, wherein the polymerase is a reverse transcriptase and wherein the guide nucleic acid sequence is an extended guide2026204466   11 Jun 2026nucleic acid sequence comprising a reverse transcription template sequence, a reverse transcription primer binding site, or a combination thereof.

185. A method for producing a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity, the method comprising:5           a) modifying a single target nucleobase in a first gene sequence or a regulatoryelement thereof in an immune cell; andb) modifying a second gene sequence or a regulatory element thereof in the immune cell with a Casl2 polypeptide, wherein the Casl2 polypeptide generates a site-specific cleavage in the second gene sequence;10          wherein each of the first gene and the second gene is a immunogenic gene, acheckpoint inhibitor gene, or an immune response regulation gene,thereby generating a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity.

186. The method of claim 185, further comprising expressing an exogenous15           functional chimeric antigen receptor (CAR) or a functional fragment thereof in theimmune cell.

187. The method of claim 186, wherein a polynucleotide encoding the CAR or the functional fragment thereof is inserted into the site specific cleavage generated by the Cas 12 polypeptide.20       188. The method of claim 187, wherein the Casl2 polypeptide is a Casl2bpolypeptide.

189. A method for producing a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity, the method comprising: a) modifying a single target nucleobase in a first gene sequence or a regulatory 25 element thereof in an immune cell; andb) modifying a second gene sequence or a regulatory element thereof in the immune cell by inserting an exogenous functional chimeric antigen receptor (CAR) or a functional fragment thereof or an exogenous functional T cell receptor or a functional fragment thereof in the second gene;30          wherein each of the first gene and the second gene is a immunogenic gene, acheckpoint inhibitor gene, or an immune response regulation gene, thereby generating a modified immune cell with reduced immunogenicity and / or increased anti-neoplasia activity.2026204466   11 Jun 2026190. The method of claim 189, wherein step b) further comprises generating a sitespecific cleavage in the second gene sequence with a nucleic acid programmable DNA binding protein (napDNAbp).

191. The method of claim 190, wherein the napDNAbp is a Cas 12b.5       192. The method of any one of claims 185-191, wherein expression of the first geneis reduced by at least 60% or wherein expression of the second gene is reduced by at least 60% as compared to a control cell of a same type without the modification.

193. The method of any one of claims 138-192, wherein the first gene is selected from the group consisting of CD3 epsilon, CD3 gamma, CD3 delta, CD4, TRAC,10        TRBC1, TRBC2, PDCD1, CD30, CD33, CD7, CD52, B2M, CD70, CIITA, CD2, andCD5.

194. The method of claim 193, wherein the first gene or the second gene is selected from the group consisting of TRAC, CIITA, CD2, CD5, CD7, and CD52.

195. The method of any one of claims 185-194, wherein the second gene is TRAC.15       196. The method of any one of claims 185-195, wherein step a) further comprisesmodifying a single target nucleobase in two other gene sequences or regulatory elements thereof.

197. The method of any one of claims 185-195, wherein step a) further comprises modifying a single target nucleobase in three other gene sequences or regulatory20          elements thereof.

198. The method of any one of claims 185-195, wherein step a) further comprises modifying a single target nucleobase in four other gene sequences or regulatory elements thereof.

199. The method of any one of claims 185-195, wherein step a) further comprises 25          modifying a single target nucleobase in five other gene sequences or regulatoryelements thereof.

200. The method of any one of claims 185-195, wherein step a) further comprises modifying a single target nucleobase in six other gene sequences or regulatory elements thereof.30      201. The method of any one of claims 185-195, wherein step a) further comprisesmodifying a single target nucleobase in seven other gene sequences or regulatory elements thereof.2026204466   11 Jun 2026202. The method of any one of claims 185-201, wherein the modifying in step a) comprises deaminating the single target nucleobase with a base editor comprising a deaminase and a nucleic acid programmable DNA binding protein (napDNAbp).

203. The method of claim 202, wherein the napDNAbp comprises a Cas9 nickase 5          or nuclease dead Cas9.

204. The method of claim 203, wherein the deaminase is a cytidine deaminase and wherein the modification comprises conversion of a cytidine (C) to a thymine (T).

205. The method of claim 203, wherein the deaminase is an adenosine deaminase and wherein the modification comprises conversion of an adenine (A) to a guanine10           (G).

206. The method of any one of claims 185-205, wherein the modifying in a) comprises contacting the immune cell with a guide nucleic acid sequence.

207. The method of claim 206, wherein the guide nucleic acid sequence comprises a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG,15       CCUACCUGUCACCAGGACCA, CUCUUACCUGUACCAUAACC,CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

208. The method of any one of claims 185-207, wherein the modifying in b)20          comprises contacting the immune cell with a guide nucleic acid sequence.

209. The method of claim 208, wherein the guide nucleic acid sequence comprises a sequence selected from sequences in Table 1.

210. The method of any one of claims 185-209, wherein the modifying in a) comprises replacing the single target nucleobase with a different nucleobase by target-25          primed reverse transcription with a reverse transcriptase and an extended guidenucleic acid sequence, wherein the extended guide nucleic acid sequence comprises a reverse transcription template sequence, a reverse transcription primer binding site, or a combination thereof.

211. The method of any one of claims 185-210, wherein the modifying in a) and b) 30          generates less than 1% indels in the immune cell.

212. The method of any one of claims 185-211, wherein the modifying in a) and b) generates less than 5% off target modification in the immune cell.

213. The method of any one of claims 185-211, wherein the modifying in a) and b) generate less than 5% non-target modification in the immune cell.2026204466   11 Jun 2026214.      The method of any one of claims 185-213, wherein the immune cell is ahuman cell.

215. The method of claim 214, wherein the immune cell is a cytotoxic T cell, a regulatory T cell, a T helper cell, a dendritic cell, a B cell, or a NK cell.5       216.      The method of any one of claims 186-215, wherein the CAR specifically bindsa marker associated with neoplasia.

217. The method of claim 216, wherein the CAR specifically binds CD7.

218. A modified immune cell with reduced immunogenicity and / or increased antineoplasia activity, wherein the modified immune cell comprises:10          a) a single target nucleobase modification in a first gene sequence or a regulatoryelement thereof; andb) a modification in a second gene sequence or a regulatory element thereof, wherein the modification is a Cas 12 polypeptide generated site-specific cleavage;wherein each of the first gene and the second gene is a immunogenic gene, a15       checkpoint inhibitor gene, or an immune response regulation gene.

219. The modified immune cell of claim 218, wherein the immune cell further comprises an exogenous functional chimeric antigen receptor (CAR) or a functional fragment thereof.

220. The modified immune cell of claim 219, wherein a polynucleotide encoding20          the CAR or the functional fragment thereof is inserted into the site specific cleavagegenerated by the Cas 12 polypeptide.

221. A modified immune cell with reduced immunogenicity and / or increased antineoplasia activity, the modified immune cell comprising:a) a single target nucleobase modification in a first gene sequence or a regulatory25 element thereof in an immune cell; andb) a modification in a second gene sequence or a regulatory element thereof, wherein the modification is an insertion of an exogenous chimeric antigen receptor (CAR) or a functional fragment thereof or an exogenous T cell receptor or a functional fragment thereof;30          wherein each of the first gene and the second gene is a immunogenic gene, acheckpoint inhibitor gene, or immune response regulation gene.

222. The modified immune cell of claim 221, wherein the modification in b) is generated by a site-specific cleavage with a Cas 12b.2026204466   11 Jun 2026223. The modified immune cell of any one of claims 218-222, wherein expression of the first gene is reduced by at least 60% or wherein expression of the second gene is reduced by at least 60% as compared to a control cell of a same type without the modification.5       224. The modified immune cell of any one of claims 218-223, wherein the firstgene or the second gene is selected from the group consisting of CD3 epsilon, CD3 gamma, CD3 delta, CD4, TRAC, TRBC1, TRBC2, PDCD1, CD30, CD33, CD7, CD52, B2M, CD70, CIITA, CD2, and CD5.

225. The method of claim 193, wherein the first gene or the second gene is selected10         from the group consisting of TRAC, CD2, CD5, CD7, and CD52.

226. The modified immune cell of claim 225, wherein the second gene is TRAC.

227. The modified immune cell of any one of claims 218-226, wherein the immunecell further comprises modification in a single target nucleobase in two other gene sequences or regulatory elements thereof.15       228. The modified immune cell of any one of claims 218-226, wherein the immunecell further comprises modification in a single target nucleobase in three other gene sequences or regulatory elements thereof.

229. The modified immune cell of any one of claims 218-226, wherein the immune cell further comprises modification in a single target nucleobase in four other gene 20          sequences or regulatory elements thereof.

230. The modified immune cell of any one of claims 218-226, wherein the immune cell further comprises modification in a single target nucleobase in five other gene sequences or regulatory elements thereof.

231. The modified immune cell of any one of claims 218-226, wherein the immune 25           cell further comprises modification in a single target nucleobase in six other genesequences or regulatory elements thereof.

232. The modified immune cell of any one of claims 218-226, wherein the immune cell further comprises modification in a single target nucleobase in seven other gene sequences or regulatory elements thereof.30      233. The modified immune cell of claim any one of claims 218-232, wherein themodification in a) is generated by a base editor comprising a deaminase and a nucleic acid programmable DNA binding protein (napDNAbp).2026204466   11 Jun 2026234. The modified immune cell of claim 233, wherein the deaminase is a cytidine deaminase and wherein the modification comprises conversion of a cytidine (C) to a thymine (T).

235. The modified immune cell of claim 233, wherein the deaminase is an5          adenosine deaminase and wherein the modification comprises conversion of anadenine (A) to a guanine (G).

236. The modified immune cell of any one of claims 218-235, wherein the immune cell comprises less than 1 % indels in the genome.

237. The modified immune cell of any one of claims 218-236, wherein the immune 10           cell is a human cell.

238. The modified immune cell of any one of claims 218-237, wherein the immune cell is a cytotoxic T cell, a regulatory T cell, a T helper cell, a dendritic cell, a B cell, or a NK cell.

239. The modified immune cell of any one of claims 218-238, wherein the CAR15           specifically binds a marker associated with neoplasia.

240. The modified immune cell of claim 239, wherein the CAR specifically binds CD7.

241. The modified immune cell of any one of claims 218-240, wherein themodification in b) is an insertion in exon 1 in the TRAC gene sequence.20      242.      A population of modified immune cells, wherein a plurality of the populationof immune cells comprises:a) a single target nucleobase modification in a first gene sequence or a regulatory element thereof in an immune cell; andb) a modification in a second gene sequence or a regulatory element thereof, wherein 25 the modification is a Cas 12 polypeptide generated site-specific cleavage;wherein each of the first gene and the second gene is a immunogenic gene, a checkpoint inhibitor gene, or an immune response regulation gene,and wherein the plurality of the population comprises an exogenous chimeric antigen receptor (CAR) or a functional fragment thereof.30      243. The population of modified immune cells of claim 242, wherein apolynucleotide encoding the CAR or the functional fragment thereof is inserted into the site specific cleavage generated by the Cas 12 polypeptide.

244. A population of modified immune cells, wherein a plurality of the populationof immune cells comprises:2026204466   11 Jun 2026a) a single target nucleobase modification in a first gene sequence or a regulatory element thereof; andb) a modification in a second gene sequence or a regulatory sequence thereof, wherein the modification is an insertion of an exogenous chimeric antigen receptor (CAR) or a5 functional fragment thereof or an exogenous T cell receptor or a functional fragment thereof;wherein each of the first gene and the second gene is a immunogenic gene, a checkpoint inhibitor gene, or immune response regulation gene, and wherein the plurality of cells with the modification in a) or b) exhibit reduced immunogenicity and / or increased 10 anti-neoplasia activity.

245. The population of modified immune cells of claim 244, wherein themodification in b) is generated by a site-specific cleavage with a Cas 12b.

246. The population of modified immune cells of any one of claims 242-245,wherein expression of the first gene is reduced by at least 60% or wherein expression 15           of the second gene is reduced by at least 60% in the plurality of cells with themodification in a) or b) as compared to plurality of control cells of a same type without the modification.

247. The population of modified immune cells of any one of claims 242-246,wherein the first gene or the second gene is selected from the group consisting of CD3 20         epsilon, CD3 gamma, CD3 delta, CD4, TRAC, TRBC1, TRBC2, PDCD1, CD30,CD33, CD7, CD52, B2M, CD70, CIITA, CD2, and CD5.

248. The population of modified immune cells of claim 247, wherein the first geneor the second gene is selected from the group consisting of TRAC, CIITA, CD2, CD5, , CD7, and CD52.25       249.      The population of modified immune cells of claim 248, wherein the first geneis TRAC,, CD7, or CD52.

250. The population of modified immune cells of claim 248, wherein the secondgene is TRAC.

251. The population of modified immune cells of any one of claims 242-250,30          wherein the plurality of cells with the modification in a) or b) further comprises amodification in a single target nucleobase in two other gene sequences or regulatory elements thereof.2026204466   11 Jun 2026252. The population of modified immune cells of claim 251, wherein the plurality of cells with the modification in a) or b) further comprises a single target nucleobase in three, four, five, or six other gene sequences or regulatory elements thereof.

253. The population of modified immune cells of any one of claims 242-252,5          wherein the modification in a) is generated by a base editor comprising a deaminaseand a nucleic acid programmable DNA binding protein (napDNAbp) to form a base editor.

254. The population of modified immune cells of claim 253, wherein the deaminase is a cytidine deaminase and wherein the modification comprises conversion of a10          cytidine (C) to a thymine (T).

255. The population of modified immune cells of claim 253, wherein the deaminase is an adenosine deaminase and wherein the modification comprises conversion of an adenine (A) to a guanine (G).

256. The population of modified immune cells of claim 254, wherein the base15           editor further comprises a uracil glycosylase inhibitor.

257. The population of modified immune cells of any one of claims 242-256, wherein at least 60% of the population of immune cells are viable.

258. The population of modified immune cells of any one of claims 242-256, wherein at least 60% of the population of immune cells expand at least 80% of 20          expansion rate of a population of control cells of a same type without themodification.

259. The population of modified immune cells of any one of claims 242-258, wherein the immune cells are a human cells.

260. The population of modified immune cells of any one of claims 242-259,25          wherein the immune cells is are cytotoxic T cells, regulatory T cells, T helper cells,dendritic cells, B cells, or NK cells.

261. The population of modified immune cells of any one of claims 242-260,wherein the CAR specifically binds a marker associated with neoplasia.

262. The population of modified immune cells of claim 261, wherein the CAR30          specifically binds CD7.

263. The population of modified immune cells of any one of claims 242-262, wherein the modification in b) is an insertion in exon 1 in the TRAC gene sequence.

264. A method for producing a modified immune cell with increased anti-neoplasia activity, the method comprising: modifying a single target nucleobase in a Cbl Proto2026204466   11 Jun 2026Oncogene B (CBLB) gene sequence or a regulatory element thereof in an immune cell, wherein the modification reduces an activation threshold of the immune cell compared with an immune cell lacking the modification; thereby generating a modified immune cell with increased anti-neoplasia activity.5       265. A composition comprising a modified immune cell with increased antineoplasia activity, wherein the modified immune cell comprises: a modification in a single target nucleobase in a Cbl Proto-Oncogene B (CBLB) gene sequence or a regulatory element thereof, wherein the modified immune cell exhibits a reduced activation threshold compared with a control immune cell of a same type without the 10           modification.

266. A population of immune cells, wherein a plurality of the population of immune cells comprises: a modification in a single target nucleobase in a CBLB gene sequence or a regulatory element thereof, wherein the plurality of the population of the immune cells comprising the modification exhibit a reduced activation threshold 15          compared with an control population of immune cells of a same type without themodification.

267. A method for producing a population of modified immune cells with increasedanti-neoplasia activity, the method comprising: modifying a single target nucleobase in a Cbl Proto Oncogene B (CBLB) gene sequence or a regulatory element thereof in a 20        population of immune cells, wherein at least 50% of the population of immune cells aremodified to comprise the single target nucleobase modification.

268. A composition comprising at least four different guide nucleic acid sequences for base editing.

269. The composition of claim 268, further comprising a polynucleotide encoding a 25 base editor polypeptide, wherein the base editor polypeptide comprises a nucleic acid programmable DNA binding protein (napDNAbp) and a deaminase.

270. The composition of claim 269, wherein the polynucleotide encoding the base editor is a mRNA sequence.

271. The composition of claim 269 or 270, wherein the deaminase is a cytidine30 deaminase or an adenosine deaminase.

272. The composition of claim 268, further comprising a base editor polypeptide, wherein the base editor polypeptide comprises a nucleic acid programmable DNA binding protein (napDNAbp) and a deaminase.2026204466   11 Jun 2026273.    The composition of claim 272, wherein the deaminase is a cytidine deaminaseor an adenosine deaminase.

274. The composition of claim 272 or 273, further comprising a lipid nanoparticle.

275. The composition of any one of claims 267-274, wherein the at least four guide5 nucleic acid sequences each hybridize with a gene sequence selected from the group consisting of CD2, CD3 epsilon, CD3 gamma, CD3 delta, CD4, CD5, CD7, CD30, CD33, CD52, CD70, and CIITA. In some embodiments, the at least 1, 2, 3, 4, 5, 6, 7, 8, or more genes or regulatory elements thereof are selected from CD2, CD3 epsilon, CD3 gamma, CD3 delta, CD4, CD5, CD7, CD30, CD33, CD52, CD70, and CIITA. In some10 embodiments, the at least 1, 2, 3, 4, 5, 6, 7, 8, or more genes or regulatory elements thereof comprise one or more genes selected from CD2, CD3 epsilon, CD3 gamma, CD3 delta, CD4, CD5, CD7, CD30, CD33, CD52, CD70, and CIITA. In some embodiments, the at least 1, 2, 3, 4, 5, 6, 7, 8, or more genes or regulatory elements thereof are selected from ACAT1, ACLY, ADORA2A, AXL, B2M , BATF, BCL2L11,15 BTLA, CAMK2D, cAMP, CASP8, Cblb, CCR5, CD2, CD3D, CD3E, CD3G, CD4, CD5, CD7, CD8A, CD33, CD38, CD52, CD70, CD82, CD86, CD96, CD123, CD160, CD244, CD276, CDK8, CDKN1B, Chi311, CIITA, CISH, CSF2CSK, CTLA-4, CUL3, Cypl lai, DCK, DGKA, DGKZ, DHX37, ELOB(TCEB2), ENTPD1 (CD39), FADD, FAS, GATA3, IL6, IL6R, IL10, IL10RA, IRF4, IRF8, JUNB, Lag3,, LAIR-1 (CD305),20 LDHA, LIF, LYN, MAP4K4, MAPK14, MCJ, MEF2D, MGAT5, NR4A1, NR4A2, NR4A3, NT5E (CD73), ODC1, OTULINL (FAM105A), PAG1, PDCD1, PDIA3, PHD1 (EGLN2), PHD2 (EGLN1), PHD3 (EGLN3), PIK3CD, PIKFYVE, PPARa, PPARd, PRDMI1, PRKACA, PTEN, PTPN2, PTPN6, PTPN11, PVRIG (CD112R), RASA2, RFXANK, SELPG / PSGL1, SIGLEC15, SLA, SLAMF7, S0CS1, Spryl,25 Spry2, STK4, SUV39, H1TET2, TGFbRII, TIGIT, Tim-3, TMEM222, TNFAIP3, TNFRSF8 (CD30), TNFRSF10B, TOX, TOX2,, TRAC, TRBC1, TRBC2, UBASH3A, VHL, VISTA, XBP1, YAP1, and ZC3H12A.

276. The composition of any one of claims 267-274, wherein the at least four guidenucleic acid sequences each hybridize with a gene sequence selected from the group30       consisting of CD3 epsilon, CD3 delta, CD3 gamma, TRAC, TRBC1, and TRBC2, CD2,CD5, CD7, CD52, CD70, and CIITA.

277. The composition of any one of claims 267-274, wherein the at least four guidenucleic acid sequences comprise a sequence selected from the group consisting of UUCGUAUCUGUAAAACCAAG, CCUACCUGUCACCAGGACCA,2026204466   11 Jun 2026CUCUUACCUGUACCAUAACC, CACCUACCUAAGAACCAUCC, ACUCACGCUGGAUAGCCUCC, ACUCACCCAGCAUCCCCAGC, CACUCACCUUAGCCUGAGCA, and CACGCACCUGGACAGCUGAC.

278. An immune cell comprising the composition of any one of claims 267-277, 5         wherein the composition is introduced into the immune cell with electroporation.

279. An immune cell comprising the composition of any one of claims 267-277, wherein the composition is introduced into the immune cell with electroporation, nucleofection, viral transduction, or a combination thereof.

280. The modified immune cell of any one of claims 86 and 88-134 having10 increased growth or viability compared to a reference cell.

281. The modified immune cell of claim 280, wherein the reference cell is animmune cell modified with a Cas9 nuclease.

282. The population of modified immune cells of claim 87 and 135-179 having increased yield of modified immune cells compared to a reference population of cells.15        283. The population of modified immune cells of claim 282, wherein the referencepopulation is a population of immune cells modified with a Cas9 nuclease.

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