Compositions and methods for mediating epitope engineering

Abstract

Provided herein are compositions and methods for genetically engineering cells (e.g., hematopoietic cells) to modify genes encoding lineage-specific cell surface antigens and to modify epitopes of the lineage-specific cell surface antigens that are recognized by agents. Also provided are methods that involve administering such genetically engineered cells to a subject, e.g., a subject with a hematopoietic malignancy, and the genetically engineered cells themselves.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit under 35 U.S.C. §119(e) of U.S. Provisional Patent Application No. 63 / 327,266, filed April 4, 2022, and U.S. Provisional Patent Application No. 63 / 424,085, filed November 9, 2022, each of which is incorporated by reference in its entirety.

[0002] Electronic Sequence Listing Reference The contents of the electronic sequence listing (V029170014WO00-SEQ-CEW.xml, size: 570,220 bytes, and creation date: April 3, 2023) are incorporated herein by reference in their entirety. [Background technology]

[0003] When a subject receives immunotherapy targeting an antigen associated with a disease or condition, such as anti-cancer CAR-T therapy, the therapy may deplete not only the diseased cells intended to be targeted, but also non-disease cells that may express the target antigen. This "on-target, off-disease" effect has been reported for some CAR-T therapeutics, such as those targeting CD19 or CD33. If the target antigen is expressed on the surface of cells required for the subject's survival or on the surface of cells whose depletion would be significantly detrimental to the subject's health, the subject may be unable to undergo immunotherapy or may experience severe side effects once such therapy is administered. Summary of the Invention

[0004] Aspects of the present disclosure describe compositions, methods, strategies, and therapeutic modalities that address the adverse on-target and off-disease effects of certain immunotherapeutic approaches, e.g., immunotherapeutic agents comprising lymphocytic effector cells that target specific antigens in subjects in need of treatment, such as CAR-T cells or CAR-NK cells. Some aspects of the present disclosure provide compositions, methods, strategies, and therapeutic modalities related to modifying an epitope of a lineage-specific cell surface antigen on a hematopoietic cell such that binding of an agent that specifically binds to the lineage-specific cell surface antigen is reduced or eliminated. In some embodiments, the epitope modification does not alter (e.g., impair) the function of the lineage-specific cell surface antigen. In some embodiments, hematopoietic cells are provided that comprise an epitope-modified lineage-specific cell surface antigen characterized by reduced or eliminated binding of an agent (e.g., an immunotherapeutic agent, such as CAR-T cells or CAR-NK cells) to the modified epitope. In some embodiments, administering such hematopoietic cells comprising the epitope-modified lineage-specific cell surface antigen, e.g., in combination with an agent, can reduce or alleviate adverse on-target and off-disease effects in a subject. Some aspects of the present disclosure provide compositions and methods for genetic modification (or gene editing) of cells using homology-directed repair (HDR). In some embodiments, the methods and compositions described herein combine sequence specificity (e.g., of the CRISPR / Cas system) with HDR-mediated gene editing to allow targeted integration of sequences from a template polynucleotide at a target sequence specified by the homology of a portion of the template polynucleotide with the target sequence. In some embodiments, the methods and compositions utilizing HDR described herein are characterized by high editing efficiency and high survival rates and / or high viability in the resulting edited cell population, e.g., an edited human hematopoietic cell population, e.g., human hematopoietic stem cells. Some embodiments of the present disclosure provide the advantage of utilizing highly efficient HDR editing to achieve targeted epitope editing, e.g., to produce modified lineage-specific cell surface antigens that retain functionality but exhibit reduced or eliminated binding to immunotherapeutic agents that target the antigen.

[0005] Accordingly, some aspects of the present disclosure provide genetically engineered hematopoietic cells, or progeny thereof, comprising a genomic modification of a gene encoding a lineage-specific cell surface antigen, wherein the genomic modification alters the amino acid sequence of an epitope recognized by an agent that specifically binds to the lineage-specific cell surface antigen, resulting in a modified lineage-specific cell surface antigen, wherein the modified lineage-specific cell surface antigen is characterized by reduced or no binding of the agent.

[0006] In some embodiments, the genomic modification alters 1, 2, 3, 4, or 5 amino acid residues of the lineage-specific cell surface antigen. In some embodiments, the genomic modification alters no more than 1, no more than 2, no more than 3, no more than 4, or no more than 5 amino acid residues of the lineage-specific cell surface antigen. In some embodiments, the genomic modification results in the deletion, substitution, insertion, or inversion of one or more amino acid residues, or a combination thereof. In some embodiments, the genomic modification results in the substitution of one or more amino acid residues.

[0007] In some embodiments, the epitope is characterized by an endogenous post-translational modification, hi some embodiments, the endogenous post-translational modification is glycosylation.

[0008] In some embodiments, the agent is an immunotherapeutic agent. In some embodiments, the immunotherapeutic agent comprises an antibody or an antigen-binding fragment thereof. In some embodiments, the modified lineage-specific cell surface antigen is not recognized by the agent. In some embodiments, the modified lineage-specific cell surface antigen is recognized by a second agent that specifically binds to a region of the lineage-specific cell surface antigen that is different from the epitope recognized by the first agent.

[0009] In some embodiments, the genomic modification does not substantially alter the function of the lineage-specific cell surface antigen. In some embodiments, the genomic modification does not substantially alter the expression of the lineage-specific cell surface antigen. In some embodiments, the genomic modification does not substantially alter the viability or growth of the cell. In some embodiments, the hematopoietic cells or their progeny retain the ability to differentiate normally compared to a reference population of hematopoietic cells, optionally a population of hematopoietic cells that does not comprise the genomic modification.

[0010] In some embodiments, the hematopoietic cells are hematopoietic stem cells (HSCs). In some embodiments, the hematopoietic cells are CD34+ cells. In some embodiments, the hematopoietic cells are obtained from bone marrow, blood, umbilical cord, or peripheral blood mononuclear cells (PBMCs). In some embodiments, the hematopoietic cells are human.

[0011] In some embodiments, the lineage-specific cell surface antigen is selected from the group consisting of CD123, CD47, CD34, CD38, CD19, CD33, CLL-1, CD30, CD5, CD6, CD7, EMR2, and BCMA. In some embodiments, the lineage-specific cell surface antigen is CD123. In some embodiments, the lineage-specific cell surface antigen is CD38. In some embodiments, the lineage-specific cell surface antigen is CD19. In some embodiments, the lineage-specific cell surface antigen is EMR2. In some embodiments, the lineage-specific cell surface antigen is CD5. In some embodiments, the lineage-specific cell surface antigen is CD47. In some embodiments, the lineage-specific cell surface antigen is CD34.

[0012] In some embodiments, the epitope is encoded by exon 3 and / or exon 4 of the gene encoding CD123. In some embodiments, the epitope is a region of CD123 bound by the murine anti-CD123 antibody 7G3, a humanized variant thereof (e.g., antibody CSL-362), or talacotuzumab. In some embodiments, the agent comprises the murine anti-CD123 antibody 7G3, a humanized variant thereof (e.g., antibody CSL-362), or talacotuzumab. In some embodiments, the epitope comprises 1, 2, 3, 4, or 5 amino acids at positions 51, 59, 61, 82, or 84 of the wild-type gene encoding CD123. In some embodiments, the genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 51, 59, 61, 82, or 84 of the wild-type gene encoding CD123, or at the corresponding positions of a homologous CD123 gene. In some embodiments, the genomic modification results in a substitution of one or more (e.g., 1, 2, 3, 4, or all) amino acids at positions 51, 59, 61, 82, or 84 of the wild-type gene encoding CD123, or at the corresponding positions of a homologous CD123 gene. In some embodiments, the one or more substitutions are conservative substitutions. In some embodiments, the genomic modification results in a substitution of an amino acid at position 51 of the wild-type gene encoding CD123, or at the corresponding position of a homologous CD123 gene. In some embodiments, the genomic modification results in a substitution of a lysine for a glutamic acid at position 51 of the wild-type gene encoding CD123, or at the corresponding position of a homologous CD123 gene.

[0013] In some embodiments, the epitope is encoded by exon 7 of the gene encoding CD38. In some embodiments, the epitope is a region of CD38 bound by the murine anti-CD38 antibody HB7, a humanized variant thereof, or daratumumab. In some embodiments, the agent comprises the murine anti-CD38 antibody HB7, a humanized variant thereof, or daratumumab. In some embodiments, the epitope comprises 1, 2, 3, 4, or 5 amino acids at positions 270-274 of the wild-type gene encoding CD38. In some embodiments, the genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 270-274 of the wild-type gene encoding CD38 or at the corresponding positions in a homologous CD38 gene. In some embodiments, the genomic modification results in the substitution of one or more (e.g., one, two, three, four, or all) amino acids at positions 270-274 of the wild-type gene encoding CD38, or at the corresponding positions in a homologous CD38 gene. In some embodiments, the one or more substitutions are conservative substitutions. In some embodiments, the genomic modification results in the substitution of an amino acid at position 272 of the wild-type gene encoding CD38, or at the corresponding position in a homologous CD38 gene. In some embodiments, the genomic modification results in the substitution of an arginine, histidine, or alanine for glutamine at position 272 of the wild-type gene encoding CD38, or at the corresponding position in a homologous CD38 gene.

[0014] In some embodiments, the epitope is encoded by exon 2 or exon 4 of CD19. In some embodiments, the epitope is a region of CD19 bound by anti-CD19 antibody B43, anti-CD19 antibody FMC63, or a fragment thereof. In some embodiments, the agent comprises anti-CD19 antibody B43, anti-CD19 antibody FMC63, tafasitamab, loncastuximab, blinatumomab, or a fragment thereof. In some embodiments, the epitope comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at positions 216-224 or 218-238 of a wild-type gene encoding CD19. In some embodiments, the genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 163, 164, 216-224, or 218-238 of the wild-type gene encoding CD19, or at the corresponding positions of a homologous CD19 gene. In some embodiments, the genomic modification results in the substitution of one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more, e.g., all) amino acids at positions 163, 164, 216-224, or 218-238 of the wild-type gene encoding CD19, or at the corresponding positions of a homologous CD19 gene. In some embodiments, the one or more substitutions are conservative substitutions. In some embodiments, the genomic modification results in an amino acid substitution at position 163 of the wild-type gene encoding CD19 or at the corresponding position of a homologous CD19 gene. In some embodiments, the genomic modification results in a cysteine ​​or leucine substitution of the amino acid at position 163 of the wild-type gene encoding CD19 or at the corresponding position of a homologous CD19 gene. In some embodiments, the genomic modification results in an amino acid substitution at positions 163 and 220 of the wild-type gene encoding CD19 or at the corresponding positions of a homologous CD19 gene. In some embodiments, the genomic modification results in an amino acid substitution at positions 163 and 164 of the wild-type gene encoding CD19 or at the corresponding positions of a homologous CD19 gene.In some embodiments, the genomic modification results in an amino acid substitution at positions 163 and 164 of the wild-type gene encoding CD19, or at the corresponding positions of a homologous CD19 gene, wherein the amino acid substitution at position 163 is a cysteine ​​or leucine and the amino acid substitution at position 164 is a phenylalanine. In some embodiments, the genomic modification results in a substitution of a phenylalanine for the amino acid at position 164 of the wild-type gene encoding CD19, or at the corresponding position of a homologous CD19 gene.

[0015] In some embodiments, the epitope comprises 1, 2, 3, 4, 5, or 6 amino acids at positions 124, 132, 146, 292, 294, 295, 296, 298, 299, 303, 304, 305, 306, 307, 308, 312, 318, 320, 328, 329, 331, 332, 335, 340, 347, 527, or 708 of a wild-type gene encoding EMR2. In some embodiments, the genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 124, 132, 146, 292, 294, 295, 296, 298, 299, 303, 304, 305, 306, 307, 308, 312, 318, 328, 329, 331, 332, 335, 340, 347, 527, or 708 of the wild-type gene encoding EMR2, or at the corresponding positions of a homologous EMR2 gene.

[0016] In some embodiments, the epitope is a region of CD47 bound by the anti-CD47 antibody B6H12, the anti-CD47 antibody 2D3, or a fragment thereof. In some embodiments, the agent comprises the anti-CD47 antibody B6H12, the anti-CD47 antibody 2D3, ligufalimab, or a fragment thereof. In some embodiments, the epitope comprises 1, 2, 3, 4, 5, or 6 amino acids at positions 117-122 of the wild-type gene encoding CD47. In some embodiments, the epitope comprises 1, 2, 3, or 4 amino acids at positions 47, 49, 52-55, or 117-122 of the wild-type gene encoding CD47. In some embodiments, the genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 31, 47, 49, 52-55, 117-122, or 124 of the wild-type gene encoding CD47, or at the corresponding positions of a homologous CD47 gene. In some embodiments, the one or more substitutions are conservative substitutions. In some embodiments, the genomic modification results in a substitution of one or more amino acids at positions 31, 47, 49, 52-55, 117-122, or 124 of the wild-type gene encoding CD47, or at the corresponding positions of a homologous CD47 gene. In some embodiments, the genomic modification results in a substitution of an amino acid at position 49 of the wild-type gene encoding CD47, or at the corresponding position of a homologous CD47 gene.In some embodiments, the genomic modification is a substitution of the amino acid histidine at position 4 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene, a substitution of the amino acid arginine at position 49 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene, a substitution of the amino acid proline at position 49 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene, or a substitution of the amino acid alanine at position 52 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene. a substitution of an alanine at the amino acid at position 53 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene; a substitution of a proline at the amino acid at position 53 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene; a substitution of an alanine at the amino acid at position 120 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene; or a substitution of a lysine at the amino acid at position 124 of the wild-type gene encoding CD47 or at the corresponding position in a homologous CD47 gene.

[0017] In some embodiments, the epitope is a region of CD34 bound by anti-CD34 antibody QBend10, anti-CD34 antibody 561, or an antigen-binding fragment thereof. In some embodiments, the genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 42, 45, 46, 47, 49, 50, 51, 54, or 55 of the wild-type gene encoding CD34, or at the corresponding positions of a homologous CD34 gene. In some embodiments, the one or more substitutions are conservative substitutions. In some embodiments, the genomic modification results in a substitution of one or more amino acids at positions 42, 45, 46, 47, 49, 50, 51, 54, or 55 of the wild-type gene encoding CD34, or at the corresponding positions of a homologous CD34 gene. In some embodiments, the genomic modification results in a substitution of the amino acid alanine at any one or more of positions 45, 46, 50, 51, 54, and 55 of the wild-type gene encoding CD34, or at the corresponding position in a homologous CD34 gene. In some embodiments, the genomic modification results in a substitution of the amino acid phenylalanine at position 46 of the wild-type gene encoding CD34 or at the corresponding position in a homologous CD34 gene, a substitution of the amino acid lysine at position 47 of the wild-type gene encoding CD34 or at the corresponding position in a homologous CD34 gene, a substitution of the amino acid glutamic acid at position 47 of the wild-type gene encoding CD34 or at the corresponding position in a homologous CD34 gene, a substitution of the amino acid phenylalanine at position 49 of the wild-type gene encoding CD34 or at the corresponding position in a homologous CD34 gene, or a substitution of the amino acid serine at position 49 of the wild-type gene encoding CD34 or at the corresponding position in a homologous CD34 gene.

[0018] In another aspect, the present disclosure relates to a method comprising administering to a subject in need thereof a population of genetically engineered hematopoietic cells described herein or progeny thereof. In some embodiments, the method of the present disclosure further comprises administering an effective amount of an agent that specifically binds to a lineage-specific cell surface antigen. In some embodiments, the subject has a hematopoietic malignancy.

[0019] In some embodiments, the agent is a single-chain antibody fragment (scFv). In some embodiments, the agent is an antibody or an antibody-drug conjugate (ADC). In some embodiments, the agent is an immune cell expressing a chimeric antigen receptor comprising the antigen-binding fragment.

[0020] In some embodiments, the immune cells are T cells. In some embodiments, the T cells express CD3, CD4, and / or CD8.

[0021] In some embodiments, the chimeric antigen receptor further comprises a hinge domain, a transmembrane domain, at least one costimulatory domain, a cytoplasmic signaling domain, or a combination thereof. In some embodiments, the chimeric antigen receptor comprises at least one costimulatory signaling domain derived from a costimulatory receptor selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, GITR, HVEM, and a combination thereof. In some embodiments, the chimeric antigen receptor comprises a cytoplasmic signaling domain derived from CD3ζ. In some embodiments, the chimeric antigen receptor comprises a hinge domain derived from CD8α or CD28.

[0022] In some embodiments, the agent comprises the murine anti-CD123 antibody 7G3, a humanized variant thereof (e.g., antibody CSL-362), or talacuzumab; the murine anti-CD38 antibody HB7, a humanized variant thereof, or daratumumab; B43; blinatumomab; FMC63, or HIB19; or the anti-CD47 antibody B6H12 or 2D3; or the anti-CD34 antibody QBend10 or 561; or the anti-CD5 antibody H65.

[0023] In some embodiments, the hematopoietic malignancy is Hodgkin's lymphoma, non-Hodgkin's lymphoma, leukemia, multiple myeloma (MM), myelodysplastic syndrome (MDS), or blastic plasmacytoid dendritic cell neoplasm (BPDCN). In some embodiments, the hematopoietic malignancy is acute myeloid leukemia, B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia, acute lymphoblastic leukemia, or chronic lymphoblastic leukemia. In some embodiments, the hematopoietic malignancy is B-cell acute lymphoblastic leukemia (B-ALL). In some embodiments, the hematopoietic malignancy is acute myeloid leukemia (AML). In some embodiments, the hematopoietic malignancy is multiple myeloma (MM). In some embodiments, the hematopoietic malignancy is myelodysplastic syndrome (MDS).

[0024] In another aspect, the disclosure relates to a method comprising genetically modifying a hematopoietic cell to introduce a genomic modification in a gene encoding a lineage-specific cell surface antigen, wherein the genomic modification alters the amino acid sequence of an epitope recognized by an agent that specifically binds to the lineage-specific cell surface antigen, resulting in a modified lineage-specific cell surface antigen, wherein the modified lineage-specific cell surface antigen is characterized by reduced or no binding of the agent, thereby producing a genetically engineered hematopoietic cell that has reduced or no binding to an agent that targets the lineage-specific cell surface antigen. In some embodiments, the method of the disclosure further comprises providing the hematopoietic cell.

[0025] In some embodiments, the genetically engineered hematopoietic cells are genetically engineered hematopoietic cells described herein.

[0026] In some embodiments, genetically modifying a hematopoietic cell comprises contacting the cell with (a) a clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) (CRISPR / Cas) system comprising a Cas nuclease and a guide RNA (gRNA) comprising a nucleotide sequence that hybridizes to a gene encoding a lineage-specific cell surface antigen in the genome of the hematopoietic cell (e.g., a sequence encoding an epitope bound by an agent that specifically binds to the lineage-specific cell surface antigen), and (b) a template polynucleotide. In some embodiments, the contacting further comprises contacting the hematopoietic cell with (c) one or both of an expansion factor and a homology-directed repair (HDR) promoting factor. In some embodiments, the CRISPR / Cas system generates a double-strand break (DSB) in the gene encoding the lineage-specific cell surface antigen in the genome of the hematopoietic cell.

[0027] In some embodiments, the template polynucleotide is a single-stranded donor oligonucleotide (ssODN) or a double-stranded donor oligonucleotide (dsODN). In some embodiments, the template polynucleotide hybridizes to a genomic sequence adjacent to a DSB in a gene encoding a lineage-specific cell surface antigen and is integrated into the gene encoding the lineage-specific cell surface antigen. In some embodiments, the template polynucleotide comprises a donor sequence, a first flanking sequence that is homologous to a genomic sequence upstream of the DSB in the gene encoding the lineage-specific cell surface antigen, and a second flanking sequence that is homologous to a genomic sequence downstream of the DSB in the gene encoding the lineage-specific cell surface antigen. In some embodiments, the donor sequence of the template polynucleotide is integrated into the genome of a hematopoietic cell by homology-directed repair (HDR).

[0028] In some embodiments, the expansion factors include SR1 and UM171. In some embodiments, the HDR-promoting factors include at least one of SCR7, NU7441, Rucaparib, and RS-1.

[0029] In some embodiments, the ssODN is 50 to 200 nucleotides in length, hi some embodiments, the ssODN is 120 nucleotides in length.

[0030] In some embodiments, the contacting comprises contacting a population of hematopoietic cells. In some embodiments, the methods described herein further comprise sorting the population of hematopoietic cells. In some embodiments, the sorting comprises selecting viable hematopoietic cells. In some embodiments, the sorting comprises selecting hematopoietic cells that have integrated donor sequences into their genome. In some embodiments, the sorting comprises fluorescence-activated cell sorting (FACS). In some embodiments, the sorting comprises selecting viable, long-term engrafting HSCs.

[0031] In some embodiments, the editing efficiency in a population of hematopoietic cells is at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 95, or at least 99%. In some embodiments, the percent viability in a population of hematopoietic cells is at least 50, at least 60, at least 70, at least 80, at least 90, at least 95, or at least 99%. In some embodiments, the efficiency of HDR is 50% or greater. In some embodiments, the efficiency of HDR is 60% or greater. In some embodiments, the efficiency of HDR is 80% or greater.

[0032] In some embodiments, the lineage-specific cell surface antigen is selected from the group consisting of CD33, CD123, CD19, CLL-1, CD30, CD5, CD6, CD7, CD34, CD38, CD47, EMR2 / CD312, and BCMA. In some embodiments, the lineage-specific cell surface antigen is CD123. In some embodiments, the lineage-specific cell surface antigen is EMR2.

[0033] In some embodiments, the gRNA comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 3, 6, 9, and 12.

[0034] In some embodiments, the first flanking sequence is homologous to a first portion of the CD123 gene, and the second flanking sequence is homologous to a second portion of the CD123 gene. In some embodiments, the first portion of the CD123 gene comprises a portion of exon 3 or a sequence proximal thereto. In some embodiments, the first portion of the CD123 gene comprises a portion of exon 4 or a sequence proximal thereto. In some embodiments, the second portion of the CD123 gene comprises a portion of exon 3 or a sequence proximal thereto. In some embodiments, the second portion of the CD123 gene comprises a portion of exon 4 or a sequence proximal thereto. In some embodiments, the first and second portions are not identical. In some embodiments, the donor sequence comprises a sequence corresponding to codons encoding 1, 2, 3, 4, or 5 amino acids at positions 51, 59, 61, 82, or 84 of a wild-type gene encoding CD123. In some embodiments, the first flanking sequence comprises a flanking sequence set forth in any one of SEQ ID NOs: 93-99. In some embodiments, the second flanking sequence comprises a flanking sequence set forth in any one of SEQ ID NOs: 93-99. In some embodiments, the donor sequence comprises a donor sequence set forth in any one of SEQ ID NOs: 93-99. In some embodiments, the template polynucleotide comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 93-99.

[0035] In some embodiments, the lineage-specific cell surface antigen is CD38. In some embodiments, the gRNA comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, and 60. In some embodiments, the first flanking sequence is homologous to a first portion of the CD38 gene, and the second flanking sequence is homologous to a second portion of the CD38 gene. In some embodiments, the first portion of the CD38 gene comprises a portion of exon 7 or a sequence proximal thereto. In some embodiments, the second portion of the CD38 gene comprises a portion of exon 7 or a sequence proximal thereto. In some embodiments, the first and second portions are not identical. In some embodiments, the donor sequence comprises a sequence corresponding to codons encoding 1, 2, 3, 4, or 5 amino acids at positions 270 to 274 of a wild-type gene encoding CD38.

[0036] In some embodiments, the lineage-specific cell surface antigen is CD19. In some embodiments, the gRNA comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 66, 69, 72, 75, 78, 81, and 84. In some embodiments, the first flanking sequence is homologous to a first portion of the CD19 gene, and the second flanking sequence is homologous to a second portion of the CD19 gene. In some embodiments, the first portion of the CD19 gene comprises a portion of exon 2 or a sequence proximal thereto. In some embodiments, the first portion of the CD19 gene comprises a portion of exon 4 or a sequence proximal thereto. In some embodiments, the second portion of the CD19 gene comprises a portion of exon 2 or a sequence proximal thereto. In some embodiments, the second portion of the CD19 gene comprises a portion of exon 4 or a sequence proximal thereto. In some embodiments, the first and second portions are not identical. In some embodiments, the donor sequence comprises a sequence corresponding to codons encoding 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at positions 216-224 or 218-238 of the wild-type gene encoding CD19.

[0037] In some embodiments, the genomic modification results in the expression of a variant form of the lineage-specific cell surface antigen that is not recognized by the agent, hi some embodiments, the genomic modification results in the expression of a variant form of the lineage-specific cell surface antigen that is recognized by a second agent that specifically binds to a different region of the lineage-specific cell surface antigen than the agent that binds the epitope.

[0038] In some embodiments, the Cas nuclease is a Cas9 nuclease. In some embodiments, the Cas nuclease is a Streptococcus pyogenes Cas9 (spCas9) nuclease. In some embodiments, the Cas nuclease is a Staphylococcus aureus Cas9 (saCas9) nuclease. In some embodiments, the Cas nuclease is a Cas12a nuclease. In some embodiments, the Cas nuclease is a Cas12b nuclease.

[0039] In some embodiments, the contacting comprises introducing the CRISPR / Cas system into the cell in the form of a preformed ribonucleoprotein (RNP) complex. In some embodiments, the ribonucleoprotein complex is introduced into the hematopoietic cell via electroporation. In some embodiments, the template polynucleotide and the CRISPR / Cas system are co-electroporated into the cell.

[0040] In another aspect, the disclosure relates to a method of producing a genetically engineered hematopoietic stem or progenitor cell, or a plurality thereof, comprising at least one nucleotide substitution in a gene encoding a lineage-specific cell surface antigen, the method comprising introducing into the hematopoietic stem or progenitor cell a guide RNA (gRNA) comprising a targeting domain that targets a nucleotide sequence within the genome of the hematopoietic stem or progenitor cell, and a base editor comprising a catalytically impaired Cas9 endonuclease fused to a cytosine (CBE) or an adenosine deaminase (CBE), thereby producing the genetically engineered hematopoietic stem or progenitor cell, or a plurality thereof.

[0041] In some embodiments, at least one substitution produces a missense variant in the gene encoding the lineage-specific cell surface antigen. In some embodiments, at least one substitution produces an alteration in the translation start site of the gene encoding the lineage-specific cell surface antigen. In some embodiments, at least one substitution produces a splice region variant in the gene encoding the lineage-specific cell surface antigen. In some embodiments, the substitution results in reduced or eliminated expression of the gene encoding the wild-type form of the lineage-specific cell surface antigen.

[0042] In some embodiments, the gene encoding the lineage-specific cell surface antigen is selected from the group consisting of CD123, CD47, CD34, CD38, CD19, CD33, CLL-1, CD30, CD5, CD6, CD7, and BCMA. In some embodiments, the gene encoding the lineage-specific cell surface antigen is selected from the group consisting of CD123, CD47, CD34, CD38, CD19, and CD5. In some embodiments, the gene encoding the lineage-specific cell surface antigen is CD123. In some embodiments, the gene encoding the lineage-specific cell surface antigen is CD47. In some embodiments, the gene encoding the lineage-specific cell surface antigen is CD34. In some embodiments, the gene encoding the lineage-specific cell surface antigen is CD38. In some embodiments, the gene encoding the lineage-specific cell surface antigen is CD19. In some embodiments, the gene encoding the lineage-specific cell surface antigen is CD5.

[0043] In some embodiments, the gRNA comprises a nucleotide sequence set forth in any one of Tables 1-13. In some embodiments, the gRNA comprises a nucleotide sequence set forth in any one of SEQ ID NOs: 1-12, 16-60, 64-84, 100-181, 195, 196, and 204-423.

[0044] In some embodiments, the catalytically impaired Cas9 nuclease is SpRY Cas9. In some embodiments, the catalytically impaired Cas9 nuclease is SpG Cas9. In some embodiments, the base editor is introduced into the cell as mRNA. In some embodiments, the base editor and gRNA are introduced into the cell via electroporation.

[0045] In some embodiments, the method further comprises sorting the genetically engineered hematopoietic stem or progenitor cells, or a plurality thereof, via fluorescence-activated cell sorting (FACS).

[0046] In another aspect, the disclosure relates to genetically engineered hematopoietic cells, the cells being obtained or obtainable by the methods described herein.

[0047] In another aspect, the present disclosure relates to a population of genetically engineered hematopoietic cells comprising a plurality of the genetically engineered hematopoietic cells described herein.

[0048] In another aspect, the present disclosure relates to a pharmaceutical composition comprising a genetically engineered hematopoietic cell described herein or its progeny, or a population of genetically engineered hematopoietic cells described herein.

[0049] In another aspect, the present disclosure relates to a method of treating a hematopoietic disorder, comprising administering to a subject in need thereof an effective amount of the genetically engineered stem or progenitor cells, cell populations thereof, or pharmaceutical compositions thereof, hi some embodiments, the hematopoietic disorder is a hematopoietic malignancy.

[0050] In some embodiments, the method further comprises administering an effective amount of an agent that targets a wild-type form of the lineage-specific cell surface antigen. In some embodiments, the agent comprises an antibody or antigen-binding fragment that binds to a wild-type form of the lineage-specific cell surface antigen. In some embodiments, the antibody is selected from the group consisting of the anti-CD123 antibody 7G3, talatuzumab, the anti-CD38 antibody HB7, daratumumab, the anti-CD38 antibody B43, blinatumomab, the anti-CD19 antibody FMC63, the anti-CD19 antibody HIB19, the anti-CD47 antibody B6H12, the anti-CD47 antibody 2D3, the anti-CD34 antibody QBend10, the anti-CD34 antibody 561, and the anti-CD5 antibody H65.

[0051] In some embodiments, the agent is an immune cell. In some embodiments, the immune cell is a cytotoxic T cell. In some embodiments, the cytotoxic T cell expresses a chimeric antigen receptor (CAR) comprising an antibody or antigen-binding fragment that binds to a wild-type form of a lineage-specific cell surface antigen.

[0052] In some embodiments, the genetically engineered stem or progenitor cells, immune cells, or both are allogeneic, hi some embodiments, the genetically engineered stem or progenitor cells, immune cells, or both are autologous.

[0053] In some embodiments, the subject is a human patient with Hodgkin's lymphoma, non-Hodgkin's lymphoma, leukemia, acute myeloid leukemia (AML), chronic myeloid leukemia, acute lymphoblastic leukemia, or chronic lymphoblastic leukemia.

[0054] The above summary is intended to describe in a non-limiting manner some of the embodiments, advantages, features, and uses of the technology disclosed herein. Other embodiments, advantages, features, and uses of the technology disclosed herein will be apparent from the detailed description, drawings, examples, and claims. [Brief explanation of the drawings]

[0055] [Figures 1A-1D]An exemplary strategy for CD123 editing to specifically target diseased cells using monoclonal antibody therapy is shown. Figure 1A shows the crystal structure of CD123 bound to the anti-CD123 antibody CSL-362 (JNJ-56022473, 7G3) (from PDB:4JZJ). Figure 1B shows the crystal structure of the N-terminal domain (NTD) of CD123 bound to the heavy and light chains of CSL-362 (JNJ-56022473; 7G3). Amino acid residues S59, E51, and R84 of CD123 are labeled as important residues for binding of the CSL-362 antibody to CD123. The crystal structure was obtained from PDB:4JZJ. Figure 1C shows a list of residues believed to be important for CD123 binding to CSL-362 (7G3), including E51, S59, P61, T82, and R84. FIG. 1D shows a diagram of an exemplary process for generating the CD123-targeting antibody talacuzumab (JNJ-56022473) from the murine anti-CD123 antibody 7G3 to the humanized form CSL-362. [Figure 2A-2B] Identification of acceptable CD123 variants. Figure 2A shows annotation of CD123 variants in the human genome using the Genome Aggregation Database (gnomAD). Figure 2B shows amino acid sequence alignments of human CD123 with various non-human primate CD123 sequences. [Figures 3A-3F]Exemplary strategies for characterization of epitope modifications and downstream mutations are shown. Figure 3A shows an exemplary modification mutagenesis strategy. Figure 3B shows an exemplary strategy for ectopic expression and screening of CD123 mutations. Figure 3C shows an exemplary microscopy analysis used to screen CD123 mutations. Figures 3D-3E show flow cytometry analysis of antibody binding to CD123-modified epitopes (i.e., CD123 mutations) (APC = allophycocyanin, PE = phycoerythrin). Figure 3D shows flow cytometry analysis of CD123 antibody clones 6H6 (left panel) and 9F5 (right panel) binding to the indicated CD123-modified epitopes using the PE fluorophore. Figure 3E shows a comparison of flow cytometry analysis of CD123 antibody clone 7G3 binding to the CD123-modified epitope using the APC (left panel) or PE (right panel) fluorophore. Figure 3F shows the percentage of PE+ only (left plot) CD123+ cells relative to the GFP+ and PE+ (right plot) cell populations as a plasmid expression control for the 6H6 / 9F5 antibody clone (top plot) and the 7G3 clone (bottom plot). [Figure 4A-4B] Flow cytometry analysis of the effect of IL3 on antibody binding to CD123 and epitope-modified CD123 is shown. Figure 4A shows that IL3 does not affect antibody binding to CD123 for either antibody clone 6H6 (left plot) or clone 7G3 (right plot). Figure 4B shows that IL3 does not affect antibody binding to epitope-modified (E51K) CD123 for either antibody clone 6H6 (left plot) or clone 7G3 (right panel). [Figures 5A-5E]Figure 5 shows the characteristics of daratumumab (Genmab / JNJ) binding to the cyclic ADP-ribose hydrolase 1 type II transmembrane glycoprotein (also known as CD38). Figure 5A shows an annotated diagram of the structural features of CD38 isoforms, indicating the daratumumab binding site. Figure 5B shows the crystal structure of daratumumab (top polypeptide) docked onto CD38 (bottom polypeptide), showing residues important for binding (central amino acid segment). Figure 5C shows flow cytometry analysis of anti-CD38 antibodies binding to CD38 wild-type and the indicated CD38 mutants. Figure 5D shows flow cytometry analysis of HB7 antibody clones (right panel) binding to the indicated CD38 mutants or CD38 wild-type compared to the HIT2 antibody clone control (left panel). Figure 5E shows flow cytometry analysis of HB7 antibody clones (right panel) binding to cells expressing the indicated CD38 glutamine 272 mutations (Q272A, Q272H, or Q272R) or CD38 wild-type compared to the HIT2 antibody clone control (left panel). [Figure 6] FIG. 1 shows a diagram of an exemplary experimental design for editing CD34+ cells using CRISPR-mediated ssODN-based homology-directed repair (HDR). [Figure 7] FIG. 1 shows a diagram of an exemplary HDR approach used to edit the interleukin-3 receptor alpha type 1 cytokine receptor (CD123) gene, showing the sites targeted by an exemplary guide RNA (gRNA) and donor oligonucleotide (ssODN). [Figures 8A-8C]Figure 8A shows flow cytometry analysis of HDR-edited CD34+ hematopoietic stem cells (HSCs) stained with anti-CD123 antibodies 7G3 (light gray peak) and 6H6 (dark gray peak), where gRNA (g31 and g29) and ssODN (ss31 and ss29) correspond to the oligonucleotides mapped in Figure 7. Figure 8A shows flow cytometry analysis of healthy cells from donor 1 treated with mock electroporation (EP) (negative control without electroporation), ss29 / or ss31 alone (negative ssODN control), Cas9g29 or Cas9g31 alone (positive control for NHEJ cleavage), or both. Figure 8B shows flow cytometry analysis of healthy cells from donor 2. Figure 8C shows quantification of the histogram representation of data in Figures 8A and 8B (y-axis shows % staining of CD123+ cells with 6H6 or 7G3 gated based on isotype control). [Figure 9A-9B] An exemplary experimental approach for editing and characterizing HDR-edited CD123 mutations is shown. Figure 9A shows a diagram of an exemplary experimental design for HDR-editing of cells from three CD34+ donors (donor 1, donor 2, and TIB-202 (THP-1 cells)). Figure 9B shows radiation-assisted amplification sequencing (RAMP-Seq) data as a quality check for DNA sequencing controls. [Figures 10A-10C]Figure 10A shows the editing results from HDR targeting of AML donor cells. Figure 10A shows a diagram of possible genomic changes after the HDR editing procedure. Non-homologous end joining (NHEJ) can result in deletions at genomic loci, "incomplete" editing can result in a combination of deletion and integration of mutations encoded by single-stranded donor oligonucleotides (ssODNs), and HDR results in site-specific changes at genomic loci using donor DNA templates for direct repair of the cut site. Figure 10B shows a summary of exemplary editing rates resulting from targeting donor cells with g29, ss29, or g29 + ss29 (G116), or g31, ss31, or g31 + ss31, and TIB-202 refers to the CD123+ control cell line. Figure 10C shows flow cytometry analysis of knockout (KO) and HDR product-bearing CD34 donor 2 cells via staining with an antibody that does not recognize the HDR-edited epitope of CD123 (antibody clone 6H6) and an antibody that recognizes the HDR-edited epitope (antibody clone 7G3). [Figures 11A-11D] Results from CD19 epitope modification are shown. Figure 11A shows flow cytometry analysis of anti-CD19 antibody clone FMC63 (right panel) binding to HEK293T cells expressing the indicated CD19 mutations compared to HIB19 control antibody (left panel). Figure 11B shows flow cytometry analysis of anti-CD19 clone FMC63 (left panel) binding to HEK293T cells expressing the indicated CD19 mutations compared to HIB19 control antibody (right panel). Figure 11C shows exemplary gRNAs for CD19 epitope modification and predicted substitution mutations. Figure 11D shows Sanger sequencing and flow cytometry analysis of Raji cells expressing CD19 epitope modifications at amino acid positions 162, 163, and / or 164 of CD19. Flow cytometry was performed using anti-CD19 antibody clone FMC63. [Figures 12A-12B]Figure 12 shows results from epitope modification of CD47. Figure 12A shows flow cytometry analysis of anti-CD47 clone B6H12 (left panel) binding to HEK293T cells expressing the indicated CD47 mutations compared to the 2D3 control antibody (right panel). Figure 12B shows quantification of flow cytometry analysis of anti-CD47 clone B6H12 binding to HEK293T cells expressing the indicated CD47 mutations. [Figures 13A-13F]

[0049] Figure 13A shows results from epitope modification of CD34. Figure 13A shows flow cytometry analysis of anti-CD34 clones QBend10 (left panel) and 561 (right panel) binding to HEK293T cells expressing the indicated CD34 mutations. Figure 13B shows flow cytometry analysis of anti-CD34 clones QBend10 (left panel) and 561 (right panel) binding to HEK293T cells expressing the indicated CD34 mutations. Figure 13C shows quantification of the flow cytometry data shown in Figure 13B. Figure 13D shows exemplary gRNAs for epitope modification of CD34 using base editors CBE or ABE in hematopoietic stem progenitor cells (HSPCs). Figure 13E shows flow cytometry analysis of binding of anti-CD34 clones QBend10 (left panel) and 561 (right panel) to CD34+ donor cells after transfection with the indicated gRNA and either CBE or control RNP (Cas9 and CD34 gRNA). Figure 13F shows quantification of the flow cytometry data in Figure 13E. [Figure 14] 1 shows the crystal structure of CD5, showing the extracellular and transmembrane domains (TMDs), as well as the binding region of the anti-CD5 monoclonal antibody clone H65 located in domain 1. [Figures 15A-15B]Results from epitope modification of EMR2 are shown. Figure 15A shows the crystal structure of EMR2 (source: alphafold.ebi.ac.uk / entry / A0JNV7) showing the EGF domain as well as the binding region of anti-EMR2 monoclonal antibody clone 2A1 located in helix 1 of the GAIN domain / GPS. Figure 15B shows flow cytometry analysis of Flag L5 control antibody (left panel) binding to HEK293T cells expressing the indicated EMR2 mutations compared to anti-EMR2 clone 2A1 antibody (right panel). DETAILED DESCRIPTION OF THE INVENTION

[0056] Some embodiments of the present disclosure provide compositions and methods for genetically engineering cells (e.g., hematopoietic cells, e.g., hematopoietic stem cells (HSCs)) to modify genes encoding lineage-specific cell surface antigens and alter the amino acid sequence of an epitope of the lineage-specific cell surface antigen recognized by an agent. Some embodiments of the present disclosure are based, at least in part, on the identification and characterization of modified epitopes that reduce or abolish binding of the agent. Some embodiments of the present disclosure provide strategies and therapeutic modalities related to genetically modified / engineered cells that express modified epitopes that are variant forms of lineage-specific cell surface antigens targeted by therapeutic agents, such as immunotherapeutics. The genetically engineered cells provided herein are useful, for example, to reduce or avoid certain undesirable effects associated with certain immunotherapeutics, such as any on-target or off-disease cytotoxicity.

[0057] Such undesirable effects associated with certain immunotherapeutic agents may occur, for example, when healthy cells in a subject requiring immunotherapeutic intervention express the antigen targeted by the immunotherapeutic agent. For example, a subject may be diagnosed with a malignancy associated with elevated levels of expression of a particular antigen that is typically not expressed on healthy cells but may be expressed at relatively low levels on a subset of non-malignant cells in the subject. Alternatively, or in addition, a subject may require removal of cells expressing lineage-specific cell surface antigens such as CD33, CD123, CD19, CLL-1, CD30, CD5, CD6, CD7, CD34, CD38, CD47, EMR2 (CD312), and BCMA. Administration of an immunotherapeutic agent, e.g., an antigen-targeting CAR-T cell therapy or a therapeutic antibody or antibody-drug conjugate (ADC), to a subject can result in efficient killing of target cells, e.g., malignant cells characterized by expressing the respective lineage-specific cell surface antigen, but can also result in elimination of non-target cells in the subject that express the antigen, e.g., hematopoietic cells characterized by expressing the respective lineage-specific cell surface antigen. This on-target and off-disease cytotoxicity can result in significant side effects and, in some cases, can completely negate the use of the immunotherapeutic agent.

[0058] The compositions, methods, strategies, and treatments provided herein address the problem of on-target and off-disease cytotoxicity of certain immunotherapeutic agents. Some aspects of the present disclosure provide engineered cells containing modifications in their genome that result in the expression of modified lineage-specific cell surface antigens that exhibit reduced or no binding to agents (e.g., immunotherapeutic agents) that specifically bind to the lineage-specific cell surface antigen. In some embodiments, such engineered cells, and their progeny, are not targeted by the agent or are targeted only to a significantly reduced extent compared to unengineered cells of the same cell type, and are not subject to the cytotoxicity, or are subject to a reduced degree of cytotoxicity, affected by the immunotherapeutic agent. In some embodiments, the engineered cells of the present disclosure are produced using homology-directed repair (HDR), which allows for the targeted integration of sequences from a template polynucleotide at a target sequence specified by the homology of a portion of the template polynucleotide to the target sequence. In some embodiments, the genetically engineered cells of the present disclosure are produced using base editing, which allows for targeted substitution, insertion, and deletion of sequences in a target sequence specified by a gRNA directed against the target sequence. Accordingly, some aspects of the present disclosure provide genetically engineered cells containing modified genes encoding lineage-specific cell surface antigens, methods of treating a subject in need thereof by administering such cells to the subject, compositions for use in the genetically engineered cells, e.g., genetic modification mixtures, methods of genetically engineering cells to contain modified genes encoding epitope-modified lineage-specific cell surface antigens, and other compositions (e.g., pharmaceutical compositions) related to any of the foregoing.

[0059] cell Some aspects of the present disclosure provide methods and compositions for genetically modifying cells, genetically modified cells produced by such methods, and methods of using the modified cells (e.g., to treat a subject in need thereof). In some embodiments, the genetically modified cells are hematopoietic cells. In some embodiments, the genetically modified hematopoietic cells are hematopoietic stem cells (HSCs) or hematopoietic progenitor cells (HPCs). In some embodiments, the methods or compositions described herein are used to genetically modify hematopoietic cells (e.g., HSCs or HPCs), e.g., in genes encoding lineage-specific cell surface antigens.

[0060] Some aspects of the present disclosure provide genetically modified hematopoietic cells and uses thereof. In some embodiments, such cells are generated by contacting cells with a CRISPR / Cas system (e.g., a Cas nuclease and / or a gRNA) and a template polynucleotide, or in some embodiments, the cells are daughter cells of cells contacted with the CRISPR / Cas system and a template polynucleotide. In some embodiments, such cells are generated by contacting cells with a preformed ribonucleoprotein complex comprising a base editor and a gRNA, or in some embodiments, the cells are daughter cells of cells contacted with the ribonucleoprotein complex. In some embodiments, the cells described herein (e.g., genetically engineered HSCs or HPCs) are capable of populating an HSC or HPC niche and / or reconstituting the hematopoietic system of a subject. In some embodiments, the cells described herein (e.g., HSCs or HPCs) are capable of one or more (e.g., all) of engrafting a human subject, producing myeloid cells, and producing lymphoid cells. In some preferred embodiments, the genetically engineered hematopoietic cells provided herein, or their progeny, can differentiate into all blood cell lineages, preferably without any differentiation bias compared to hematopoietic cells of the same cell type, but without the respective HDR-mediated genomic modifications. In some embodiments, the genetically engineered cells, e.g., genetically engineered HSCs, are autologous to a subject, e.g., a subject being treated for a disease, e.g., cancer, an autoimmune disease, or a genetic disease. In some embodiments, the genetically engineered cells, e.g., genetically engineered HSCs, are derived from a subject with cancer, an autoimmune disease, or a genetic disease, or a subject at risk of developing cancer, an autoimmune disease, or a genetic disease (i.e., autologous cells). In some embodiments, the HSCs genetically engineered using the methods of the present disclosure are obtained from a subject other than the subject to whom the cells will be administered and are referred to as allogeneic cells. In some embodiments, the HSCs are derived from a donor with an HLA haplotype that matches the HLA haplotype of the subject.Human leukocyte antigens (HLA) encode major histocompatibility complex (MHC) proteins in humans. MHC molecules are present on the surface of antigen-presenting cells and many other cell types and present self- and non-self-antigen (e.g., foreign) peptides for immune surveillance. However, HLA is highly polymorphic, resulting in many distinct alleles. Different (foreign, non-self) alleles are antigenic and can stimulate robust and harmful immune responses, particularly in organ and cell transplants. HLA molecules recognized as foreign (non-self) can lead to transplant rejection. In some embodiments, it is desirable to derive HSCs from a donor with the same HLA type as the patient to reduce the incidence of rejection.

[0061] The HLA loci of a donor subject can be typed to identify the individual as an HLA-matched donor for that subject. Methods for typing HLA loci are clear to those skilled in the art and include, for example, serology (serotyping), cell typing, gene sequencing, phenotyping, and PCR. The HLA from the donor is considered "matched" with the HLA of the subject if the HLA loci of the donor and the subject are identical or sufficiently similar so that no adverse immune response is expected.

[0062] In some embodiments, the genetically engineered hematopoietic cells of the present disclosure comprise a genetic modification proximal to a PAM sequence, e.g., a PAM sequence in a gene encoding a lineage-specific cell surface antigen (e.g., a sequence encoding an epitope bound by an agent that specifically binds to a lineage-specific cell surface antigen). In some embodiments, the genetic modification comprises incorporation of a donor sequence. In some embodiments, incorporation of the donor sequence results in an insertion or substitution mutation. In some embodiments, the donor sequence is inserted 5' to the PAM sequence, e.g., a Cas9 PAM sequence. In some embodiments, the donor sequence is inserted 5' to the PAM sequence. In some embodiments, the donor sequence is inserted 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides 5' to the PAM sequence. In some embodiments, the donor sequence is inserted 1-10, 1-8, 1-6, 1-4, 2-10, 2-8, 2-6, 2-4, 4-10, 4-8, 4-6, 6-10, 6-8, 8-10, 10-20, 15-20, 16-20, 17-20, 18-20, 19-20, 16-19, 17-19, 18-19, 16-18, or 17-18 nucleotides 5' of the PAM sequence, e.g., 2, 3, or 4 nucleotides 5' of the PAM sequence. In some embodiments, the donor sequence is inserted 3' of the PAM sequence, e.g., the Cas9 PAM sequence. In some embodiments, the donor sequence is inserted 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 nucleotides 3' of the PAM sequence. In some embodiments, the donor sequence is inserted 1-10, 1-8, 1-6, 1-4, 2-10, 2-8, 2-6, 2-4, 4-10, 4-8, 4-6, 6-10, 6-8, 8-10, 10-20, 15-20, 16-20, 17-20, 18-20, 19-20, 16-19, 17-19, 18-19, 16-18, or 17-18 nucleotides 3' of the PAM sequence, for example, 17, 18, or 19 nucleotides 3' of the PAM sequence.

[0063] In some embodiments, the genetically engineered hematopoietic cells comprise a genetic modification corresponding to the incorporation of a donor sequence (e.g., from a template polynucleotide described herein) into a gene encoding a lineage-specific cell surface antigen in the hematopoietic cell. In some embodiments, the genetic modification corresponds to a position where the donor sequence differs from the sequence of the gene encoding the lineage-specific cell surface antigen. In some embodiments, incorporation of the donor sequence results in modification at 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bases (e.g., 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, or 9-10 bases) in the gene encoding the lineage-specific cell surface antigen. In some embodiments, integration of the donor sequence results in an insertion of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bases (e.g., 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, or 9-10 bases) in the gene encoding the lineage-specific cell surface antigen.In some embodiments, integration of the donor sequence results in the substitution of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 bases (e.g., 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4, 1-3, 1-2, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 2-4, 2-3, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 3-4, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, 5-6, 6-10, 6-9, 6-8, 6-7, 7-10, 7-9, 7-8, 8-10, 8-9, or 9-10 bases) in the gene encoding the lineage-specific cell surface antigen. In some embodiments, integration of the donor sequence results in modifications at several positions in the gene encoding the lineage-specific cell surface antigen that correspond to up to 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% of the length of the donor sequence. In some embodiments, integration of the donor sequence results in the insertion of several bases into the gene encoding the lineage-specific cell surface antigen that correspond to up to 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100% of the length of the donor sequence. In some embodiments, the donor sequence is between 1 and 100, 1 and 80, 1 and 60, 1 and 40, 1 and 20, 1 and 15, 1 and 10, 1 and 9, 1 and 8, 1 and 7, 1 and 6, 1 and 5, 1 and 4, 1 and 3, 1 and 2, 5 and 100, 5 and 80, 5 and 60, 5 and 40, 5 and 20, 5 and 15, 5 and 10, 5 and 9, 5 and 8, 5 and 7, 5 and 6, 10 and 100, 10 and 80, 10 and 60, 10 and 40, 10 and 20, 10 and 15, 20 and 100, 20 and 80, 20 and 60, 20 and 40, 60 and 100, or 60 and 80 nucleotides in length. In some embodiments, the donor sequence is 100 or less, 90 or less, 80 or less, 70 or less, 60 or less, 50 or less, 45 or less, 40 or less, 35 or less, 30 or less, 25 or less, 20 or less, 15 or less, 14 or less, 13 or less, 12 or less, 11 or less, 10 or less, 9 or less, 8 or less, 7 or less, 6 or less, 5 or less, 4 or less, 3 or less, 2 or less, or 1 or less bases in length.In some embodiments, the donor sequence is 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, 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 bases in length. In some embodiments, integration of the donor sequence results in the modification of no more than 1, no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, or no more than 10 bases in the gene encoding the lineage-specific cell surface antigen. In some embodiments, integration of the donor sequence results in the substitution of no more than 1, no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, or no more than 10 bases in the gene encoding the lineage-specific cell surface antigen. In some embodiments, integration of the donor sequence results in the insertion of no more than 1, no more than 2, no more than 3, no more than 4, no more than 5, no more than 6, no more than 7, no more than 8, no more than 9, or no more than 10 bases in the gene encoding the lineage-specific cell surface antigen.

[0064] In some embodiments, integration of the donor sequence into the genetically engineered hematopoietic cell alters the amino acid sequence of an epitope of a lineage-specific cell surface antigen, where the epitope is recognized by an agent that specifically binds to the lineage-specific cell surface antigen. In some embodiments, the integrated donor sequence comprises one or more mutations relative to the wild-type and / or naturally occurring sequence of the gene encoding the lineage-specific cell surface antigen. In some embodiments, the donor sequence comprises an artificial sequence or a heterologous sequence. In some embodiments, integration of the donor sequence generates a restriction nuclease site or a unique sequence tag in the gene encoding the lineage-specific cell surface antigen of the genetically engineered hematopoietic cell. In some embodiments, integration of the donor sequence into the gene encoding the lineage-specific cell surface antigen of the genetically engineered hematopoietic cell generates one or more silent mutations along with non-silent mutations (e.g., one or more silent mutations that involve an alteration in the amino acid sequence of the epitope). In some embodiments, the one or more silent mutations are adjacent to another mutation described herein (e.g., adjacent to an alteration in the amino acid sequence of the epitope). For example, in some embodiments, the genetically engineered hematopoietic cells include a genetic modification corresponding to an alteration of the amino acid sequence of the epitope, e.g., a single nucleotide point mutation, and one or more silent mutations adjacent to the alteration (e.g., mutation). Accordingly, some aspects of the present disclosure provide genetically engineered hematopoietic cells that include a genetic modification corresponding to the incorporation of a donor sequence described herein, e.g., a donor sequence described herein.

[0065] It will be appreciated that upon engraftment of donor cells into a recipient host organism, for example, the relative levels of engrafted donor cells (and their progeny) and host cells within a given niche (e.g., bone marrow) are important to the physiological and / or therapeutic outcome of the host organism. The level of engrafted donor cells or their progeny relative to host cells within a given tissue or niche is referred to herein as "chimerism." In some embodiments, the cells described herein (e.g., HSCs or HPCs) are capable of engrafting into a human subject and do not exhibit any difference in chimerism compared to hematopoietic cells of the same cell type but that do not contain a genomic modification that results in expression of a variant form of a gene product (e.g., containing a mutated epitope). In some embodiments, cells described herein (e.g., HSCs or HPCs) capable of engrafting into a human subject exhibit no more than a 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% difference in chimerism compared to hematopoietic cells of the same cell type but that do not contain a genomic modification that results in expression of a variant form of a gene product (e.g., comprising a mutated epitope).

[0066] In some embodiments, the genetically engineered cells provided herein contain only one genomic modification, for example, a genomic modification that results in the expression of a variant form of a gene product (e.g., containing a mutant epitope). In some embodiments, the genomic modification is a modification to a gene encoding a lineage-specific cell surface antigen. It will be understood that the gene editing methods provided herein can result in a genomic modification in one or both alleles of a target gene. In some embodiments, genetically engineered cells that contain genomic modifications in both alleles of a given locus are preferred.

[0067] In some embodiments, the genetically engineered cells provided herein comprise two or more genomic modifications, e.g., one or more genomic modifications, in addition to a genomic modification that results in expression of a variant form of a gene product (e.g., comprising a mutant epitope). For example, in some embodiments, the genetically engineered cells comprise a modification to a gene encoding a lineage-specific cell surface antigen and one or more additional genomic modifications, e.g., a modification to a second gene proximal (e.g., adjacent) to the modification to the gene encoding the lineage-specific cell surface antigen or one or more silent mutations.

[0068] In some embodiments, the genetically engineered cells provided herein comprise a genomic modification that results in expression of a variant form (e.g., comprising a mutated epitope) of a gene encoding a lineage-specific cell surface antigen. In some embodiments, the modification alters the amino acid sequence of the epitope recognized by an agent that specifically binds to the lineage-specific cell surface antigen. In some embodiments, the genomic modification does not substantially alter (e.g., impair, extend, or enhance) the function of the lineage-specific cell surface antigen. In some embodiments, the modified lineage-specific cell surface antigen has at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of the activity of a wild-type or naturally occurring (i.e., unmodified) lineage-specific cell surface antigen that does not contain the altered epitope, such as a cell that has not been subjected to a gene editing method described herein (e.g., HDR-mediated gene editing, base editing).

[0069] In some embodiments, the genomic modification does not substantially alter (e.g., increase or decrease) expression of the lineage-specific cell surface antigen. In some embodiments, the modified lineage-specific cell surface antigen is expressed at a level that is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of the expression level of a wild-type or naturally occurring (i.e., unmodified) lineage-specific cell surface antigen that does not contain the altered epitope, such as in cells that have not been subjected to a gene editing method described herein (e.g., HDR-mediated gene editing, base editing).

[0070] In some embodiments, the genomic modification does not substantially alter (e.g., increase or decrease) the viability of the engineered cell. In some embodiments, the engineered cell has a level of viability that is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of that of a corresponding wild-type cell or other similar cell that does not contain the genomic modification, such as a cell that has not been subjected to a gene editing method described herein (e.g., HDR-mediated gene editing, base editing).

[0071] In some embodiments, the immune effector cells are lymphocytes. In some embodiments, the immune effector cells are T lymphocytes. In some embodiments, the T lymphocytes are alpha / beta T lymphocytes. In some embodiments, the T lymphocytes are gamma / delta T lymphocytes. In some embodiments, the immune effector cells are natural killer T (NKT cells). In some embodiments, the immune effector cells are natural killer (NK) cells. In some embodiments, the immune effector cells express a chimeric antigen receptor (CAR). In some embodiments, the immune effector cells do not express a CAR and / or do not express any transgenic proteins, e.g., except for lineage-specific cell surface antigens, except as provided by the genetic modifications described herein (e.g., except as modified using the methods using HDR or base editing described herein).

[0072] In some embodiments, the genetically engineered cells provided herein are hematopoietic cells, e.g., hematopoietic stem cells, hematopoietic progenitor cells (HPCs), hematopoietic stem or progenitor cells. Hematopoietic stem cells (HSCs) are cells characterized by pluripotency, self-renewal, and / or the ability to generate and / or reconstitute all lineages of the hematopoietic system, including both myeloid and lymphoid progenitor cells, which further give rise to myeloid cells (e.g., monocytes, macrophages, neutrophils, basophils, dendritic cells, erythrocytes, platelets, etc.) and lymphoid cells (e.g., T cells, B cells, NK cells), respectively. HSCs are characterized by the expression of one or more cell surface markers, e.g., CD34 (e.g., CD34+), and the absence of cell surface markers associated with lineage commitment, which can be used to identify and / or isolate HSCs. In some embodiments, the genetically engineered cells described herein (e.g., genetically engineered HSCs) do not express, express reduced amounts of, or express variant cell surface markers that are not recognized by immunotherapeutic agents that target one or more cell surface markers typically associated with HSC identification or isolation, yet are capable of self-renewal and can generate and / or reconstitute all lineages of the hematopoietic system.

[0073] In some embodiments, the populations of genetically engineered cells described herein comprise a plurality of genetically engineered hematopoietic stem cells. In some embodiments, the populations of genetically engineered cells described herein comprise a plurality of genetically engineered hematopoietic progenitor cells. In some embodiments, the populations of genetically engineered cells described herein comprise a plurality of genetically engineered hematopoietic stem cells and a plurality of genetically engineered hematopoietic progenitor cells.

[0074] In some embodiments, the genetically engineered HSCs are obtained from a subject, such as a human subject. Methods for obtaining HSCs are described, for example, in International Publication No. WO 2017 / 066760, which is incorporated herein by reference in its entirety. In some embodiments, the HSCs are peripheral blood HSCs. In some embodiments, the mammalian subject is a non-human primate, a rodent (e.g., a mouse or rat), a cow, a pig, a horse, or a livestock animal. In some embodiments, the HSCs are obtained from a human subject, such as a human subject with a hematopoietic malignancy. In some embodiments, the HSCs are obtained from a healthy donor. In some embodiments, the HSCs are obtained from a subject to whom immune cells expressing a chimeric antigen receptor will subsequently be administered. HSCs administered to the same subject from whom the cells were obtained are referred to as autologous cells, whereas HSCs obtained from a subject other than the subject to whom the cells will be administered are referred to as allogeneic cells.

[0075] In some embodiments, the population of genetically engineered cells is a heterogeneous population of cells, e.g., a heterogeneous population of genetically engineered cells comprising different mutations, e.g., different mutations in a gene encoding a lineage-specific cell surface antigen. In some embodiments, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, or at least 95% of the copies of the gene encoding the lineage-specific cell surface antigen within the population of genetically engineered cells comprise a mutation introduced by the genome editing approach described herein. As an example, the population of genetically engineered cells can comprise multiple different mutations in the gene encoding the lineage-specific cell surface antigen (e.g., multiple different mutations that alter the amino acid sequence of an epitope of the lineage-specific cell surface antigen), with each mutation in the multiple contributing to the proportion of copies of the gene in the population of cells that have the mutation.

[0076] In some embodiments, expression of the modified gene encoding the lineage-specific cell surface antigen in the engineered hematopoietic cells is compared to expression of the unmodified gene in a reference hematopoietic cell (e.g., a wild-type counterpart, an otherwise similar hematopoietic cell that does not contain the modification, or a mock-engineered hematopoietic cell (e.g., a hematopoietic cell contacted with Cas9 and a scrambled gRNA that does not effectively localize Cas9 or base editors to the gene, or a hematopoietic cell contacted with a targeted gRNA in the absence of Cas9 or base editors).

[0077] In some embodiments, the cells described herein (e.g., hematopoietic cells, e.g., hematopoietic stem cells) are characterized by reduced or no binding of an agent that specifically binds to the lineage-specific cell surface antigen. In some embodiments, the cells described herein comprise a modified lineage-specific cell surface antigen that is not bound by an agent that specifically binds to the lineage-specific cell surface antigen (i.e., the unmodified lineage-specific cell surface antigen) or that has reduced binding to an agent that specifically binds to the lineage-specific cell surface antigen (i.e., the unmodified lineage-specific cell surface antigen). In some embodiments, the cells are characterized by reduced binding of an agent that specifically binds to the lineage-specific cell surface antigen compared to binding of the agent to wild-type hematopoietic stem cells or other similar cells that express the genomic modification without (not including) the modified lineage-specific cell surface antigen. In some embodiments, cells with reduced or eliminated binding of an agent to the lineage-specific cell surface antigen are resistant or immune to targeting by immunotherapeutic agents that specifically bind to the lineage-specific cell surface antigen. In some embodiments, the genetically modified cells produced by the methods described herein comprise a genetic modification that modifies an epitope on a lineage-specific cell surface antigen, reducing or eliminating binding of an agent that specifically binds to the lineage-specific cell surface antigen compared to wild-type cells or cells that do not comprise the genomic modification. In some embodiments, the genetically modified cells can be advantageously administered to a subject to treat cancer, autoimmune disease, or genetic disease, allowing for co-administration of immunotherapeutic agents that may otherwise target (e.g., reduce the effectiveness of) the modified cells. Lineage-specific cell surface antigens are readily apparent on a variety of cell types.In some embodiments, the lineage-specific cell surface antigen is selected from BCMA, CD19, CD20, CD30, ROR1, B7H6, B7H3, CD23, CD33, CD38, C-type lectin-like molecule-1, CS1, IL-5, L1-CAM, PSCA, PSMA, CD138, CD133, CD70, CD5, CD6, CD7, CD13, NKG2D, NKG2D ligand, CLEC12A, CD11, CD123, CD45, CD56, CD30, CD14, CD66b, CD41, CD61, CD62, CD235a, CD146, CD326, LMP2, CD22, CD52, CD10, CD3 / TCR, CD79 / BCR, EMR2 (CD312), and CD26. In some embodiments, the lineage-specific cell surface antigen is selected from CD33, CD19, CD123, CLL-1, CD30, CD5, CD6, CD7, CD38, EMR2 (CD312), and BCMA. In some embodiments, the lineage-specific cell surface antigen is selected from CD7, CD13, CD19, CD22, CD25, CD32, CD33, CD38, CD44, CD47, CD56, CD96, CD117, CD123, CD135, CD174, CLL-1, folate receptor b, IL1RAP, MUC1, NKG2D / NKG2DL, TIM-3, and WT1. In some embodiments, the lineage-specific cell surface antigen is selected from CD123, CD38, CD19, CD33, CD34, CD47, CLL-1, CD30, CD5, CD6, CD7, EMR2 / CD312, and BCMA.

[0078] In some embodiments, the cells described herein comprise a genomic modification in a gene encoding a lineage-specific cell surface antigen. In some embodiments, the lineage-specific cell surface antigen is CD123, CD38, CD47, CD34, CD5, or CD19. In some embodiments, the lineage-specific cell surface antigen is CD123 or CD38. In some embodiments, the lineage-specific cell surface antigen is CD123. In some embodiments, the lineage-specific cell surface antigen is CD38. In some embodiments, the lineage-specific cell surface antigen is CD19. In some embodiments, the lineage-specific cell surface antigen is CD34. In some embodiments, the lineage-specific cell surface antigen is CD47. In some embodiments, the lineage-specific cell surface antigen is CD5. In some embodiments, the lineage-specific cell surface antigen is EMR2.

[0079] CD123 (also known as interleukin-3 receptor alpha or IL3Rα) is a type I cytokine receptor that binds IL3. IL3 is a pleiotropic cytokine that regulates the function and production of hematopoietic and immune cells (see, e.g., Testa et al. Biomarker Research volume 2, Article number: 4 (2014)). Dysregulated expression of IL3 is associated with various cancers, including myeloma (see, e.g., Lee et al. Blood (2004) 103(6):2308-2315). In some embodiments, hematopoietic malignancies are characterized by cells that express (e.g., overexpress) CD123. Dysregulated expression of CD123 has been associated with various hematopoietic malignancies, such as hairy cell leukemia, acute myeloid leukemia, blastic plasmacytoid dendritic cell neoplasm, and systemic mastocytosis (e.g., Del Giudice et al. Hematologica (2004) 89(3):303-308, Muñoz et al. Hematologica (2001) 86(12):1261-1269, Angelot-Delettre et al. Hematologica (2015) 100(2):223-230, Alayed et al. American Journal of Hematology (2013) 88(12):1055-1061, Paradanani et al. Leukemia (2016) 30(4):914-918, Testa et al. Biomarker Research (2014) 2:4, and Lamble et al. Journal of Clinical (See Oncology (2022) 40(3):252-261.) In some embodiments, CD123 is expressed on hematopoietic cells, e.g., hematopoietic stem cells and / or hematopoietic progenitor cells.

[0080] CD38 (also known as cyclic ADP-ribose hydrolase) is a transmembrane ectoenzyme glycoprotein involved in cell adhesion, signal transduction, and calcium signaling (e.g., van de Donk et al. Blood (2018) 131(1):13-29). In some embodiments, the hematopoietic malignancy is characterized by cells that express (e.g., overexpress) CD38. In some embodiments, CD38 is expressed on hematopoietic cells, e.g., hematopoietic stem cells and / or hematopoietic progenitor cells.

[0081] CD19 is a type I transmembrane glycoprotein containing two extracellular Ig-like domains and a conserved C-terminal cytoplasmic tail. It is typically expressed on the surface of human B cells and hematopoietic stem and progenitor cells that are predisposed to differentiate into the B cell lineage. CD19 is required for B cell survival, development, and differentiation and forms a multimolecular signaling complex on the cell surface. CD19 has also been identified as a regulator of neoplastic growth and cell proliferation in B cell cancers. The gene encoding human CD19 is 7.41 kilobases long and contains at least 15 exons, four of which encode the extracellular domain. Multiple alternatively spliced ​​mRNA transcripts derived from the CD19 gene have been detected. In addition to its expression on B cells and hematopoietic cells that are predisposed to differentiate into B cells, CD19 expression has also been associated with some hematopoietic malignancies.

[0082] EGF-like module-containing mucin-like hormone receptor-like 2 (EMR2), also known as CD312, is an approximately 90 kDa protein (isoform dependent) of 823 amino acids in the EGF seven-transmembrane (TM7) family of adhesion G protein-coupled receptors (GPCRs) with high levels of homology to CD97. EMR2 forms heterodimers and binds to chondroitin sulfate B via its EGF-like domain 4, mediating cell adhesion, granulocyte chemotaxis, degranulation, and pro-inflammatory cytokine release in macrophages. See, e.g., Kuan-Yu et al. Front. Immunol. (2017) 8:373. Without wishing to be bound by any particular theory, EMR2 is expressed on myeloid cells with highest expression in granulocytes, macrophages, and Kupffer cells. The ADGRE2 gene, located on human chromosome 19, encodes human EMR2 and classically contains 19 exons, but due to alternative RNA splicing, there are many isoforms with different numbers of EGF domains. The dominant isoform in whole blood contains 17 exons. See, for example, Safaee et al. Onc. Rev. (2014). 8 (242): 20-24.

[0083] CD5 is a member of the scavenger receptor cysteine-rich (SRCR) superfamily and functions as a signaling transmembrane glycoprotein involved in tyrosine phosphorylation of intracellular effector proteins. CD5 plays several functions in T and B lymphocyte receptor signaling and immune system regulation (see, e.g., Burgueno-Bucio et al. Journal of Leukocyte Biology (2019) 105(5):891-905). CD5 contains three SRCR domains that act as a receptor controlling T cell proliferation. CD5 is primarily expressed on thymocytes and mature T lymphocytes. Additionally, CD5 expression on B lymphocytes is associated with poor prognosis in large B-cell lymphoma (see, e.g., Tagawa et al. Cancer Research (2004) 64(17):5948-5955). The gene encoding human CD5 is located on chromosome 11 and contains 12 exons.

[0084] CD47 is a transmembrane integrin-associated protein belonging to the immunoglobulin superfamily and is involved in the increase in intracellular calcium concentration that occurs upon cell adhesion to the extracellular matrix. CD47 binds to various ligands, including thrombospondin-1 and signal-regulatory protein alpha, and functions in processes such as apoptosis, proliferation, adhesion, and migration. CD47 also plays a role in immune and angiogenic responses, including the regulation of phagocytosis by macrophages (see, e.g., Brown and Frazier, Trends in Cell Biology (2001) 11(3):130-135). CD47 is widely expressed across various human tissues and in solid tumors and hematological malignancies (see, e.g., Jiang et al., Journal of Hematology & Oncology (2021) 14:180). Human CD47 is located on chromosome 3 and contains 13 exons.

[0085] CD34 is a transmembrane phosphorylated glycoprotein belonging to the single-pass transmembrane sialomucin protein family that functions as an intercellular adhesion molecule. Therefore, CD34 is an important adhesion molecule required for T cell entry into lymph nodes and for hematopoietic stem cell attachment to bone marrow extracellular matrix or stromal cells. CD34 is highly expressed on hematopoietic stem and progenitor cells, as well as endothelial cells. Furthermore, CD34 is commonly found to be expressed on the cell surface of hematopoietic cancer cells (see, e.g., Sydney et al. Stem Cells (2014) 32(6):1380-1389; Nielsen and McNagny. Journal of Cell Science (2008) 121(22):3683-3692; Lanze et al. Journal of Biological Regulators and Homeostatic Agents (2001) 15(1):1-13; Sutherland and Keating. Journal of Hematotherapy (2009) 1(2):115-129). CD34 is located on chromosome 1 and contains eight exons.

[0086] Due to the shared expression of CD33, CD123, CD19, CLL-1, CD30, CD5, CD6, CD7, CD34, CD38, CD47, EMR2 / CD312, and / or BCMA on both normal and healthy cells (e.g., healthy hematopoietic cells), and being antigens expressed on malignant cells, therapeutic targeting of CD33, CD123, CD19, CLL-1, CD30, CD5, CD6, CD7, CD34, CD38, CD47, EMR2 / CD312, and / or BCMA may result in depletion of the healthy hematopoietic cell and / or progenitor cell pool.

[0087] In some embodiments, the cells described herein comprise a genomic modification that results in a mutation in a gene encoding a lineage-specific cell surface antigen. In some embodiments, the mutation in the gene encoding the lineage-specific cell surface antigen alters one or more amino acids in the lineage-specific cell surface antigen. In some embodiments, the one or more amino acids are part of an epitope recognized (i.e., bound by) an agent that specifically binds to the lineage-specific cell surface antigen. In some embodiments, the epitope is part of a domain of the lineage-specific cell surface antigen, e.g., the extracellular domain or a subdomain thereof.

[0088] The modification of one or more amino acids may include one, two, or all of substitution, insertion, or deletion. For example, the modification may include substituting an amino acid listed herein with a different amino acid. As a further example, the modification may include deleting an amino acid listed herein. As a further example, the modification may include inserting one or more amino acids at a position listed herein or as part of a deletion of an amino acid listed herein.

[0089] In some embodiments, the mutation in the gene encoding CD123 alters one or more amino acids associated with an epitope of CD123. In some embodiments, the epitope of CD123 is a portion of CD123 bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-CD123 antibody. In some embodiments, the agent comprises an anti-CD123 antibody or a portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T-cell engager). For example, the agent can be the anti-CD123 antibody 7G3 or a variant thereof (e.g., a humanized variant, e.g., the antibody CSL-36). In some embodiments, the agent is an anti-CD123 agent, e.g., talaxuzumab. In some embodiments, the epitope of CD123 is one or more amino acids of a protein domain (e.g., the extracellular domain) or an amino acid encoded by an exon or combination of exons of the gene encoding CD123. In some embodiments, the epitope of CD123 comprises one or more amino acids encoded by exon 3 of the gene encoding CD123. In some embodiments, the epitope of CD123 comprises one or more amino acids encoded by exon 4 of the gene encoding CD123. In some embodiments, the epitope of CD123 comprises one or more (e.g., two or more, three or more, four or more, or all) amino acids at positions 51, 59, 61, 82, or 84 of a wild-type gene encoding CD123, or at the corresponding positions of a homologous CD123 gene.

[0090] In some embodiments, the mutation in the gene encoding CD123 comprises a substitution of an amino acid at position 51 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, a lysine is substituted for the amino acid at position 51 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, a glycine is substituted for the amino acid at position 51 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, the mutation in the gene encoding CD123 comprises a substitution of an amino acid at position 59 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, a phenylalanine is substituted for the amino acid at position 59 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, a cysteine ​​is substituted for the amino acid at position 59 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, the mutation in the gene encoding CD123 comprises a substitution of an amino acid at position 61 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, a leucine is substituted for the amino acid at position 61 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, the mutation in the gene encoding CD123 comprises a substitution of an amino acid at position 82 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, an alanine is substituted for the amino acid at position 82 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, the mutation in the gene encoding CD123 comprises a substitution of an amino acid at position 84 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, a glutamine is substituted for the amino acid at position 84 of wild-type CD123 or at the corresponding position in a homologous CD123 gene. In some embodiments, an alanine is substituted for the amino acid at position 84 of wild-type CD123 or at the corresponding position in a homologous CD123 gene.

[0091] In some embodiments, the mutation in the gene encoding CD123 creates an amino acid sequence change that corresponds to the amino acid sequence of a CD123 ortholog. In some embodiments, the mutation substitutes an amino acid in human CD123 with an amino acid at the corresponding position in an orthologous CD123, e.g., a non-human primate CD123. In some embodiments, the mutation inserts or deletes one or more amino acids in human CD123 to correspond to the sequence of an orthologous CD123, e.g., a non-human primate CD123. In some embodiments, the mutation alters the amino acid sequence in a manner that corresponds to an allowable genetic variant identified by one or more genome sequence comparison algorithms, e.g., gnomAD (e.g., Gudmundsson et al. arXiv:2107.11458v3, e.g., gnomad.broadinstitute.org / ), or in a manner that corresponds to a position characterized by multiple allowable genetic variants. In some embodiments, mutations to CD123 corresponding to the amino acid sequence of a CD123 ortholog, or at positions characterized by multiple permissible genetic variants, reduce or eliminate binding of immunotherapeutic agents that target CD123 while preserving some or all of the structure, expression, and / or functionality of CD123, providing cells expressing CD123 that are less likely to be targeted by anti-CD123 immunotherapeutic agents or that are not targeted at all (e.g., functional CD123). In some embodiments, the modifications result in missense variants of CD123.

[0092] In some embodiments, the mutation in the gene encoding CD38 alters one or more amino acids associated with an epitope of CD38. In some embodiments, the epitope of CD38 is a portion of CD38 bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-CD38 antibody. In some embodiments, the agent comprises an anti-CD38 antibody or a portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T-cell engager). For example, the agent can be the anti-CD38 antibody HB7 or a variant thereof (e.g., a humanized variant). In some embodiments, the agent is an anti-CD38 agent, e.g., daratumumab. In some embodiments, the epitope of CD38 is one or more amino acids of a protein domain (e.g., the extracellular domain) or an amino acid encoded by an exon or combination of exons of the gene encoding CD38. In some embodiments, the epitope of CD38 comprises one or more amino acids encoded by exon 7 of the gene encoding CD38. In some embodiments, the epitope of CD38 comprises one or more (e.g., two or more, three or more, four or more, or all) amino acids at positions 270-274 of the wild-type gene encoding CD38, or at the corresponding positions of a homologous CD38 gene.

[0093] In some embodiments, the mutation in the gene encoding CD38 comprises a substitution of an amino acid at position 270 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, an alanine is substituted for the amino acid at position 270 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, the mutation in the gene encoding CD38 comprises a substitution of an amino acid at position 271 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, the mutation in the gene encoding CD38 comprises a substitution of an amino acid at position 272 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, a histidine is substituted for the amino acid at position 272 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, an arginine is substituted for the amino acid at position 272 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, an alanine is substituted for the amino acid at position 272 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, the mutation in the gene encoding CD38 comprises a substitution of an amino acid at position 273 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, the mutation in the gene encoding CD38 comprises a substitution of an amino acid at position 274 of wild-type CD38 or at the corresponding position in a homologous CD38 gene. In some embodiments, a phenylalanine is substituted for the amino acid at position 274 of wild-type CD38 or at the corresponding position in a homologous CD38 gene.

[0094] In some embodiments, mutations in the gene encoding CD38 create an amino acid sequence change that corresponds to the amino acid sequence of a CD38 ortholog. In some embodiments, the mutations substitute an amino acid in human CD38 for an amino acid at the corresponding position in an orthologous CD38, e.g., a non-human primate CD38. In some embodiments, the mutations insert or delete one or more amino acids in human CD38 to correspond to the sequence of an orthologous CD38, e.g., a non-human primate CD38. In some embodiments, the mutations alter the amino acid sequence in a manner that corresponds to an allowable genetic variant identified by one or more genome sequence comparison algorithms, e.g., gnomAD, or in a manner that corresponds to a position characterized by multiple allowable genetic variants. In some embodiments, mutations to CD38 corresponding to the amino acid sequence of a CD38 ortholog, or at positions characterized by multiple permissible genetic variants, reduce or eliminate binding of CD38-targeted immunotherapeutic agents while preserving some or all of the structure, expression, and / or functionality of CD38, providing cells expressing CD38 (e.g., functional CD38) that are less likely to be targeted or are not targeted at all by anti-CD38 immunotherapeutic agents.

[0095] In some embodiments, the mutation in the gene encoding CD19 alters one or more amino acids associated with an epitope of CD19. In some embodiments, the epitope of CD19 is a portion of CD19 bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-CD19 antibody. In some embodiments, the agent is the anti-CD19 antibody FMC63 or HIB19. In some embodiments, the agent comprises an anti-CD19 antibody or portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T-cell engager). In some embodiments, the agent is an anti-CD19 agent. In some embodiments, the epitope of CD19 corresponds to amino acids of a protein domain (e.g., the extracellular first or second Ig-like domain or a non-Ig-like domain) or amino acids encoded by an exon or combination of exons of the gene encoding CD19. In some embodiments, the epitope of CD19 comprises amino acids encoded by one, two, three, or all of exons 1, 2, 3, or 4 of CD19. In some embodiments, the epitope of CD19 comprises amino acids encoded by exon 2 of CD19. In some embodiments, the epitope of CD19 comprises amino acids encoded by exon 4 of CD19.In some embodiments, the CD19 epitope is selected from the group consisting of amino acids 216-238, 216-236, 216-234, 216-232, 216-230, 216-228, 216-226, 216-224, 216-222, 216-220, 216-218, 218-238, 218-236, 218-234, 218-222, 216-220, 216-218, 218-238, 218-236, 218-234, 218-222, 216-220, 216-218, 218-238, 218-236, 218-234, 218-222, 216-220, 216-228, 216-226, 216-224, 216 ...2, 216-220, 216-218, 218-238, 218-236, 218- 32, 218-230, 218-228, 218-226, 218-224, 218-222, 218-220, 220-238, 220-236, 220-234, 220-232, 220-230, 220-228, 220-226, 220-224, 220-222, 222-238, 222-236, 222-234, 2 22~232, 222~230, 222~228, 222~226, 222~224, 224~238, 224~236, 224~234, 224~232, 224~230, 224~228, 224~226, 226~238, 226~236, 226~234, 226~232, 226~230, 226~228, 228~ In some embodiments, the CD19 epitope comprises amino acids 163 and / or 164 of CD19.

[0096] In some embodiments, the mutation in the gene encoding CD19 creates an alteration in the amino acid sequence that corresponds to the amino acid sequence of a CD19 ortholog. In some embodiments, the modification comprises a substitution of an amino acid at position 218 of wild-type CD19 or at the corresponding position in a homologous CD19. In some embodiments, the modification comprises an insertion of one or more amino acids at position 224 of wild-type CD19 or at the corresponding position in a homologous CD19. In some embodiments, the modification comprises a substitution of an amino acid at 218 of CD19 and an insertion of one or more amino acids at position 224 of wild-type CD19 or at the corresponding position in a homologous CD19.

[0097] In some embodiments, the modification comprises an amino acid substitution at positions 163 and / or 164 of wild-type CD19 or the corresponding positions of a homologous CD38. In some embodiments, the modification comprises an amino acid substitution at position 163 of wild-type CD19 or the corresponding position of a homologous CD38. In some embodiments, the modification comprises an amino acid substitution at position 164 of wild-type CD19 or the corresponding position of a homologous CD38. In some embodiments, the modification comprises an amino acid substitution at positions 163 and 164 of wild-type CD19 or the corresponding positions of a homologous CD38. In some embodiments, a leucine is substituted for the amino acid at position 163 of wild-type CD19 or the corresponding position of a homologous CD19. In some embodiments, a cysteine ​​is substituted for the amino acid at position 163 of wild-type CD19 or the corresponding position of a homologous CD19. In some embodiments, a cysteine ​​is substituted for the amino acid at position 163 and a phenylalanine is substituted for the amino acid at position 164 of wild-type CD19 or the corresponding position in a homologous CD19.

[0098] In some embodiments, the modification results in a missense variant of CD19. In some embodiments, the modification results in an alteration in a splice region of CD19.

[0099] In some embodiments, mutations in the gene encoding CD19 create an alteration in the amino acid sequence that corresponds to the amino acid sequence of a CD19 ortholog. In some embodiments, the mutations substitute an amino acid in human CD19 for an amino acid at the corresponding position in an orthologous CD19, e.g., a non-human primate CD19. In some embodiments, the mutations insert or delete one or more amino acids in human CD19 to correspond to the sequence of an orthologous CD19, e.g., a non-human primate CD19. For example, in some embodiments, histidine 218 is replaced with arginine, which corresponds to the rhesus monkey CD19 sequence at that position. As a further example, in some embodiments, an amino acid (e.g., serine) is inserted at position 224 of human CD19, which corresponds to the rhesus monkey CD19 sequence at that position. In some embodiments, mutations to CD19 that correspond to the amino acid sequence of a CD19 ortholog reduce or eliminate binding of CD19-targeted immunotherapeutic agents while preserving some or all of the structure, expression, and / or functionality of CD19, providing cells expressing CD19 (e.g., functional CD19) that are less likely to be targeted or are not targeted at all by anti-CD19 immunotherapeutic agents.

[0100] In some embodiments, the mutation in the gene encoding EMR2 alters one or more amino acids associated with an epitope of EMR2. In some embodiments, the epitope of EMR2 is a portion of EMR2 bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-EMR2 antibody. In some embodiments, the agent comprises an anti-EMR2 antibody or a portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T-cell engager). In some embodiments, the agent is an anti-EMR2 agent. In some embodiments, the epitope of EMR2 corresponds to amino acids in a protein domain (e.g., the extracellular domain) or encoded by an exon or combination of exons of the gene encoding EMR2. In some embodiments, the epitope of EMR2 includes amino acids encoded by one, two, three, four, or all of exons 6, 10, 11, 14, and 18 of EMR2. In some embodiments, the epitope of EMR2 comprises amino acids encoded by exon 6 of EMR2. In some embodiments, the epitope of EMR2 comprises amino acids encoded by exon 10 of EMR2. In some embodiments, the epitope of EMR2 comprises amino acids encoded by exon 11 of EMR2. In some embodiments, the epitope of EMR2 comprises amino acids encoded by exon 14 of EMR2. In some embodiments, the epitope of EMR2 comprises amino acids encoded by exon 18 of EMR2.

[0101] In some embodiments, the mutation in the gene encoding EMR2 creates an amino acid sequence change that corresponds to the amino acid sequence of an EMR2 ortholog. In some embodiments, the modification comprises an amino acid substitution at any one or more of positions 124, 132, 146, 292, 294, 295, 296, 298, 299, 303, 304, 305, 306, 307, 308, 312, 318, 320, 328, 329, 331, 332, 335, 340, 347, 527, or 708 of wild-type EMR2, or at the corresponding position in a homologous EMR2.

[0102] In some embodiments, the modification results in a missense variant of EMR2. In some embodiments, the modification results in an alteration in a splice region of EMR2.

[0103] In some embodiments, the mutation in the gene encoding EMR2 alters one or more amino acids associated with an epitope of EMR2. In some embodiments, the epitope of EMR2 is a portion of EMR2 that is bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-EMR2 antibody. In some embodiments, the agent comprises an anti-EMR2 antibody or a portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T cell engager). For example, the agent can be anti-EMR2 monoclonal antibody 2A1 (Thermo Fisher), or a variant thereof (e.g., a humanized variant), Q9UHX3, OASA01861, AB_2738756, NLS6381, ab75190, MAB4894, or A100,000. Additional anti-EMR2 antibodies will be apparent to those skilled in the art. See, for example, WO 2017 / 087800 A1, Chang et al. FEBS Letters. (2003) 547(1-3): 145-150, Yona et al. FASEB J. (2008). 22(3): 741-751.

[0104] In some embodiments, mutations to EMR2 that correspond to the amino acid sequence of an EMR2 ortholog reduce or eliminate binding of immunotherapeutic agents that target EMR2 while preserving some or all of the structure, expression, and / or functionality of EMR2, providing cells expressing EMR2 (e.g., functional EMR2) that is less likely to be targeted or not targeted at all by anti-EMR2 immunotherapeutic agents.

[0105] In some embodiments, the mutation in the gene encoding CD5 alters one or more amino acids associated with an epitope of CD5. In some embodiments, the epitope of CD5 is a portion of CD5 bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-CD5 antibody. In some embodiments, the agent comprises an anti-CD5 antibody or a portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T-cell engager). For example, the agent can be the anti-CD5 monoclonal antibody H65, or a variant thereof (e.g., a humanized variant). In some embodiments, the agent is an anti-CD5 agent (e.g., zolimomab). In some embodiments, the epitope of CD5 is one or more amino acids of a protein domain (e.g., the extracellular domain) or an amino acid encoded by an exon or combination of exons of the gene encoding CD5.

[0106] In some embodiments, the CD5 epitope comprises one or more (e.g., two or more, three or more, four or more, or all) amino acids at positions 35-133 of the wild-type gene encoding CD5, or at the corresponding positions of a homologous CD5 gene.

[0107] In some embodiments, the modification of the epitope of CD5 comprises an insertion, deletion, substitution, or inversion of one or more amino acids (e.g., 1, 2, 3, 4, or more) occurring at positions 35 to 133 of wild-type CD5. In some embodiments, the alteration results in a missense variant of CD5 occurring at one or more (e.g., 1, 2, 3, 4, or more) amino acids occurring at positions 35 to 133 of wild-type CD5.

[0108] In some embodiments, the modification results in a missense variant of CD5. In some embodiments, the modification results in an alteration in a splice region of CD5.

[0109] In some embodiments, the mutation in the gene encoding CD5 creates an amino acid sequence change that corresponds to the amino acid sequence of a CD5 ortholog. In some embodiments, the mutation substitutes an amino acid in human CD5 with an amino acid at the corresponding position in an orthologous CD5, e.g., a non-human primate CD5. In some embodiments, the mutation inserts or deletes one or more amino acids in human CD5 to correspond to the sequence of an orthologous CD5, e.g., a non-human primate CD5. In some embodiments, the mutation alters the amino acid sequence in a manner that corresponds to an allowable genetic variant identified by one or more genome sequence comparison algorithms, e.g., gnomAD (e.g., Gudmundsson et al. arXiv:2107.11458v3, e.g., gnomad.broadinstitute.org / ), or in a manner that corresponds to a position characterized by multiple allowable genetic variants.

[0110] In some embodiments, mutations to CD5 corresponding to the amino acid sequence of a CD5 ortholog, or at positions characterized by multiple permissible genetic variants, reduce or eliminate binding of CD5-targeted immunotherapeutics while preserving some or all of the structure, expression, and / or functionality of CD5, providing cells expressing CD5 (e.g., functional CD5) that are less likely to be targeted or not targeted at all by anti-CD5 immunotherapeutics.

[0111] In some embodiments, the mutation in the gene encoding CD47 alters one or more amino acids associated with an epitope of CD47. In some embodiments, the epitope of CD47 is a portion of CD47 bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-CD47 antibody. In some embodiments, the agent is an anti-CD47 B6H12 antibody or a 2D3 antibody. In some embodiments, the agent comprises an anti-CD47 antibody or a portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T-cell engager). For example, the agent can be an anti-CD47 antibody or a variant thereof (e.g., a humanized variant). In some embodiments, the agent is an anti-CD47 agent. In some embodiments, the epitope of CD47 is one or more amino acids of a protein domain (e.g., the extracellular domain) or an amino acid encoded by an exon or combination of exons of the gene encoding CD47.

[0112] In some embodiments, the epitope of CD47 comprises one or more amino acids at positions 117-122 of CD47. In some embodiments, one or more amino acids at positions 117-122 of CD47 are deleted. In some embodiments, amino acids at positions 117-122 of CD47 are deleted. In some embodiments, amino acids at positions 117, 118, 119, 120, 121, and / or 122 of CD47 are deleted, or any combination thereof. In some embodiments, the epitope of CD47 comprises one or more amino acids at positions 52-55 of CD47. In some embodiments, one or more amino acids at positions 52-55 of CD47 are deleted. In some embodiments, amino acids at positions 52-55 of CD47 are deleted. In some embodiments, amino acids at positions 52, 53, 54, and / or 55 of CD47 are deleted, or any combination thereof.

[0113] In some embodiments, the mutation in the gene encoding CD47 comprises a substitution of an amino acid at position 31 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, a methionine is substituted for the amino acid at position 31 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a substitution of an amino acid at position 47 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, a histidine is substituted for the amino acid at position 47 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, a glycine is substituted for the amino acid at position 47 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a substitution of an amino acid at position 49 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, an arginine is substituted for the amino acid at position 49 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, a proline is substituted for the amino acid at position 49 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a substitution of an amino acid at position 52 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a deletion of an amino acid at position 52 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a substitution of an amino acid at position 53 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, an alanine is substituted for the amino acid at position 53 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a deletion of an amino acid at position 53 of wild-type CD47 or at the corresponding position in a homologous CD47 gene, hi some embodiments, the mutation in the gene encoding CD47 comprises a deletion of an amino acid at position 54 of wild-type CD47 or at the corresponding position in a homologous CD47 gene.In some embodiments, the mutation in the gene encoding CD47 comprises a deletion of an amino acid at position 55 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a substitution of an amino acid at position 120 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, an alanine is substituted for the amino acid at position 120 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, the mutation in the gene encoding CD47 comprises a substitution of an amino acid at position 124 of wild-type CD47 or at the corresponding position in a homologous CD47 gene. In some embodiments, a lysine is substituted for the amino acid at position 124 of wild-type CD47 or at the corresponding position in a homologous CD47.

[0114] In some embodiments, mutations in the gene encoding CD47 create an amino acid sequence change that corresponds to the amino acid sequence of a CD47 ortholog. In some embodiments, the mutations substitute an amino acid in human CD47 for an amino acid at the corresponding position in an orthologous CD47, e.g., a non-human primate CD47. In some embodiments, the mutations insert or delete one or more amino acids in human CD47 to correspond to the sequence of an orthologous CD47, e.g., a non-human primate CD47. In some embodiments, the mutations alter the amino acid sequence in a manner that corresponds to an allowable genetic variant identified by one or more genome sequence comparison algorithms, e.g., gnomAD (e.g., Gudmundsson et al. arXiv:2107.11458v3, e.g., gnomad.broadinstitute.org / ), or in a manner that corresponds to a position characterized by multiple allowable genetic variants.

[0115] In some embodiments, mutations to CD47 corresponding to the amino acid sequence of a CD47 ortholog, or at positions characterized by multiple permissible genetic variants, reduce or eliminate binding of CD47-targeted immunotherapeutic agents while preserving some or all of the structure, expression, and / or functionality of CD47, providing cells expressing CD47 (e.g., functional CD47) that are less likely to be targeted or not targeted at all by anti-CD47 immunotherapeutic agents.

[0116] In some embodiments, the mutation in the gene encoding CD34 alters one or more amino acids associated with an epitope of CD34. In some embodiments, the epitope of CD34 is a portion of CD34 that is bound by an agent, e.g., an immunotherapeutic agent. In some embodiments, the agent is an anti-CD34 antibody. In some embodiments, the anti-CD34 antibody is clone QBend10 or 561. In some embodiments, the agent comprises an anti-CD34 antibody or portion thereof, e.g., an antibody-drug conjugate (ADC), a chimeric antigen receptor (CAR), or a multispecific antibody (e.g., a bispecific T cell engager). In some embodiments, the epitope of CD34 is one or more amino acids of a protein domain (e.g., the extracellular domain) or an amino acid encoded by an exon or combination of exons of the gene encoding CD34.

[0117] In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 42 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, a glycine is substituted for the amino acid at position 42 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 46 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, an alanine is substituted for the amino acid at position 46 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 47 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, a lysine is substituted for the amino acid at position 47 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, a glutamate is substituted for the amino acid at position 47 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 49 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, a proline is substituted for the amino acid at position 49 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, a serine is substituted for the amino acid at position 49 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 50 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, an alanine is substituted for the amino acid at position 50 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, a proline is substituted for the amino acid at position 50 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 51 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, an alanine is substituted for the amino acid at position 51 of wild-type CD34 or at the corresponding position in a homologous CD34 gene.In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 54 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, an alanine is substituted for the amino acid at position 54 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, the mutation in the gene encoding CD34 comprises a substitution of an amino acid at position 55 of wild-type CD34 or at the corresponding position in a homologous CD34 gene. In some embodiments, an alanine is substituted for the amino acid at position 55 of wild-type CD34 or at the corresponding position in a homologous CD34 gene.

[0118] In some embodiments, the mutation in the gene encoding CD34 creates an amino acid sequence change that corresponds to the amino acid sequence of a CD34 ortholog. In some embodiments, the mutation substitutes an amino acid in human CD34 for an amino acid at the corresponding position in an orthologous CD34, e.g., a non-human primate CD34. In some embodiments, the mutation inserts or deletes one or more amino acids in human CD34 to correspond to the sequence of an orthologous CD34, e.g., a non-human primate CD34. In some embodiments, the mutation alters the amino acid sequence in a manner that corresponds to an allowable genetic variant identified by one or more genome sequence comparison algorithms, e.g., gnomAD (e.g., Gudmundsson et al. arXiv:2107.11458v3, e.g., gnomad.broadinstitute.org / ), or in a manner that corresponds to a position characterized by multiple allowable genetic variants. In some embodiments, mutations to CD34 corresponding to the amino acid sequence of a CD34 ortholog, or at positions characterized by multiple permissible genetic variants, reduce or eliminate binding of immunotherapeutic agents that target CD34 while preserving some or all of the structure, expression, and / or functionality of CD34, providing cells expressing CD34 (e.g., functional CD34) that are less likely to be targeted or are not targeted at all by anti-CD34 immunotherapeutic agents.

[0119] How to edit cells Some aspects of the present disclosure provide compositions and methods for generating the genetically engineered cells described herein, e.g., genetically engineered cells that comprise modifications in their genomes that result in alterations in the amino acid sequence of an epitope of a lineage-specific cell surface antigen, or the expression of a variant form of the lineage-specific cell surface antigen that is not recognized by an agent (e.g., an immunotherapeutic agent) that targets (e.g., specifically binds) the lineage-specific cell surface antigen. Such compositions and methods provided herein include, but are not limited to, suitable strategies and approaches for genetically engineering cells, for example, by using RNA-guided nucleases, such as CRISPR / Cas nucleases, and suitable RNAs that can bind to and target such RNA-guided nucleases to suitable target sites within the genome of the cell, resulting in genome modifications that result in alterations in the amino acid sequence of an epitope of the lineage-specific cell surface antigen, or the expression of a variant form of the lineage-specific cell surface antigen that is not recognized by an immunotherapeutic agent that targets the lineage-specific cell surface antigen.

[0120] In some embodiments, for example, the engineered cells described herein (e.g., engineered hematopoietic cells, such as engineered hematopoietic stem or progenitor cells or engineered immune effector cells (e.g., B cells or T cells)) are generated via genome editing techniques, including any technique capable of introducing targeted changes, also referred to as "editing," into the genome of a cell.

[0121] One exemplary suitable genome editing technique is "cellular editing," which involves the use of RNA-guided nucleases, e.g., CRISPR / Cas nucleases, to introduce targeted single- or double-stranded DNA breaks into the genome of a cell, which triggers cellular repair mechanisms such as, for example, non-homologous end joining (NHEJ), microhomology-mediated end joining (MMEJ, sometimes referred to as "alternative NHEJ" or "alt-NHEJ"), or homology-directed repair (HDR), which typically results in an altered nucleic acid sequence at or immediately proximal to the site of the nuclease cut (e.g., via a nucleotide or nucleotide sequence insertion, deletion, inversion, or substitution). See Yeh et al. Nat. Cell. Biol. (2019) 21:1468-1478, e.g., Hsu et al. Cell (2014) 157:1262-1278, Jasin et al. DNA Repair (2016) 44:6-16, Sfeir et al. Trends Biochem. Sci. (2015) 40:701-714. In some embodiments, genomic modifications are introduced into cells using HDR, e.g., as described herein.

[0122] Another exemplary suitable genome editing technique is "base editing," which involves the use of base editors, e.g., nuclease-damaged or partially nuclease-damaged RNA-guided CRISPR / Cas proteins fused to deaminases, which target and deaminate specific nucleobases, e.g., cytosine or adenosine nucleobases of C or A nucleotides, resulting in a change from a C nucleotide to a T nucleotide (also changing a G nucleotide on the opposite strand to an A nucleotide) or from an A nucleotide to a G nucleotide (also inducing a G nucleotide on the opposite strand to a C nucleotide), via cellular mismatch repair mechanisms. Base editors or "BEs" that catalyze the conversion of a C to a T nucleotide may be referred to as "cytosine base editors" or "CBEs," and base editors that catalyze the conversion of an A to a G nucleotide may be referred to as "adenosine base editors" or "ABEs." See, for example, Komor et al. Nature (2016) 533:420-424, Rees et al. Nat. Rev. Genet. (2018) 19(12):770-788, Anzalone et al. Nat. Biotechnol. (2020) 38:824-844.

[0123] Yet another exemplary genome editing technique involves "primed editing," which involves the introduction of new genetic information, e.g., modified nucleotide sequences, into specifically targeted genomic sites using catalytically impaired or partially catalytically impaired RNA-guided nucleases, e.g., CRISPR / Cas nucleases fused to engineered reverse transcriptase (RT) domains. The Cas / RT fusion also contains a nucleic acid sequence encoding the desired edit and is targeted to the target site in the genome by a guide RNA, which can function as a primer for the RT. See, e.g., Anzalone et al. Nature (2019) 576(7785):149-157.

[0124] The use of genome editing techniques is typically characterized in some embodiments by the use of a suitable RNA-guided nuclease that is catalytically impaired or partially catalytically impaired, for example, for base editing or prime editing. Examples of suitable RNA-guided nucleases include CRISPR / Cas nucleases. For example, in some embodiments, a suitable RNA-guided nuclease for use in the methods of genetically engineering cells provided herein is a Cas9 nuclease, such as an spCas9 or saCas9 nuclease. For another example, in some embodiments, a suitable RNA-guided nuclease for use in the methods of genetically engineering cells provided herein is a Cas12 nuclease, such as a Cas12a nuclease. Exemplary suitable Cas12 nucleases include, but are not limited to, AsCas12a, FnCas12a, other Cas12a orthologs, and Cas12a derivatives, such as the MAD7 system (MAD7™, Inscripta, Inc.) or Alt-R Cas12a (Cpf1) Ultra nuclease (Alt-R® Cas12a Ultra; Integrated DNA Technologies, Inc.). See, e.g., Gill et al. LIPSCOMB 2017. In the United States: Inscripta Inc.; Price et al. Biotechnol. Bioeng. (2020) 117(60):1805-1816.

[0125] In some embodiments, the engineered cells described herein (e.g., engineered hematopoietic cells, such as engineered hematopoietic stem or progenitor cells, or engineered immune effector cells) are generated by targeting an RNA-guided nuclease, such as a CRISPR / Cas nuclease, such as a Cas9 nuclease or a Cas12a nuclease, to a suitable target site within the genome of the cell under conditions suitable for the RNA-guided nuclease to bind to the target site and cleave the genomic DNA of the cell. In some embodiments, the engineered cells described herein (e.g., engineered hematopoietic cells, such as engineered hematopoietic stem or progenitor cells, or engineered immune effector cells) are generated by targeting a base editor, such as a CBE or ABE, to a suitable target site within the genome of the cell under conditions suitable for the base editor to bind to the target site and cleave the genomic DNA of the cell. A suitable RNA-guided nuclease can be targeted to a specific target site within the genome by a suitable guide RNA (gRNA). Suitable gRNAs for targeting CRISPR / Cas nucleases according to some embodiments of the present disclosure are provided herein, and exemplary suitable gRNAs are described in more detail elsewhere herein.

[0126] In some embodiments, a gRNA that binds to a gene encoding a lineage-specific cell surface antigen described herein (e.g., CD123 gRNA, CD38 gRNA, CD5 gRNA, CD47 gRNA, CD34 gRNA, EMR2 gRNA, or CD19 gRNA) is complexed with a CRISPR / Cas nuclease, e.g., a Cas9 nuclease, a base editor. Various Cas9 nucleases and base editors are suitable for use with the gRNA provided herein to effect genome editing according to some aspects of the present disclosure, e.g., to create a genomic modification in a gene encoding a lineage-specific cell surface antigen. Typically, the Cas nuclease or base editor and gRNA are provided in a form and under conditions suitable for the formation of a Cas / gRNA complex that targets a target site on the genome of a cell, e.g., a target site within a gene encoding the lineage-specific cell surface antigen (e.g., a target site in a sequence encoding an epitope bound by an agent that specifically binds to a gene encoding a lineage-specific cell surface antigen). In some embodiments, Cas nucleases that exhibit the desired PAM specificity are used to target the Cas / gRNA complex to a desired target domain of a gene encoding a lineage-specific cell surface antigen. Suitable target domains and corresponding gRNA targeting domain sequences are provided herein.

[0127] In some embodiments, a Cas / gRNA or base editor / gRNA complex is formed, e.g., in vitro, and a target cell is contacted with the Cas / gRNA complex, e.g., via electroporation of the Cas / gRNA or base editor / gRNA complex into the cell. In some embodiments, the cell is contacted separately with a Cas protein or base editor and a gRNA, and a Cas / gRNA or base editor / gRNA complex is formed within the cell. In some embodiments, the cell is contacted with a nucleic acid, e.g., DNA or RNA (such as mRNA), encoding the Cas protein or base editor, and / or a nucleic acid encoding the gRNA, or both.

[0128] In some embodiments, the genetically engineered cells provided herein are generated using a suitable genome editing technique characterized by the use of a Cas9 nuclease. In some embodiments, the Cas9 molecule is of or derived from Streptococcus pyogenes (SpCas9), Staphylococcus aureus (SaCas9), or Streptococcus thermophilus (stCas9). Additional suitable Cas9 molecules include Neisseria meningitidis (NmCas9), Acidovorax avenae, Actinobacillus pleuropneumoniae, Actinobacillus succinogenes, Actinobacillus suis, Actinomyces spp., Cycliphilus denitrificans, Aminomonas paucivorans, Bacillus cereus, Bacillus smithii, Bacillus thuringiensis, Bacteroides sp., Blastopirellula marina, Bradyrhizobium sp., Brevibacillus laterosporus, Campylobacter coli, Campylobacter jejuni(CjCas9), Campylobacter lari, Candidatus puniceispirillum, Clostridium cellulolyticum, Clostridium perfringens, Corynebacterium accolens, Corynebacterium diphtheria, Corynebacterium matruchotii, Dinoroseobacter shibae, Eubacterium dolichum, gamma proteobacterium, Gluconacetobacter diazotrophicus, Haemophilus parainfluenzae, Haemophilus sputorum, Helicobacter canadensis, Helicobacter cinaedi, Helicobactermustelae, Ilyobacter polytropus, Kingella kingae, Lactobacillus crispatus, Listeria ivanovii, Listeria monocytogenes, Listeriaceae bacterium, Methylocystis spp., Methylosinus trichosporium, Mobiluncus mulieris, Neisseria bacilliformis, Neisseria cinerea, Neisseria flavescens, Neisseria lactamica, Neisseria meningitidis, Neisseria spp., Neisseria wadsworthii, Nitrosomonas spp., Parvibaculum lavamentivorans, Pasteurella multocida, Phascolarctobacterium succinatutens, Ralstonia syzygii, Rhodopseudomonas palustris, Rhodovulum spp., Simonsiella muelleri, Sphingomonas spp., Sporolactobacillus vineae, Staphylococcus Suitable Cas9 nucleases include those derived from or derived from Bacillus lugdunensis, Streptococcus sp., Subdoligranulum sp., Tistrella mobilis, Treponema sp., or Verminephrobacter eiseniae. In some embodiments, catalytically impaired or partially impaired variants of such Cas9 nucleases may be used. Additional suitable Cas9 nucleases and nuclease variants will be apparent to those of skill in the art based on this disclosure. The present disclosure is not limited in this respect.

[0129] In some embodiments, the Cas nuclease is a naturally occurring Cas molecule, hi some embodiments, the Cas nuclease is an engineered, altered, or modified Cas molecule that differs, e.g., by at least one amino acid residue, from a reference sequence, e.g., the most similar naturally occurring Cas9 molecule, or a sequence in Table 50 of WO 2015 / 157070, which is incorporated by reference in its entirety.

[0130] In some embodiments, a Cas nuclease belonging to Class 2 Type V of Cas nucleases is used. Class 2 Type V Cas nucleases can be further classified as Type VA, Type VB, Type VC, and Type VU. See, e.g., Stella et al. Nature Structural & Molecular Biology (2017). In some embodiments, the Cas nuclease is a Type VB Cas endonuclease, such as C2c1. See, e.g., Shmakov et al. Mol Cell (2015) 60:385-397. In some embodiments, the Cas nuclease used in the genome editing methods provided herein is a Type VA Cas endonuclease, such as Cpf1 (Cas12a) nuclease. See, e.g., Strohkendl et al. Mol. Cell (2018) 71:1-9. In some embodiments, the Cas nuclease used in the genome editing methods provided herein is a Cpf1 nuclease from the genus Provetella or Francisella, the genus Acidaminococcus (AsCpf1), Lachnospiraceae bacteria (LpCpf1), or Eubacterium rectale. In some embodiments, the Cas nuclease is MAD7™.

[0131] Both naturally occurring and modified variants of CRISPR / Cas nucleases are suitable for use according to aspects of the present disclosure. For example, dCas or nickase variants, Cas variants with altered PAM specificity, and Cas variants with improved nuclease activity are encompassed by some embodiments of the present disclosure. In some embodiments, the Cas nuclease is a variant with reduced PAM sequence specificity. In some embodiments, such gRNAs are referred to as "PAMless" or "near PAMless." In some embodiments, the Cas nuclease is a SpRY nuclease. See, e.g., Walton et al., Science. 2020 Apr 17;368(6488):290-296, incorporated herein by reference.

[0132] Without wishing to be bound by any particular theory, some features of some exemplary, non-limiting suitable Cas nucleases are described in more detail herein.

[0133] Naturally occurring Cas9 nucleases typically comprise two lobes: a recognition (REC) lobe and a nuclease (NUC) lobe, each of which further comprises domains described, for example, in WO 2015 / 157070, e.g., at Figures 9A-9B therein (which application is incorporated herein by reference in its entirety).

[0134] The REC lobe contains an arginine-rich bridge helix (BH), a REC1 domain, and a REC2 domain. The REC lobe is thought to be a Cas9-specific functional domain. The BH domain is a long alpha-helical and arginine-rich region, encompassing amino acids 60-93 of the S. pyogenes Cas9 sequence. The REC1 domain is involved in recognizing repeat:anti-repeat duplexes, for example, of gRNAs or tracrRNAs. The REC1 domain contains two REC1 motifs at amino acids 94-179 and 308-717 of the S. pyogenes Cas9 sequence. These two REC1 domains are separated by the REC2 domain in the linear primary structure, but assemble in the tertiary structure to form the REC1 domain. The REC2 domain, or a portion thereof, may also play a role in recognizing repeat:anti-repeat duplexes. The REC2 domain encompasses amino acids 180-307 of the S. pyogenes Cas9 sequence.

[0135] The NUC lobe contains a RuvC domain (also referred to herein as a RuvC-like domain), an HNH domain (also referred to herein as an HNH-like domain), and a PAM-interacting (PI) domain. The RuvC domain shares structural similarity with members of the retroviral integrase superfamily and cleaves single strands, e.g., non-complementary strands, of target nucleic acid molecules. The RuvC domain is assembled from three separate RuvC motifs (RuvC I, RuvC II, and RuvC III, often referred to in the art as the RuvCI domain, or the N-terminal RuvC domain, RuvC II domain, and RuvC III domain) at amino acids 1-59, 718-769, and 909-1098 of the S. pyogenes Cas9 sequence, respectively. Similar to the REC1 domain, the three RuvC motifs are linearly separated by other domains in the primary structure, but in the tertiary structure, the three RuvC motifs assemble to form the RuvC domain. The HNH domain shares structural similarity with HNH endonucleases and cleaves a single strand, e.g., the complementary strand, of a target nucleic acid molecule. The HNH domain is located between the RuvC II-III motifs and includes amino acids 775-908 of the S. pyogenes Cas9 sequence. The PI domain interacts with the PAM of the target nucleic acid molecule and includes amino acids 1099-1368 of the S. pyogenes Cas9 sequence.

[0136] Crystal structures have been determined for the naturally occurring bacterial Cas9 nuclease (see, e.g., Jinek et al., Science (2014) 343(6176):1247997) and S. pyogenes Cas9 with a guide RNA (e.g., a synthetic fusion of crRNA and tracrRNA) (Nishimasu et al., Cell (2014) 156:935-949, and Anders et al., Nature (2014) doi:10.1038 / naturel3579).

[0137] In some embodiments, the Cas9 molecules described herein exhibit nuclease activity, resulting in the introduction of a double-stranded DNA break at or directly proximal to the target site. In some embodiments, the Cas9 molecule is modified to inactivate one of the endonuclease's catalytic residues. In some embodiments, the Cas9 molecule is a nickase, generating a single-strand break. See, e.g., Dabrowska et al. Frontiers in Neuroscience (2018) 12(75). It has been shown that one or more mutations within the enzyme's RuvC and HNH catalytic domains can improve Cas9 efficiency. See, e.g., Sarai et al. Currently Pharma. Biotechnol. (2017) 18(13). In some embodiments, the Cas9 molecule is fused to a second domain, e.g., a domain that modifies DNA or chromatin, e.g., a deaminase or demethylase domain. In some such embodiments, the Cas9 molecule is modified to eliminate its endonuclease activity.

[0138] In some embodiments, the Cas nucleases or Cas / gRNA complexes described herein are administered with a template for homology-directed repair (HDR), e.g., as described herein. In some embodiments, the Cas nucleases or Cas / gRNA complexes described herein are administered without an HDR template.

[0139] In some embodiments, Cas9 nucleases are used that are modified to enhance the specificity of the enzyme (e.g., to reduce off-target effects and maintain robust on-target cleavage). In some embodiments, the Cas9 molecule is an enhanced specificity Cas9 variant (e.g., eSPCas9). See, e.g., Slaymaker et al. Science (2016) 351(6268):84-88. In some embodiments, the Cas9 molecule is a high-fidelity Cas9 variant (e.g., SpCas9-HF1). See, e.g., Kleinstiver et al. Nature (2016) 529:490-495.

[0140] Various Cas nucleases are known in the art and are available from a variety of sources, and can be engineered / modified to modulate one or more activities or specificities of the enzyme. The PAM sequence preferences and specificities of suitable Cas nucleases, such as suitable Cas9 nucleases, such as spCas9 and saCas9, are known in the art. In some embodiments, the Cas nuclease is engineered / modified to recognize one or more PAM sequences. In some embodiments, the Cas nuclease is engineered / modified to recognize one or more PAM sequences that are different from the PAM sequences that the Cas nuclease recognizes without engineering / modification. In some embodiments, the Cas nuclease is engineered / modified to reduce the off-target activity of the enzyme.

[0141] In some embodiments, Cas nucleases are used that are further modified to alter the specificity of their endonuclease activity (e.g., to reduce off-target cleavage, decrease endonuclease activity or lifetime within cells, increase homology-guided recombination, or reduce non-homologous end joining). See, e.g., Komor et al. Cell (2017) 168:20-36. In some embodiments, Cas nucleases are used that are modified to alter the PAM recognition or preference of the endonuclease. For example, while SpCas9 recognizes the PAM sequence NGG, some variants of SpCas9 containing one or more modifications (e.g., VQR SpCas9, EQR SpCas9, VRER SpCas9) can recognize variant PAM sequences, such as NGA, NGAG, and / or NGCG. For another example, SaCas9 recognizes the PAM sequence NNGRRT, while some SaCas9 variants containing one or more modifications (e.g., KKH SaCas9) can recognize the PAM sequence NNNRRT. In another example, FnCas9 recognizes the PAM sequence NNG, while FnCas9 variants containing one or more modifications (e.g., RHA FnCas9) can recognize the PAM sequence YG. In another example, a Cas12a nuclease containing substitution mutations S542R and K607R recognizes the PAM sequence TYCV. In another example, a Cpf1 endonuclease containing substitution mutations S542R, K607R, and N552R recognizes the PAM sequence TATV. See, e.g., Gao et al. Nat. Biotechnol. (2017) 35(8):789-792. In another example, SpG Cas9 endonuclease recognizes the PAM sequence NG (also referred to as "PAM-flexible" PAM). In another example, SpRY Cas9 endonuclease recognizes the PAM sequence NRN or NYN (also referred to as "PAM-less" PAM) with greater efficiency, where R is A or G and Y is T or C. See, e.g., Liang et al. Nat. Comm. (2022) 13:3421; Walton et al. Science (2020) 368(6488):290-296.In some embodiments, the base editor (e.g., ABE or CBE) comprises an SpG Cas9 endonuclease. In some embodiments, the base editor (e.g., ABE or CBE) comprises a SpRY Cas9 endonuclease.

[0142] In some embodiments, base editors are used to create genomic modifications that result in the expression of variants of genes encoding lineage-specific cell surface antigens that are not targeted by immunotherapy. Base editors typically comprise a catalytically inactive or partially inactive Cas nuclease fused to a functional domain, e.g., a deaminase domain. See, e.g., Eid et al. Biochem. J. (2018) 475(11):1955-1964; Rees et al. Nature Reviews Genetics (2018) 19:770-788. In some embodiments, catalytically inactive Cas nucleases are referred to as "death Cas" or "dCas."

[0143] In some embodiments, the catalytically inactive Cas molecule has reduced activity, e.g., a nickase (nCas). In some embodiments, the endonuclease comprises a dCas or nCas fused to an adenine base editor (ABE), e.g., an ABE evolved from the RNA adenine deaminase TadA. In some embodiments, the endonuclease comprises a dCas or nCas fused to a cytosine base editor (CBE), e.g., a CBE evolved from a cytidine deaminase enzyme (e.g., APOBEC deaminase, pmCDA1, activation-induced cytidine deaminase (AID)).

[0144] Examples of suitable base editors include, but are not limited to, BE1, BE2, BE3, HF-BE3, BE4, BE4max, BE4-Gam, YE1-BE3, EE-BE3, YE2-BE3, YEE-CE3, VQR-BE3, VRER-BE3, SaBE3, SaBE4, SaBE4-Gam, Sa(KKH)-BE3, Target-AID, Target-AID-NG, xBE3, eA3A-BE3, BE-PLUS, TAM, CRISPR-X, ABE7.9, ABE7.10, ABE7.10*, ABE8, ABE8e, xABE, ABESa, VQR-ABE, VRER-ABE, Sa(KKH)-ABE, CBE, CBE1, CBE2, CBE3, CBE4, and CRISPR-SKIP. Additional examples of base editors can be found, for example, in U.S. Publication No. 2018 / 0312825A1, U.S. Publication No. 2018 / 0312828A1, and WO 2018 / 165629A1, which are incorporated by reference in their entireties.

[0145] Some aspects of the present disclosure provide guide RNAs that are suitable for targeting, e.g., RNA-guided nucleases provided herein to suitable target sites within a cell's genome, to generate modifications in the cell's genome that result in the expression of a variant form of a gene encoding a lineage-specific cell surface antigen that is not recognized by an immunotherapeutic agent that targets the lineage-specific cell surface antigen.

[0146] The terms "guide RNA" and "gRNA" are used interchangeably herein and refer to a nucleic acid, typically an RNA, that is bound by an RNA-guided nuclease and facilitates specific targeting or homing of the RNA-guided nuclease to a target nucleic acid, e.g., a target site in the genome of a cell. A gRNA typically contains at least two domains: a "binding domain," sometimes also referred to as a "gRNA scaffold" or "gRNA backbone," that mediates binding to the RNA-guided nuclease, and a "targeting domain," that mediates targeting of the gRNA-bound RNA-guided nuclease to the target site. Some gRNAs contain additional domains, such as a complementarity domain or a stem-loop domain. The structures and sequences of naturally occurring gRNA binding domains and their engineered variants are well known to those of skill in the art. Some suitable gRNAs are unimolecular molecules containing a single nucleic acid sequence, while other suitable gRNAs contain two sequences (e.g., a crRNA and a tracrRNA sequence).

[0147] Some exemplary suitable Cas9 gRNA scaffold sequences are provided herein, and additional suitable gRNA scaffold sequences will be apparent to those skilled in the art based on this disclosure. Such additional suitable scaffold sequences include, but are not limited to, those listed in Jinek, et al. Science (2012) 337(6096):816-821, Ran, et al. Nature Protocols (2013) 8:2281-2308, WO 2014 / 093694, and WO 2013 / 176772.

[0148] For example, the binding domain of a naturally occurring spCas9 gRNA typically comprises two RNA molecules: a crRNA (in part) and a tracrRNA. Variants of spCas9 gRNA have been engineered that comprise only a single RNA molecule, including both the crRNA and tracrRNA sequences covalently linked to each other, e.g., via a tetraloop or via a click chemistry-type covalent bond, and are commonly referred to as "single guide RNAs" or "sgRNAs." Because naturally occurring Cas12a guide RNAs comprise a single RNA molecule, suitable gRNAs for use with other Cas nucleases, e.g., Cas12a nuclease, typically comprise only a single RNA molecule. In some embodiments, suitable gRNAs can be unimolecular (having a single RNA molecule) or modular (comprising more than one, typically two separate RNA molecules), sometimes referred to herein as sgRNAs.

[0149] Suitable gRNAs for targeting target sites in genes encoding lineage-specific cell surface antigens can contain multiple domains. For example, in some embodiments in which Cas9 nuclease is used, the unimolecular sgRNA may contain, from 5' to 3': a target domain corresponding to a target site sequence in the CD123 gene (e.g., a target site in or proximal to exon 3 and / or exon 4); a first complementarity domain; and a binding domain; a second complementarity domain (complementary to the first complementarity domain); a proximal domain; Optionally, include a tail domain.

[0150] Each of these domains is now described in more detail.

[0151] The gRNAs provided herein typically contain a targeting domain that binds to a target site in the genome of a cell. The target site is typically a double-stranded DNA sequence that includes a PAM sequence and a targeting domain on the same strand as and immediately adjacent to the PAM sequence. The targeting domain of a gRNA typically includes an RNA sequence that is similar to the sequence of the targeting domain, sometimes with one or more mismatches, but typically corresponds to the targeting domain sequence in that it includes RNA instead of a DNA sequence. Thus, the targeting domain of a gRNA base-pairs with the sequence of the double-stranded target site that is complementary (with full or partial complementarity) to the sequence of the targeting domain, and thus the strand that is complementary to the strand containing the PAM sequence. It will be understood that the targeting domain of a gRNA typically does not include a PAM sequence. It will be further understood that the location of the PAM can be 5' or 3' of the targeting domain sequence, depending on the nuclease employed. For example, the PAM is typically 3' of the target domain sequence for Cas9 nucleases and 5' of the target domain sequence for Cas12a nucleases. For illustrations of the location of the PAM and the mechanism of gRNA binding to a target site, see, e.g., Figure 1 in Vanegas et al., Fungal Biol Biotechnol. 2019;6:6, incorporated herein by reference. For additional illustrations and descriptions of the mechanism of gRNA targeting RNA-guided nucleases to target sites, see Fu Y et al., Nat Biotechnol 2014 (doi:10.1038 / nbt.2808) and Sternberg SH et al., Nature 2014 (doi:10.1038 / naturel3011), both of which are incorporated herein by reference.

[0152] The targeting domain can comprise a nucleotide sequence corresponding to the sequence of the targeting domain, i.e., the DNA sequence immediately adjacent to the PAM sequence (e.g., 5' of the PAM sequence for Cas9 nuclease, or 3' of the PAM sequence for Cas12a nuclease). The targeting domain sequence typically comprises 17-30 nucleotides and can either perfectly match the targeting domain sequence (i.e., without any mismatched nucleotides) or contain one or more, but typically no more than four, mismatches. Because the targeting domain is part of an RNA molecule, a gRNA typically contains ribonucleotides, while a DNA targeting domain will contain deoxyribonucleotides.

[0153] An exemplary illustration of a Cas9 target site comprising a 22 nucleotide targeting domain and an NGG PAM sequence, and a gRNA comprising a targeting domain that corresponds perfectly to the targeting domain (and thus base pairs with perfect complementarity to the DNA strand complementary to the strand comprising the targeting domain and PAM) is provided below: [ka]

[0154] An exemplary illustration of a Cas12a target site comprising a 22 nucleotide targeting domain and a TTN PAM sequence, and a gRNA comprising a targeting domain that corresponds perfectly to the targeting domain (and thus base-pairs with perfect complementarity to the DNA strand complementary to the strand comprising the targeting domain and PAM) is provided below: [ka]

[0155] In some embodiments, the Cas12a PAM sequence is 5'-TTTV-3'.

[0156] While not wishing to be bound by theory, it is believed that, at least in some embodiments, the length of the targeting domain and its complementarity with the target sequence contribute to the specificity of the interaction of the gRNA / Cas9 molecular complex with the target nucleic acid. In some embodiments, the targeting domain of a gRNA provided herein is 5-50 nucleotides in length. In some embodiments, the targeting domain is 15-25 nucleotides in length. In some embodiments, the targeting domain is 18-22 nucleotides in length. In some embodiments, the targeting domain is 19-21 nucleotides in length. In some embodiments, the targeting domain is 15 nucleotides in length. In some embodiments, the targeting domain is 16 nucleotides in length. In some embodiments, the targeting domain is 17 nucleotides in length. In some embodiments, the targeting domain is 18 nucleotides in length. In some embodiments, the targeting domain is 19 nucleotides in length. In some embodiments, the targeting domain is 20 nucleotides in length. In some embodiments, the targeting domain is 21 nucleotides in length. In some embodiments, the targeting domain is 22 nucleotides in length. In some embodiments, the targeting domain is 23 nucleotides in length. In some embodiments, the targeting domain is 24 nucleotides in length. In some embodiments, the targeting domain is 25 nucleotides in length. In some embodiments, the targeting domain completely corresponds to a targeting domain sequence provided herein or a portion thereof, without mismatches. In some embodiments, the targeting domain of a gRNA provided herein contains one mismatch to a targeting domain sequence provided herein. In some embodiments, the targeting domain contains two mismatches to a targeting domain sequence. In some embodiments, the targeting domain contains three mismatches to a targeting domain sequence.

[0157] In some embodiments, the targeting domain comprises a core domain and a secondary targeting domain, e.g., as described in WO 2015 / 157070, which is incorporated by reference in its entirety. In some embodiments, the core domain comprises about 8 to about 13 nucleotides from the 3' end of the targeting domain (e.g., the 3'-most 8 to 13 nucleotides of the targeting domain). In some embodiments, the secondary domain is positioned 5' to the core domain. In some embodiments, the core domain completely corresponds to the targeting domain sequence or a portion thereof. In other embodiments, the core domain may comprise one or more nucleotides that are mismatched to corresponding nucleotides in the targeting domain sequence.

[0158] In some embodiments, for example, a Cas9 gRNA is provided. In some embodiments, the gRNA comprises a first complementarity domain and a second complementarity domain, wherein the first complementarity domain is complementary to the second complementarity domain and has sufficient complementarity to the second complementarity domain to form a double-stranded region, at least in some embodiments, under at least some physiological conditions. In some embodiments, the first complementarity domain is 5 to 30 nucleotides in length. In some embodiments, the first complementarity domain comprises three subdomains, from 5' to 3': a 5' subdomain, a central subdomain, and a 3' subdomain. In some embodiments, the 5' subdomain is 4 to 9 nucleotides in length, e.g., 4, 5, 6, 7, 8, or 9 nucleotides in length. In some embodiments, the central subdomain is 1, 2, or 3 nucleotides in length, e.g., 1 nucleotide in length. In some embodiments, the 3' subdomain is 3 to 25, e.g., 4 to 22, 4 to 18, or 4 to 10, or 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length. The first complementarity domain can share homology with or be derived from a naturally occurring first complementarity domain. In one embodiment, it has at least 50% homology with a first complementarity domain of S. pyogenes, S. aureus, or S. thermophilus.

[0159] The sequences and arrangements of the above domains are described in more detail in WO 2015 / 157070, which is incorporated by reference in its entirety, including pages 88-112 thereof.

[0160] The binding domain may serve to link the first complementary domain to the second complementary domain of the unimolecular gRNA. The binding domain can link the first complementary domain and the second complementary domain covalently or non-covalently. In some embodiments, the linkage is a covalent bond. In some embodiments, the binding domain is or includes a covalent bond interposed between the first complementary domain and the second complementary domain. In some embodiments, the binding domain comprises one or more nucleotides, e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides. In some embodiments, the binding domain comprises at least one non-nucleotide linkage, e.g., as disclosed in WO 2018 / 126176, the entire contents of which are incorporated herein by reference.

[0161] In some embodiments, the second complementarity domain is at least partially complementary to the first complementarity domain, and in one embodiment, has sufficient complementarity to the second complementarity domain to form a double-stranded region under at least some physiological conditions. In some embodiments, the second complementarity domain can include a sequence that lacks complementarity with the first complementarity domain, e.g., a sequence that creates a loop from the double-stranded region. In some embodiments, the second complementarity domain is 5 to 27 nucleotides in length. In some embodiments, the second complementarity domain is longer than the first complementarity domain. In one embodiment, the complementarity domain is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length. In some embodiments, the second complementarity domain comprises three subdomains, in the 5' to 3' direction: a 5' subdomain, a central subdomain, and a 3' subdomain. In some embodiments, the 5' subdomain is 3 to 25, e.g., 4 to 22, 4 to 18, or 4 to 10, or 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 nucleotides in length. In some embodiments, the central subdomain is 1, 2, 3, 4, or 5, e.g., 3 nucleotides in length. In some embodiments, the 3' subdomain is 4 to 9, e.g., 4, 5, 6, 7, 8, or 9 nucleotides in length. In some embodiments, the 5' and 3' subdomains of the first complementarity domain are complementary, eg, perfectly complementary, to the 3' and 5' subdomains of the second complementarity domain, respectively.

[0162] In some embodiments, the proximal domain is 5-20 nucleotides in length. In some embodiments, the proximal domain shares homology with or can be derived from a naturally occurring proximal domain. In one embodiment, it has at least 50% homology with a proximal domain from S. pyogenes, S. aureus, or S. thermophilus.

[0163] A wide range of tail domains are suitable for use in gRNAs. In some embodiments, the tail domain is 0 (absent), 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 nucleotides in length. In some embodiments, the tail domain nucleotides are derived from or share homology with sequences from the 5' end of a naturally occurring tail domain. In some embodiments, the tail domains comprise sequences that are complementary to each other and form a double-stranded region under at least some physiological conditions. In some embodiments, the tail domain is absent or is 1-50 nucleotides in length. In some embodiments, the tail domain can share homology with or be derived from a naturally occurring proximal tail domain. In some embodiments, the tail domain has at least 50% homology / identity to a tail domain from S. pyogenes, S. aureus, or S. thermophilus. In some embodiments, the tail domain includes nucleotides at the 3' end that are relevant for in vitro or in vivo transcription methods.

[0164] In some embodiments, the gRNAs provided herein are For example, in the 5' to 3' direction, a targeting domain (corresponding to a targeting domain in a gene encoding a lineage-specific cell surface antigen, e.g., a sequence encoding an epitope described herein), and a first strand comprising a first complementarity domain; For example, in the 5' to 3' direction, optionally, a 5' extension domain; a second complementarity domain, the proximal domain, and and optionally a second strand comprising a tail domain.

[0165] In some embodiments, any of the gRNAs provided herein contain one or more chemically modified nucleotides. Chemical modifications of gRNAs have been previously described, and suitable chemical modifications include any modifications that are beneficial to gRNA function and do not measurably increase any undesirable properties of a given gRNA, such as off-target effects. Suitable chemical modifications, for example, are those that make the gRNA less susceptible to endo- or exonuclease catalytic activity, and include, but are not limited to, phosphorothioate backbone modifications, 2'-O-Me-modifications (e.g., at one or both of the 3' and 5' ends), 2'F-modifications, substitution of the ribose sugar with the bicyclic nucleotide-cEt, 3' thioPACE (MSP) modifications, or any combination thereof. Additional suitable gRNA modifications will be apparent to those of skill in the art based on the present disclosure, including, but not limited to, those described in, for example, Rahdar et al. PNAS (2015) 112(51) E7110-E7117 and Hendel et al., Nat Biotechnol. (2015); 33(9): 985-989, each of which is incorporated herein by reference in its entirety.

[0166] For example, the gRNAs provided herein may comprise one or more 2'-O-modified nucleotides, e.g., 2'-O-methyl nucleotides. In some embodiments, the gRNA comprises 2'-O-modified nucleotides, e.g., 2'-O-methyl nucleotides, at the 5' end of the gRNA. In some embodiments, the gRNA comprises 2'-O-modified nucleotides, e.g., 2'-O-methyl nucleotides, at the 3' end of the gRNA. In some embodiments, the gRNA comprises 2'-O-modified nucleotides, e.g., 2'-O-methyl nucleotides, at both the 5' and 3' ends of the gRNA. In some embodiments, the gRNA is 2'-O-modified, e.g., at the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, and the third nucleotide from the 5' end of the gRNA. In some embodiments, the gRNA is 2'-O modified, e.g., the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA are 2'-O-methyl modified. In some embodiments, the gRNA is 2'-O modified, e.g., the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA are 2'-O-methyl modified. In some embodiments, the gRNA is 2'-O modified, e.g., the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA are 2'-O-methyl modified. In some embodiments, the nucleotide at the 3' end of the gRNA is not chemically modified. In some embodiments, the nucleotide at the 3' end of the gRNA does not have a chemically modified sugar.In some embodiments, the gRNA is 2'-O modified, e.g., the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA are 2'-O-methyl modified. In some embodiments, the 2'-O-methyl nucleotide comprises a phosphate linkage to the adjacent nucleotide. In some embodiments, the 2'-O-methyl nucleotide comprises a phosphorothioate linkage to the adjacent nucleotide. In some embodiments, the 2'-O-methyl nucleotide comprises a thioPACE linkage to the adjacent nucleotide.

[0167] In some embodiments, the gRNAs provided herein may comprise one or more 2'-O-modified and 3'-phosphorus modified nucleotides, such as 2'-O-methyl 3' phosphorothioate nucleotides. In some embodiments, the gRNA comprises a 2'-O-modified and 3'-phosphorus modified nucleotide at the 5' end of the gRNA, such as a 2'-O-methyl 3' phosphorothioate nucleotide. In some embodiments, the gRNA comprises a 2'-O-modified and 3'-phosphorus modified nucleotide at the 3' end of the gRNA, such as a 2'-O-methyl 3' phosphorothioate nucleotide. In some embodiments, the gRNA comprises a 2'-O-modified and 3'-phosphorus modified nucleotide at the 5' and 3' ends of the gRNA, such as a 2'-O-methyl 3' phosphorothioate nucleotide. In some embodiments, the gRNA comprises a backbone in which one or more non-bridging oxygen atoms are replaced with a sulfur atom. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, for example, 2'-O-methyl 3' phosphorothioate modified at the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, and the third nucleotide from the 5' end of the gRNA. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, for example, 2'-O-methyl 3' phosphorothioate modified at the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, e.g., 2'-O-methyl 3' phosphorothioate modified at the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, e.g., 2'-O-methyl 3' phosphorothioate modified at the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA. In some embodiments, the nucleotide at the 3' end of the gRNA is not chemically modified.In some embodiments, the nucleotide at the 3' end of the gRNA does not have a chemically modified sugar. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, for example, 2'-O-methyl 3' phosphorothioate modified at the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA.

[0168] In some embodiments, the gRNAs provided herein may comprise one or more 2'-O modifications and 3'-phosphorus modifications, such as 2'-O-methyl 3'-thio PACE nucleotides. In some embodiments, the gRNAs comprise a 2'-O modification and a 3'-phosphorus modification, such as 2'-O-methyl 3'-thio PACE nucleotides, at the 5' end of the gRNA. In some embodiments, the gRNAs comprise a 2'-O modification and a 3'-phosphorus modification, such as 2'-O-methyl 3'-thio PACE nucleotides, at the 3' end of the gRNA. In some embodiments, the gRNAs comprise a 2'-O modification and a 3'-phosphorus modification, such as 2'-O-methyl 3'-thio PACE nucleotides, at the 5' and 3' ends of the gRNA. In some embodiments, the gRNAs comprise a backbone in which one or more non-bridging oxygen atoms are replaced with a sulfur atom and one or more non-bridging oxygen atoms are replaced with an acetate group. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, for example, 2'-O-methyl 3' thio PACE modified at the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, and the third nucleotide from the 5' end of the gRNA. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, for example, 2'-O-methyl 3' thio PACE modified at the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, for example, 2'-O-methyl 3' thio PACE modified at the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, e.g., 2'-O-methyl 3' thio PACE modified at the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA. In some embodiments, the nucleotide at the 3' end of the gRNA is not chemically modified.In some embodiments, the nucleotide at the 3' end of the gRNA does not have a chemically modified sugar. In some embodiments, the gRNA is 2'-O modified and 3' phosphorus modified, for example, 2'-O-methyl 3' thio PACE modified at the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA.

[0169] In some embodiments, the gRNAs provided herein comprise a chemically modified backbone. In some embodiments, the gRNA comprises a phosphorothioate bond. In some embodiments, one or more non-bridging oxygen atoms are replaced with a sulfur atom. In some embodiments, the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, and the third nucleotide from the 5' end of the gRNA each comprise a phosphorothioate bond. In some embodiments, the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA each comprise a phosphorothioate bond. In some embodiments, the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA each comprise a phosphorothioate bond. In some embodiments, the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA each comprise a phosphorothioate linkage. In some embodiments, the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end, the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA each comprise a phosphorothioate linkage.

[0170] In some embodiments, the gRNAs provided herein comprise a thioPACE bond. In some embodiments, the gRNA comprises a backbone in which one or more non-bridging oxygen atoms are replaced with a sulfur atom and one or more non-bridging oxygen atoms are replaced with an acetate group. In some embodiments, the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, and the third nucleotide from the 5' end of the gRNA each comprise a thioPACE bond. In some embodiments, the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA each comprise a thioPACE bond. In some embodiments, the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end of the gRNA, the nucleotide at the 3' end of the gRNA, the second nucleotide from the 3' end of the gRNA, and the third nucleotide from the 3' end of the gRNA each comprise a thioPACE bond. In some embodiments, the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA each comprise a ThioPACE bond. In some embodiments, the nucleotide at the 5' end of the gRNA, the second nucleotide from the 5' end of the gRNA, the third nucleotide from the 5' end, the second nucleotide from the 3' end of the gRNA, the third nucleotide from the 3' end of the gRNA, and the fourth nucleotide from the 3' end of the gRNA each comprise a ThioPACE bond.

[0171] In some embodiments, the gRNAs described herein comprise one or more 2'-O-methyl-3'-phosphorothioate nucleotides, e.g., at least 1, 2, 3, 4, 5, or 6 2'-O-methyl-3'-phosphorothioate nucleotides. In some embodiments, the gRNAs described herein comprise modified nucleotides (e.g., 2'-O-methyl-3'-phosphorothioate nucleotides) at one or more of the three terminal positions and at the 5'-terminus and / or at one or more of the three terminal positions and at the 3'-terminus. In some embodiments, the gRNAs comprise one or more modified nucleotides, e.g., as described in WO 2017 / 214460, WO 2016 / 089433, and WO 2016 / 164356, which are incorporated by reference in their entireties.

[0172] The gRNAs targeting genes encoding lineage-specific cell surface antigens provided herein can be delivered to cells in any suitable manner. For example, various suitable methods have been described for delivering CRISPR / Cas systems, including RNPs containing gRNAs linked to RNA-guided nucleases. Exemplary suitable methods include, but are not limited to, electroporation of RNPs into cells, electroporation of mRNAs and gRNAs encoding Cas nucleases into cells, various protein or nucleic acid transfection methods, and delivery of coding RNA or DNA via viral vectors, such as retroviral (e.g., lentiviral) vectors. Any suitable delivery method is encompassed by the present disclosure, which is not limited in this respect.

[0173] The present disclosure provides a number of CD123 target sites and corresponding gRNAs that are useful for targeting RNA-guided nucleases to human CD123. Table 1 below illustrates preferred target domains in the human endogenous CD123 gene that can be bound by the gRNAs described herein. The exemplary target sequences of human CD123 shown in Table 1 are, in some embodiments, for use with Cas9 nucleases, e.g., SpCas9. [Table 1]

[0174] The present disclosure provides exemplary CD123-targeting gRNAs that are useful for targeting RNA-guided nucleases to human CD123. Table 2 below illustrates preferred targeting domains for use in gRNAs that target Cas9 nucleases to the human endogenous CD123 gene. The exemplary target sequences for human CD123 shown in Table 2 are, in some embodiments, for use with Cas9 nucleases, such as SpCas9. [Table 2] [Table 3]

[0175] A representative DNA sequence of the CD123 gene is provided by NCBI gene ID: 3563 and is shown below.

[0176] A representative mRNA sequence for CD123 is provided by NCBI reference sequence number: NM_002183.4 and is shown below: [ka]

[0177] A representative amino acid sequence of CD123 is provided by NCBI reference sequence number NP_002174.1 and is shown below: MVLLWLTLLLIALPCLLQTKEDPNPPITNLRMKAKAQQLTWDLNRNVTDIECVKDADYSMPAVNNSYCQF GAISLCEVTNYTVRVANPPFSTWILFPENSGKPWAGAENLTCWIHDVDFLSCSWAVGPGAPADVQYDLYL NVANRRQQYECLHYKTDAQGTRIGCRFDDISRLSSGSQSSHILVRGRSAAFGIPCTDKFVVFSQIEILTP PNMTAKCNKTHSFMHWKMRSHFNRKFRYELQIQKRMQPVITEQVRDRTSFQLLNPGTYTVQIRARERVYE FLSAWSTPQRFECDQEEGANTRAWRTSLLIALGTLLALVCVFVICRRYLVMQRLFPRIPHMKDPIGDSFQ NDKLVVWEAGKAGLEECLVTEVQVVQKT (SEQ ID NO: 15)

[0178] The present disclosure provides several CD38 target sites and corresponding gRNAs that are useful for targeting RNA-guided nucleases to human CD38. Table 3 below illustrates preferred target domains within the human endogenous CD38 gene that can be bound by the gRNAs described herein. The exemplary target sequences for human CD38 shown in Table 3 are, in some embodiments, for use with Cas9 nucleases, such as SpCas9. [Table 4-1] [Table 4-2]

[0179] The present disclosure provides exemplary CD38-targeting gRNAs that are useful for targeting RNA-guided nucleases to human CD38. Table 5 below illustrates preferred targeting domains for use in gRNAs that target Cas9 nucleases to the human endogenous CD38 gene. The exemplary target sequences for human CD38 shown in Table 5 are, in some embodiments, intended for use with Cas9 nucleases, such as SpCas9. [Table 5-1] [Table 5-2] [Table 6-1] [Table 6-2]

[0180] A representative DNA sequence of the CD38 gene is provided by NCBI gene ID: 952 and is shown below.

[0181] A representative mRNA sequence of CD38 (transcript variant 1) is provided by NCBI reference sequence number: NM_001775.4 and is shown below: [ka] [ka]

[0182] A representative amino acid sequence of CD38 is provided by NCBI reference sequence number NP_001766.2 and is shown below: MANCEFSPVSGDKPCCRLSRRAQLCLGVSILVLILVVVLAVVVPRWRQQWSGPGTTKRFPETVLARCVKY TEIHPEMRHVDCQSVWDAFKGAFISKHPCNITEEDYQPLMKLGTQTVPCNKILLWSRIKDLAHQFTQVQR DMFTLEDTLLGYLADDLTWCGEFNTSKINYQSCPDWRKDCSNNPVSVFWKTVSRRFAEAACDVVHVMLNG SRSKIFDKNSTFGSVEVHNLQPEKVQTLEAWVIHGGREDSRDLCQDPTIKELESIISKRNIQFSCKNIYR PDKFLQCVKNPEDSSCTSEI (SEQ ID NO: 63)

[0183] The present disclosure provides a number of CD19 target sites and corresponding gRNAs that are useful for targeting RNA-guided nucleases to human CD19. Table 7 below illustrates preferred target domains in the human endogenous CD19 gene that can be bound by the gRNAs described herein. The exemplary target sequences of human CD19 shown in Table 7 are, in some embodiments, for use with Cas9 nucleases, such as SpCas9. [Table 7]

[0184] The present disclosure provides exemplary CD19-targeting gRNAs that are useful for targeting RNA-guided nucleases to human CD19. Table 8 below illustrates preferred targeting domains for use in gRNAs that target Cas9 nucleases to the human endogenous CD19 gene. The exemplary target sequences for human CD19 shown in Table 8 are, in some embodiments, for use with Cas9 nucleases, such as SpCas9. [Table 8] [Table 9]

[0185] A representative DNA sequence of the CD19 gene is provided by NCBI gene ID: 930 and is shown below:

[0186] A representative mRNA sequence of CD19 (transcript variant 2) is provided by NCBI reference sequence number: NM_001770.6 and is shown below: [ka] [ka]

[0187] A representative amino acid sequence of CD19 is provided by NCBI reference number NP_001761.3 and is shown below: MPPPRLFFLLFLTPMEVRPEEPLVVKVEEGDNAVLQCLKGTSDGPTQQLTWSRESPLKPFLKLSLGLPG LGIHMRPLAIWLFIFNVSQQMGGFYLCQPGPPSEKAWQPGWTVNVEGSGELFRWNVSDLGGLGCGLKNRS SEGPSSPSGKLMSPKLYVWAKDRPEIWEGEPPCLPPRDSLNQSLSQDLTMAPGSTLWLSCGVPPDSVSRG PLSWTHVHPKGPKSLLSLELKDDRPARDMWVMETGLLLPRATAQDAGKYYCHRGNLTMSFHLEITARPVL WHWLLRTGGWKVSAVTLAYLIFCLCSLVGILHLQRALVLRRKRKRMTDPTRRFFKVTPPPGSGPQNQYGN VLSLPTPTSGLGRAQRWAAGLGGTAPSYGNPSSDVQADGALGSRSPPGVGPEEEEGEGYEEPDSEEDSEF YENDSNLGQDQLSQDGSGYENPEDEPLGPEDEDSFSNAESYENEDEELTQPVARTMDFLSPHGSAWDPSR EATSLGSQSYEDMRGILYAAPQLRSIRGQPGPNHEEDADSYENMDNPDGPDPAWGGGGRMGTWSTR (SEQ ID NO: 87)

[0188] The present disclosure provides numerous EMR2 target sites and corresponding gRNAs that are useful for targeting RNA-guided nucleases to human EMR2. Tables 10 and 11 below illustrate preferred target domains in the human endogenous ADGRE2 gene encoding EMR2 (CD312) that can be bound by the gRNAs described herein. The exemplary target sequences of human EMR2 shown in Tables 7 and 8 are, in some embodiments, for use with base editors, such as CBEs or ABEs.

[0189] Tables 10 and 11. Exemplary target site sequences for human EMR2 are provided, as well as exemplary gRNA targeting domain sequences useful for targeting such sites. For each target site, the first sequence represents the DNA targeting domain sequence, the second sequence represents its reverse complement, and the third sequence represents an exemplary targeting domain sequence of a gRNA that can be used to target the respective target site. [Table 10-1] [Table 10-2] [Table 10-3] [Table 10-4] [Table 10-5] [Table 11-1] [Table 11-2] [Table 11-3] [Table 11-4]

[0190] A representative amino acid sequence of full-length EMR2 is provided by UniProtKB / Swiss-Prot accession number Q9UHX3-1 and is shown below. MGGRVFLVFLAFCVWLTLPGAETQDSRGCARWCPQDSSCVNATACRCNPGFSSFSEIITTPMETCDDINECATLSKVSCGKFSDCWNTEGSYDCVCSPGYEPVSGAKTFKNESENTCQDVDECQQNPRLCKSYGTCVNTLGSYTCQCLPGFKLKPEDPKLCTDVNECTSGQNPCHSSTHCLNNVGSYQCRCRPGWQPIPGSPNGP NNTVCEDVDECSSGQHQCDSSTVCFNTVGSYSCRCRPGWKPRHGIPNNQKDTVCEDMTFSTWTPPPGVHSQTLSRFFDKVQDLGRDYKPGLANNTIQSILQAL DELLEAPGDLETLPRLQQHCVASHLLDGLEDVLRGLSKNLSNGLLNFSYPAGTELSLEVQKQVDRSVTLRQNQAVMQLDWNQAQKSGDPGPSVVGLVSIPGMG KLLAEAPLVLEPEKQMLLHETHQGLLQDGSPILLSDVISAFLSNNDTQNLSSPVTFTFSHRSVIPRQKVLCVFWEHGQNGCGHWATTGCSTIGTRDTSTICRCTHLSSFAVLMAHYDVQEEDPVLTVITYMGLSVSLLCLLLAALTFLLCKAIQNTSTSLHLQLSLCLFLAHLLFLVAIDQTGHKVLCSIIAGTLHYLYLATLTWM LLEALYLFLTARNLTVVNYSSINRFMKKLMFPVGYGVPAVTVAISAASRPHLYGTPSRCWLQPEKGFIWGFLGPVCAIFSVNLVLFLVTLWILKNRLSSLNSE VSTLRNTRMLAFKATAQLFILGCTWCLGILQVGPAARVMAYLFTIINSLQGVFIFLVYCLLSQQVREQYGKWSKGIRKLKTESEMHTLSSSAKADTSKPSTVN (SEQ ID NO: 424)

[0191] A representative amino acid sequence of ADGRE2 encoding EMR2 is provided by NCBI reference sequence number NG_047146.1. TACTTACTTATTTTCTGAGACAGAGTCTCACTCTGTGGCCCAGGCTGGAGTACAGTGGCGAGAGCTCACT GCAACCTTGAACTCCTGGGCTCAAGAGATCCTCCCACCTCAGCCCCGTGAGTAGCTGGGATTCCAGGCAC ATGCCATCACTCCTGGCTAATTTTTGCATTTTTTGGTAGAGATGGAGTTTTACCATGTTGCCCAGGCTGG TCTCGATCCCCTGGGCTCAAGGGATCCTCCCGCCTCGGCCTCCAAAAGTGCTGAGATCACAGGCATGAGC CACTCTGCCTGGCCGATTTTATCTTCCTTGATACAAACTCTTTTCAGAATTCTCTTTGTTTTTGCTTTGG TCCCTGATAAAATCACATGTGAATATAACCGCAAAACACCACATGCCTGCATTACATGGCGTTACGTTAA ACAACCATCATTTTGGCTGGGCGCAGTGACTCATGCCCGTCTCTACTAAAAATACAAAAAAATTAGCCTG GCGTGGTGGTGTGCACCTGTAATCCCAGCCACTTGGGAGGCTGAGGCAGGAGAATGGCTTGAACGCGGGG GGCGGAGGTTGCAGTGAGCCGAGATTGTGCCACTGCACTCCTGCCTGGGCGACAGAGCAAGAGACTCTGT CTCAAGAAAACAAAACAAAACAGCCATCATTTTAGGAAACATTCATATGTGGAAACTACTATGTGTATTG TTCACAATTTGCGGGAATAAAGAACTTTGCTTTGCAAAAAATATGTAAATTAAAAAATAAAAGGGAAGGA AGGCATACCCATGAGCCTCCCTCATCACCAAGAACTGAGACATGACCTTGTTCCCATGCTCCCATCTTTA TCTTGCTGATTACCACCACTCTGAATATTCTTTGCACCTTATTCCTTTGCTCAGTTACTTATTTTTTTAT TTTATTTATTTTTGAGACAGGGTCTTGCTCTGTCACCCAGGCTGGAGTGCAGTGGTATGATCACAGCTCA CTGCCACCTCGAATTCCTGGGCTCAAGCCATCCTCCCACTTCAGCCTCCTGAGTAGCTGGGACCACAGGT GCGAGTCACCACACCTGGCGAATTTTTTATTTTTATTGTAGAGATGGAGGTCTCATTATGTTGCCCAGGA TGGTCTCGAACTCCTGGGCTCAAGCAATCCTCCCGTCTCAGCATCCCCAAGTAGCTGGGATTACGAGCAT GAACCACTGCGCCCGGCCCAGCCTTGTTTATTTTTATTTATTTATTTATATTTATGTATTTTTATTTTAT TTTATTTTATTTAGACGGAGTCTCACTCTTGTCTCCCAGGCTGGAGTGCAGTGGCGCAATCTCAGCTCAC TGCAGCCTCTGCCTCCCGGGTTCAAGTGATTCTCTTGCTTCAGCCTCCCAAGTAGCTGGGATTACAGGCA CCCACCACCATACCCGGCTAATTTTTGTACTTTTAGTACAGACGGGGTTTCACCATGTTGGCCAGGCTGG TCTTGAACTCCTGACCTCAGGTGATCTGCCCACCTCGGCCTCTCTGGGATTACAGGCATGAGCCACCGCG CCCGGCAGCCTTGATTATTTTTTAAAGTCTTCTTGCATATGTTTGTATCCTGAAACCAAATCTTTTATTA TCTTTGTATCTCAGCTTTTTTGCAAGGTGGCTTTTTCCTATGGGGCTTTGGAGACTTTTTCAAGATTCTA GTATGGAATCTGTCATGCTGCAATTTAAGATTTTTTTCTCCTGACTTCACTCAGTTTTTAAATTTAAAAA AGGAGTGTGGCTAGGCATGGTGGCATGCACCTGTAACCCCAGCATTTTGGGAGGCTGAGGTGGGCGGATC ACTTGAGGGTCAGGAGATCGAGACCAGCCTGGCCAACATGGTGAAACCCCGTCTCTACTAAAAAATTAGC TGGGCGTGGTGGTGTGTGCCTGTAATCCCAGCTTCTAGGGAGGCTGAGGCAGGAGAATTGCTTGAAGCTG GGAGGCGGAGGTTGCAGTGAGCCGAGATCTCGCCACTGCACTCTAGCCTGGGTAACAGGGCAAGATTCCG TCCCCCTCACACACACACAAAAAAAGTGAAAAGTTAGCCAGGCATGGCAACATATGCCTGTAGTCCCAGC TACTTGGTAGGCTGAGGCAGGAGGATCACATAAGCCCAGGACGTGCAGGTTGCCTGAAGCCATAATTGTG CCACCGCCACTGTGCTCCAGCGTGGGCAACAGAGACCCTGTCTCTAAAATAATAACAATAAATTATACTA ATATAATTAATAATACCATTATTATTGTTGGTTGGATAATCTTGGGTTGCTTCTTTTTTTTTTTTTTTTT TTTTTGAAACGGAGTCTGGCTCTATCGCCCAGGCTGGAGTGCAATAACGCGATCTCAGCTCACTGCAACC TACATCTCCTGGGCTCAAGCAATTCTCCTGCTTCAGCCTCCCAAGCAGCTGGGATTACAGGTAATCTTGG GTTACTTCTTAGCTGAGCAACCTGGGGCCAGGGGCCCCAAACCCCCAGGTTCCAGTTCCCCTTTTGACAA ATGAGGACCATGTTCGCACCCCCCATCATTGCAGAGGTTGTCCAGAAGCTGCAAAATAGGTCAGCAGGAC TGAGACTTCTGGACAAAGAGAAGGCGGAGAAACATAGGCAGACACCCCGGCAGGGACCAGATCAGCGAGG CTGAAGGAGGCTTCCTTTAGCTGGGCGGCAGCGGAGTGGAGTGGCAGGAGTACTACTAATTTGCATGGTT TACCATATAGGGCCCCACTCAGCACAGGCTCTGTGCTGGTGCTCTGCAGAGAAAAAGACAAAAGTCACTC AGGGGGCTGACAGCCTAATGCGGCTCGAGGGCAGATGGTTGCTAGGGAAGAAGGGAGATGGTGAGAAGTA TTGGAGAGACTCAAGAAAGCCGCCAGGAATGGTGACTCACGCCTGTAATCCCAACGCTTTGGGAGGCTGA GGCGCGTGGATCATTTGAGGACAGGAGTTCGAGACCAGCCTGGCCAACATGGTGAAACCCTGTCTCCACT AAAAATACAAAAATTATCTAGATGCGATGGCAGGTGCCTATAATCCCAGCTACTCGGGAGGCTGAGGCAG GAGAATTGCTTGAACCCGGGAGGCAGAGATTGCAGTGAGACGAGATAGCACCACTGCACTCCGGCTTGGG TGACAGAGCAAGACTTCGTCTCAGAAAAAAGTAATAATAAATAATAATAATAATAATAAAATAAAAATTT AAAAAGAATGTTTCAGGCCGGGCATTGTGGTTCACGCCTGTAATCCCAGCACTTTTGGGATGCCAAGGCA GGCAGATCACTTGAGGCCAGAAGTTCGAGACCAGCCTGACCAACATAGCGAAACTCCATCTCTGCTAGAA ATACAAAAATTAGCCAGGCGTGGTGGCTTGCACCTGTAATTCCAGATACTTGGGAGGCTGAGGCAGGAGA ATCTCTTGAACCCAGGAGGCAGAGGTTGCTGTGAGCTGAGATCACATCACCACACTCCAGCCTGGGTGAC AGAGCGAGACTCTGTCTCAAAATAAATAAATGAATGAACAAATAAATAAAATAAAATAAAAATACAAAAA TTAGCCAGGTGTGGTGGTGGGCACCTGTAGTCCGAGCTACTCTGGAGGCTGAGGCAGGAGAATCGCTTGA ACCCGGGAGGCGGAGGTTGCAGTGAGTCGAGATGGCGCCACTGTACTCCAACCTGGGCAACACCAGAGTC TGTCTCTAAAATACAGTAAAATAAAATTAAATTAAGAAGAGTCGATGATCACCATGAAGGGGTGTAGGGA CAATCCCTGGCATCAGGAAGTGCCACTACCTCCCTGAGATTCTGCAGACGCCAGCTGTCTGACCTGCCCG TCTGATCTGCAGAGAAGGGACGTTGAGCCCTGGGGTTCTAGCTGGAGTTTCTGGCTTTCTGAGAAAAAAA AGGCAAAATCCCTCCAGACCTGGTCTGGCCCTCTCTCCTTCCAGCTTCTGCTGGTGTTCACTCCCCTCCC CCACAGCCCAGGCCACCCCCACAGGAGCCCGGGACAGGAAGGGGTAAATTCTAGGACTGTGGATGTTTGA TTATTCCCAGGACAATAGTTCTCTGACTTCAGTCATTAGTGACTTTTTTCTCCATCGCTGCAATCTCCCC CTTTGAACAACTGAACTATATTTTCCTTAGCATTCATCACTTCTGCATCTTGAATTTACTCATGTAACAA GGAGACTTGATTTCGGCATCCTCACTGAAAAACCCTCCTCATTGCCCCAGATAGTAGGGAAACCTAAAAA TAAATATAAATATGAGGAACACAAAACCAAGTCCTTGAATTCTGGCTGGATACAGACACCTGCTAAGCCT CAGGGTCACGGAGGCGCTAAAGACATGTTAGTGCCCAGTGGAGACTTTCCCCTGGGCTCATCGCAGGGAC TCCAGGGCTGGGCTGCCGGCCTGTGGATCTGAGTCTCAGCTGGGTGAGTCACTTGTGCCTCAGTTTCCTC CTCTGTCAAATGGGAATGACAACATCATTGCAGATGTTTTAAGGAGTAAATGAGTTAATATAGATTAAAA AAAAAAAGCACTTAGGACCAGACCTGGTACAAATTAAGTGCTCCATAAATAATAGCATTTTCTTGGCAGG ATAGCGAGATCTCGTCTCTACAAAAAAAAAAAACACTTAAGGAGAAAACTAGCCAGACGTGATGGTGCAC AGCTGTAGTCCCAGCTCCTCAGGAGGCTGAGGCAGGAGGATCCCTTGAGCCCTGAAGTTTGAGGCTGCAG TGAGCTGTGATGGTGACCCTGCACTCCAGCCTGGGTGACAGAGTAAGATCTCTTAAAAATATATATAGCG TTTTAAAAAATGATTACGTACCTCAAGAGACAAGCTAATCATATGATTTATCCAGAAAGGAAAGAACCCT CCTATCACATGATTCTCTGTTGATTAACGAAGTGCCCAGGGGCCTCTGAAATATAAGTTTCCCACAGGAG GAAACTGGCTTAGAAAGACCAGAAAGATCCGGGGGGTCTGCTCTGTGCGGTGAAGCTTCTCTTCTTGGCA CCTGCCTGGCATCGGAAGAGGGCCCCTTCTCCCTCCCTGGGCTTTTATGTGGACACTGTAATGCCTCAGT TTTCTTTCTTTCTTTTTGTTTTTGACACAGGGTCTCACTCTATCACCCAAGCTGGAGTGCAGTGGCACCA TCTTAGCTCACCGCAGCCTCAAACTCCCCAGCTCGGGTGATTCTTCCTGCCTCAGCCTCCTGAGTAGTAG CAGCTGGGACCACAGACGTCTGCCACCAAGCCCAGGTAATTTTTTTTCTTTTTGAAGAAACAGAGCTTTG CCATGTTGCCCAGGCTGGTCTCAAACTCCTGAGCTCAAGCGATCCTCCTGCCTCGGCCTCCCAGAGTGCT GGGATCACAGGTGTGGAGAAATAGGAACGCTTTTACACTGTTGGTGGGAATGTAAATTAGTTCAACCATT GTGGAAGATAGTGTGGTGATTTCTCAAGGATCCAGAACTAGAAATACCATTTGACTCAGCGATCCCATTG CTGGGTATATACCCAAAGGATTATAAATCATGCTACTATAAAGACATATGCACATGTATGTTTATTGCGG TACTATTCACAATAGCAAAGACCTGGAATCAACCCAAATGTCCATCAATGATAGACTGGATTAAGAAAGT GTGGCACATATACACCAGGGAATACTATGCAGCTATAAAAAAGGATGAGTTCATGTCCTTTGCAGGGACA CAGATGAAGCTGAAAACCATCATTCTCAGCAAACTAACACAGAAACAGAAAACCAAACACCACATGTTCT CACTCATAGGTGGGAACTGAGCAATGAGAACACCTGGACACAGGGCGGGAAACATCACACACTGGGGCCT GTCGGGGGGTGGGGGCCTGGGGGAGGGATTGCATTAGGAGAAATACCTAATGTAAATGACGAGTTGATAG GTGCAGCAAGCCAACATGGCACATGTATGCCTATGTATCAAACCTGCTCGTTGTGCACATGTACCCTAGA ACTTAAAGTCTAAAAAATTTAATTAATTAATTAATTAATTAATTTAAAAAGAAAGAAAGAAACCGAGCTT CGCCATGTTGCCCAGGCTGGTCTCAAACTCCTGAGCTCAAGCGATCCTCCTGCCTCGCCCTCCCAGAGTG CTGGGATCACAGGCGTGAGCCTCTGTACCTGGCCTCAGTTTCCTCATCTGGAAAACAGCAGGGGAAGATA CTGCCAGCTGGGACGCAGGCTCTGCCAACTCTGGCATGCCAAAGCTCTGGGCACAGGGTGAGCCCTACCA AAGTGGGAGCCCAGAGCTATTTCTGACAAGGCCCCTGCGTGCCTGCCCAGGGAAGAGCCAAGTGGCCCCA GGCGAGACCGCCCCGCCCGGCTGCCACACCTTCCCCTGCACAGCAGCCAGCCTGGAACACACAGAGGCCA GAGGGTTGGTCAGAGCCACATGGTGGAAACTCCAGCAGAGGGATTTTTAAAGCAGGTTGCACTTCATTCT TGCTGAGCGAGTACACGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGTGCAGCGCCCCTGGTTC TGTGTTTTTATTGTGCTTTCCTGCTGGCTTCCAGCTGCACCGCCAGTTCCGGGGAGGGCCCTGGGCCAGC GGCTGTCCGCCCCCCCTCCTTTATAAAGTCCTGGCCTCGGGACAGCCCGCACAGCTGCCCAGCCTGCGGA GACGGGACAGCCCTGTCCCACTCACTCTTTCCCCTGCTGCTCCTGCCGGCAGCTCAGCTGGAACCATGGG AGGCCGCGTCTTTCTCGTCTTTCTCGGTAAGTACTTTGGGGCCCCGCTGGGGGTGGTAGCGAGGAAGCTC CAGCGGGACCCCTTGGATGCGTCTGAGAAACGGGAGGCGCCGCTGGTGTCTGTGGGGCCGGCGATGGAGA AAGCCACATGGTCAAGCAGGCACTCACGGGCACACGGCGAGAGGGGCTGGTGGTGCTTGGGCACCCCCAC ACCCCGGCAGGAACTCCAGGGTGCTCTCGGAGCCTGCTACCCTGCCCGGAGGGATCCTCAAGCTTTCTAC TGGAGCCAGTTTGGCATCTGCTCCATAAGAGGGGAAACTGAGGCACGGGGGAAGGAAAGGGGCAAAAAGT GAGTGCTAGGACCCCAGAGCCACTGCTCTTAATATAGCGCATAGAAAGACATGGGCGGGGGGCGGGGAAA ATAGAGGCCAGGACTTCTCCACTGCAGGGTACCCCCAGCTTCTCCCTTCTTTCTCTCTCTCTCTCTGTCT TTCTCCTTCTGTCTCTCTTTCAGTCTCTCTATCTCCCTAATCTCCGTTTTCTCTCTACCTCTGCCTCTTT GTCTCTCTCTCTGGTGAACAACAAGGAGATCCCTAGGTCCCCATCTCTAAAGCCCCTGGTGACCTGAGGA GGGTCCCCCAGGACATGCCTTGGAAAGGGGAACTTCCTGAGTGCTGGGAACCGCAGCGCCACATGCACCC CAACAGGCGGCCACAGCGTTTTCCTGTGGGAGGGGCGCCCGGGGTGAAATATCACCACTGGGTTTCAGGA TGGGCCGCCAGGGCTGGCGGCGGTTGTGGTGTGTGTGCACCTGTGTCCCTGAGGAAGCATGCGGTTCCAG GGCATCTTAGAGCCTGGGGGATCCAGCGCCATCATTTTTTTTTTTTTTTTTCTGAGGCGGAGCCTTGCTC TGTCGCCCAGGCTGGAGTGCAGTGGCACGATCTCAGCTTGCTGCAACCTCCGCCTCCTGGGTTCAAGCGA TTCTCCTGCCTCAATCTCCCAAGTAGTTGGGTAACAGGAGTCTGCCACCACGCCCAGCTAATGTTTTGTA TTTTTAGTAGAGACGGGGTTTCACCATATTGGGCAGGCTGGTCTCGAACTCCCGACCTCAGATGATCTGC CCACCTTGGCCTCCCAAAGTGCTGGGATTACAGTCCCGAGCCACCGTGCCTGACCATGCCTTCATTTTTA TCTAAAGAAATGAAGATGGCAGTAAATGCTCAGGCACACCAGACAGCCCCCAAGTCAGAAGAGCGGCAGC TGGAGCTGAGACCCCCACCAGGCTCATGGCCCTTTCCTACTCCTCAGTTCCTTAAACCCCACCCCCAAGC CAAGCTAGGGAGGCTGAGGCAGGAGGATCGCTTGAGGCCAGGAGTTCAAGATCAGCCTGGGCAACAGAGC AAGACTCTGTCTCTAAAATAACTTTTTAAATTATTTTTAATTTTTTTGGCCAGGCGTGATGGCTCATGCC TGTAATCCCAGCACTTTGAGAGGCCAAGGCGGGCAGATCACCTGAGGTTAGGAGTTCAAGACCAGCCTGG CCAACATGGTGAAACCCCATCTCTACTTAAGAAATACAAAAATTAGCTGGGCATGGTGGGTCATGCCTGT AGTCCCAGCCACTCAAGAGGCTGAGGCAGGAGAACCGCTTGAACCCAGGAGGTGGAGATTGCAGTGAGCC AAGATTGCACCACTGCATTCCAGCCTGGGCAACAAAGGGAGACTCCATCTCAAAAAAAAATAAATAATAA AAATAAATAAATCATTTTAAAATTTTTGAGCTGGGAGCCGTGGCTCACACCTGTAATCCTAGCACTTTGG GAGGCCAAGGCCGGCAGATCACCTGAGGTCAGGAGTTCGAGACCAGCCTGGCCAATATGGTGAAACCCCA TCTCACTAAAAATACAAAAAATTAGCCAGGCACCATGGTGGGTGCCTGTAGTCCCAGCTATTTGGGAGGT TGAGGCAGGAGAATTGCTTGAACCCAGGAGGTAGAGATTGCAATTAGCCGAGATCGCACCATTGCACTTC AGCCTGGGCGACAGAGCAAGACCCTGTCTCAAAAAGAAAGAAAGAAAATTTAAAATATGGCGGTACTCAC CTGTGGTCCCAGCTACTCGGGAGGGTGAGGCAGGGGGATCGCTTGAGCCTAGGAATTGGAGGCTGCAGTG AGCTATAATCGCATCACTGCACCCCAGCCTGGGTGACCCAGAGAGACCCTGTGTCAAACAAAACAAAACA AAACCCACCCCCATGAGGACAGGGTGACAGACTCTCTGGCTTTGAGCATCCGAGTGGGACGTGACATCTG CTCACTCTGTTTGAACGATGGGCAGGATCTGAGCCTGAGAGGGGTCAGCATCCTCCTCCCACAAAGTGCA TCACCCTTACGCCTCCTTTCCCACCCTGGGATCCCCTCTGACCCCCTTTCCTTTCTCTGTTGCAGCATTC TGTGTCTGGCTGACTCTGCCGGGAGCTGAAACCCAGGACTCCAGGGGTGAGTCTGCTGGGAAGCAGAAAG CACAGTCCACAGCCAGAGCCTGGGGAGGGTCCTGGACCCCCGCCCAGCCCCCTTCAGCCCAGGGAAAGAG AGGGCTCGTGCACGGGAAACTCAGCGCTCTGCCCCATCTCCCCCAGTGCCCCCTTTTTGTGTATTCCCTT ACCCCTCACCTTCTGACCGTGCTCCCTGCTCTTGCAGGCTGTGCCCGGTGGTGCCCTCAGGACTCCTCGT GTGTCAATGCCACCGCCTGTCGCTGCAATCCAGGGTTCAGCTCTTTTTCTGAGATCATCACCACCCCCAT GGAGACTTGTGACGGTACAGAGGCTTGAGGGCAGCGCAGGGGACATCTGCGATTATGAGGCATAGCCCAG TGCCAGTGGGGGACAGAGGTTGTTGTGAGGGGCCACAGCCTTACCTTCCAGACTATCATCATGGCCAGAG AAAAGAGAAAGAGGGCAGGTGTGGTGGCTCCCAGCACTTTGGGAGGCAGAGGCGGGTGGATCATTTGAGG CCAGGAATTCAAGACCAGCCTGGGCAACATGGTGAGACCCTGTCTCTACAAAAAATACACAAAATAGCCG CGCATGGTGGCACACATCTGGAGTCCCAGCTACTCAGAAGACTGAGGCGGGAGGATTGTTTGAGCCCAGG AGTTTGAGGTTGCAGTGAGCTATGATTGCACCATTGCACTCTAGCCTGGGCAACAGAGTGAGATCCTGTC TCAGGAAAAAGAAAGAAACAAAGAAAGAGGCCAGGTGTGGTGGCTCATGCCTGTAATCCCAGCACTTTGG GAGGGATTGTCTGGGCAACAGGAGCAAGACCCCATCTCAAAAAAAAAAAAAAAAGACAGAAAGAGAGAAG GAGAGAAAGAAAAGAGAAGAAAAGAAAAAGAGAGAGAGGGAGGGAGGGAAAGAAAAAGGAAAGGAGGGAG GGAGGGAGGGAGGAAAGAAGGAAGGAAGGAAGGGAGGAAAAAGGAAAAAATATGAAGGGGGAGAAGTAAG AAGTGAATAGGCATGGCTTCCTGGAGAGAGAGAAGCTGCGTGCTCAGGAATCTGGAGTCTGTGCCTCAGT TTACCGTTAAGACTGGGAAGGGGGTACATTCTGGCCATAGGTTTTTTTTTCTCCCTTATTTTTCTTTTTT CTTTTTTTTTTTGTTTTTTTTTTTTTTTTTTGAGATGGAGTCTCACTCTGTCGCTCAGGCTGAGGAGTGC AGTGGCGCGATCTCGGCTCACCGCAACCTCCACCTCCCAGGTTCAAGCGATTCTTCTGCCTCAGCCTCCC GAGTAGCTGGGATTACAGGCATGCACCACCATGCCCGGCTAGTTTTTGTATTTTTAGTAGAGATGGGGTT TTGCCCTGTTGGCAGGCTGGTCTCGAACTCCTGACCTCAAGTGATCTGCCGGCCTCGGCCTCCCAAAGTG CTGGGATTACAGGTATGAGCCACTGCACCCGGCTGCCATGGGGTTTAAATTTCTACAAAAAGACTGGGGA TGGGGATATGGCGCTTCCTGTCCATCCCCCACCTCAGCACTGTCCACTGTGTCCAAATCTAGGATGCCAG CCAGAGAACTGAGATCAAGTGTGGTCTTCAGGAATGGCCTGCTCCAGCTGCAGCATAGCCTGTGCAGTGC AACTTGAGATCCTTTCCAAAAGATACTAAGCAGCAGGCCCCATGTTGGGGAGTCAGGACGGGGGTCTCTG CCCTGACGGAACTCACATCTTTGGGAGATGACCTCCTGTCCTGTTGTGTTCCAGACATCAACGAGTGTGC AACACTGTCGAAAGTGTCATGCGGAAAATTCTCGGACTGCTGGAACACAGAGGGGAGCTACGACTGCGTG TGCAGCCCAGGATATGAGCCTGTTTCTGGGGCAAAAACATTCAAGAATGAGAGCGAGAACACGTGTCAAG GTAAGAACCACCCCACATCCTCCATCACCACTGTCCATGAGGTTTGGGGTCACCAGAGCTGTTTCTGCAG CATCCAGGGAGCAGGTACCCAAGTATAGGTTCAGTTCCTGGAGTCTGAGATGAGACAGGTGTGCATGTAC CTGTTTCGCCAGCCCAGAGAGGCAGCGTGGTGTGAAGGCCAAGGGAAAACTCTGGGACCAGCTACCTGGG CCCAAATCCTGGCTTTGCCACCTGTTACCTAAATGATATTACCAACATCACTACTGCTCTTAGAGCCTTG GTTTACCCATCTATAAAATGAGGATGATAATAATGGCCTACTCCTATGCCTTCTAGGAGGACCAAGGAAA GAATAGGTGCTTCTAACAGAGCCCAGCACACAGCAGGTGCTACAGGCATCTTCATTCTTTAAACGTTATT ATGCACTGTAATCCCAGCACTTTGGGAGGCCGAGGCGGATGGATCACGGGGTCAGGAGATCGAGACTATC CTGGCCAACATGGTGAAACCCTGTCTCTACTAAAAGTACGAAAATTAGCCGGGTGCAGTGGTGGGTGCCT GTAATCCCAGCTACTCAGGAGGCTGAGGCAGGAGAATCACTTGAACCAGGGAGTCGGAGGTTTCAGTGAG CGGAGATCGTGCCACTGCACTCCAGCCTGGTGACAGTGCAAGACTCCATCTAAAATAAATAAATAAATAA ATAAATAAATAAATAAATAAGCAATGGATGCCTTTTAGGAGTTGTTCAGACAAGCTGGGCTCCTGTGGGG CCACACTCCTGGCTGGCACTCAGAAGAAATCTATTTCGACACAGCCTGCCTCATCCCTTGCCATGGTCTT ATTCCACACTGCAGCTCCAGCCCATGAGAAGCCAGGCCCCCATGCATCACTCTGCTTTGCCTTCTTTCCA GTGGGAACTAGTTGGGTGCGGGAGGGAGCTTCAGACATCTGCAAGGCCAGATGGGAACCAGTAGCAGGTG GACAGAGAAGGGAGATTAACACAGAGTACACAGAGTGGGAAGGGAAGGTGAGCAAAGGCAGGAAATGAAG AAGCTGAGGCTGAGCCAAGGGTCCCAGGGGACATTAAGGGGGTTCAGCAGACCTGCCCTGCTGTAAAGAG AAAAGTGGCAGTGCTCCTGTTCCCAGCCCTGTCCCTCCTGTTCTCCCTGCAGTCAGTAGAGCCTCCCTGG TGGGAGGAAAGTGAGGACAGAGCAGAGGTGTACGTTCTGAGAGCACGCAGCTGCATAGAGCCCAGCAGGG GGGCTGGTTCACAGGGGTCCCCTGTGAACAGATGCTACATTGCATGCACATATCTTGACCAAAGGACAGA GCCTTCCTGTGAATTAGAAAAAGGACTCTCCGGGCCAGGCACAGTGGCTCACGCCTGTAATCCCAGCACT TTGGGAGGCTGAGGCGGGTGGATCACCTGAGGTCAGGAGTTCAAGACCAGCCTGACAAACATAGTGAAAC CCCATCTCTACTAAAAAATACAAAAGTTAGCCAGGCATAGTGGTGAGCACCTGTAATCCCAGCTACTCAG GAGGCTGAGGCAGGAGAATCACCTGAACCCAGAGGCAGAGGTTGCAGTGAGCCAGGATCTCAATATTGCA CTCCAGCCTGGGTGACAGAGCAAGACTTTGTCTCAAAAAAAAAAAAAAAAAAAAAAAAAAAAGAAAAGAA AGAAACAAAAAGAAAAAGAGAAACAGAAAAGCCTCTCCTCTCCAGGTAGACACAGCCCCATTTGGGGAAT GGAGAGGTTTTCAGCCTCTGCTCACCGCCTTGGCTGGGCATCCATATGTAGTCAGCAACCTACACAACTG TACGTGGTGGTCATGGGTCCTTGTCAGTGAGAGCCTCTGTTTTAGTCTATGTTGTGTTGCTATAAAGGAA TACATGAGGCTGGATCATGTATAAAGAAAAGAAGCTGATTTGGCTCATGGTTCTGCAGGCCATGTAAGAC ACATGGCTCCAGCATCTGCTTCTGGTGAGGGCCTTAGGGAGCTTCCACTCTTGGCAGACGGGAAAGGGGA AGCTGGCATCCCATGGCAAGAGGAAGGAAGCAAGAGAGAAGGGAGGAGATACCAGGCTTTTTCCAACAAG CAGTTTTCATGGGAACTAACAAGGCGAGAACTCACTCATGACCCAAGGACAGCACCGAGATGTTCATGAG GGATCTGCACTCAGGACCCAAACACCTCCCACGAGGCTCCATCTCCAACACTGGGGATCACATTTCAACG TGAGATTTGGAGGGGACAAATATCCAAACTGTATCACCGCATGCACAGGTGGTGTTTGTAATTATTGAGG AGGTGTGTTGCCATGTTAGAAACAGGCCAGCACTGGCCGGGCGCAGTGGCTCATGCCTGTAATCCTAGCA CTTTGGGAGGCCGAGGTGGGCGGATCACAAGGTCAGGAGATCGAGACCATCCTGGCTAACATGGTGAAAC CCCGTCTCTACTACAAATACAAAAAAATTAGCTGGGCGTGGTGGTGGGTGCCTGTAGTCCCAGCTAATCA GAGACTGAGGCAGGAGAATGGCATGAACCTGGGAGGCGGAGCTTGCAGTAAGCCGAGATGGCACCACTGC ACTCCAGCCTGGACGACAGAGCGAGACTCCGTCTCCAAAAAAAAGAAAAGAAAAGGAGAGAAGAGAAGAG AGAAAAGAAAAGAGGCCAACACTGAGAGAGGGGGCTTAGTTGGTGGATAGGCAGAGACACAGATGAGCAG GAATGAAGTGGGAGAGGGCTCTCATGGACCCCATCCGCACAGAACAGCCTGCTAGTTCGGAGCAGGACCT GCCTCTTGAAGCCTCAGCTCAGGGAAGGGCTCCGTGGGGCCCTGTCTGCATCCCCCTCACCCCTCAGCAT TCTTGTTTTTTTTGTCTACCTGGTGTCACAGGCCTCATAGGCAGGAGACCTGGGGCCACTCCCTCAGCTC CTTGCCCCACTTACTCCAGGCATCACAGGGCCGTCCCAAACACCTGGTCACCTGTCGCTATCCAAACCTC AAGGTTCAGAACCCTCCACAGAGGGTCACTTCCAAAAGCTCAGAACATCCCCCTGCCCATCTGGCATCTT ATTTGTTTGTTTTGTTTTGTTTTATTAGTAAAGATGGGGGGGGGGTGTCTCACCATGTTGTCCAGGCTGG TCTCGAACTCCTGGCCTCAAGTGATCCGCCCGTCTCAGCCTCCCAAAGTGCTGGGATTGCAGGCGTGAGC CACCATGTCCGACCTGGTATCTTATTTGTACAAAACAAAAAGGGGGCTGGCACTCAGGATATTCGATGAG CTCACTGCGTGGTGCATTCCAGCGCAGTATCCATTCTCTCTGCTATCATCAGCGACTGCCAGGATGAGAC AGTCTCCACACAGCTCTGCTCACCTGGAACTCTCTCTGATCCCCCAGCTCATCATGGACCCAGGCACCCT GTCCAGAAATGAAGGAGGAGGGAGATTTGAAGGCAGGGCCACTGTCCCCAGTCGAACCTGCCACTGGCCA ACAACCCCACCTGGAGAAGATGCCTCGTGGAGCCCTGACCTCTTTTTCTGCCACTCAGTCTCTGTCCCTT TCACTGAGCGAGACTGCCCTGACCCTTCCTGATAGTTTTGGTCCACCTGAGTGCACTGGAGGATGTTGTG GCTAACGGTGGGCACAGTGACATGGACTCCCCATACTTGGAGAGTTGAGTGAATGGAAGAGGCACTAGTG TGGACCCAATGGCAAGAGAGGTTAACATCAGGGCCATTGCTCCAAGCCTGATCCTCTCCCAGGAGGTCAA CCCAGATCCCATGGGAGTGGAACCATTCAGAAATAGGTCTGTGTCCCTGGTGGAATGCCAAGAGATGAGA TGACAGATACACCTCCATCTTTGCCCATCAGCATCTGTGCCCCTCTCCAGCCACTAAACCTTCACCCAGG GCCAGTCTTTCTCAGCTATGGATTCTCACTACATCCTGCCATTTCCAAGACACCTTCCTGAGGATCAAAA AAAGGCACCTTCACAGCCCTGGGGCAACCCAGCTGTATGTCGGTCCCCACTGTCCACCCTGGCCTGGGAC CCACTGTCCAGGCAGCCCTGGCCTATGCAGCACCAGACTCATGCTCCCCTGGGGACTGGCTCAGGGGCCA GTCTTGGTCCATGTGCCTGGAGCTGGGCTCTCAGAGCTAGTGTGGATGGGAGAGCACCATCCTGCTAGGG CCACCGGGATTCTGATAACTCAGCCAGTGAGCAGCTGTCCTGGGGAAATCAAGGCCACCAGAAGCAGATG CTGGAGCCATGCATCTTACACAGCCTGCAGAACCATGAGCCAAATAAACTTCTTTTCTTGCAACACAAAA CGGACTAAACCAGAGGGTTCTCACTGAGAAGGACCCATGACCACCATGTACAGTTGCATAGGCTGCTGAC TGCATATGGATGCCCAGCCAAGAGGGTGAGCAGAAGCTGAAAACCTGCCTACTCTCCATTCCTCAAATAT GACTGTGACTACTTGAAGAGGAGATGCCTTTTTCCAATTCACAGGAAGGCTCTGTCCTTTGGCCCAGGAC TGCCACTGGGAACCAAGACAGGTCCTACCTGGGCAAGCAGCATGCCCCCTGCACTTGGTCCCAAGAAAAT CCCACCCTGACCTTTGTCCTGAATCCTCAGGCTGGGACATCCCTGCCCCAGCTGGGAGCTGCCCAGGGAG GCAGGGGAGATGGGGAGGACTCAGATGCTTCCAGCCATATCTGAACAACAGAAGGCCTGGGCCACCATGC TCACCATGGGACACCTCTGTGTGGATTAGAAAAGGGAGGAGATGGCCAGGCATGGTGGCTCATGCCTGCA ATCCCAGCACTTTGGGAGGCCGAGGTGGGCTGATCACGAGGTCAGGAGATCAAGACCATCCTGGCTAACA CGGTGAAACCCCGTCTAATACTAAAAATACAAAAAATTAGCCGGGCATGGTGGCGGGCGCCTGTAGTCCC AGCTATTCGGGAGGCTGAGGCAGGAGAATGGCGTGAACCCGGGAGGCGGAGGTTGCAGTGAGTTGAGATC AGGCAATTGCACTCCAGCCTGGGTGACAGAGCAAGACTCCGTCTCAAAAAAAAAAAAAAGAAAGAAAGAA AAGAAAAGGGAGCAGACAGTGGTGCCTTCCAGATGCTTCCGTGCCAGGAAACATGGCGGACCCTCAGCCC CGAAAGCTGCTCACAGCTACGGGCATTCTTCACCCTCTCCTTCCTCTTGCAGATGTGGACGAATGTCAGC AGAACCCAAGGCTCTGTAAAAGCTACGGCACCTGCGTCAACACCCTCGGCAGCTACACGTGCCAGTGCCT GCCTGGCTTCAAGCTCAAACCTGAGGACCCGAAGCTCTGCACAGGTAGAGGCCCCAGGAAGACGCTGTGA GGCTGGACGGGAGCTGGGGATGGAGCTGAGTCAGGTCCTCCAAAGCAGCCGAGGAGGAGGGAGAAGATCC GCAGGTTCCCACAAGGTCAAGGACCTGCTAAGCCCCTGCCTAAGGATTCACCTCCCAGGAAGGACTCGCC ACACGGCAGGGAGGCGGCAGGGCCTTCAGGGCTTGGAGTGCCCTTGTGGGCCCCCAGACCTCACCCCTTC CTCATCTGTCACGTGCGAGTGGAAGAAGGTGTCACTTCCAGTTCTAAGAAGGGGAACCTCCACCATAGTG AGGGAGGGAGCAGGAGAGAGATTTTAGTCAGAAAGTCCAGCTCCCATGACTCAGTTTCCCTTTTGGTTTC ACCTCCTATAAACTGGGGACCATGGTCCCTGCTGTGCCCATCTGGTAGGAAGACCATCATAAGATGCTTT GAAGGTGAAGTTGAAGGTCACCAAGTGGTGGCACGTGCAATAATGGTCTTCAGTCTGAGGCAATGAAAAG ATAGGGGAGTGGGTGCAGTGAGTGGAGGGCAGGCCCAGGCTGGTCAGGGACAGGGTTTGACCCTCTGGCT TTGTCCTCAGATGTGAATGAATGCACCTCCGGACAAAACCCATGCCACAGCTCCACCCACTGCCTCAACA ACGTGGGCAGCTATCAGTGCCGCTGCCGCCCGGGCTGGCAACCGATTCCGGGGTCCCCCAATGGCCCAAA CAATACCGTCTGTGAAGGTCGAGAGCTCAGATCCCACGTTCCCAGAGACCCACAAACATCTGATCACATG TTCAACGGCGCCCACACAAACCAAGCAGAATGAGCGCTGGAGGCGCCCGACTGTGTCAGGCGTTCATTCT TCTGAGGCTAGATGAGAAAAGAGCAAGGGTCCTGCGGAAGGAGCTGGGGTACTGAGGGGGGAGGCTCAGG GGGACCCCAGGCAACAGCTGATGACTCACTGGGAGGAAGGCGTTTCACCATATTCATAACCTGCTCATCT GCACGGGGCCCACCTGCTGTGCCCAGGCCTCTCCACGCTTCCATAACCCAGCGTCCACCTCTCCAAGGGG GGCACTAATGCCGGGAGGAACGAGCTGGGGGCACAGACAGGAGACAGGACCCTCTCCAGGCTGGGACAGG ACCTGACCCCCTTCTTCCTGTCCTCAGATGTGGACGAGTGCAGCTCCGGGCAGCATCAGTGTGACAGCTC CACCGTCTGCTTCAACACCGTGGGTTCATACAGCTGCCGCTGCCGCCCAGGCTGGAAGCCCAGACACGGA ATCCCGAATAACCAAAAGGACACTGTCTGTGAAGGTATGACCTGGCCCTAGAAGCTCCCCACCCCCAGCA CACACACTGACACGCTCCCGCCTAATGAGCCGCTTGTCTTGTTCCCTACAGATATGACTTTCTCCACCTG GACCCCGCCCCCTGGAGTCCACAGCCAGGTGAGTGGCCCCCACAGGGACGAGGCGGCGGGAACTCCATCC ACACAGCACTGCATCCGTCTCCTTGTTCTAAACTTCCCACCCGCCGTCCAGGCTCTCTGACCCCCACATC TCCTCTCTCTGCAGACGCTTTCCCGATTCTTCGACAAAGTCCAGGACCTGGGCAGAGACTACAAGCCAGG CTTGGCCAATAACACCATCCAGGTAAGGACAGGACCCAGGGCAAGGGGGCGAGGCAGGAAGGTGGCTGCA TCCCACAGAGGCCTGGGGCAGTTTGGGTCTGGGAGGGGACAGGACCCAATGCAGTGGGTGCTGGTCTGAC TCCCAGCATCTTTCAGGGCTGGTGGAAGCTAATGGACACCCAAGTGCACTTAATATCTTTCCTCTTGCTT TCCCTGGACTTTGGGTTTAGGTCAGCATTGATATTGCACAGAAGTGTTGGCCTCTGAGGGGCCATAACCA GAGTCAAGGATCACCTAGGGAATCCCATCCAATGTCATTTTTTTTTTAGACGGAGTCTCACTCTGTCGCC AGTCCAGAGTTTAGTGGCCCGATCTCAGCTCACTGCAGTCTCTGCCTCCCGGGTTCAAGCGATTCTCCTC CCTCAGCCCCCTGAGTAGCTAGGATTACAGGCACACGCCACCACGCCCGGCTAATTTTTGTATTTTTAGT AGAGACGGGGTTTCACCATGTTGGCCAGGATGGTCTCGATCTCCTGACCTTGTGACCCACCCTCATCGGC CTCCCAAAGTGCTGGAATTAGAGGCGTGAGCCACCGCGCCCAGCCGCCAATGCCATCTTCATCCCCCAGA TAGACAGTCTCTAGGATCTGTTCCCTGGGGCTGAGCGGTTGGAGTCTTCATGCGGGCCCTCTGGCCCATG GCTCACTAGGTCTGTGTCCACATCCCTCCAGAGCATCTTACAGGCGCTGGATGAGCTGCTGGAGGCCCCT GGGGACCTGGAGACCCTGCCCCGCTTACAGCAGCACTGTGTGGCCAGTCACCTGCTGGATGGCCTAGAGG ATGTCCTCAGAGGCCTGAGCAAGAACCTTTCCAATGGGCTGTTGAACTTCAGTTATCCTGCAGGCACAGG TAGGTCCCTGGGTCTGCCCCAGACTCCAGCTCTGCATGTTTTCTGTCTCCTTCCTTTCCCAGTCCCACCA GAGCCAAGTGACCACACCTGTATATCAGTGTTACTCTCATCGACAAACTAAAATAAGAGATTAAAAAGAA ATACATACCAGTCTGGACAAGATGGCAATACTCAATCTCTACAAAAAAAAAAAAATCTTTTAAATTAGAC AGATGTGATGCACAACTGTGGCCCCAGCTAGTTGGGAGGCTGAGGTGAGAGGATCGCTTGAGCCCAGGA GGTTGAGTCTGCAGTGACTATGATTGCATCACTGTACTCCAGCCTGGTTCACAGGGCAAGACCCCATCT CAAAAAGAGAAGACGAAGAGGAGGAGGAGAGAAGAGAGAGAGAGAGAGAGGAGGTGAAGAGA GGAGGAGGATAAAGAGGAAAAAGGAGGGAAGAGGGCAGGGGGAGGACAGAGGAGGAGACAGGA GGAGAGGAGGGGAGGAGGAAGGAAGGAAGGAAAGAGGATGGGAGGAGGACAAGAGGAGGAGAGCGGGAGG AGGAGGACAGGATGGGAGGAGGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAGAATGGGA AAGAGAGGAGGAGGAAAAGGAGGAGGAGGAGGAGGAGGAGAGAGAAGGAGGAGGAATGGAGGAGG AGAGGAGGCAGAGGAGATAGGAGGAGGAGGAAGGAAGGAAGGAAGGAAGGAAGGAGGAGGAGGAGGAGG AGACGAGAAAGAGCGGAGGAGAGGAGGAGGAGGAGGAGGAGAGAGACACAGGAGGAGGAGGAGGTAGAGGA GAAGGAAGAATAGAAAAGAGGAAAGGAAAGGAAAAAGAAATGCATAAATAACATGCATTTTCATTGTAAT ATGAATTATCAGCAGTAAATAATTATGATGAAGAATTTAAAATAACAAAATAGAAATCACTTCCAAAGAG AACAATTTATTTAATTTGGAATACTATAAATTAAATATCATTGATCTCTAAGTATGATTTCTTCTTGTT TGTTTTTGTTTGTTTTGTTTTGTTTTGTTTTGTTTTTTTGAGACAGAGTCTCACTCTGTCGCCCAGGCTG GAGAGCAGTGACGTGATCTCGGCTCACTGCAAGCTCCGCCTCCTGGGTTGACAGCATTCTCCTGTCTCAG CCTCCCGAGTAGCTGGGACTACAGGTGCCCGCCACCACGCCCAGCTAATTTTTTGTATTTTTAGTAGAGA CGGGGTTGCACCGTGTTAGCCAGGATGGTCTCTCGATCTCCTGACCTTGTGCATAATCCACTTGCCTCGG CTTCCCAAAGTGCTGGGATTACAGGCGTGAGCCACCACGCCCGGCCTGCATAATAATTTTTAAAGCATGA AAATGCATATAGCATTAATTTTATTAAGATAAAATTCACATAACATCAAGTTCACCACTTTAAAGTGTCT GTTTCACGTGGGGCGTGCCTGTAATGCCAGGCGCTCACACCTGTAATCCTAGCACTTTGGGAGGCTGAAA TGGGTGGATCACTTGAGGTCAGGAGTTCAAATCCAGCCTGGCCAACATAGTGAAACCCCATCTCTACTGA AAATACAAAAATTAGACAGGCGTGGTGGTGCACACCTGTAATCCTAGCTACTCAAGAGGCTGAGGCAGGA GAATCACTTGAGCTTGGGAGGCAGAGGTTGCAGTGAGCCAAGTTTGCACCACTGCACTCCAGACTACGTG AGACAGAGTGCCACTCCATCTCAAACAAAAAAATTAAAATTAAAAATAAAAATAAGAAAAAATTTTAAAA GCGTGTGTTTCGGTAGAATCTAGTATATTCAGAAAGTTATACAACCGTCACCTCTATTTCTAAAATGCTT GCATCAGCCCAAAAGTGAACACTGCACATGGTAAGCAACCACTCCTCATTCCTCCCTTCCTCCAACCCAT GGCAGCCGCTAATCTGCTTTCTGTTTCCATGGATTTTCCTAATCAGGATATCTCACATAAATGGAATCAT ACAATATGTGACCATCTAGTCTAGCTCCTTTCACCTAGCATAATGGTTTTGAGGTTCATGCATGCCATAG CATTTATCAATATTTCATTCTTTTTCATGGCTGAATACTATTCTATTGTATGAATACATCACTTTTTTTT TTTTTATCCATCCATCCATTGATGGACATTTGGGTTGTTTCCACCTTTTGGCTGTTGACAAAAAGAGTCA AACTCTGTAAAATATTTGAAGAGATTTATTGTGAGCTAAATATGAATGTATGACTGACCATGGCCCGTGA CACAGCCCTCAGGAGCCCCTGAGAACATATTCCCAAGTGAAGTGGTGTTGTTTTCTGGGGTAATACCTGA GGTTCATTGCCTCATGCCAAGAAAATTAAGGACACAGACACACACCAGGAGTGAGTTTATGAGTGGAGGT TTAATAGGCAAAAGAAAGAGAAAGGAGAACAACTCTCTCTCTTGTGAGAGAAAGGAATGCCCAAATGGGA CTTCCCACCCATGCCAGAGGGCACCGGATTTTATAGACAGGCTTGAGGAGGCAGTGTCTAATTTACATAG GGCCCAAAGATTGGTTGGAACAGGTGTGACGTTTACATAAGGTGCAGGGAAGCTGGCTGCCCCACCCTAA TCTTATTATGCAAATGGAGTCTTCGCCAGCTCCTTACTGTATACGTGGTTGGCAAAGAGATGGGAAGATG GAGCCGCCATTTTCAACATGCCTAGTCCCAGGTGGCCTTTTCCTATTGGCACAGCTGCCAGCATTCACCT GTGCAGGCTTCCAGCTTGCTTGCCTATGTCTGCAGCTCGATTTTACAGGCTGCTCTTTGTTAGAAAAAAA AAAAATGATATAGGGGCTGATTTTCATTAAAAGGAAAGACTTACCCAGGACTTCCTTACCCACACTCTCT GCCTAAATAATTCCTTTTTAACTTCTATATCACAAGGTAGTCGTGGTACAGCTTGTTGTGATTGTTGTTG TTGTTGTCTTCTGTTGGAGACAGAGTCTTGCTCTGTCAACCAGGCTGGAGTGCAGTGGTGCGATCCTGGC TCACTGTAGACTCCACCTCCCGGGTTCAAGCGATTCTCCTGCCTCAGCCTCCCAAGTAGCTGGGACTACA GGCGCCCGCCACCACGCCCGGCTAATTATTTTGTGTGTTTTCAGTAAAGACAGGGTTTCGCCATGTTAAC CAGGATGGTCTTGATGTCCTGACCTCGTGATCCGCCTGCCTCGGCCTCCCAAAGTGCTGGGATTACAGGC GTGAGACACCACGCCTGCCCCCGCAACTTGGTTTTATACATTTTAGGGGGACATGAGACATCAATCAAAT ATGTTTAAGCTATACATTGGTTCAGTCCAGAAATGCAGGACAATTTGAAGTGGAGGTGTGGGGCGGGTGC TTCCAGATTATAGGTAGATTTAAGCTTTTCTGATTGGCAGTTGGTTGAAGGAGTTATTATCCATAGAAAA GAATGTCTGGAATACGATAAGGGGCTGTAAAGGCTGAAGTTGTATCATCCAGATGAACTCTTCAGGTATC AGGCTTCAGAGAGAATAGATGGTAAATGTTCCTTATTAAGCTTCAGGTCTGTGTTATGTTAAATGCTGGT CAGCTTTTCCTCAACTCCAAAAGGGAGGAGGGCATGACACATGTCAGACCCCCCACTTCCCACCATAGCC TGAACTAGTCTTTCCAGTTAAATTTAGAGTGCCCTGGCCAAAAAGGAAGTCCATTCAGATGGTTTTGAGG GGGGCGGGAGAGGACTTCGAATTTTTTTTTTTTGTTTAGATGGCTGGTGTGAATAGTGCTCTTATGAATA TTAGTGGACACATTTTGGGTTGAATAACTGTCTTCAGTTCTTTGGGGTATATACTAAGGAGTGGAATTGC TGGGTGATATGGTAATTCTATGTTCAGCTTTTGGAGGAACCACCAAACTTTTTCCAAAGCAGCTGCATCA TTCTAATTCCCCACCAGAAACATACACGGGTTATCCTCTCCACACCTTCAGCAAAACTTGTTACTTCCGT TTTCTTTTGTTCTGTTTTTAATTGCTTTTATTGATACATAAATCTTACATATTTATGGGTACATGTGATA CTTTGTTGCTTGCCTAGAATGTGTAACGATCAAGTCAGGGTATCTCAGGTATCCACCACTTTGAGTACTT ATCATTTGTATGTGTTGGGAACAATTGAAGTCCTCTTTTGTAACTACTTTGAAATACATCATACAGTCTT GTTAATTATAGTCACTCTGCTCTGCTGTCAAACAATAGAACTTGGGCCGGGCACGGTGGCTCACGCCTGT AATCCCAGCACTTTGGGAGGCCGAGGTGGGAGGATCACGAGGTCAGGAGATCGACACCATCCTGGCTAAC AGGGTGAAACCCCATCTCTACTAAAAATACAAAAAATTAGCCAGGCGTGGTGGCGGGTGCCTGTAGTCCC AGCTACTCGGGAGACTGAGGCAGGAGAATGGCGTGAACCTGGGAGGCGAAGCTTGCAGTGAGCCAAGATG GTGCCACTGCACTCCAGCCTGGGTGACAGAGTGAGACTCCGTCTCAAAAAAAAAAAAAAAAAAAAATCTG CTTAATAAGCGGAGAGGTGGCTGGCTTGGTGGCTCACACCCGTAATCCTAGCACTTTGGGAGGTCAAGGC AGGCAGATCACCTGACATCAGGAGTTTGAGCCCAGCCTGGCCAATATGGTGAAACCCTGGCTATTAAAAA TACAAAAATTAGCTGGGCTTAGTGACACGTGCCTGTAATCCCAGCTACCTGGGAAGCTGAGGCAGGAGAT TCGCTGGAACCCAGGAGGCAGAGGCTGCAGTGAGCTGAGATCGTGCCACTGCACTCCAGCCTGGGTGACA GAGTGAGACTCCGTCTAAAAAAAAAAAATATATATATATATATGTATAACTTATACTTTTTGTCTAACTG TATGTCTCTACCTGTTAACTCACCTCTCTTCATCCCAGTGATACAGGAGTGAAAAAGAAATTATTTAGGC AGTTAGCAAGGGTCAGAGAGTCCTCAGTAAGGTTTCCCTTTTAATAAAAACCAGCCCCCAAATCATTTCT TTTCTAATAACGAGCAGCCTGAAAATTCGAGCTGCAGACATAGAAAAGCAAGCTGGAAGCTTGCACGGGT GAATGCCGGCAGCTGTGCCAATAGGAAAAGGTTACCTGGGGGCCCGACATGTTCAACGTGGGGGCTCCCT CTTCCTTTATCTTTGTCAACCACGTGTACAGTACAGAAGCAGGCAACGTGGAGCCAGCCAGGTAGAGAAT CCATTTGCATAATAAAAGATTAGGCTGGGGCAGCCAGTTTCTTCATGCTGTATGTAAATGGCTCACCTGT TCAGACCAATCTTTAAGCCTTATGTAAATCGGACACCGCCTACCCAAGCTCATCTATAAAACTTCGTGCA TTTCACCACGCAACCGGAAGACCCACTTGGGAGCCCCTGTCTCTCTGCAGGAGAGGGAGTTATTCTCTTT TTCCTTTTTTTTTTTTTGAGACGGAGTCGTGCTGTGTCCCGCAGACTGCAGTGCAGGGGCGCGATCTCGG CTCACTGCAAGCTCCGCCTCCCGGGGTCACGCCATTCTCCTGCCTCAGCCTCCGGAGTAGCTGGGACTAC AGGCGCCCACCACCATGCCCAGCTAATTTTTTTGTATTTTTAGTAGAGACGGGGTTTCACCGTGTTATCC AGGTTGGTCTCGATCTCCTGACCTCGTGATCCTCCCGCCTCAGCCTCCCAAAGTGCTGGGATTACAGGCG TGAGCCACTGCGCCCAGCCAGAGCTATTCTTTTTTTCTTTCGCCTATTAAATCTCCACTCTTAAACTAAC TTCTTATGTGTCCACATCCTCAATTTCCCTGGTGTGAAACAGTGAAACTTGGGTATTTACACCACTTCAC TAGTGAGATGATACCTCATTGTGGTTTTGATTTGCCCTTCCCTGATAACTAATGATGTTTAACATCTTTT CATGTGTTTGCTCACCATTTGCATGTCATTTTTGGAGAAATGTCTATTCAAATCCTTTGCCTATTTTTAA TTGTGCTGTTTGTCTTTTGGTTGTCCAGTTGTAAGCATTCTTTAAACATTCTGATAGTAGATCATCTGAG ATGAATGATTTGCAAAATATTTTCTCCCGTTCTGTAGATTGTCTTTTCATTTTCTTGTCTTTTCACTTTA ATGAATAGAAGTTTTAAATCTTGAAGACATTTATTTTATCTAATTTTTCTTTTGTTGCATGTGCAATTGG TGTCATATTTAAGAATTCATTGACAGGGGGTGGTGGCTCACGCCTGTAATCCCAGCACTTTGGAAGGCCA ATGTGGGCGGATCACCTGAGGTCAGGAGTTTGAGACCAGCCTGGCCAACAAGGCGAAACCCTGTCCTTAC TAAAAATAAAAATAAAAATAATTAACCAGATGTGTTGGCACATGACTGTAATCCTAGCTACTCAGGAGGC TGAGGCAGGAGAATTGCTTGAACCTGGGACACGGAGGTTGCAGTGAGCTGAGATTGTGCCACTACACTCC AGCCTGGATGACAGAGCAAGACTCTGTCCCAAAAATAAAAATAAAAAAAGAATTCATTACCGCATCCAAG ACCAGGAAGATTTACCCCTGTTTCCTCCTAAGTGTTCTAGTTTTAGCTTCTAAGCTTAGGTCACTGGCTT ATTTTGAGTTAATTTTTGTTTGTTGTGTGAGGGTAGGGTTAAACTTCCTTATTTTCATGTTACTATCCA GTAATCCCAGCACTATTTGTTGAAGAGATTATTCTTTCCCCCATGTGTATAGTCTTGGCACCCCTTGTTGA GAATCAACTGACCATAAGATGGATGGGTTTATTCCTGGACTCTCAATTCTATTCTGTTGTGTCCTTATGC CAGTATTACTGTCAGTTTGTAGTAAGTTTTGGAATCAGGTAGTGTGAATTCTCCAACCTTCTTGTTCTTC TTCAAGATTGTTTTTGGGTATTTGGATTCCTTACAACACCATATGAATTTTAAGATGAACTTTTCTTATT CTGCAAAGTAATAATAGGCTAAAATATTGATAGGGATTGCATTGAGTCTCTAGACCACTTTGGGGGAGTA TTGTTATCTTAACAATAGAAAGTCTTCCAATCCATGAACATAGGCTGTCTTTCCATTTAGAGAGGGCCTT TTATTTCTTTCAAACACTTTTGTAGGCCAGAAGCAGTGGCTCACGCCCAATATTGTCAGAGGCCACCAAG GTGGGAGAATTGCTTGAGCCCAGGAGTTCAAGCCCAGCCTTGGAAATATACTGAAACATGATCTCCACAA AAAATTTAAAAATTAGCTGGCTGTGGTAGTGAGCACCTGTAGTGCCAGCTACTTGGGAGGCTGAGGTGGA AGGATTGGTTGAGCCCAGGAGACTGCAGTGAGCTATGATCATGCCACTGCACTCTAGCCTGACCAACAGG TGAGAATTTGTCTCAAAAAAAAAAAAAATACTCAAAAAACTTTCCTATAAGTCTTGAGTCTCCTTCGTAAA ATGTATTCCTAACATTTTTGCTGAAAAATTTTTGCATCTATGTCCATAAGAATATTGGTCTATAATTTTA TTTTCTTGTGATGTCTTCATCTGGTTTTGTTATCAGAGCAATATTAGACTCATAAAATTACTTAAATAGT GTTTCCTCCCTCTTCCATATTTTTAAAAAGGTAAAAGATTCATGTTCATTCTTTCTTAAGCTATTGGTAGA ACTCACCAGTGAAGTCATCTGGTCTGGGCTGTTAGTTTTGAGAGGTTTTGATTACTGATTTAATCTGTTA TAGGTCTGTTCAGATCTTCTATTTCTTCTTGAGTTGATTTCAGTGGTTTGTGTGTCACTAGGAATGTGTC CATTTCATCTAGGTTACCTACTTTGTAGATATACAGTTGCTCACACTATTCTCTTCCGATATTTTGTAAC TTCAGACTATATATGTATATAGTATATATATTAAGTCATAGTATATCAACCACTGCAGGCACGATTCCTA ACCTAACCCCCCTTATGTAATTCCCACAACTCATAGAATTGTCCCTGGAGGTGCAGAAGCAAGTAGACAG GAGTGTCACCTTGAGACAGAATCAGGCAGTGATGCAGCTCGACTGGAATCAGGCACAGAAATCTGGTGAC CCAGGTAATGGCTGAGATGGGGAGGTGGTCAGTGAAGCTTCATGGTGGGAAAAGATGCCCAAGAATGTGA AAAATAAGAAATTTAAAGATTAAAGGGAAGTGTGGGATGTTTGGAGTGGGGCTGTAAGAGGAGAGTCCAG CTGAGGAGGAACGGACCTTGTTGGGGACCCTGAGATATTAACCGTGGCCTTGTCCTGCAGGATGTGATAC ACATAGAACTAAAGATTGAAGGAAAGTGTGGGAGGGGTGAAGCCAAAGGAAGGTAGCCCATCTGAGTAGA AAGTTCTGTGCAGGGCAGAGGAACTCTGGTGTGGACCCAGAGATTCAACCTACTCCAAGATTCAACCTTG GCCTCGCCCTGCAGGCCCTTCTGTGGTGGGCCTTGTCTCCATTCCAGGGATGGGCAAGTTGCTGGCTGAG GCCCCTCTGGTCCTGGAACCTGAGAAGCAGATGCTTCTGCATGAGACACACCAGGGCTTGCTGCAGGACG GCTCCCCCATCCTGCTCTCAGATGTGATCTCTGCCTTTCTGAGCAACAACGACACCCAAAACCTCAGCTC CCCAGTTACCTTCACCTTCTCCCACCGTGTGAGTGCTGGTGGAGTTGGTTGGTGGGTGAATAGTCTGAGT CCGGGCATAGCCTTGCTGCTCAGCTCAGCCCTGGGGCTCAGGGGTCTCTGTTATGGGTACATTATCTCCC CAGGAAAGTCAGTCCTTCCCAAGCCGGCTTTGGGTAAGCATTTCTGAGCATCCGACCCACCAGCTCACAC CTGTGTTCTTTTTTTTATTGAGACAGAGTCTCGCTGTGTCAGCCAGGCTGGAGTGCAGTGGCACGATCTC AGCTCACTGCAGTCTCCGCCTCCCAGGTTCGAGCGATTCTCCTGCCTCAGCCTCCCTAATAGCTGGGACT ACAGGTGTGTGCTACCACGCCCAGCTAATTTTTGTATTTTTTTTTTTTTTTTTTTTTTTTAGTAGAGATG GGGTTTCACCATGTTGGCCAGGCTGGTCTCAAACTCCTGACCTCAAGTGATCCACCTGCTTCAGCCTCCC AAAGTGCTGGGATTACAGGCGTGAGCCACCATGCCCAGCCCACACCTGTGTTCTGTTCCTGCAGTCAGTG ATCCCGAGACAGAAGGTGCTCTGTGTCTTCTGGGAGCATGGCCAGAATGGATGTGGTCACTGGGCCACCA CAGGCTGCAGCACAATAGGCACCAGAGACACCAGCACCATCTGCCGTTGCACCCACCTGAGCAGCTTTGC CGTCCTCATGGCCCACTACGATGTGCAGGTGAGACCCTTAGGAGGGGATGCACTCTGCATTTATTGCCGT GTAACAAATCCCCAGAGACGTAGCAGCCTTTTAAAAAAATATATGCATTATCTCACAGCTCTGGGTCAGC AAGCTGGCATAGCAAGATGGCTTTTCTGCTCAGGGTCTTACAAGACTGAAATCATCATGTCACCCCACCC CCAGGGCTGCCATTTCATCTGAGACTTGAGGTCCTTCTCCAAGCTCCCTGGTTGCTGGCAGAATTCAGTT TCTTGTGATTGTAGGACTGAAGTCTCACTTTCTTAGCTGTCAGGAAGGCATCACATTCAGTTCCTAGAGG CCACTCATATCCCTTCTCACATGACCCTCTGGCAAACTTCTAGCATTCCTATCTGGAGGACAAGGAAAAA AATTTCCTCACGCAGAATCCCTCTTAAGCTTTGTATTAGTCGAGGTTCTCTAGAGGGACAGAACTAATGG AATAGATAGATAGATAGATAGATAGATAGATAGATAGATAGATAGATAGATAATTTCTGCCTGCTTATAT TCTAGCCACGCTGGCAGCTGATTAGATGGTGCCCACCCAGATTAAGAGTGGATCTGCCTTTCCCAACCCA CTGACTCAAATATATACATATATAGAGATATATAGAGAGAGATCTATATATATAGATAGATATAGATATA TATAAAGGGGAGTTTATTAGGTATTAACTCACATGATCACAAGGTCCCACGATAGGCCGTCTGCAGGCTG AGGAGCAAGGAGAGCCAGTCCGAGTCCCAGAACTGAAGAACTTGGAGTCTGATGTTCAAGGGCAGGAAGC ATCCAGCACAGGAGAAAGATGTAGGCTGGGAGGCTAGGCCAGTCTCTCTTTTCACATTTTTCTGCCTGCT TATATTCTAGCCACGCTGGCAGCTGATTAGACAGTGCCCACCCAGATTAAGAGTGGATCTGCCTTTCCCA GTCCACTGACTCAAATGTTAATTTCCTTTGGCAACACCCTGATAGACACACCAGGATCAATACTTTGTA TCCTTCAATCCAATCAAGTTGACACCAGTATTAACCGTCACAAGTTTTGAATCTGATTCTCCAGGAAGA GCCCAGAGCTTATAAAGACTCATCTGATTAGGTCTGGCCCACCCAGGGTAAACTCTGCCTTTTTTTTTTT TTTAGACAGGGTCTCTGTCACCCAGGCTGGAGTGCAGTGGCACAGTCATAGCTCACTGCAGCCTTGACCT TCTGGGTTCAAGGGATCCTCCTGCCTCAGCCTCCTGAGTAGCTGGGACTACAGGCATGCACCACCATGCC CGGCTAATTTTTGTATTTTACTAGAGATGGGGTTTCACCAAGTTGGCCAGGCTGGTCTCGAACTCCTTA CCTCAAGTAATCCACCCGCTCGGCCTCCCCAAAGTGCTGGTATTACAGGCAGGAACCACCACACCTCGCC CACCTGACTTTCTTATCTCATCACTTAGAGTAGATCCAAGGGACATCATCATCATATCCTGCGGGAGAAA GGGCCAAACCACCCTTTTGTTTTCCATGACAGGAACATCTTATCAATATCCTCCCAGGCAGCAAGCCATA CCGCCCAGCCCCTCCCGCCAGACCTGTAATTACCCAGCCTGTAAGCGGCAGTGGGTTCTGGCACGAAG CTAGCTCCCCCCTCCACAAGTCTCCTGCTGGACATAAACCTGCATTGCTGTAGAGCTGCCAACTCTCTGT CTTTCTTTAACTCTCACTTTCCCTTCAAAACCTAACGGGCCCACACCTGGAATCTCTGTGCTTTGGGAGG CTGAAACGGGAGGATCACTTGAGCCCAGGAATTCGAGACCAGCCCTGTGCAACACAGAAAGACCCTGTCTC TACAAAAATAAAAAAAATTAGGACGGGTGTGGTGGCTCACGACTGTAATCCCAGCACTTTGGGAGGCC GAGGCGGGTGGATCATGAGGTCAGGAGTTCAAGACCAGCCCTGGCCAACATGGTGTGACCCTGTCTCTACT AAAAGTATAAAATTAGCCTGGCATGGAGGCACGTGCCTGTAATCCCAACTACTTGGGAGGCTGAGACAC GAGAATCGCTTGAACCGAGGAGACAGAGGCTGCAGTGAGCAGAGATCGCACCACTGCACTCCAGCCTGGG CGACAGAGCAAGACTGTCAAAAAAAAAATTAGCAGGGCATGGTGCCATACATGACTATAATGCACACCC GTTGTCACCTACATGACTACGCCTGTATTCCTAGCCACCCATGAGGCTCGGGTGACACCTTCGCAATTA AATTAAATTGGTGACATCTTCACAAATTTACCTGGCTCCCTCATTCTTGAATGTGCTATGCCAAAGTAAA AATTTTAAATGGTGGTGGGGTCATTACCCCAGAGCAGGAAAGCATCACTGTGAACTCTTCTTTTCCGGGT CCCAGGAGGAGGATCCCGTGCTGACTGTCATCACCTACATGGGGCTGAGCGTCTCTCTGCTGTGCCTCCT CCTGGCGGCCCTCACTTTTCTCCTGTGTAAAGCCATCCAGAACACCAGCACCTCACTGCATCTGCAGCTC TCGCTCTGCCTCTTCCTGGCCCACCTCCTCTTCCTCGTGGCAATTGATCAAACCGGACACAAGGTATTGA CAGCGGTGTTCCAGAGGGCTCCCTTCCTTGACGCAGGACACGCTGGCTCATGGAATGATGCGGCATTTGA TTCCTTAATATAATAGCCCGGCTGGGCGTGGTGGCTCATGCCTGTAATCCCAGCACTTTGGGAGACTGAG GTGGGCGGATCACCTCAGGTCGGGAGTTTGAGACCAGCCTGACCAACATGGAGAAACCCCGTCTCTACTA AAAATACAAAATTAGCCAGGCATGGTGGCGCATACCTGTAATCCCAGTTACTCAGGAGGCTGAGGCAAGA GAATCTCTTGAACCCAGGAGGTGGAGGTTGTGGTAAGCCGAGATCGCACCATTGCATGCCAGCCTGGGCA ACAAGAGTGAAATTCCGTCTCAAAAAAAAAAAAAAAAAAATATATATATATATATATATGTGTGTGTGTG TGTGTGTGTATACGTATATATACACGTATATATATATAGGCCCTTGAGTCCTGGGGAATATTAGAAGAGA ACACCACCAATTTCCCATTGCCTGACTCAACTACACAGCAAATATTCTCTATAATGCCATCAAATATCTC AGAATTCTCCCCAACAACATGGTACAGGAGAAGCCATGGGCCACACTCAAATCTGATCGCCGCTCTGGGC GTCTTTTACTTCTCCGCCAGGTGCTGTGCTCCATCATCGCCGGTACCTTGCACTATCTCTACCTGGCCAC CTTGACCTGGATGCTGCTGGAGGCCCTGTACCTCTTCCTCACTGCACGGAACCTGACGGTGGTCAACTAC TCAAGCATCAACAGATTCATGAAGAAGCTCATGTTCCCTGTGGGCTACGGAGTCCCAGCTGTGACAGTGG CCATTTCTGCAGCCTCCAGGCCTCACCTTTATGGAACACCTTCCCGGTTAGTGCAAATTCTCACAATCCT TATCTTCTCCGGCATAACCATGGCCATTGCTAGAACCTAGATAGCATTTTTATGACACCTGCTTTTAATT AGATCAGTAGGGATTTTAGGCTCTATTGGTTGGATTACACTGCAGCTCCTTATTCTTAGCTATTGAGTTT ATTTCAATTACATTTCAAGCTAGGCATTGTAGAAGAGGGTTTTCGGAAATTTTCTGAATGAAATAAAGGG CAAGCCTTTGGAAAGCACGAATAGGACAAAGACACTATGTCATAATCCCTAGGGCATCTTTCTGCTAGAA GATTATAATAATTAAGGAGGGTCAGGCTCAGTGGCTCGCACCTGGAATCCCTGTGCTTTGGGAGGCTGAC ACAGGAGGATCACTTGAGCCCAGGATCTTGAGACCAGCCTGGGCAACACAGCAAGACCCCTTCTTTACAA AATTTAAAAAATTAGCAGGGCCTACTGGCACACACCTGTAGTCCTAGCTACTCAGGAGGCTGAGGTGGGA GGATCGCTTGAGCCCAGGAGTTCGAAGTTGCAGTGAGCTATGATTGCACCACTGCACTCCAGCCTGAGCA AGAGAGAAAGACCTTGTCTCTAAAAAATAATAATAATAGTTAGGGATAATGCATTTAAATCTGTCTCACT CLASSAGAGATGAATCAAAGTGCTCAAGACAGCATTTTTTAAAAAATCTAACCTACTTTATCTAAAT CTTTTAGTACTCCAATTTTTCATTTGAAAGTGAAACTGATTTGATAACTAAAATTTGATTG TCATAGTTAAGATTTATGAGATGCTTACTTTGTGCCAGGTACTTTTCTAGCTAGTTTGTTTGTTTGTTTG TTTGCCAATTGCACTGGTTGAAACCTCCAGGACACTGTTGACTAGAACAGCGAACAATTGTCTTGCTC CTGTACTTAGGGAGAAAAGCATTGACTCTGTCCTCATTAATGTATGATGTTAGCTGAGGATTTTTCATAGA TACCCTTTTTCAGGTTAAAAATTCTTTTCTATTCCTCATGTGTTCGGTATTTTGATGAAGAAAGAGTG TTTGATTTTGTCAAATGCTTTTCCGAAGCATAATTTTTAATTATATGGGGTAAAAATTATATGATCAC ATAATTAGATAATCATATATATATATATATTAGATTATTTATTATTATTTATATATATATA ATCTATGTATTATTATTTATATATATATATATTTATATATATACTATTATTATTATTATG WINDOWNGATTACHTAGTACHTAGTACHTAGTACTATTAGTAGTACTATTACKGTACHTAGTACHTAGTA WINDOWNGROUNDGATTATTATGATTACTGATTATTATTACTGATTACTGATTACT PATCHTATTATTATTTATTATGTATTACTATATTATTATTACTATTACTATATTATTATGTATTACT ATGAATAATTACATAACATAGCTATAAATTATGTAACTATGAATAATTAACATAACATAGCTATAAT AATTAGTGACTATGAATAACATAGCTATAAATAATTTGTAACTATGAATAATTACATAACACAG CTATAAATAATTTGTACTATGAATAATTACATAACATAGCTATAAATAATTTGTACTATGAATAAT TACATAACATAGCTATAATAATTTGTACTATAATAATTACATAACATAGCTATAATAATTATTATA ACTATAAATAATTACATAACATAGCTATAAATAATTTGTACTAAATAATTATAACATAGCTATA AATAATTATAACTATAATATACATAACATAACTATAATATTATATAACCATAAAATAATTATTAT TACATACTATAATATATATATATATAATATATATATATATATATATATAACT FATHERTATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAATTATATATATATATATATATATATATATAATTATATATATATATATAATTATATAATTATATATATATATATATATATATATAATTATATATATATATATAATTATATATATATATATATATATATATAATATTATATATAATATATATATAATAATTATATATAATATAATAATATAATATATATATATAATATAAATAATAATATATATAATATAATAATAATAATAATAATAAATAAaaaaaaaaaaaaaaaaaaaaaaaaaaa Data thanucay AATATGATCACATTAAATTGATAATCATATATTATCATATCATTATTATATTATTATTATATATAT CATATAATTACATATAATTATAAGCATAATTATTATGCTTCCTTAATTCTTTATTTTTAATACAC ATTAATAGAATAAGGATTAAATTTATGATCACATTAATTATTATGGTTATAATTTATGGTT ATATATGATTAATTATTACCTTTTGAGTCCTGGGGAATATTATATGGTTATATATGGCTAATTTATATGGT TATATAATTATAATGTGATCTTATAATTATATGATTACATAATTATATAACTATAATGTGATCATATAAC TTTAATCCTTATTCTATTAATATGTGTATTACATTGATGGATGATTAGATGTTAAACTAGCCTTACATT CCTGGGATAATTACCACCTGGTCAGGGTATATAATCCTTTTTAATATGTTGCAGGACTTGGTCGGCTAGTA TTTTTTTGAGGATTTTTGCATTCATTTCATAAGGGATTTTGGTCTGCAGTTTTCTTTTCTTGTGATGTC TATTTCTGGGTTGGTGTCAGAGTAACACTGGCCTCGTAGAAGATTTGGAAAGTGCTTCCTCTTTTCCG TTTTTGGTTTTGTGAACTATTAACACTAATCCTCCAAAAGTTAACCATAGAGCTACCATATGACCCAAGGT ATATACCCAAAACAAATGAAAACGATGTCCACAAAAAAACTTGTACATGATGCAGCCATAAAGAATAAGG AGATCGGCCGGGTGCGGTGGCTCACACCTGGAATCCCAGCACTTTGGGAGGCCGAGGCGAGCAGATCACA AGGTCAAGAGATTGAGACCATCCTGGCCAACATGGTGAAACCCCGTCTCTACTAAAAATACAAAAGTTAG CTGGGCGTGGTCGTGGGTGCCTGTAATCCCAGCTACTTTGGGAGGCTGAGGCACAAGAATCGCTTGAACCT GGGAGGTGGAGTTTGCAGTGAGCCAAGATGGCTCCATTGCACTTCAGCCTGGCCAGAGCGAGACTCCA TCTCAAAAAAATAAAATACAAGATCATGTCCTTTGCAGCAATATGGATGGAGCTGGAGGCCATTATTCT AAGTGAATTAATGCAGGAACAGAAAATCAAATACCGCATGTTCTCACTTATAAGTAGGAGCTAAACATTG GGTACCCATGGACACAAACAAGGAAACTTCAGACACCGAAGCCTGCTTGAGGGTGGAGAGTGGGAGGAGG GCGAGGATAAATAAACGACCTATAGAGTACTGTGCTGATGACCAGGGTGACGAAATAATCTGTACACCAA ACCGCCGCAACATGCATTTTACCTACATAACAAACCTACAAAATGTACCTCTGAACCTAAAATAAACGTT TAGAAAAATGTTGTACATGAATGCTCATAGAAGCAGTATTTAGAATAGTCAAAAATTGGAAACAATGCCT ATCACTGATGAATGAATAAACAAAACGTAATATATCCACAATGAAATGTTATTCAGCCATAAAAAGGAAC GTTAAGAGACCATTCAGTCATTATTTATTTATTTAGAAACACTCTGTCACCCAGGCTGGAGTGCAGTGGT GCCATCTTGGCTCACTGCAACCTCTGCTCTCTGCTTCCTGGGTTCAAGCAATTCTCATGCCTCAGCCTCC CGAGTAGCTGAGATTACAGGCATGCGCCACCACACGCAGCTCATTTTTTGTGTGTTTTTAGTAGAGAAAG GATTTCACCACACTGGGCAAGCTGGTCTCAAACTCCTGATATCAAATGATCCACCCGCCTCGGCTCCCAA AGTGCTGGGATTACAGGTGTGAACCACTGCACCTGGCCTCATTCAGCCATTTTAAAATGCTGCAATATAG GTCTTGCTCTGTTGCCCAGGCTGGAGTGCAGTAGTGCAATCACAGCTCACTGCAGTCTCGACCTCCTGGA CTCAAGCGATCCTCCCACCTCAGTCTCCCAAGTAACTGGGAATACAGGTGTGAGCCACCACACCCGGCTA TTTTTTTATTTTTTGTAGAAATGGGGTCTTGCTATGTTGCTCAGGCTGGTCTCAAGCTCTTGGTCTCAAG CAATCCTCCCACCTCGGCCTCCCAAAGTCCTGGTGTGAGCCACTGTGCCTGGCACAACCTTCCATTTTTG TTTCATTGTTTGAATGTGTCTGATTGTGTGTATCTTCCTTGAGTTGAAATTTTGGGGATCTAAGACCGTG TTCATTTTCTCCACCACTGTATCTTCAGAGCCAACCACAGACCCTATCACTAAAGAAACACTTAATATTG TTGACGTAGGAAGGAAGTGAGGAAGTGGTGAGGAGGGATGGAATAGATTTACATAACTGTTCCATATCTC CAAAAACTGATGCATTTTTTTTAATCTCTCTTTTTTCAGCTGCTGGCTCCAACCAGAAAAGGGATTTATA TGGGGCTTCCTTGGACCTGTCTGCGCCATCTTCTCTGTGAGTGACATCATCTGAGCATCCTCGCTGCCCC GCTGAGGGTCATTAGAAAAACAAGATAATAAAGATGGTGACAAGAGCAGGAATGTCCCCTGGGTTGTCAT GGGTTGGGTTAGGTTTCCTAAGACCACTCTTGAATCTTCAAACTCAACACTGTTACTCAATATTTAAGAA AAAGACTTACGTGTATAATTCATATTAGACAGGCTAGGATGAAGAGATAATGGGCTTCAGGCCCGGTGTG GTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGCAGGCAGATCACTTGAGGTCAGGAGTTCA AGACCAGCCTGGCCAACATGGTGAAACCCTGTCTCTACTAAAAATACAAAAATTAGCCAGGCGTGGTGGT GGGTGCCTGTACTCGGGAGGCTGAGACAGGAGAATCGCTCGAACCCGGGAGGCAGAGGTGGCAGTGAGCT GAGATTGCACCGCCACACTCCAGCCTGGGCAACAGAGCAAGACCCTGTCTGAAAAAAAAAAAAAAAGAGA TAACAGGTTTCAAAATCTATTCAACAAATCTTCTGTGGTTTTCTGAAGGTGAATTTAGTTCTCTTTCTGG TGACTCTCTGGATTTTGAAAAACAGACTCTCCTCCCTCAATAGTGAAGTGTCCACCCTCCGGAACACAAG GTAAGATGGAGAAGGGGGTGATAACCACACAGACATGTATGGTTCCCAAGAGCCAAGCCATGTGCCTCAG CAGCCACTGAGGTCCCTTCTCAGGGTCTTTTGAAGGGACCCAGAAAAAACAGACGCAGCCATCCTTCTAT GGCAGCCATCACAGATTCAGTGGTATCCTGTTGAGACCCCATGCAACATGTTTTGTCCATGTCTAATCTC AGTTTGTGAATCCCAGAGCAACAGGAAGAATCTTCATTGACTGGTAAATGGTATATATACAAAATAGGAT ACGATTCGATCACAAAACAGAATGAAACCATGTCATTTGCAGCAACATGGATGAAACTGGAGGTCACTCT CTTAAGTGAAATAAGCCAGGCACAAAAAGACAAATACCACAGGTTGTCACTTACATAGAGAGGAGCCGAA GAAGCTGATCTTGTGGAGGCAGAAAGTAGAATGATAAAAGTTAGCCGGGCATGGTGGCAGGCACCTGTAG TCCCAGCTACTCGGGAGGCTGAGGCAGGAGAACCGCTTGAACCCAGGAGGCGGAGGTTGCGGTGAGCCGA GATCACACCACTGCACTCCAGCCTGGGCAATAGAGTGAGACTCTGCCTCAAAACTAAAAAATAAAATAA AGTAGAATGATAGATTCCAGAGTCTGGAAAGGGTATGTGAGTGGGCAAGGCGATGAAGAGGTTGGTTA ATGGGTACAACATACAGTCAGATAGAAGGAATAAGTTATAATATTGGGCAGAGTAGGGTGACTCTAG TTCACAATATGTTGTATATTTCAAATAGCTAGAGGGGAGAATTTGAAGTGTTCCCAACACATACAAATG GTAGACACGCGAAGTGACGGATACCCCAAATACTCTGACTTAATCATGACACACTCTATGCATGTTAACA AAATATCACATATGTCCCCACAAAGATGTACAAACATTACGAATCAATAAACTAAAAGGTTAAAAACTTC TTTGGACTTTTGTCAGCAGTCCTATAACAAAAGCACAAAGAAGGCCGGGCATGGTGGCTCATGCCTGTAA TCCCAGCACTTTGGGAAGCTGAGGGGCGGGGCAGATCACAAGGTCAGGAGTTCGAGACCAGCCCTGGCCA AAGAGACCAGCCTGACCAACGTGGTGAAACCCCGTCTCTACTAAAAATACAAAAATTAGCTGGGCGTGGT GGCGGGTGCCTGTAATCCCAGCTACTCCAGAGGCTGAGGCAGGAGAATTGTTTGAACCTGAGAGGCAGAG GTTGCAGTGAGCCGAGATTGCGCCACTGCACTCCAGCCTAGGTGACAGAGCAAGACTCCAACTCAAAAAAA AAAAGTGCAAAGAACACGTACATTTTCTTGCCTGTTCCCACAGCCCAGTCTGCACCTTCATACCGTACCC TCAGTTCAGGCTCCTCATCACTTCTCATGCAATCCTCCCTCTGGGAAAATCCTAATCAACATTAAATCCT TTACGGGGGCCAGGCATGGTGGCTCACGCCTGTAATCCTAACACTTTGGGAGGCCGAGGTAGGCGGATCA CTTGAGGTCATGAGTTCAAGACCAGCCTGGCCAACATGGTGAAACCCCATCTCTACTAAAAATACAAAAA TTAGCCAGGCATGGTGGTGGGCGTCTATAATCCCAGCTACTCAGGAGGCTGAGGTGGGAGAGTCACTTCA CCTGGGAGGCAGAGGTTGCAGTGAGCCAAGATCACGCCACTGCACTCCAGCCTGGGTGACAGAGGGAGAT TCCATCTCAAAAAAATAAATAAATAAAAATAAAAATAATTCCTCTACAGGGTTTGAGCAGTTTGTTGTTC CAGGCACCCTTGTGCAAAATTAATGAACTTTCTTCTAGATTCCTATCATTATCCTTTCCAAATTTTGTGT GTCATGCCTGGTTGTATTGGTGATTTGAAGAGACTGTGTAGTTCATGAACACAGAAATCTCTTTGCATAT TCAAAAATAATCCTTTTTTTTTTTTTGAGACAGAGTCTCGCTCTGTCACTCCCAGGCTGGAGTGCAGTGG CTCTATCTCGACTCACTGCAACCTCCGCCTCCTGAGTTCAAGCAATTCTACTGCCCCAGCCTCCTGAGTA GCTGGGATTACAGGCGCCCCCCCGACCACACCTGGCTAATTTTTGTATTTTTTTAGTAGAGACCAGGTTT CACCATGTTGGTCGGGCTGGTCTCGAACTCCTGACCTCAGGTGATCTGCCTGCCTTGGCCCCCCAAAGTG CTGGGATTACAGGCATGAGCCACCGTGGCCAACCCCAGGTGGCATTTCTACTGTGGGCTATAGTTGACAG TAAATATCTAAGCTGACCACAGCCCAGTCTGAGAAATCATCTACAGGGGAGGGCAGGGGACTCCATGGGC ACAGATGAAATAGCCTTGATTCTCTTTCTCTGGGTGCATCACCAGCCTCCTTTCTTACTCAATCCAGGAT GCTGGCATTTAAAGCGACAGCTCAGCTGTTCATCCTGGGCTGCACGTGGTGTCTGGGCATCTTGCAGGTG GGTCCGGCTGCCCGGGTCATGGCCTACCTCTTCACCATCATCAACAGCCTGCAGGGTGTCTTCATCTTCC TGGTGTACTGCCTCCTCAGCCAGCAGGTACCACTGCCCAGCTCCCACCCAGGACTCTTCCTGTCCTCACT GCTCTCTGTGAGCTGACCCAGTACACACTTTGCCTCTGCAGGTCCGGGAGCAATATGGGAAATGGTCCAA AGGGATCAGGAAATTGAAAACTGAGTCTGAGATGCACACACTCTCCAGCAGTGCTAAGGCTGACACCTCC AAACCCAGCACGGTAAGATCACGCATTGCTCCAGAGCACTTCACTAACCGACCCACCTGAGGAGCATGTG CCTATCACACAAGGAAACCTGGGAATACAGCAGGCAATGCCCTAGAAAGGCTCGCATCTGAGTACGCCTT GACTCATTAACCATTAGCAATGATCTCAGTTTAAATGTTTTTTTTTAATCAGTCATAGCCTGTCATCCCA GCATCACTGTCATCCCAGCATTTGGGAGGCCTAGGCAAGAGGATCACCTGAGGCCAGGAGTGCAAGATGA CCCTGGGCAACATAGCAAGATCCCATCTCTACAAAAAATAAAAATAAAAATTAGCCAGACATGGTGGCAT ACACCTGTAGTCCCAGCTACTCCTCGGGAGGCTGAGGTGGGACAATCACTTGAGCCCAGTAGGTTGAGGC TGTAGTGAGCCATGATCATGCCACTGCACTCCAGCCCAGGCAACAGAGTGAGACCCTATCCCAAAAATTA AAAAAAAAAAAAAAATCCTTGGCTCTTACTCCTTAGGGAACTGTGCTTAGTTTGACTCTCACACACAAAC ACTCCTATTGGACACCTACTATGTGCCATGCATGGTTAACTAAATGAGCAGGAATTGATCTGAAGCAGTG ATGCACATTCTACCTAGAAGCATTTACCCTATCCCTTTCCTCTACATACAAGTCTATCTCACAGTCACTT GGCACCACCACCCAATAAGGCAAGCATGGGAGGAAATGCTGTACATATTTATGAATTAAACAGAAATAAG TGTTAACTGGGATTTAGGCTTTCTTAACAAGTCTGCCTCTACCAACTACCACTTAGTAGTGGGTAGTGGG CAGTGCCTCTACTCAGCACCCTCTACCCACTGCAAAACTCAGAAAGAAAAGCAGTAATGGGGCCAGCGAT GGTGGCTCATGCCTGTAATCCCAGCACTTTAGGAGGCTGAGGCAGGACAACTGCTTGAGGCCAAGAGTTC AAGACTAGCCTGAGCAACCCAGTGACACCTCGTCCCTATAAAGCATTTAAAAATTAGCTGAGGGTGGTTG TGTGCATCTGTAGTCCCAGCAACTCAGGAAGCTGAGGTGAGAGGATCACTTGAGCCCAGGAGTTTGAGGC TGCAGTGAGCTATGATTGTGCCACTGCATTTCAGCCTGGATGACAGCAAGACCTCTTCTCTAGAAAATAA TACAAATAACAAAATTAAATAAATAAAGTTTTAAAAACACACTGGGCAACCTGGGAATCAGCGGGTGATG CCCTAAGAGGGTTCACATTTGAGCGACTCCTTGACTCATTAACCATTAGCAATGATCTCAGTTTAAATGC TTTTTTATAAATCAGCCAGGAACAGTGGTTCAAGCTTGTAATGCCAGCACTCTGGGAGGCCAAAGCAGGA AGATCACTTGAGCCCGGGACAACTTGGGGATCACTTGAAGCCATATTATCCTATGCCTCAATGGCATGGG ATAATTCATGACATTGATGACTAATAACCAACATTTATTAATGTAGACATACAAAGTTCTGTTGTATTTT TCTCACCTCTTCTTCCATCTTCTCCAGATTATATGTGTAATTCTAACCCCATTTCACTGATGAGCAAAGT AAGACTACTTGGATTTGTGTCAAATGTACAGAGTCTCGGCCAGGCACAGTGACTCATGCCTGTAATCCCA GCACTTTGGGAGGCCGAGGCAGGTGGATCACAAAGTCAGGAGATCGAGACCATCCTGGCTAACACGGTGA AACCCCATCTCTACTAAAAATACAAAAAAAAAAAAAAAATAGCCAGGCGTGGTGGTGGGCGACTGTAGTC CCAGCTACTCGGGAGGCTGTGGCAGGAGAATGGTGTGAACCCAGGAGGCAGAGCTGGCAGTGAGCCGAGA TCGCGCCAATGCACTCCAGCCTGGGCAACAGAACGAGACTCTGTCTCAAAAAAAAAAAAAAAAAAAAAAG AAATGTACAGAGTCTCTTCAGCCAGTGAGATTTAGGCCTCTTGAGTCTTGCTTCACATTTAAAGAGCCCA TTCAGGGCCAGGCACCGTGGCTCATACCTATAATCTTAGCACTTCAGGAGGCCAAGGCAAGCAAATCACC TGAGGCCAGGAGTTCGAGACCAGCCTGGCCAACATGGCAAAACTCCATCTTTACTAAAAATACATAAATT AGCCAGGCATGGTGGTGCACACCTATAATCCCAGCTACTCAGGAGGCTGAGGCACAAGAATTGCTTGAGC CGGGGAGGTAAAAGTTGTGGTGAGCCAAGATTGCCCCATGCCACTGTACTCCACCCTGGGCAACACAGCA ATACTCTGCCAAAAAAAAAAAAAAAAAAAACAGCCTATTCAGCCTGGGCAATGTAACGAGACCCCATCTC TATAAAAAAGTTTTAAAAATTCGCTGGGCGTGGTGGCACATGCCTGTAGTCCCAGCTACTTGGGAGGCGA GAGGATCGCTTGAGCCCAGGAGTTGGAGACCAACCTGTGCAACATGGCAAAATCCCATCTCTACACAAAA TATAAAAACTAGCTGGGCATGGTGGTGCATGCCTATAATCCCAGCTGCTTGGGAGGCTGAGGCAGGAGGA TCACTTGAGCCCAGGAGTTCAAGGCCACGGTGAGCTACGATTATGCCACTGCACTACAGCCTGGGTGACA GAGCAAGACCCCTTGTGTAAAGTAAATACATAAATAATAAATTTTTTAAAGTCTTGAAAAGAAGCCCACC ATGCTGCCAGGCACTTTTTCCACATTTGCATTTCTCTCATTACCTTTTTCCTTTTTTATTTTTCTTTTTT TTTTTTTTTTGAGACGGAGTTTCGCTCTTATTACCCAGGCTGGAGTACAATGGTGCTATCTCAGCTCACC GCAACCTCCGCCTCCTGGGTTAAAGCGATTCTCCTGCCTCCACCTCCCAAGTAGCTGGGATTACAGGCAT GTGCCACCAAGCCTGGCTAATTTTGTATTTTTACTAGAGACGGGATTTCTCCATGTTGGTCAGGTTGGTC TCAAATTCCCGACCTCAGGTGATCTGCCCGCCTCAGCCTCCCAAAGTACTAGGATTACAAGCGTGAGCCA CCACACCTGGCCTCATTACCTTTTTCATCCAACCTGCACAATCCCTGTTTCCAACACAGGATCTTCTATT TCCTTTTCTGTTCTCACTAAATGCCCTCACCACCTATGTCACACACACACACACACACAAATCCAAGTCA TCATGATGACACATTGTAAACTCCATGCTCTCAATCATACAACCAAACATGACCATCCACCTTCCACATT CCCATATTATTGACTGTCCATCCTCTTTTTTTTTTTTTAGACAGAGTCTCACTCGCTCTGTCACCCAGGC TGGAGTGCAGTGACACCATCTGGGCTCACTGCAAACTCTGCCTCCAGGGTTCAAGTGATTCTCGTGCCTC AGCCTCCCGAGTAGCTAGGATTACAGGCACCCGCCACCACGCCTGGCTAATTTTTGTATTTTTAGTAGAG ACAGGGTTTCACCATGTTGGCCAGGCTAGTCTTGAACTCCTGGCCTCAAGCCATCTGCCCGCCTTGGCCT CCCAAAGTGCTGGGATTACAGACGTGAGCCACTGCGCCCAGTCCATCCCTGTCTTTGACTCTCCTACAGC CTTCATACTTTATAAAGACCTTGATGATTACATTGGGTTCACCAAGACAATCCAGGGCAATTTTCCCATC TCAAGATCCTTTTCATAACCGTATCTACAAAGTCTCTTTCTCCATGTCAGGTAGCATTTACAGGTTCTGG GGATTAGGGATGTGAACCTCTTTGGGGGGTCATTATTCTGTCTCTCATACCCATATAGTTTGGGTTTACC TATTGTAGAATCATCAAAACCTGCCTCATATTGTAGATATTTGCATAAATATTTATCACCCTTTCACCCC ACACAGCCACTAGAAATGATATATACACATTGAAGAAAGGAAACATGTACTTTTGTGTAGGGAATTAGAG TTAGGTCATCAGATAGCCAACAAAGGCTTCTGAAATTTAAGGTGTGTGATCAGATATTTCTACCAGCTAC TGACAAAGGACCAAGAAAATCCATTGCATTACTTAATGTCATTTTATCTTCTACTGTTTCCTAAAACTTG TTATCTCTGGTGTTTGAAAATGTATGTGCCTTGGCCGGACACGCTGGCTCACACCTGTAATCCCAACACT TTGGGAGGCCAAGGCGGGCAGAGCACTTGAGGTCAGGAGTTCGAGACCAGCCTGGCCAACATGGCAAGAC CCCACCTCTACTAAAAACACAAAAACTAGCCAGGCATGGTGGTGGATGCTTGTAATTCCAGCTACTTGGG AGGCTGAGGCACAAGAATCACTCGAACCCAGGAGGCAGAGGTTGCAGTAAAGCCAAGATCGAGCCACTGC ACTCCAGCCTGGGTGACAGAGTGAGACTATGTCTCAAAAAAAAAAAAAAAAAGAAAAGAAAAGAAAAGAA AATGTGTGTCTTGCTCTTATTCTGCAGTCTCTGCCATGGCTTTCTGCAGTCTTGTTTCATAAAAACAAAA TGTCTTGTGATCACTACTGAATAATCTCCTGAAGCACAATATTTATTAATCTCTTGAGCACATATTAATT ATGCATTAAATACTAACTATATAAGCAAGTGAAGCTGACCATCTCGAACCATAGAAATTACACTGAGTTT CCTATGAACCTAGGAATAAATAACTCATATTGGAGATGGAAAAGATATGGAAAAAAGGAATTCAGGCTGG GCGCAGTGGCTCACGCTTGTAATCCCAGCACTTTGGGAGGCCGAGGCAGGCAGATCACTTTAGCTCAGGA CTTCAAGACCAGCCTGGGTAACATGGCAAAATTCCGTTTCTACAAAAAGTATAAAAATTAGCCAGGCATG GTGGTGCATGCCTGTAGTCCCAGCTACCGGGAGGCTAAGATTGGAGTATTGCTTGAGCCCAGGAGGTCAA GTATCCAGTGAGCTATGATCCCATCACTGCACTCCAGCCTGGGCAACAAAGCAAGACCCCTGTCTCAGGG GAAAAAAAAGGAATTCAGTGCTTTTTCCTTTAAATCTTATGTGGTTAACAATAGATAACACTTCTAACAT TTCATTAATTTCCCACCTGGCTTCTTTTATGACTCCACATTTAAAGCAACATTACACAAACCCTGGAGCT CCGAGGTCAAGACATACATTTGGAAACAGATAGATCTTTTTAAATGTTTTTTAATTGATCCATAATAGAC GTACATAGTTTCTAGGTACATGTGATCATTTGCTACACTCATATAAGCAAATCAGGGTAACTAAGATATC TATCACCTTAAATACTTATCTTTTCTTTGTACTAATAACATTCAAATTATTCTCTTCTAGCTAGTTTGAA ATGTATGGTAGATTAATGTTAACTATAATCACCCTACTTCAGATACAACTTTTTTTTTTTTTTTTGAGAG GGAGTTTTACTCTGTCGCCCAGACTGGAGTACAGTGGCACCATCTTGGCTCACTGTGACCCCCACCTCCC AGATTCAAGCGATTCTCCTGCCTCAGCCTCCCGAGTAGCTGGGACTACAGGCGCCTGCCACCAAGCCCAG CTAATTGATTATTATTATTACTACTGTATTCTAGTAGAGACAGGGTTTCACCATGTTGGCCAGGCTGGTC TCAAACTCCTGACCTCAAGTGATCTGCCCGCCTCGGCCTCCCAAAGTGCTGGGATTACAGGTGTGAGTCA CCACACCTGGCCCAGATAGATACTTTACTGCAACACATTTACTTCCTCTTAACTTTGGTAACTTGGGGTT TAGGAGGCATACAAAGATGGGGCTTCTGAGAAAGAACAGGATGGAGCAAAATGAAAGCTGATATGGCCTT TTGATGTTGCCCTGACAAATACATATGGTAAGAAAACCTTATGATTAAAGACTTCTTGGCCAGGCATGGT GGCTCACACCTAGAATTCCAGCATTTTTGGGAGGTTGAGATGGGAGGATCACTTGAGCCCAGAAGTTCAA CAGCAGCTTGGGCTATACAGTGAGACCCCATCTCTACAAAAAATTAAAATTAAAAAATGAGACAGGAGTG GTGGCACATGCCTGTAGTTGCAGCTACTCGGGAGGCTGAGGCAGGAGGATCACTTGAACCTGGGAGTTGG AGGTTGCAGTGAGCAGAGATGGTGCCACTGCACTCCAGACTGGGTGACAGAGCAAGGCCCTGTCTCTAAA AAGAAAAAAAAAAAAAAGACTTCTTAAATTTCTTTTTTTAATAAATTTTATTGTGTATACACACAATTG AGGTTTAACCCATGAAGTTATGGGATACATATAGATAAAAATGGTTATCTATATGTGAAGCAAATTAA TATATCTATCATCTCACATAGTTACTTATTTTTGTGGCCACAAGCAGCTATAATCTACTTATTTAACTAAT TTCTTATTTCTTTTTTCAAAATGGAAATTATAGGTTTCTTTCTATAGTCATATATATTTCTAAAATATATA TATATTTCTAAAATATATAGATATTAAATATATATATTCTTAAAATATATAGATATTAAATATCTATATATT TCTAAATATATAGATATTAAATATCTATATTTTCTAAATATCTATATTTCTAAATATCTATACTTCT AAATATCTATATTTCTAAAATATCTATATATTTCTAAAATATCTATATTCTTAAAATATCTATATTTTCTAA ATATCTATATATTCTTAAAATATCTATATTTTCTAAATATATATATTTAGAAATATCTATGTATTTCTAA ATATCTATATATTCTTAAATATCTATATACTTCTAAATATATCTATGTTTCTAAATATATTTAGAAATAT ATATATATTCAGGCCAGCTCAGTGGCTCACACCTGTAATCCCAGCACTTTGGGAGGCTGAGGCAGGCAG ATCACCTGAGGTCAGGAGTTCAAGGCCAGCCTGGCCTACGTGGTGAAACCCTGTCTCTACTAAAAATACA AAATTAGCCGGGTGTGGTGGCACATGCCTCTAATCCTGGCTACTCTGGAGGCTGAGACAGGAGAATTGCT TGAACCTGGGAGGCGGGGGTTACAGTGAGTCGAGGCTGCGCCATTGCACTCCAGCCCAGGCAACAAGAGT GAAACTCTGTCTCAACAACAAAATACATACATATATATATATTTCAGGTTAACTAGAAAAATCTTCTGAA TAAGATCTTCCCTCTTTGCCCGTGGAAAATCTGAACAATCTTTGAGCCATCTAGAGGGGAAAGAAAAGAC TTTGTTCTGTGTGTTTCAAGAAATTCACCATGTCAGCAATATGAAGGATGTTATGGAAGGCGTGCTAGGC ATTCAATTCCTGCAGAAACCGGAAATCTTCCATGCCCTGCAATGTGCTCATCAAACTCTCAGCATATGGA CGGCCAGCTGTGGCCCATATCTTGGTCACTCTGAAGCACAATATTTATGAAGCTATAGAACGTTAAGACC TCTTTCACAGCCTCTCCTTCCTACAAAGACTCCTCCAAATCTTAAAATGAAGCAGGAAAACGAGCCTAAG AGGACTTTCATACCGACAACATCTGAAAGGACTAGAATGTTCACACCACGATCTGGATTTCTTAATTTTT TGTTTTTGTTTTTGTTGTTCTCTAGTTCTACGGGTTTGATTATTTAGTCATGTGAAAAATATTGATTACT CACACATAGATCAAGAGAGACACGGCTCCTGCCTTCATGGAGCTTTTAGGGGAAAATGAAGTGGCTCTTG CAGCTAGAGTTGACTCAGAAGCCGAAATTCCTAGAAATCAGGTTTCTACTGCTAGGCAATTGAAGTATAA ACTATTTTATAAACACTGTCTTCTTTCGTCTTCACACCAACATGCAGAAAAGTTTCTAATCTCAGATCGG GGATGTGCAACAAATTCCATTTCAAAGGAATGACCTGCAAAACTCCTAAATATTCCAAGCAAATGCCCTT AACCCTGTCTGTTATCTGCTTTCCTTGAACAGAAATTCTACATGACCATAAAACCTCGAAGATGGGTATG GCACAGTTCATGCCCTGTAATCCTAGCACTTTGGGAGGGTGAGGCAGGAGGATGGCTCAAGCCCAGGAGT TTGAGACCAGTGTGGGCAACAGAGTGAGAACCATCTCTACCCAAAAAAAAAATTAAAAATTAGCCAAGCA TGGTGATGATATAGGAGTTAAGGAGAAATCATTTAGGCAAATAGCAAGGGTAGGAAGTCCTCAGTAAGGT TTTCCATTTAATGAAAAGCAGCCCCCAAAATCATTTTCTTTTCTAACAAAGAACAGCCTGTAAAATCGAG CTGCAGACATAGACAAGCAAGCTGGAAGCTTCCACGGGTGAATGCCGGCAGCTGTGCCAATAGGAAAAAG CTACCTAGACTAGGCATGTCCAAAATGGCGGCTCCAAGTTCCCTTCTCTTTGCCAGCCATGTGTACAGTA AAAAGCAGGCAACATAGTGTCAGCCAAAGCTCATTTGCATAATAAGATTAGGGTGGGGTGGCCAGCTCAC ATAGGGGTAGGCCCTAGGTAAATCAGACACCGCCTTCTCAAGCCTGTCTATAAAATCTGGTACACTATGA CGAGGGTCAGATTTCCCATTCAGACGCCCCTCTCCCATGCAAGAGAAAGAGCTGTTCTCCTTTCTCTTTC TTTTGCCTATTAAACCTCTGCTCCTGGCCAGGCACAGTGGCTCACGCCTATAATCCCAGCACTTTGGGAG GCTGAGGTGGTCAGATCACCTAAGGTCAGGAGTTCAAGACCAGCCTGGTCAACATGGTGAAATCTTGTCT CTAGTAAAAATACAAAAATATATGAAATCTCACATAGATGATAATATTAAGTTCCAAAAGCAACTCAACC TGGTAGATTCTAATTTTTTTTGAGGCAGGGTCTTGCTTTGTCACCCATGCTGGAGTACAATGGCACAAAC ACTGCTCACTGCAGCCTCGACCTCCCAAGGCCTAAGCAATCCTCCTGCCTCAGTCCCCCTCCAGGTATTT GAAACTACAGGTGTGTACCACCACACCGGGCTAATTTTTGTATTTTTTGTAGAGACGTGGGTCTCACTAT GCTGCCCAGGCTCAGGTCTTAATCTCCTGAGCTCAGGCAATCCGCAGGCCTCAGCCTCCCTAAGTGCGGG GATTACAGGCTTGAGCCACTGCACCTAGCCTCTATTTGTTTTACAAAAGAGAAATTGAGATCCTGAATGT TAAGTGACTTGCCTGAGGCCATCCCACTAACAGGAGCCAGGGTTAGGATTCAAACCCCATCCAACTGGTC CCAGAGCTGGAGCTTCTTGCACTGCCCTACACTACCTACCATCTCCATCCTCTGGGCACCTTTTTATAAG AACCAAAACATTACAGAGCATTGCTTTGTCAACTCAGCTGGGAACATTTCCCAGTGCAACTCACATTTTT CACTGCTCTGTGCCTGTCCGTATAAGCTCAATGAGTATTGATTTAGGGGCTTTGGAGAACTTTGAATGCT ACCCCCCAAGTAACCATTGTTGGCAACCTGGTACCTCTACTTTTAGCCATTTCTCCTTCTCTATAAATAG TGCAGAAGTAACCCACTTGGTAACAGGCATCCTTGCCAAGCCTCCACCACTAGGTCAGTGTAAGAATTAA AGAAAGAGGAAAGAAACACAAAAAGTGGCTTGATGGTTAAGACAGGTTTATTTTAGAGAAAACACACCTG AGAGGGGCTGCTGGCTGAATTAGGTTAGAGTCTTTTCTACAGACTAAGAGTGTTTAAGGATTTAGGGTGG GAGAGTTTCTTAGAGGCTTGGACTGCTTCTGTGTTTTTTTTGTTGTGCTTATATGGGAGGGAGAGTGGTG TGTTTGCTTTTATACATTTTTCTGCAGCTGTAGGCATACCCCCCAAGTCTGCTTTTAGCTTCCCTATTTT AGTGCACCTGGAGGGAAAGGAATGTGCTTATTAAGGCCCACTGTTTTACTGGGGCCCATTGTATGAGGGT GAAGTTTGGCAGTTACCCAAGAGACTTTTCCTCCACCTTCCTCTGTGCCCGAGCTGTTTTATCTGCATTT TACTGTCTGCTTTTTTTGGCTGCTTATAGTTTTTAAAAAAGTAATTTCCTTAAATCCAGAAGGCTAAAAA TGAAGCTGAAACTTAAAGTGGCGGTGTTTGTCCAAAATAACGGGGCTCCTGCTCTGCCAGTCAGTACCCT CAAGTCACTCCTGATCCTCAACCTCCATGCCTAAGGCTGGTTCAAGAGACCACATAATATCTGCCTTTTA TTACATACATGATGGGTGCATGGGATTCTGCGTGCCCTTTGCTTGATATAGACTGCTAAGGTGAGATGGG GAATATCAGAGTCAGCTGCTGCTTGAGGAAGCAGAACACACAGCTGGAGGCTTGGAACATGTGGGTCCCT ATGAGTGTAGAGCCCATATCCCCATAGAGTCTACCTAGAGCAGGGGTCGCCAAATGTTTTCTTAAAGAGC CTGATAGTGTATATGTTAGGCTTTGTGAGCCAGGTATTTACAGCAACTCAATTCTACCACTGTGGTATGA AAACAGCTATAGACAATCATAAATGAATGATCATGGCTATGTTTTAATAAAACTTTACAGACACTGAACT TGAACTTCCATTGTGATATGAAAACAGCTATAGACAATCATAAATGAATGATCATGGCTATGTTTTAATA AAACTTTATGGACACTGAGCTTGAACTTCATATATCAGTCATGTGACACAAAATATCATTCTTCTTTTTA TTTGTTTTCACCCATTTGAAAAATGTAAAAACTATTCTTAGCTGTACAGAAACAGATGGTGGGTCAGATT TGACTCACTGCCCATAGTTTCCAGACCATGATGTTCAGGTTCATGGAAGCACTTCACTCTACACTAATTT ATTCATCCATTCATCACATATGTGATGAGCACATACAGACACACCTCAGAGATATTGCAGGCTCCACTCC AGATCACCACAATAATGTGAATATTGCAATAAAGCAAGTCATACACATTTTTTGGTTTCCCAGTGTGCAT AAAAGTATGCTTACCCTATACTGTAATGTGTGCAATAGCATTATGTCTTTTTTTTAATGTACATACTTTA ATTTAAAAATAGTTTATTCCTAAAAAATGCTCAAGCAGGGACACAAAGTGAGCACATGCTGTTGGGAAAA TGGCACCTACAGACTTGCTCGACACAGGGCTGCCATAAACGTTCAATTTGTAAAAAATGCAATATCTGTG AAGGGCAATGAAATGAGGTATTCCTATACTCTCGTGCAAGGCATAATGCTAAATTGAGAGAAACTAAAAC ATATAAGACAGACATGATCCTACATCCAGGAATATTGATGATTTGTAAGATGGAAAATGAGGAGGAAGGA AAGGTGCAGTGCAATGGACTCTGTGGTCTGGGGGACACACAGCCCCTTTAAGGACATGGCAGAAGTGAAA CACAGACGAGGTTGGGGAAGGAAGGTCAGAGGAAACTCTAGGGTCAGCTGATGGCAGCATCCATTTGCCC CCATATCTTCACTTTTTAAATTTATTAAGTCCATTCTCTTTATTTATGATGGAAATAATGTTAATTTGCT CATGGGTTGTGTTTGAATATTAAGTAAGAAAATAAAGATTAAGCTCTGGTATGTAGTAAGAAACCCAAAG ATAAACAAGAGTTAAGGGGCAAGTAGGCAGCAGAGTTTCCCATCTCATTTCTCCAGAATTATATCATCCA GTTTGAAACATGAAATACGGCTTTAAGGTAGAGTTTGGCTTCTCAGCTGGATTTTAAGTACAAGACCACG CCCACCAAAGACAGCAGACAAAGGAGCTAGGGTACACGTTTGAAGAACCCTCTCCCTCCAGCTCTCCACC GTGGGGGTCCACTGTGAGCAGTTTCCCTGGCTGTTTTCCCCCTTTCTGCATCTAACAGTCATTGCACGGC TGGGGAAACAAAGACCCACACAGGCCTTAGTCTCCTTGCGGGGAACCTACAATCCGGAAGCCAGAAGTAT GTGACTAAGCTCCCGGAAGCCGGAAGTTAAGGAGCCCGGATGTCACGGAACTGGAAGTTTGAAAGTCCAA GGCCACTGACCTGCAAGCTTCAGAATCTGCAGATTTGGAGTCTTCAAGCCCCAGTAGTGATCCTTTGATA TCCAGAGGTTGGATTCTAGCGATCTGAACATGCAAGAGAGTACTGGATGTGGAAATTCTTCCAAATTAGT TGGGTGGCTGAAAAGGGAATTATAAGGAGGAAGAATGAGGGAAAGAGAAGAGAAATTGAATGAATAAAGG GGCCAATCCTTGGGGGGAGGGGGCAGGAGGGGCGGTGCTTGAGACCAGGGCAGCATGTAAGGGACTGCCT ATGAATGTCCTAAATATCTGCTATTTAGGGCATTCTGGGATGGGAGTTTGAAAGACTGATGGTGGTGTGT TTTGCAGCCACATGAATGCAGCTGGAACCATCATCCTAAGTGAATTAACTTAGAAAACCAAATACCGCAT GTTCTCACCTGTAAGAGAGAGCTAAATATTGGGTACACGTGAACATAAAGATGGGAGCATTAGACAATGG GGACTACTAGATGGAGGAGGGAGGGAGGAGGGCAAGGGTTGAAAAACTACCTATCAGGTACTATGCTCAC TAACTGGATTCATTTGTACCCCAAACCTCAGCATCATGCAATATACTCTTGTAACAAACCTAAACGTGTA CCCTCAGAATCTAAAAATAAAAGTTGAAATTAAAAAAATCACTGTTGATAGACATTTGGGTTGTTCCCAG TTTGGGTTATTACAAACTGCTTTGCTAGCTTTAACTTTTTTTAAATAATAAACTCTTAAATTTACAGTGC AAGAAATACAGAAAGAGAGAGGAAGACAAAGAAAAAGAAAGGAGATAGAGAAAGGAAAAGCAAAGAAAGA AAAAGAAAAAAGTGAAAGAAAGAGAGGAAGGGAGGAAGGAAAGAAGGAAGGAAGGACAGACTGATGGTGG GATAAGAGAGAAGGGGCTGGGAAGAACTAAGTGGAACATTCTGAAAATGAAGACAGTGGCCGGGCACGGT GGTTCACACCTGTAATCCCAGCACTTTGGGAGACTGAGGCAGGCAGATCACCTGAGGTCAGGAGTTCAAG ACCAGCCTGACCAACATGGTGAAACCCCATCTCTACTAAAAATACAAAAATTAACCAGGCATGGTCACGG GCACCTGTAATCCCAGCTACTCAGGAGGCTGCGAGGCAGAAGAATCACTTGAACCTGGGAGGCAGAGGTT GCAGTGAGCAGAGATTGTGCCATTGTACTCCAGCCTGGGTGACAGAGCGAGACTCCATCTCAAAAAATAA AAAATAAAAATAACTCTCTTCTCTCTATTTTCCTTCAATAAATTCTCTTTTTGGCTAAAGTAGTCATTTT AGCCAAAATCTTTCTCTCAAGACAACAAAGAACTGAGGATTTCTGCCCTTTCCAGTATACAACTAGCAGG GATCCCACTGGCATTTTTCTTTGGACAGATAAAATTGGATGTATTTATCAAGAGCAACATGATGTTTAAT GGATGTATACATTGGGGAATAACCAGATCTAGCAAATTAATGTATGCCTCGTCTCATATAGTTATTATTT GGTGGTGAGAACAATTTACATCCACTCTCTTAGCATTTTTTAAGAATATAATTTATTATTAACTATAGTC ACCATGCTCTACAATTGATCTCCTGAACTTCTTCCTCCTGTCTAACTGAAATTTGTATCCCACCGGGCGC AGTGGTTCACACCATAATCCCAGCACTTTGGGAGGCTGGGGTGGGCAGATCGCTTCAGCCCAAGAGTTCA AGCCCAGCCTGGGCAAAATGGAGAAACCCTAACTCTACAAAAAATATTAGCCAGGCATGGTGGTGCATGC CTGTGGTCCCAGCTACTCTGGAGGCTGAGGAGGAAGGATCACTTGAGCCCAGTAGTTTGAGGCTGCAGTG AGCTGTGACCACACCACTGCACTCCAACCTGAACAACAATTAGACCCTGTCTCAAAAAACAAAAAAAAAA AGTGTATCGTTTGACCAACATCTCCCCAACCCAAGCTACTGGCATTTAGTTAATAGTCCAGGGATGCTGC TCAACAGTCTTCAGTGCATAGGGCAGTTCCTACAGCAAAGAATTATCTGGCCAATAAGGTCACAGTTGAG AAACCCTGACTGTAGGATGCAATGAATCTCTCGGTTTCTCTTCAAAGGTTCAGTCTGTTAACTTCCTTGT CTCTATGCCTCCCTGCCCCAAGTTACTATAAACAGCCTTCCCGTCGGCACTAATCAATAACTCACATCTG TTCCCTCGGTCACGCATTCTGCACCCGTTCCACCCCTCAAAACCACACACATACCCTTCAAAACTGCACG TCCCACCATTGTAACTCACGTCCCCTTCCCCCTTCTTTATTTGAGAGAAATATTCGCAAGTAGCCAATCG GGTCAGCTTACATTGTGCCGACCCCAGCCCATGGGGGAAGGACATGGAGGCAGGGATTGCGTTAAGGATA TAAAACCCCCCTGGCCTCCTTTGTTCTCTGTGCTCTTGGATCTTGATTGACGTGGGTGGCACCCTTCTGC AGAAGTAAACTGCCTTGCTGAGAGAACTTTTGCCTCAGTGCTGGTTTCACTTTGCAGCACCCAGCATTTA TTGCTAGAGCATTTTTATATTCAACATGACCTAAGGTGTAAAGGGATTCACATGGAAGTTTTTGCCATTT TCTTCATCCTTTCCATTGTCGTCATAGGTCACTCATTCCCAGGTCTAGAGAAAGTTTGATGCAGTAACTG TTGTTTGAGATGAGTGGCTTCACCTGCTCATCTTTCTATTGAATACATTTATAAGTGATGTGCTTCAAGA AATTCTTCAAAGCTAAACATAAGACAAAGTTTCTACATATCCTAGCAAGATAACATGACCATTTAAAAAC TGATATGAAGCTACAGAAGAGACCTCAGGGATGAGAGGAGAAGACCACCCCCACAACTTCAGCCAGACCT CAGAGAAACAGACTGTCATCACACCCTAGCATAAGAAGAGACACTAAGCAGCGAGGTTTCTGTCTTCAGC CTGTAGATAGAAAAAATATTCGAAGAATTATCTCATGCCTTTACCAAGTCTCAGCTCAAATTTTGCAGCT TGAGTATTCTGAAAAAAAAATTTTTTTTTGAGACGGAGTCTCGCTCTGTCGCCCAGGCTGGAGTGCAGTG GCACAATCTCGGCTCACTGCAAGCTCCGCTTCCCAGGCTCACGCCATTCTCCTGCCTCAGCCTCCCGAGT AGTTGGGACTACAGGCGCCCACCACCACACCCGGCTAATTTTCAGTAGAGATGGGGTTTCACCGTGTTAG CCAGGATGGTCTCGATCTCCTGACCTCGTGATCCACCTGTCTCAGCCTCCCAAAGTGCTGGGATTACAGG CGTGAGCCACCGCACCTGGCCAAATATATTTTTAAGTACTGGAAATTTTTATAAACTCTGAATTCCAAGG TAAGATAGCTCAGTTTACTTTATAAAGGTGTACCTGCATGCATGTATACATAAGCCATTGATTTTCCACC ATTGTGGGCAAGAGGGTATAAGGTGAGACTTAACATTAAATGGGATTAAAGGAAATGATTAGGATACTGG GACTCTTAAGTTTGTAAATCACTGCCTCAGATATTTGGTTATATTTAACTAATAGGTACATGATATAGTT TGGATGTTTGTCCCCTCCAAATCTCACACTGAAATGTAATCCCCAGTGCTGGAGGTGGGGCCTGGTGGGA GGTGTTGGATCATGGGGGCAAATCGCTCATGAATGGTTTAGTAATCCCTTTGCTGTAAGTCAGTTCTTGC TCTGGTAGTTCACACCAGATCAGGTTGTTTAAAAGAGTATGGAACGTCTCTACTCTGTCTCTTGCTCCTG GTCTCACTATGTGATGTGCCTGCTCCCGCTTTGCTTTCTGCTGTGATTACAAGCTTCCTGAGGCCCTCAC CAGAAGACAAGCAGATGTTGGTACCATGTTTGTGCAGCCTGCAAAACAGTGAGCCAATTAAACCTCTTTT CTTGGGCCGGGCTTGGTGGCTCACACCTGTAATCCCAGCATTTTGGGAGGCCAAGGCGGGCGGATCACAA GGTCAGGAGATCGAGACCATCCTGGCTAGCACGGTGAAACCCTGTCTCTACTAAAAATACAAAAAAATTA GCCAGGCGTGGTGGCGGACACCTGTAGTCCCAGCTGCTCAGGAGGCTGAAGTGAGAGAATGGTGTGAACC CAGGAGGCGGAGCTTGTAGTGAGCCAAGATTGCGCCACTGCACTGTCTGGGCAACAAAGCAAGACTCCAC CTCAAAAAAAAAAAAAAAAAAAAAAAAGCCTCTTTTCTTTATAAATTACCCAGCCTCAGGTATTCCTTTA TAGCAACGCAAGAACAGGCTAATACAGAAAATTTCTGAAAATGTGAAAGCAGCTTTAGAACTGTGTAACA GGCAGAAGTTGGAAGAGTTTGGAGGGCTCAGAAGGAAGAAGGAATATGAGAGAAAGTTTGGAACTTCTTG GAGACTGGTTAAAGGGTTGTGACCAAAATACTGGTAGAGATATGGACAGTGAAGGTCAGGCTGAGGAGGT CTCAGATGAGGAACTTACTGGGAACTGGAGCAAAGGTCAGCCTTGTTATTCCCTGGCAAAGGACCTGGCA GCATTGCGTCTATGTCCTAAGGCTTTCGGGAAGTTTGAACTTAATAGTAATGACTTAGAGAGTGTCTGGT GGGAGAAATTTCTAAGCAGCAAAGTGTTCAAGGAGAGGCACGGCTGCTTCTAACAGCTTACAGTCAGATG TGGGAGCAAAGGAATTAAAGTTGAACATTATATTTAAAAAGGGAAGCAGAGCATAAAAACTTGGAAAATT CTCAACCTGGCCATGCGGTAAAGAAGGAAAGAGCATTTTCGGGGGAAGAATTTAAGAGGGCTGCAGAACC ACCACTTGCTGAAGAGACTAACGTGACTAAAAGGGAGCCAAGTGCTAAGATCCAAGACAAAAGGAAAAAG GCTTTGAAGGCATTTCGGGGATCTTCCAGGCCACCTCTCCCATCACAAACCCAGAAGCCTAGGAGAAAAG AATGATTTAGCGGGCCAGTTCCAGGGCCCCGCTGCCCTGTGCAGCCTTTGGACACCATTCTCCACGTTCC AGCCACTCAAGGCTCCAGCCTTGGCTCAAAGGGGCCCAGGTATAGCTTGGGTTGCAGCTCTGCAGGTTGC AAGCTGTAAGCTTTGGTGGTTCCAGGTAGTATTAAGCCTGCAGATGCTCAGATTGCAAGCATGAAGGAGG CTTAGTGGCTTCCACCTAGATTTCAAAGGATGTATGGAAAAGCCACGGTGCCCAGGAAGGAGACTGACAT TGGGGCATAGCTCCCACAGAGGAACTCTATTAGGATTGGATTCCCCCGTACCCCCACAAGAATCCCTACC AGGGCACTGCCTAGTGGAGCTGGAGGGCCACCTCCCTCCAGACCCCAGAATGGTAGATTCACTGGCAGCT TGTGCCCTCTGCCTGAAAAACCTGCAGGCACTCAACTCCAACCCATAAGAGCAGCCATGGGGGTTGCACC CTGCAAAGCCACAGGGCTGGAGCTGCCCAAAGCCTTGAGATCCCATATCTTGCACTAGTGTGTCTTGACG CAGGACATGGAGTCAAAGGAGATTTTGGAGCTTTAAGATTTAATGACTGCTCTTCTGGGTTTACAACTTG CATGAGGTCTGTTGCCTTTTTCTTTTGGCCTGTGTCTCCCTTTTGAAATGAGAATGTTTAACCAGTGCCC ATACCACCATCTTCTCTTGGAAGTAAATAACATGTCTTTGATTTCACAGGCTCACGGGGAAGGAATTCAT CTTCAGATGAGACTTTTGACTTGGGACTTGGGACTTTTGATTGAGTTGATGGTGGAACAACTTATGACTT TGGGGAGGTATTTAGAAGGGATGATTGTATTTTGCAATTTGAGAAGGTATTGAGATTTAAGGGGCAGGGG CAGAATAATATAGTTTAGATGTTTGTCTCCTCCAAATCTCTTGTTGAAATGTAATCCCCAATATTGGAGA TGGGGACTGGTGGGACGTATTCGGATCATGGGGGCAAATCCCTCATGAATGGCTTAGTGTCATCCCCATG GTGCTGTCAGTTCTCACTCTAGTAGTTCATGCGAGATCTGGTTGTTTAAAAGAGCATGCAATCTCCCCTG CCTGCTCGCTCCCATTCTCATCATGTGATGTTCCTGCTCCTGCTTCACTTTTTGCCATGATTGTGAGCTT CCTGAGGCCCTCGCCAGAAGCTGAGCAGAGGCTGGTGCCATGCTTGTAGAGCCTGCAGAACCGTGAGCCC GTTAAACCTCTTTTCTTTATAAATTACCCAGTCTCAGGTATTCCTTTTTAGTAATGCAAAAACAGACTAA CACAGTACATCTCTCCCCTAAGGTGCTTGGTATTTGCCTACATTTATGAAGAACATAGCATTAAATACAG CACCAAATAGGTGCTCAATATTTTCACGGAAAGAATTAATGCACATGTGTAAGAATCAAGCAGCACAGTC ACCTAATGAGGAATAGCCAAATTTTTAAACAAAGTTTTTCTTTTTGAGACAAAATTTCACTCTTGTTGCC CAGGCTGGAGTGCAATGGCATGATGTCGGCTCACTGCAACCTCCGCCTCCTGGGTTCAAGCAATTCTCCT GCCTCAGCCTCCAGAGTAGCTGGGATTACAGGCTCCCGCCACCACACCCAACTAATTTTTGTATTTTTAG TAGAGATGGAGCTTCACCATGTTGGCCAGGCTGGTCTCGAGCCCCTGACCTCAGGTGATCCACCCGCCTC GGCCTCCCAAAGTGCTGGGATTACAGGTGTGAGCCACTGCGCCCGGCTTAAACAAACTTTTTAATGGAGA AAATGTCATCCAATAAATCAACCAAGTGTGACCCATCCTCGTGTGAGCCATCCTAGAACATCACCCACTG TGGACCAGGTTGCCAGTGTTATAACCATGTGCTCCCCAAGTGAATCAGTAAGAGAAGCTATGTGGGGAAG AGCAATGTATGATCTTACTGTGCTAGGTTTGAAGGCATCAGGCACAGCCCTTCTAAAGTGTGTGTATTTC TCAGACCTAGCTCTTCTCATTCTGATCTTTTTGAACCATTTCTTGTTTTGTCCCCAGACCTGCTAGAGGC AAAGCATCTTCTGGGTCAGCTCATTAAGATAGGCAAGGACAAGAAGAGTCTTGAGTGGGACCTAGACAGT GGCACCTGTTGACTGAAAAGGCAGTACACCAAAAGATGGAGACACCCTGCAGGGTGTTGATGATGGTGAA GAGATAGGCAAGAGCATAGCAACTGGGCCCACCTGCAGAATGCCCAAACACCAGGTGCAGCCCAAGATGA ACAGCTGATCTGTCACCTTGAATGTCAGCATCCTGGATTGGATGAGAAAAGAGGCCCATAATATACCCAT AATAACAAAGGCTATTTCATCTCCATCCCATGGATATCTCTGCCCTCTCCTGTTGGTCATTTCTCACACT TGAGTTTTGATCAACCTAGATCTTTACTGTTGATAATTCCCCACAAAAGAAATGCCATTTGGATCCAGGA AAGCATTTTAACTTGGACCAAGGAGAGTAGTGACACTTCATAATATGGGTTGCAAAAGAAGTGTGATGAT CAACACAAAAAAACAGTTGCATTTCTATGCACTAACAATGAACAACCCAAAAAGGAAATTGAGAAAACAA ATCCATTTACAACAGCATTAAAAACAATAAAATACTTTGGAATTAACTTAACCAATGAGATGAAGAACCT GTACACTGAAAGCTAAAAAGCATTGCTGAAAGAACTTAAAAGAGACACAAATAGACATACTATGTTCACG GGCTGAATGACTTAATATTTTTGCAGAAACAGAAAAATATCCATCCTAAAACTCATATGAAATCTCAAGA ATACCAAACCAAGAAGTCAAAACCATCTTGAAAAAGAACAGAGTTGGAGGTCTCACACCACACTTATTTC AAAATTTATTGTAAAACTACAGTAATCAACACAATACGGTAATGGCATAAAGATAAGACATATAGACCAA TGGAATCGAGGAGGGAGCCCAGAAATAAAACCTCACATTTATAGTCAAATGAATTTTGACAAGGGTGCCA AGACCATTCAATGGGGAAAGGACAGTCTTCAACAAATGGTGCTGGGAAAACTGGATATGCACATGCAAAA GAATGAAGTTGGACTCTTTTCTTACTCCATATATAAAAATTAACTTGGCTGGGCATGGTGGCTCACACCT GTAATCCCAGCACTTTGGGAGGCTAAGGCAGGCAGATCACAAGGTCAGGAGATCAAGACCATCCTGGCTA ACATGGTGAAACCCCATCTCTACTAAAAATACAAAAAAATTAGCCAGGCATGGTGGTATGCGCCTGTAGT CCCAGCTACTCAGGAGGCTGAGGCAGGAGAATCACTTGAACCGGGGAGGCGGAGGTTGCGGTAAGCCAAG ATTGCGCCATTGCACTCCAGCCTGGGCAACACAGCAAGACTCCGTCTCAAAAACACAAAAACGAACAAAC AAACAAACAGAAATTAACTCATAATACATCAAAGACCTAAATGCAAGAGCTGAAACAATAAGACTCTTAG AAGAAAACATAGGGGGTAAGCCTCACGACATTAGATTTGCAATGATTTTTTGGATATGACATCTAAAGAA AACAAAAAATATCTATAAGTTGGACTACACCAAAATCTTAGACTTCTGCATATCAAAGGATACAATTAAT AAAGTAAAAAGGTAACCCCTAGAATGGGAGAAAATATTTGCAAATCATATATCTGATAAGGGGTTAATAT TCAGAATATATAAAGAACTTCCACAACTCAACAACAACAAAAAACTAATTTAAAAATGGTCAAAGGACTT AGACATTTTTCCACAGATGACATACAAATGGTCACAACTAAGAAGGAAAAGCCCAACGTGTGTTCTTCCA GCCTCTGCTTCTGTCATGTTTGTCAACACTCCATTGGCCAAAGAAAGTCACATGACCAAGCTCGTCTTCA AGGTTTGGAGAAATAAACTCCACCTCTTGATAGGAGGAGCTGCAAAGTCACATGGAAAAGACATATACAT AGAGGGATAGAAGAAGTCATTGTGGCAACCATCTACCACTGCATCATTACTT

[0192] The present disclosure provides several CD5 target sites and corresponding gRNAs that are useful for targeting RNA-guided nucleases to human CD5. Table 12 below illustrates preferred target domains in the human endogenous CD5 gene that can be bound by the gRNAs described herein. The exemplary target sequences of human CD5 shown in Table 12 are, in some embodiments, for use with base editors, such as ABEs. [Table 12-1] [Table 12-2] [Table 12-3] [Table 12-4] [Table 12-5]

[0193] A representative DNA sequence of the CD5 gene is provided by NCBI gene ID: 921 and is shown below. GAGTCTTGCTGATGCTCCCGGCTGAATAAACCCCTTCCTTCTTTAACTTGGTGTCTGAGGGGTTTTGTCT GTGGCTTGTCCTGCTACATTTCTTGGTTCCCTGACCAGGAAGCAAAGTGATTAACGGACAGTTGAGGCAG CCCCTTAGGCAGCTTAGGCCTGCCTTGTGGAGCATCCCCGCGGGAACTCTGGCCAGCTTGAGCGACACG GATCCTCAGAGCGCTCCCAGGTAGGCAATTGCCCCAGTGGAATGCCTCGTCAGAGCAGTGCATGGCAGGC CCCTGTGGAGGATCAACGCAGTGGCTGAACACAGGGAAGGAACTGGCACTTGGAGTCCGGACAACTGAAA CTTGGTAAGACTAGTCCTTGGAACTTGCCCACTCCATTTGAGTGGAAGCATGGCCCAATCACCCATGGCA TGCCTTTATCAGCACTTTGGTTTTGGTTTTGGTTTTGACTTGGTTTGAATCGCTTGACAGGACTGGTCTT GGGAACTTGCCTACTCCATTTGAGTAGAAGCGTGTCCTGATCACCCACAGTGTGCCTGTACCAGCACTTT GGTTTTTGTTTTTGACTTGTCTTGGATTGCTTGATACTTTGGTTTTGGTTTTGACCTGGCTTGGATTTCT TGATACTCTGATTTTGGTTTTGATTCTGGCTTGGCGTAAACTGTAAAAGTGTGTGTGTGCCCTTTTTACC CGTTCTTTGTTTTGTGGTGTGAGCATGGTGTTTTGTCTCGAGGAAACATGGGTCAGACACAAAGTAAGCC TACCCCACTAGGAACTATGTTGAAAAATTTCAAAAAGGGATTTAAGGGAGACTCTGGAGTCACTATGACA CCAGGAAAACTTAGAGCTTTGTGTGAGATAGACTGGCCAGCATTAGAGGTGGGTTGGCCATCGGAAGGAA GCCTGGACAGGTCCCTTGTCTCGAAGGTATGGCACAGGGTAACCTGTAAGCCAGGGCACCCAGATCAGTT CTCACATATAGATTCTTGGTTACAGCTAGTTTTGGACCCCCCGCAGTGGTTAAGAGGACAGGCAGCAGCA GTACTAGTAGCAAAGGACAGTTAGTTAAGGAAGGTTTCTCGATCCACCCGCCGAGGGAAGTCAGCACCA AAAGTCCTGTCCAACCCAAACACCAGAAGAGTCACAGCAGGAATTGGTACCAGCAGTATCCCCTCCTTATC AAGAGGAAGGGCTCCCCACTCCTGAGCCCACAGCACCTACAACTCCCACCAGATAACCACACCCCAGACCA CCCAGAGTAGAAAAGGGAAGTGAAGCCACGGGAGAAACTCCTCCCTTGAAAGCTCGCTTACAGCCCA AGACTGGAATCCAAATGCCCCTGAGAGAGCAGTGATATACTGGGGTAGATGAGGACGGACACATGGTGGA AAGGCGTGCCTTTGTGTATCAACCTTTTACCTCTGCTGACCTCCTCAATTGGAAAAATAATACTCCATCT TACACTGAAAAGCCTCAAGCTTTAATTGACTTGCTCCAAACTATTATACAGACTCATAATCCTACTTGGG CTGACTGCCACCAGCTGCTCATGTACCTCTTTTAATACAGGTGAAAGGTGAAGGGTGCTCCAGGCGGCAAC TAAGTGGCTAGAGAAGCACGTCCCAGCCAATTACCAAAACCCCCAAGAATATATAAGAATTCAGCTGCCA GGAACAGACCCCCAATAGGACCTGAACGAGGGACCAGACATGGAGAGGCTAAGACGGTACCGTGAGGCAT TAATAGAAGGTCTAAGGAAAGGGGCTCAAAAGGCTACAAATGTAAATAAGGTCTCTGAGGTCCATCCAAGG AAAAGAGGAGAGTCCAGCACAATTCTATGAAAGACTGTGTGAGGCTTACTGTATGTACACTCCTTTTGAT CCAGATAGCCCTGAGAATCAGCCCATGATTAACATGGCCTTACTTAGTCAAAGCACGGAATATATCAGGA GAAAATTGCAGAAACAGGCTGGGTTTGCGGGTATGAATACCTCACAGTTACTGGGGTTTGCGGGTATGAA TACCTCACAGTTACTGGAAATAGTCAATCAAGTGTTTGAGAATGCAACAAGCCACAGAGAAAGCCGTAAG GAAGGCGAACGCCAGGCTAGGCGAAACGCCGACTTACTGGCCATGGCCATTATGGGAATTCCCCCGAAAG GAGAGGGAAAGGGGGGTTCTGGGAAGAATACCCAGTCTAATCTCCCATGCTTGCAATGTAACCAATGCGC CTGTTGTAAGGAAGTAGGGCATTGGAAAGATAAGTGTCCCCAACTGAAGGAAAAGCAAGGTGATTCAGAA CAAAAGACCTCAGATAAAGATGAGGGAGCTTTGTTCAATCTGGCTGAAGGGCTACTGGACTGAAGGGGAC CGGGCGCAAGCACCCCCAAGGAGCCCAAGGTCAGGATTACAACTGGGGGCAAGGACGTTAAGTTTTTGGT TGATAGTGGTGCTGAACCTTCAGTAGTGACCACCCCGGTCACCCCCTCATCCAAGAAAACCATTGATATA ATCAGAGCAACAGGAGTTTCCACTAAGCAGGCTTTCTGTCTACCACGGACCTGCTCGGTGGGGGGACATG AGATAGCTCACCAGTTCTTGTACATGCCTGACTGTCCCTTGCCCTTGCTGGGAAGAGACTTGCTTAGCAA GTTGAGAACCACCATCTCCTTTACAAAGCAGGGCTCTTTACAGCTAAAGTTACCAGCAACAGGAGTTATC ATGGCCCTTACGGTCCCCCGGGAAGAAGAATGAAGACTTTTTCTAACCAAGCCAGGCCAAGAGATAAAAC CAGCTCTAGCTAAGCAATGGCCCCAAGTATGGACAGAGGATAATCCTCCAGGACTGGTGGTCAGCCAAGC CCCTGTACTCATAGAAGTTAAGCCTGGGGCCCAACCAATTAGACAAAAGCAGTATCTGGTTCCCAGAGAA GCTCTCGAAGGAATCCAGGTTCCTCTCAGGCGCTTGACAGCCTATGGAATTATAGTTCCTTGCCAGTCTC CACGGAACACCCCCCTCCTACCTGTCCCTAAACCAGGGACCAAGGACTACTGGCCAGTACAGGACTTGGT CCGAGGAGAAGTTTACAAACCCTCTCCTGAGCCAGTGTTTCAAACTTTCTAAGATGAACTCAATGTGCCA GTACCACAAATTCCAGGAAAAACAAGAAATTTGTTTTTGCAATTAGCCGAGCACGTAGCCCAGTCTCTCA ATGTCACTTCATGTTATGTATGCGGAGGAACTGTAATGGGAGGTCAATGGCCATGGGAAGCCCAAGAATT ACTACCTACAGACCCAGTTCCTGATGAATTCTTGGCTCAAAAGAATCACCCTGATAATTTCTGGGTCCCA AAAGCCTCAATTATTGGACAATATTGCATAGCTAGAGAAGGAAAAGAATTCACTCACCCCGTAGGACGAC TTAGTTGTCTGGGACAGAAACTGTATAATGGTACCACAGAAACAGTCACTTGGTGGAGTTCAAATCACAT AGAGAGGAATCCATTTAGTAAATTCCCAAAGTTGCAAACCGTGTGGACCCACCCGGAGTCCCACCAGGAC TGGACAGCCCCCACTGGATTATACTGGATATGTGGGCATAGAGCTTACGCCAAATTACCTGACCACTGGG CAGGTAGTTGTGTTATTGGCACTATTAAACCATCTTTCTTCCAACTGCCCATAAAAACAGGTGAACTCCT GGGCTTCCCTGTCTATGCTTCCCGCAAAAAGAGAAGCATAGCTATAGAAAATTGGAAAGATGATGAATGG CCGCCCCCTGAGAGAATCATACAATATTATGGGCCTGTTACTTGGGCACAAGACGGCTTGTGGGGATACC AGACCCCCATTTACGTGCTCAACCGAATCATACGGTTACAAGCTGCCTTAGAAATAATCACTGATAAAAT CAGCAGAGCCTTGACTATTCTGGCCTGGCAAGAAACTCAGATGAGAAATGCTATCTCTCAAAATAGATTG GCTCTCGACTACTTGCTAGCAGCTGAAGGAGGGGTCTATGGGAAATTTAACCTTACTAATTGCTGTCTAC ACGTAGATGATCAAGGACAAGTAGTTGAAGATATAGTTAGAGATAGGACAAAACTGGCACATGTGCCCGT GCAAGTGTGGCATGAATTTGATCCTGGGGCCATGTTTGGAAAATGGTTCCCAGCACTAGGAGGATTTAAA ACTCTTATAATAGGAGTTTTAATAGTAATAGGAACCTGCTTACTGCTCCCTTGTTTGCTACCTGTACTTC TTCAAATGATAAAAAGCTTCATCGCTACCTTAGTTCACCAAAATGCTTCAGCCCAAGTGTACTATGTGAA TCACTATTGATCTGTCTTGCAAGAAGACATGGTAGTGAAAATGAAAGTGAGAACTCCCACTCATGAGTGA GATTCTCAAAAGAGGGGAATAAGGGAGGAGACCACCCCTCATACTGTCTTATGCCCAATTTCTGCCTCCA AAGAAAGAAGAAGTAAAAACTAAAAGGCAGAAATGAAATCTGCTCGGCACCACACCTGGGCTTGGTAGTT AAAGATCGACCCCTGACCTAATCTGTTATGTTATCTATAGATTACAGACATTTTATAGAAAAGCACTGTG AAAATCCCTGTCCTATTCTGTTCCATTCTAATTATCAGTGCATGCAGCCCCCAGTCATGTACCCCCTGCT TGCTCAATCGATCATGACCCTCTCAAGCAGACTCCCTTAGAGTTGTGAGCCCTTAAAAGGGACAGGAATT GCTCACTCAGAGAGCTCAGCTCTTGAGACAGGAGTCTTGCTGATGCTCCCGGCTGAATAAACCCCTTCTT TCTTTAACTTGGTGTCTGAGGGGTTTTGTCTGTGGCTTGTCCTGCTACAGGTCTAAGTGGAATCTAAGTG GAGGCATCACTGAGCCAGCCTCTCCAGCCCAGGGATGGGGAGGCCCCAGGCCCTAGTGGACCTGGACCAG TGATCACGTCCTTGAACTACTTCCTACCTGCCTGGGGTGGCCCAGAGTACAGGAAGCCCAAGGAGCCAGG AGCTTTCTCATCTGGGAGCACAAAGGATGTGAGGAAAAAACGGAACCCAGCCCAGCCCAGGGAAACACAG GGACTTTCCCCTGCCCCGGGCTTCCCATCTTCTTTGCAGAGACCCTGCCTCTTCTTCAAGGGGGATGATT CCAGACCCTCTAAGAGCAACAGGGACACGGAGCTCACATACATACCCCACATCAACATGGAGATCACACA CCTCAACACGGAGATCACACACATCAACATGGAAATCACACACACATATCGAATGGGGATTACACACATC AACGTGGAGCTCATACATACACCCCAACATGGAGATCACACCCACAACACAGAGATCACACAAGTCAACA TGGAGATCACACACACACATCAACATGGAAATCACACACACACATCAACATGGAAATCACACACATCAAC ATGGAGATCACACACACACATCAACATGGAGATTACACATACATCAACATGGAGATCACACACACATCAA CATGGAGATCACACACACACATCAACATGGAGATCACATTCGCACACATCAACATGGAGATCACACACAC ACATCAACATGGAGATTCACACATATCAACATGGAGATCACATTCACACACATCAACATGGAGATCACAT TCACACACATCAACATGGAGATCACAAACACACATCAACATGGAGATCAAATTCACACATCAACATGGAG ATTACACACACATCAACATGGAGATCACATTCACACACATCTACATGGAGATCACATTCACAACACAAAC ATGGAGATCACACACACATATATCAAATGGGGATTACACACACATCAACCCCAACATGGAGATCACACAC ACAGCACAGAGATCACACATGTCAACATGGAGATCATACACACACATCAACATGGAGATCACACACACAT CAACATGGAGATCACACACACATCAACATGGAGATCACACACACTCGTCAACATGGAGATCACATTCACA CACATCAACATGGAGATCACACACACACATCAACATGGAGATCACACACACACGACATGGAGATCACACA CACACATCAACATGGAGATCACTCACACACATCAACATGGAGATCACACACACATCAACATGGAGATCAC ATTCACACACATCAACATGGAGATCACACACACACAACATGGAGATTACACACACACATCAACATGGAGA TCACACACACATCAACATGGAGATCACACACACACAACATGGAGATCACACACACACATCAACATGGAGA TCACACACACATCAACATGGAGATCACACATACATCAACATGGAGATCACACACACACGACATGGAGATC ACACACACACATCAACATGGAGATCACTCACACACATCAACATGGAGATCACACACACACATCAACATGG AGATCACATTCACACACATCAACATGGAGATCACACACACACAACATGGAGATTACACACACACATCAAC ATGGAGATTACACACACAACATGGAGATCACATTCACACATATCAACATGGAGATCACATTCACACACAT AAACATGGAGATCACACACACACATCAACATGGAGATCACACACACATCAACATGGAGATCACACACACA ACATGGAGATCACACACACACATCAACATGGAGATTACACACACATCAACATGGAGATCACATTCACACA CATCAACATGGAGATCACAAACACACATCAACATGGAGATCACACACACACATCAACATGGAGATCACAT TCACACACATCAACATGGAGATCACAAACACACATCAACATGGAGATCACACACACACATCAACATGGAG ATTACACACACATCAACATGGAGATCACATTCACACACATCAACATGGAGATCACACACACATCAACATG GAGATCACACACACATCAACATGGAGATCACACACACACATCAACATGGAGATTACACACACATCAACAT GGAGATCACATTCACACACATCAACATGGAGATCACACACACACATCAACATGGAGATCACACACACACA TCAACATGGAGATTACACACACATCAACATGGAGATCACATTCGCACACATCAACATGGAGATCACACAC ACACAACATGGAGATCACACACACACATCAACATGGAGATTACACACACATCAACATGGAGATCACACAC TCATCCACATGGAGATCACACACACACATCAACATGGAGATCACACACACATCAACATGGAGATCACATT CACACACATCAACATGGAGATCACAAACACACATCAACATGGAGATCACACACACACATCAACATGGAGA TTACACACACATCAACATGGAGATCACATTCACACACATCAACATGGAGATCACACACACACATCAACAT GGAGATCACACACACACGTCAACATGGAGATCACATTCACACACATCAACATGGAGATCACATTCACACA CACAAACATGGAGATCACACACACATATATCAAATGGGGATTACACACACATCAACCCCAACATGGAGAT CACACACACAACACAGAGATCACACAAGTCAACATGGAGATCATACACACACATCAACATGGAAATCACA AACACACAACATGGAAATCACACACACATATCAACATGGAGATCACACACACACATCAACATGGAGATCA CACACACATCAACATGGCGATCACAATCACACACATCAACATGGAGATCACACACAACACAGAAATCACA CACGTCAACATGGAGATTACACACACATCAACATGGAGTTCACACACACATCAACATGGAGATCACACAC ACTCAACATGGAGCTCACACATACACCTCAACATGGAGATCACACAACACAGAGATGACACACACAACAC AGAGATCACACAAGTCAACATGGAGATCTCTCTCTCTCTACACACACACACACACACACACACACACACA CACACACACACTGGCACAAGCCCAGCATTCATGGCGCAGCCATCTCTTGTACTTGCTAGCAAAACACCAC CTGTGGAAAGTCAGCTCAAACTTCCAACCTCATCCCTCACAAAGGGGGAATGGAGGGTTTGGTTTTGGCT TTCACTGGAGTCTGCAACAAGAACTGGCATCATGCTGCCCATTTCCCGCCTCTCCCCACCCAGACCCCTG CCTCAGGGACGCCTGTCCTCAGCCCAGCCCTCAGCTGCAGCCAGGCCTTCAGCCTCCGTAACCCCCGCTC AGGGTCCCCACCCCCTGCAGCCCTGTCCCTCCAGGATGCATGGCCTTGTCCTGTGTGGGGGTGGCCGAGA GCACTGCCCCAGCCCTGGGTACCTTGGGCAGGAAGCTGGCAGAGGCCAGGGCTGCCATTCAAACAGGGGC AGGTGGTTTTGCCAGGAGGAAGTTGACAGTTCAACTTCAAACATGGGTGACGCAGGCCCCACACTGCCTG CTCCCCGTCCCACCCCTCCCTGAGCACGCCACCCCGCCCTCTCCCTCTCTGAGAGCGAGATACCCGGCCA GACACCCTCACCTGCGGTGCCCAGCTGCCCAGGCTGAGGCAAGAGAAGGCCAGAAACCATGCCCATGGGG TCTCTGCAACCGCTGGCCACCTTGTACCTGCTGGGGATGCTGGGTGAGTACCCCTCCCAGGTGTCCTGCG AACACCCGGGCTCGCTCCAGTGCAAGGAAGGAGTTCCCAGTTTTACCCAAGGCTGACTCTGGGATCCACA TGTCAGCCCTCTGGAGCGTTGTGGAGATTTGGGGCCACTGGGATCCCTGCCTGCCCCCACTAAGCCGCAG CTTGGCCCTCTGTCCTGCATGTCCCACCCGCCAGGAGCACAACCTTGCCTCTCTCATGCGCTGTTGAGAA CCCTGCTTTACCCTTCCAGTGCAAGAGAGACTGCAGGGGGGACCCGCATTTGATGGGGCCCAGACAACTT GATTCCTAGGCTGAGTTGGATTTTAGCAGAGCATTCAGGCCTCCCTCTGCGAGGTCCCCCACTGACAGCC CAGCCTTTACTTGGTCGCCTCCAGAGACATGGAAACTCGCCGTCTCCGAGGCAGCTCTGATGATGCTCTG GACAGACAGTGGGCCGAGGCAGGGTGGAGCCGCCTCTGAGGACGATGTCTGAGCAAATGTCAAGGACTCA TAGCGGAAGTCCTAGGCCCTCTCATGCCCAGTCCTGGGTCAGTCCCAGAGGGGCCCAGCCAGGCCGGCTG GGATGAGTACACGCTGGGCCTGCTCTCACTCCAGCCCTGGGTTGTAAGCCCAACCCTGTGTGACCTCGGA TAGGTCCCCAAGCCTCTCCAGGCCTTGATCTCCCCATTTATGAAATGTGGGGGTTGGGGGTGTGAACTGG ATGAGGCCTGACCCACTTCCTGAGGATTTAGAAGGTCTGGCAGGGTGACAGAGGTGCGTGACTCTGGGCT GGAAACAGGAGGATGCGAGCAGCTTCCAGGCTGGCAGGGAGAGGGTCCGGAGGCCTTGCCTGCCTGAGCA GGGAGCCTCAGCAGCCTTCTAGGGAAACCGGGTGTGCAGGTGTGAGTGTGGGGGAATGCATGTGTGTGTG TCTGTGTGCATGTGTGTGAGAACTGTGTGTGGGGGGAATGTGTGAGTCTATGTGTGTCTGTGTGAGTCTGT GTGTACGTGTGTGAGAACTGTGTTGGGGGGTAATGTGTGACTCTGTGTGTGAGTCTGTGTGTGAGTCTCT GGGCATGTGTGTGAGTACTGTGTGTAGGGGAATGTGTGAGTCTGTGTGTGAGTCTGTGTGTGAGTCTCTG GGCATGTGTGTGAGTACTGTCTGGGGGGGAATGTGTGAGTCTGTGTGTGAGTCTCTGGGCATGTGTGTGA GTACTGTGTGGGGGGGAATGTGTGAGTCTGTGTGTGAGTCTGTGTGTGAGTCTCTGGGCATGTGTGTGAG TACTGTGTGTGGGGGGGGAATGTGTGACTCTGTGTGTGAGTCTCTGGGCATGTGTGTGAGTATTTTGTGG GGGGGAAAGTGTGAGTCTGTGCGTGTGAGTCTGTGTGTGAGTCTGTGTTCATGTGTGTGAGAACTGTGTA TGGGGGAATGTGTGAGTCTGTGTGTGAGTCTGTGTGTGAGTCTCTGGGCATGTGTGTGAGTACTGTGTGG GGGGAATGTGTGAGTCTGTGTGTGAGTCTGTGTGTGAGTCTGTGTGTGGGAGTCTGTGCGCATGTGTGTG AGAACTGTGTGTGTGTGCTTTCATGAGCACAATGCGGCCTCCAAAGGCAGGATTATCTCCCCAACACATG TGACAATGGCACCTGTGCACACATGTTCACATGGTTCTTCTCACATGTAACAACAACAGGTACAAAATGC CTTCTGTCCCAAAGTAAGAGGCCTGAGACAGACCTCTGGCTGGCTTCAGGGACCCTGCTAAGGAGGGACC AGGCCCTTAAACCCCCCACCTGCTCCTCACCTCTGGCCTCAGACTTCATTGAGATAGGGCGGCTCCCAGC TGAGTATGGGAACCCGTGTCCTGCTCTAGGGCCCTCCCAACTGAATCCCATCATGGGTGAGGCTCCTGAT GGCACCAGGCAAGTCCCCTGGCCCAACACAGGCCACAGTCAGGAAGACTTGGAGGTAGCCCCTTCACCCC AGTATGGGGACCTCCTTGAACCCCAGGAATACTCAGTTCCCAGTGTCTGAGTGAGCCGGTGGGTACCAAG CATTCCCTTTGTTCAGTGGGTAACGGTTTACAAAGCACTTTTAAAGCCATTTTCTGCTGCAAGCTTCACA ACAGCCAATGAAGTAGGTGGGCCCATTGGCCCCATTTTACAGTGGGGAAAAGCGAGGCCCAGGAAGGCCG AGTGCAAACCCAGGGTGCAGCAGCTGGTGCACAGCCGAGCTGACCCCAGCCCAGCATTCCTGCCTTCACA GAGGCCCCTCCCTTAGCTAAAAGGGAAGGCCACCAAAGGCACTTCCGTGTGCAGGAAAATGGCCCTGGGA CACAACCTCCACCAGCAGAGATACAGCCACCAGAGCTGGGGACACCAAGGGCCAGGGTTGCTGGAGTCAA ACAGTGCAGGCACCCTTTGAGCCATTCCAAGGAGAGAGGCAGCAATGCCAGAGCCCTGGGGACACCACAG CCTCCTGGCTGGCATCGGGGCTTGGGGACCTTTGGATCCTGAGGCATGAAGATGCCCCCAGCAGGCCTGG GTGCTAGTGACAGGCCAGGGCTTCCCACAGTTGCTCCCCTCTCAGCCCCAGCCTGCCTGGACAGCTGCTG GCAGTGAGGCGGGGAAGCGGCTCTAACACAAACTGCACGTCCCTGACTGGCCTAGCCGATGCGACTCTGA AAGCCAGCACTGGGACGTGGGAAAGTCATGGACTCTGCAGTTTGAATTCCAGTCTGCCACTTACCAACTG GGCGACCTTGGACTAGTTCTGTAAGCCTCAGTTTTCTCATCCGTAAACTGGGTGCAACAGTACCTCATAG AGGCTTATAAGGATTCAATGAATCAATGCACACAGAGCTTAAGTACAATCCTGGCATATAGTATAGTAAG TACTCAATAAACAATAATAACAGCAACAACTATGATTATTATTATTACCAGGGTCAGCAAGAAACAATGC GAGGTAGGAGCCAATGGGGAGGAGGGTCTTGCCAGAGGCCTGCACAGCCGTCAATCTCTCTACCCCTCAA TCCTCTTGCACAGGCTATTTATTCACTAATTCATTCATTTTTCATCTACAAATATTATTGAGCATCTCCT GTGCATCCAGCTCTGTGCAAAACACTGGGGATACAGGGACCAAGGCAGGAGTCCCTGCCCTCAAGGATTT GATCAGCTAACAGGAGAGCTGTTAGAGAACACAAGTAAACAAGCACCAAAGACCGGGCATGGTGGCTCAT GCCTGTAATCCCAGCACTTCGGGAGGCCAAGACGGGTGGATCACCTGAGGTCAGAAGTTCAAGACCAGCC TGGCCAACATGGTGAAACCTCATCTCTACTAAAAATACAAAAATTAGCCGGGAGTGGTGGCACGCACCTG TAATGCCAGCTACTCAGGAGGCTGAGGCAGGAGAACCGCTTGAACCTGGGAGGCGGAGGTTGCGGTGAGC CGAGATCGCGCCATTGCACTCCAGCCTGGGCAACAGAGCAAGACTCCATCTCAAAAAAAAAAAAAAAAAA AAGGCACCCGAAATATTTACACATTTTGAAATACTGAGTGCAATGAGGAGAACAGCAAGTTCCTCAGGCA CAATACAGTCTGCACTAGGATAGGAGGACATCCCTTTGGCTGGGCAGGCAGGGAGGGCCTCTCTAAGGAG GCGAGGTCTCATCTGAGAATCCGAGACCTGAATGAGGGACGGAGCCAGCCAGGCAGAGAAAAAAGGGAAG AGGGTCCCAGGCAGAGCGGATGGCAGGGCAAAGGCAAGGGGGAAGGACAGGCCTGAGAGGGTGGGGCTCC TACTGGGAGCAGGGCAGCAGGCCCACTGTCCCAGGCTGGGCGAGGCCCTGGTTGGATCATCCTCCTCGAA GACCCAGAGCTCCCCACCTCCAGCAGGGAATGAAGGGACCTGGGGGCCCAGGAAGGGACCTGGCCTTCCT GGGTCACTCTGTCACCATCAGTTCAGAGGGGTTTTCCCTGCAGTGACCAGGCGGCGAATTCAGCTCAACC TGACAAAATTGATTGAACATCTATTCTGTGTGTGTGTGTGAAATTGAACATCTACTGTGTGTGTGTGTGT CCATGTGCTGTGTTTGCGTATGAACACATATGTGCACAGCCCTGTGGAGGGGAGCTTTGGAGCAATGCAG ACCTAGTTTGATTTCTGCTTGGCCACCCACAAGTGAACATCTCTCAGGGTCTCATCTATAAAAATAGGGG TTCCTTGCTGTGTCCACCTCAAAAGATCACTGCAATAATTACATCGGGTTTATTCAGAGACTAGGTATGG ATTGAACACCTACTATGTGCCAGATGCTGTTCTAAGTGCTGAGGACACAGCAGTGAACAAAACGGTGGAA AATCCCTGCCCTGGTGGATCTGGCATAATAGTCAGGGTGTGATGGGGAGACAAACAAGACACAAGCTAAG CTAGTGATTATGCGAATACTAGACGGTCATGAGAGGTGTGGAGACAGACAGATAGAGGAGGGCTGGGACC GGAAGCCTTGGTGGGGTGTGAGGTACCTGGGGAGGATTCAGCGGCTCTCATGAGAAGATGATATTTGAGT CAATATTTGAAAGAAGGGAGTGAGCCCACCGTGTGGGACCTGAGGAGAAGCGTTGCAGGCAGAGGGGCCT CCAGCTGCAGGTCAGTGGGGGATCGATGAGGCAGCCACGAGGGCCACGGATGCCCCCGTGAGGACTTGGG CTTTTTCTCCCATGCACTGGACAGCCACACAGGAGCAGAGAGGTGAGCATAACTCTCACCTCTCCAGGTC CTCAAGTGCATCTTGAACCCCAGGCCAGATGCATTTGGAACTCCAGAAGCTTTCAGATTGTAGAAGGAAA TACTGTGTACATACTGTGTGCTCTTAACACACGCAGTGAGGCCTGGGGCAGCACCCTGCGTTCAACACAT TCCCATTTCTGCAGTGAAACATGAATTTGCACACTTAGAGGGATGTAAAGACAGCAAACACCCTCCCACC AAATGCATTAGAACAAGACTTTTAGATTCTGGATTTTGAATTGCAGATAAGAGACTGTGGGCCTGGATCA CTTCTTCTATAGCTACCACTATCACCGTGACCAGCATCATCATCACCATCATGTACCTTTCAAGGCCTGA CTTTCATGGTGTCCAGAGGGACCCTAATACTCCTCTCCGGGGGACAGATACGGTAGGAATAGCGTAGGAA ATGGGTGGTGGCAGTCTGCAGAGACGACAAGGGTCCCATGCACTGCCACAGATTCCCACAGAGACATCTG GAATGAACCCCTCCCTCCTTCTGGCCCCAGTTTCCCCCATCTGTACTCCAGAGCAAGCAGGCTCTGGGGC CTCCCTGGGGTAGACTCCCACAGGCCCTGTCAGGTGATGGAGACAGGGTTTCTCTCCTGACTCTACCACC AGCTAGCTGTGTGACCTTGGGCAAGTCACTCAGCCTCTCTGAACCTCGGTTTCCTGATCTGAAAATGGAG GTGAGCATGGTCCCTTCTTCACAGGGCTGTGGGTAGAAAGAACAAGTCTAGTGCATGAGAGAGCTTTGGG GTAGGTAAAGCTCTAAACAAAACCAAAGCCTTGTCTCTGTGACAGCAGAGACACCGTATATGGTGCGAAA CTCATGCAACAGCAGACTACTAGTGGGAGAGAGAAATAACAATTCAAACAGCAAAGGTCATTCTGCCCAC GACCCACTGTCCGCTCACTGGGACAGGCCCTGGGTGAGCCAGATTAGAGACCTGAATCTGCTGTTCCTTC CTGCGGGCTCCATTCCCAGGCCTCTCCTGGTGTAAGCCCAGTGTGACTCTATATGTGTTTTGCACATAAA AGCCGCCTAATCATAAACCAATCATTTCATCTGTGCTCAACCCAAGAAAGGGAATTTTGGTCCTTTACGA CCTTCGTTTCCCTAAAGCCCTGATACAGCTCCAACTCTACAGATGACTGTCGTTTCAGAAAAGAGGGGCT TCTGACCTTTCAAGTCAGCCAACATATAGGATTCGAGGAAGGAAAATCCAGAATCCTACCCAAAACACAG GCAGATCTATTGTATTTCATGGACATCGATGGCTGTAGCTATGGGGAGGTCTGTGGACTGTTAGAAGGAA TCACTGTTTACGGGAAAAAGGGAACACCTGACTTGCAAACTACACTAAATCAGTCATAGGTGTTTTATCT GTGGAACAAAGTGGTGATAGCACCCCGTCTTCTGAGGCTCTAGAAAGAAATGGGGTCTTCTGTGAAAAAT CCTTGTCCTGGGGTCTGCACAGGGTAAGGATGCTGTCATTTTATTATCATCGGTCATTATCACAGGCCTG TGAATCCGGGTGCCTGCCAAACCCTAAGCAGGCCACACCCTTGCCTCCATTTCCCAGGGTGCAGGGGCGG CAGGACCTAGGAGTGAGCCAGAGGCAGGGGCCAGGAGTGCAGGTTACAAAGCCAGCCGCCCTGGGCTGAA TTCTGGCCCTGCCGAGTTCTCACTCCCATGCACCACGACAGCCTCCTCCTCTGTAAACTGGGACCCATCA TGCAAGTCAACAAATATTCATCTGTAATTCCAAAGCCCAAAAAACATTTGTAAGTTTCTGGCAAATGCAT TTGGTGCAAATCCTGACCTGAACGAATGGGAGGCTATTTATAATCTTGACACATCCCCCTTAGTGTGAAA CTGCATATGTTTCGCTGCAGACACTGCCATGTGTTTGAAGCCAGGTCTCTCCACACACATGGCGACCGTA AGGTGCCAGCGAGGCGTGGACGAAACAGAAGTCAAGGGCCACCGCCACTGCCTGGGGCCTCCAGAGATGC AGCCAGTGGAGCCAGGATGTCCTATTTTTCAAAAACGCCTGGATCTTGAAATGTGTTTAAAAAGCAAAAC AAAATTAAAAGAAACCTTTTTTTTTTTAATTTGGCTCAAAAAATTTTTAAACACTGTGCAGAGCAAATAA AACTCATGGGATGCCGTAATGAGAGCCAAAGTGAGAATTATCCTGAATACTGTGGGGGCAAGAGGAATGA GAGGGACAGGGAGGAGGATATCCTCACCAAGACCTCAGGGCAGGCAGGGCCAAAAATATTGTCTCTGCCA ATGCAAAGACAAGAGACGGGAGCTCCAGGGGTGTGATAAGCCGTCTGACTTCTTTAGCCCTGAATTTTGC CTGGGTTTGCCCAGTCCAGGCCCTAAATTGTGGGAGCTGCAAGGGGAGGTGGGCAGGAGAAGGAAGGGGT CTTCAGAGAATTCTCATCCGTAGCCCCAGTTGCTCGGGGTGAGGGGTGGCTGGCGGATAAGGAAACGGCA TAGCACTTCCAGGCCTCGGGGGCCTCCTGGTCTGGAGTCTCGGGGATTCTGTCCACAGTTTTGGCAATAG CAGGAGCCTCAGGCTTCATGAGGCTAGGAAGGATGGTGCAGGAGTAAGCTGTAGAGCCCCCAGAGGGGGT CAGGGAGACCCTGAACACAGACCGCAGGGCTCAGGGGATGGAGGGTGCTGGTCTGACCCCTGTGAGAAGC AACTAAAAAATAGAAATAGGTTATGGACCCTCAAAACCTCAATGTTCATTCTGCTCCCTAGATTATAAGT GAAAAACAAACAAGCAAATAAAAAAAAAACCTGAACATATTCATGGACAGGCTGAAAGAATATACACAAC ACACTCAGGAAGCTTCCAAGTCAGCTGTACATCTCACCGCCCTCCCTCCACCAGCTGCTGAAATCCATCA GGAGCTAAAGATTATAGATCACAGGTGTTTGAACCAGGGGGAAGGTGAGGGACTGTGGAATCTAACTTCC TTACTTTACAGATTAAGAAACTGAGCCCAGAGAGGCTGAGTGGCTTGCCAAGGTCACACAGCACTTCATA GCTAAGTCTTCTCACTGCATGTTTTACTGACTGAACGGAATCACAAGTTCATTCCCTTTGATCAGGATTG CACACCAGGGTGGTGCAGGAATCAATGAAACCTGGGGCAGGCCATCATGGGCTTCCTGCTGGAGGGAAGG GAGGCAGTTTGGGCCTCACTGGCAAAAGCTCAAGACTCAGTGCCAGGCGGCTAGGTTTGAGTTCGGGCCC CAGCACTCATTAGCTGCTTGGACAAGTCATTTAACCCCTCTCAAGTTTCCTGGTCTGTAAAATCGGGACC AAAATAATAGCACCTACTTCATAGCACTGTGGTGAGGACTGGGAGAGTTAATGTGTAAAAGGCCCGGAAG AGTGAAAACTTTTAAAGTGTTGGTGATGGTGACAGTGATGGTGATGATGATGGTGACGGTGATGGTGATG ATGATGGTGACGGTGATAGAGATGGAAGGAGAGGAGAGGAACTGCTGCTCATCTGGGGACCAGACAGGCA CCCAGAGGAAGGTTCCAGGGCGACACTCTAGAATGCATAAATATCTTTTTTATTAAAGCATTTGTTAGAG CATTTTTTAAAGAATTTCACTTGGTTCTCCAGCTTGTTCAGTCTTTCCTGAGAATGTTCTCTGAGCAAGA CCTAGGCTGTTCTGTGTGGAATCTGCCAAGATTGTCAGGCCCAGTCCCTGTCCCCAGGCCTTCAGGGTCT GGCTGCACGAGCGCTTCCAGGTCAGACCCAGGTGGTGTTCATCTCTGCATCTCCAAAGCTGGCACTCTGG AGGAGCAATTGGGTGGCTGGGAGCAGAGAGCATGGGTTTTGGAGTCAGACACATGGTTCGACTCCACCTG CCAGCCATATTACCCGGGACAGGGCATTTGCCCTCTCCTGACATTTATACCAGGAACGTACTCAACTCTC GTGATGGGGCTGTGTGGAAGATTAACAACAATATCTGTGATGAACCATACCGTGGGCTAAGTATGCAGCA GGTGCTCAACAGTTGGCAGCTGCTGTTAATAATGAACGTTGAGTATGATGACTCGGGAAATAGAGATGCT CCTTGACCTACAGTGGGGTTGCCTCTCAATAAACCCATTGTAAGCTGAAAACATCCTAAGTCAAAAATTC ATTTAATACCCCGACAAACCCATTGCAAACTTGAAAAGTCGTAAGTTGAACCACCCTAAGTCTGGACCAT CTGCGCGAAGTTTCCAAGTTAGGAGAAGGAAGTGGTGAGCCCCATGGCTTGGCTCTCTGTGCTCCTGGAC AGGGAATCTTCCCACTGGGAAAGCTCCCCGAGGGTCTGCTGCCCTGGGCACAGCTTCCCCCAGCCCACCC TACTCCACAGATCAGTAGCACCAGCTCTGAGCACTAGGCCAGGAAGCACTGGGCTAAGCCCTTTACACCG ATTAGCTCATCTCATCTTCACAACGGCTCTAGAAAGCTAGTCCCATGCTCATCCTCATGCTATGAGTGAG GAAATCGTGGCTTGGAGGCACAGAGACAGGACAGACGTGCTCCCCAGACTCTATAGGCAGGAAGTGGCAG AGCTGATACTCAGAACTGGAGCAGTAGAAAGTCATCGGTGACTGAAAACCCATCCTGAGCTGTGTAGCAC TGAAGGGCAGATATTTGGACTCCCACTCCCCACAGGTGGCACTTCAAGCTGCCTTAGCCCTCGAGACTTA GCCAGAGGGATGACTCCTTCAAGGCAGACTGCCCTGCTCCTGATGGGCAGCTGGGATGAGAGGGAGGTGG GAATGAGAGGTCAGGAGTGAGAGGTGAGGGCAGGACAAAAGGGCCTCCTTCCTGATTTCTCTCGCTAGCT GGACATCTCCCTGATGGCCTCCTGAGGCTGCCCTGCCCACTGCAATAGATCAGATGATTGGCTGGGCGCG GTGGCTCATGCTTGTAATCCCAGCACTTTGGGAGGCTGAGGGGGGTGGATCACCTGAGGTCAGGAGTTCA AGACCAGCCTGGCCAACATGGTGAAACCTCATCTCTACTAAAAGTACAAAAATTAGTCGGGCGTGGTGGT GCATGTCTGTAGTCCCAGCTACTCGGGAGGTTGAGGCAGGAGAATTGCTTGAACCAGGGAGGCAGTGGTT ATAGTGAGCCAAGATCATGCCACTGCATTCCAGACTGGGCAAGAGAGGAAGACTCTGTTTCAAAAAAAAA AGAAAAAATTGATCAGACGATTGACAACTAGACCATCCTTTTCCACCCAGGCCAGAGCTCTGTACACTTC CCTGTTGGTGAGAATAACCTTCCCCTTCCAAGGCCCGTGACTCCTAGAGCAACAGCCACGAAAGGATCTT GCAGATTCAGGGGAAACTTATCACATGGCCATGATGACTTCATTACAAAGAGTCTTGATTCGGGGACAAG GATGAACTGTGGAGCACATCACCCCTTAAAATTCAACCGCCAGCTTGAGAACACTCAAGCAAGAACAGCT TTTTCAAAGCACACCTATTTTTTAAGAGACGGGGTCTTGCTATGTTGTCCAGGCTGGACTCGAACTCCTT GGCTCAAGTGATCTTCCTGCCTCAGCCTGCTGAGTAGCTGGGACTACGGCCTTGAGCCATTGGCCCAGCT AAACACACCTATTTTTTAACACCAAGCAAAAATTTCTCACAAACAATTTGAACACAGCCTTTCACTATTA ATATATAAAGCAGTCTCCTTGACCCAGTGCCGTAAATCCAAGAGTAAACCCAAGTCGGACAGATGGAAAT CAAAATACCGTACTTGCACCTAACTGATCACGCTAGGTGGTGAATACAGAGGTTAAGCTTTGAAGCCACG TGAGCTTGCTACTTCACCTCGTCCCCACCCAAATTAAACCTCTTCCCCAGCCACAGATGAGCTGCTCTGC TGAGAGCCCCGCAATGGGCAACTTTGCCCCCACTCCACAGTGAGACCAGGCTACAAGGAGGCAGAGCTTC CACGTGGTCCACATGGGGGAAGGCTGGTTCCCTGCAGACCCGTGTTCCCTGGGCCTGTCGCCTTGGCTCC CTTGTCAACTAGCTGGCTACACTACTGTGGGCAGGTCACGCCATCTCTTTGAGCTTGGTCTTCTCCAGAA ATGGGCAGCTGTTAAAGATAAGTGGCTGTCATTCTTATCCTGCCTTGGGAATGCTAGAAAGTAGCAAATG TTCTCTTGGTTCCCTTTCTTGCTCATTTCTTTCTTTCCTTTTTTTTTTCTTGTAAAGACAGAATCTCACT CTCTCACCCAGTCTGGAGTATAGTGGTGCAATCTCAGCTCACTGCAGCCTCCACCTCCCAGGTTCAAGCA ATTCTCCTGCCTCAGCCTCCAGAGGAGCTGGGATTACAGGCATGCATCACCACGCCGAGCTAATTTTTGT TATTTTTAGTAGAGACAGGGTTTCACCATATTGGCTAGGCCAGTCTCAAACTCCTGACCTCAAGTGATCC CCCCACCTCGGCCTCCCAAAGTGCTTGGATTATAGGCATGGGCCACCGTGCCCGGCCCCTTTCTGGATCA TTTCTCCATTCTCTAGAACTAAGGAGCTCAAACTTTTTTGAGTGGAAACATGCGGGACATTTTTACATTG CAACCCCCAACACACAAAAAGTGATCAGTGCATTCTGACATTCTCTTTTTTATTTATTTAAAACAAAACA AACAAAAAAAAACAGGGTCTTACTCTGTCACCCAGCTAGAGTGCTGTGGCATAATCATAGCTCACTGCAG GGTCCAACTTCTGGGCTCAAGCGGTCCTCCCACCTTGGCCCCCTTAAGTGCTGGGATTGCAGTCATAAGC CACCGTGCACGGCCTCTCTATTCTACTCCATTTCATTTTTAAAAAAATTGATGGCCTGAGTGCTGTGGCT CACACATGTGAATCCCAGCACTTCAGGAAGCTGATGCACGAGGATCACTTGAGCCCAGGAGGTTCAAAACC AGCCTGGACAACATAGTGAGACCCCATCTCTACATACACACACACACATACATGCATACATACATACATACA CACATACATAGTAAAAAAATTAGCCAGGCATAGTGGCTCATGCCTGTAATCCTAGCTACTCAAGAGGCTG AGGTGGGAGGATTCTTTGAGCCAGGGAGGTTGAGGCTGCAGTGAGCTGTGATCATGCCACTGCACTCCAG CCTGGGTGAGGAGAGTGATATCCTGTCTCAAAAAGTAATAATAATAATAATTGATGGTTACATTATTACA AAGGTTAGCATGAGGGAATTTGAGGTAGGGAGTAATGGAACTGGGTGTATACCTTGATTGTGATGGTGGTC ACACACATCTATATATGTGACATTCACGGAATCATGCAGTAAAGAAAAATCAATTTCACGTCTGTTCATT TTAAAAGTAACGTTTTTTAAGAGAAAAAAAATCGATAGTTGCAGCCCACTAGATAGAATTCATATCACT CAGGGGTTCCAACCTGGAGTATGAAAATTCCTGTCCCTAAAACCCATGATAGTGGATAGGGGGAGGCAGA AAGGGCCATTGCTCGGGCTGTGGGTGGGTGAGCTGGGGAGAAGGGAGAGAGTGGGAGGTTTCACTTCCTG ACCCTCCTCTCTTCTTTCTGCAGTCGCTTCCTGCCTCGGACGGCTCAGCTGGTATGACCCAGGTAAGGAA GAGCCACATGGAGAAAGGCCTGGGGCAGGGGGAGAGTGGGGCTGTGGTTTTCATCAGGCCATCGGGGACCT CTCGATGAAGCCATCACTTCTGCCAGAGTGAACCCCACCCTATAGAGAGAGTGAACCCCAGCATACACAC AGGCACATAGATGCAGACACTGCACATTAAGATGCTCACATGCAGGTGGGTGCCCTCGACAGCCGTAAAT CACCCACAAATGCCAGATCTCATGATAATTATTATGACCCGCTCACCATGCACAGAAGACATCCCAGCTC ATAAATGTACCTTGCAAAGTCTTATTTCCCACCCAATCCTGACAGATGCTCCATGGTCAAAGATGTTTAG AGCGGAGTCTGCAGAGAGAGGCCGCAGACTGATGGTAAAGTGTGTGGAACGTCCAGCCTTAGACGTTGGA GTTTAGTCGTAGAGGCTGTTTCCCAAATAGGGTTCCATGGAGCATGTTGGACAAAGGGCAGGCAACACCG CATCTCCCACTGAAGACTGACAGTGTACACCGGCCCAGTAAAGGATCTGAGAAATCCTGCAGCGAAGAAA TACATGACACTTTTTAACCCAGCACTTTGCACCCTTATTTCACCACAGACCTCTCCTTCACATTGACTGG GATAACATCGTGCTGAATGTACCTGGGGAGAATTGTGGCTGAAGGGTCTTGAGATGCCACTGTGCAGCCA GAGGGGGTGACAGGGGCCTTGGCAGGCCCTAAGCACGATCTGGGCCCTGCACTGACATCCTAGCTCCAAG CTTTGGCACACTGCTTGACCTCTTGAGCCCGTTTCCCCAGCTGTGAAATGGGAAGAAAATCTCTTGCCTG GTGAAGCGGCAACGAGCCAGCTCGCTGGGCGGACAGCCTGCAAATCACCCCACCAATGTTAGGTGTTGAG ACGCGCTCGTTCTCCCTCTAACCAGCCAAGTGCCCTTGGCGAGCCCTGGCCCTGAGCTTTGGCCCCAGAG TGTCCTCGTCGGAAGAAAAGGGTAGCCTGACCCGTGGTTCCCGACCAGGGAAAACCTCCCCCTCAGAATC ACCTGGGAGAGCCTTTCACATTTCTAAAAATTTCTTGAATCAGTGACACATTCATCCAGCTCAAAAATTT AAAAAAATATAAAAAGATCTGTAGTGAAGAGTCTCACTTCCATCTCTATTCCCGTCTATCCCAGTCGCCC CCGACGTCGACCCCAACACCCCAGGTAATGACTACGTTTATTTGTTTCTTGCATATTCTTCAAGGAGCTC TTTATGCAAATACACACACACACACCCCACACAGACCCCACACACACACCACACATACACACAACCACAC CACACACACACACCACCACAAACACCACACACACACCGCACACCACATACACACCCCACACACACCACCA CACCACACACACACACCACCACACACACACCCCACACCACACACACACACCACACACACCACACACACAC CACACACACCACATGCACACTACACACCACACACCACACACATACCACACACACACCACACACACCACAC ACACCCCACACACACCACACACACCACACACACACACTACACACACCACATACACACCACACACACCACA TGCACACTATACACACCACACACACACCACATACACCACACACACACCACACACACCACGCACCACCACA CACACACACCACCTGCACACTACACACACCACACACACATACAACACACACAACACACACACCACCACAC ACACATCACACACACCACACACACTACACCACACAGACACACACACCACACACACACATATATTTTTCCC CTCATGGAGTTTGATAAAATCACACAAGACACATGTCTACCCATGACCCAGCAATTCCATTCCTAGGTAT TTACCCAAGAGAAATGGAAACGCCCACACAAAGACCTGTATGTGAATGTTCACAGCAGCCTTATTCATAG TAGGCAAAACCCGGAAACAAATCAAATGGTGAATGGAAAACAAATTGGCATGTCCGTGTAGTGAACTATC ATTCGGCAATAAAAAGGAATGAACTACTTATGCGTCCAGTGACATAGCTAAATCTCAAATACATGCTAAG CGAAAAAAGCCAAATCAAACAAAATAAAATCCATGCTGTATGAGTCCATGTCTATGACATTGTAGCACAG GCAAAACTCATCTTTTGTGACAAAAAGCAGATCAGTGTTTGCCTGAGGCTAGATTCAGGGAAATTAACTA TAAAGGGGCCTTAGGGAACTTTCTAGATGATGGAAATGTTCCATATCTAGATTATGGTGGTGGTTACATG CATGTATACATTTGTCAAATTCCATCGAACAGTATCCTTAAAATGAGTGCATTTCACTGTATGTAAAATT ACCTCAATAAAATTTTTTTTTTTTGAGATGGAATCTTGATCTGTTACCCAGGCTGGAGTGCAGTGGCGCG ATCTTGGCTCACTGCAACCTCTGCCTCCCGGGTTCAAGCAATTCTCCTGCCTCAGCCTCCCGAGTAGCTA GGATTACAGGCGCCTGCCACCACACCCAGCTAATTTTTGGTATTTTTTCTTTTTTTAGTAGAGATGGGGT TTCACCATGTTGGCCAGGCTGGTCTCAAACTCCTGACCTCAGGTGATCTGCCCGCCTCAGCTTCCCAAAG TGCTGGGATTACAGGCATGAGCCACCACACCTGGCCTAAAATTAATTTTTAAAGATCTCTTAAAAAGCAG ACACCAGCCCCAATCTCAGACCCCTTGAGACAGAATTTCCAGGACAGGGGCCATCCTGCTGGACAGTGGG TGCCGAGAACACCTTGCCCATTTATCTGAGCTCCCTTCTGACTCTGAAATCTGGAGCCCCACCCTCCTGG GTCTAGCTTCGGGGCTGCCTGGGTCAGGGTCCTCTGGGAAGCCCCTGCAGTGCCCCAGAAGGGACGAAGC TCACAAGGGGCAAGGCAGGCAGCCCACGGGGCAGGAGGGAGCTCAACTGGGCGTCCTAGGGAGAGGGCAG TGAGGGGTGCCAGTGGGGAACCCCTCCCAGCCTGACCCCCACCACACCTTTCTGACCCCCAGATTTCCAG GCAAGGCTCACCCGTTCCAACTCGAAGTGCCAGGGCCAGCTGGAGGTCTACCTCAAGGACGGATGGCACA TGGTTTGCAGCCAGAGCTGGGGCCGGAGCTCCAAGCAGTGGGAGGACCCCAGTCAAGCGTCAAAAGTCTG CCAGCGGCTGAACTGTGGGGTGCCCTTAAGCCTTGGCCCCTTCCTTGTCACCTACACACCTCAGAGCTCA ATCATCTGCTACGGACAACTGGGCTCCTTCTCCAACTGCAGCCACAGCAGAAATGACATGTGTCACTCTC TGGGCCTGACCTGCTTAGGTGGGTAACTAGCCAGCCACACGGGCACCCTGGGCCTGGGCGCCAGCCCCGA GGAGACTGCCCGAGGCCTGTGATCTAGGGTCTGAGCAGGCTGGTGGAAGGGGTGGGGGGACCCCAGTTTA TAACCACTCCCCAAGACACATACCCAGGAGGGGGACTGGAAGGGGCCAGCACCCATCTGTAGGATGGCAA TGGAGGACCTAGTTCTGCCAATCACTGACTTCATCGTCGCCTCTGAACCTCCATTCTCCCATCTGTGAAG TGGGGTGGTACTTCCCGCCTCGCAGGAGGCTTAGAGACAACGTGTGGGTCAAGTGGACCTGGTGTGCCAA GCGGCACTCATGCCAGGAGCTCCTGGTCCTCTCAAGGCTGCTGGCTGCCCCCGGCCCTCCCCACACCACC CATTCCTCCCTCACCAGAGTGTCTCATTGCAGAACCCCAGAAGACAACACCTCCAACGACAAGGCCCCCG CCCACCACAACTCCAGAGCCCACAGGTAAGAGGATTCTGAACCCCCCACAGGGAGTCAGAGCTAGCAAAT AAAAACCCAGGATGCCCAGTTACATTGGAATTTCTGACAAAGGTGGAAATGTTTAGTATTGGTGTGTTCT ACGCAATATTTGGGACCCCATCACCTCCCAAGGCTAAGCGTTAGTCAGTAGTTGTCCACAAGTTGGGGCC AAACAGCAAGGAGTGCCCAGGAAGCCCTCGGCGCTCAGGGTGGCTCCCCCTCCTGCTCTCTCCTCTCCTA GCTCCTCCCAGGCTGCAGCTGGTGGCACAGTCTGGCGGCCAGCACTGTGCCGGCGTGGTGGAGTTCTACA GCGGCAGCCTGGGGGGTACCATCAGCTATGAGGCCCAGGACAAGACCCAGGACCTGGAGAACTTCCTCTG CAACAACCTCCAGTGTGGCTCCTTCTTGAAGCATCTGCCAGAGACTGAGGCAGGCAGAGCCCAAGACCCA GGGGAGCCACGGGAACACCAGCCCTTGCCAATCCAATGGAAGATCCAGAACTCAAGCTGTACCTCCCTGG AGCATTGCTTCAGGAAAATCAAGCCCCAGAAAAGTGGCCGAGTTCTTGCCCTCCTTTGCTCAGGTAAGTG AGACCTGGCCAAGCCCCATGACACCTTCTGCTGCCCTAGGTGGGGTCACAGAGCATCCCAGAAGGTCAGG GAACATGTGTGCAGCACAGGGCACTATGGAGAATACAAGGGAAGTGGAGGCCTGGTCTTGGCCTCTAAGA GGTAACAAGGGTTGGGGTGGGGAGGATGCATCCACACTCAATGCCTTGGTAATCTCTGCAAAGCTACACA CCCCAAGCCCAAAGGAACCGCTGGCTCAGCTGCCACATGGGGAAGGGGAGTCGGCAGGACTTCCAGGAGG AGGCAAGGTCTCTTCGTGGGTCTGGGGGAGAAAAAAAAATGTCTAGGTAGGAATTATGGCTTGTGCCAAA GATGGGGCACTTGGAGGGGTGGGCCTTGCAGGAAGGGCATTTGTAAATCCCAAGAGTTGGCATTGCAGCA ATGGTGAGAGGTGGGAGGTCCCTGAGGCCCGGGAGTCAGAGGACAGCAGGCAAAGAGAGAAACAGAGACA GCCCCCAAGAGGGCTCTGCAGAGGGTCCTGCGCCGTGTGCTGACAAGGTAAAGAGAATGATGCCGCTGTG TGGTTGAGGGGAGGAGGCAGGTGAGACTGAGTCTGTGAGACCCCGGTGGGCCAAACCAGGATCAATGAAG AAAACTAAAGGGAAGAAACCACAGCTGAAACGAAGGGAGAAACGGAGAAACTTCGCAGTACGCCTCCCTC CGGAAGTAGTGGGTTCCCATGGCCGGAAGTGTTCTCAGTGAAGTCCACACTGTACTTACCGAGAGCTTAC TTCATGCAGGGACCATTCCTGGACATTCTAAATTTTTCACATGGCCTTAGGAGGTGGGTGTTATTATTAT CATCCCTTTTTACAGTGAGGAGGCTGAGGTTTAGAGAGGCTAAACCTCTCGCCCAGTACTATGGAGCACT TGCTGGGATTTCTGTTCAAGCAGGGTCACTCTAGAGCCTGTTATCTGAGCCTTGGTGACTCAGTGTGGTC CTGGAACAGCAGCAGCTGGTTAGAAATGCAGGTGCTCAGGCCCCAGCCTGGACCTTCCGAATCAGAATCC ACTTTTCAGCAAGGTCCTCAGTGATTTATGTGCACAAATCTAGGAATCACTGCTCGGGGCCACCTGGCTG TCCTGCAAGCACGCTGGGTACCTGGTGTAAATTAGTAAAATGCTGCCCCTCCCCCAGCAGAAGCAGCTCC CAAGGGGCTCTTGGCCTGATGAGGAGAGCAGGGGCTGGCTCGCAGTCCCACCGGCCCCTCAGCTACTATC CTTCCTGCAGGCTACAACCTGTAAAACCATACCCAGGGTTCTTGCCTTTGAAGCTTCTTCTAAATAGATT TACTAGGGCTTTGAAGGTGGATGTTGCTATGGGAACTGGGCAAAGCTGGTGAGATCACAACTCTCTCTAA ACTGAGCATGTGTGGGCCTCAGCTACAGACTCCCAGGCCCCACCCTGGCAGGCAGAAATAAATGTGTGCC TCTGTTGAGCCAGGCCCCACTTGCTGAAAGCCTGGCCCTCAGCGTTCAGTGCCTGGGCAGCACGATCTGT CCCAACTGTGAACTGCCAGGACTCAGGGCAGTAGGTCTTTCCAACCCCCAGGCCTCAGTTTCCCTCTTCT GAAAAGTCCTCCTTCTTCCCAAAGTGCAGCGGGATAATGACAAGATGTTTCCAAGTGAGAAGTGGACCTA GGTGGAGCCTATTTCCCACGCTCTTCTCTTATCTCCACCTGGCATGGAGCCGACTCCCTGCTGGTGGCTG CCAACAGCCTCCCTGCAGAAAGCGCCTATGCTGCTGTGTCTCAGGATGGTGCTGGCAGCTGCCCAGAGGC ACCAGAGACAGTGCTGGGTCAGGGGAAGAGCCACAGAGGGTGCCAGAAGGAGGGAAGGGCAGAAAAGAAG GCTGCCTCTCCTGGTGGTCCCACACAGTGCCTTCCCCTAGCCAGGGCCCCGATTTGCCAGTGGCATGGGG CCCCAGGAAGCAGCACACAGGCTCAGGTTCACACTTGCACCCTCCTTTCCCATTGCTTCCCCTCTCAGGT TTCCAGCCCAAGGTGCAGAGCCGTCTGGTGGGGGGCAGCAGCATCTGTGAAGGCACCGTGGAGGTGCGCC AGGGGGCTCAGTGGGCAGCCCTGTGTGACAGCTCTTCAGCCAGGAGCTCGCTGCGGTGGGAGGAGGTGTG CCGGGAGCAGCAGTGTGGCAGCGTCAACTCCTATCGAGTGCTGGACGCTGGTGACCCAACATCCCGGGGG CTCTTCTGTCCCCATCAGAAGCTGTCCCAGTGCCACGAACTTTGGGAGAGAAATTCCTACTGCAAGAAGG TGTTTGTCACATGTGAGTTGGCCACAGCCCACAGTGGGTGGAAGCAGTTACTACTTTACCTCTAGGACCC GGTCAGGGTGCATGTCTCTAAAGGGAAGCCCTGGGGAGCTAGACAGAGAGTCCCAGAGACCCAGAAAGGA TGGAGACAGGGAAATTGGAAGGCTAGGGAGCAAGAGTACTCAGACAGCAATGGGCATGAACAATAATAGG AGACAGGGCAAGCAACAGGCAGATCACTGTCAGGCAGCTGATGAGTCATAGAACTTCTCCTTCTACCTTC TACTTTTGCAAGGTGGAAGCCAGAAAATCAAATGGTAGATAGCACATATTTGCTACATGAATGGTAACTA AACGGGAAGTAAAAGAGAGAAATGGATGGATGGATGGATGGATGGATTGGTGGGTGGGTGGGTGGGTGGG TGGATGGATGGATGGATGGATGGATGGATGGATGGATGGATCGGTGGGTGGGTGGATGGATGATGGATGG ATGGATGGATGGATGGATGGATGGATGGATGGATGGATCAGTGGGTGGGTGGATGGATGGATGGTTGGAT GGATGGATGGATGAGTGGACTGATGGATGGGTGAGTGGATGGATGGATGGATGGATGGACAGAAGGGCGG ATGGACAGATGGATGGATGGATCAAGAATAGAGGCCCCGCTGGCTATTGGTATGCAATCATATATTGGTT TCTGATGGTATATGATGGCAAGGTGATCACTAATAACTGGCAAATGGTAGAAAATTAAATAGCTTATAAC AAGTGTGGCAAATGATAGCTAGATGATCAACTGTTAGATGACAAATGGTGGGTGGGTGAGTGGGTAGCAG GGAATCCTAGAGATAGATGGAGGGATAGCCAAATAGATCATTGCCTTCCGTGCATGCTGGTGAATTTCTC AGATGAGCAAGGATGACGGCGGAAGCCAGGCAGCCAGCAGCTTCCCTCCCACAGTTTTTCTCCCCCCAGG ACTGGGACTGACCTAACTCTTCCTCCTTCCCCAGGCCAGGATCCAAACCCCGCAGGCCTGGCCGCAGGCA CGGTGGCAAGCATCATCCTGGCCCTGGTGCTCCTGGTGGTGCTGCTGGTCGTGTGCGGCCCCCTTGCCTA CAAGAAGCTAGTGAAGAAATGTAGGTGTCACGGCCCTGAGTGGCTCCGTTCCCACGTGCAGAGACTGGAG GGGCTGCACTAGAGTCCTCCGGAGGAGGGGTCATGCCTCCAGAGAGCTGGGCACGCAGGACACCTCTGCT TCACCACCCAGCCTTCCCCTCTCCTGCCCATTAGCCATTTTCTGCCCCAAGTAACAGAAACACAAAGTAA ACAAAATTGCAATTGCTGATTTGTCCATCTCTGGAGCCCAGCTGAGACCCACTCCAGGTCAGAGAGAAGG TGTGGCCATTCCCAAGGCTGTGGCAGGCCCAGAGGGTGTTCAGTACAGCCTCTGATGGTAGGAGACACAC ATCCCAGGCCTAGCCCCACCACTTACCACTGCCCAAGATGCTGATTGGGACACTTCCTCTCTCTGACCCT CAGTTTCTTCATCTACCAAATGGGCAATCGCACACCTTGTGAACGCGGGGGGTTGTTCTCCTTGCCTTGC AGAACTTAAGAGGAAATTATTCAGACAAAGAAGTGAGAGGGGAGAGATTGGGAACAGAGGGCCCAGCAGG ACAGTGTTAGGTCCTAGTGCCTGCCCAGGTCTAGTTCTAAACGCCTGTGGTCTTTTCTCCTGGGGAGCCG GGCACCTCTACACCATCCTCGGCCTGCTCCTTCGGCACAGGATGGCTGCAATTCCCTTCCCGAGCAAGTT AGGAAACTGAGGCCTACGAGAGACTCCAGGGGGCAGAGCCCAGCAGCCCTGCCCTCAGCTGCACCTGCCG CCACCATCTCCCTCCCAGGCCCAGCCCCATCCCCACCCCTGCCTGCCCCCACCCATACCTGCCCCCACCA CTCTGATGTGCCTTTCTTGTCTCTTGCCCAGTCCGCCAGAAGAAGCAGCGCCAGTGGATTGGCCCAACGG GAATGAACCAAAACAGTAAGTGTCCCCGGAGGCAGGAGCCTCCCTCAGGCCCTGGACAGAGGCCATGGAG GCAGAGTCGAGGCTCTCTGCTGACCACAGACGGAGCCTGTGGCTGCTTGAAGCCTCTGATCTCCACGGTG CTTGTCTGACATGTGACTGTGGCCAACAGACGTCCCCACACCAGGGAGGGCGGAGGCTTCTTGATGGGCT GATGGCTGAGCGGTTTCTACCGCTGAAGCCTCAGAGTGTAGAAACATCTCATGGTATCCTTTCTGTCCCT CGCCCCATCTCTAGGGTCCCCAGCAGCCCTCACCTCCTCCAGCAGTATTCCCCAACACAGTAGCCAGTGG ATGTTAACAGGCAGCACTCACAAGAGGCCTCCCTGATCAACAGATCTGGAAAACCATGAGTTAGGCACAG CTAAGCATAGCTATTTCAGGACTTATCAGAACCTTAATGGGCATTTTTGTATTGCCCATCTCCAAAGCAA GGATCCATTAAGAAGGGCTGTCCAAGTTATTTTCATCATGGAACAACTTACAGGGCCAGTATTAGGACAT TTTGGTGCAGCCAGCCCAACACCATCCCTGGTCTTCATCCTCCCTCCTTCCAGCCACAGTCAGAGACACC CTAATGGATGGAGACATGGCGCTCTTCCTGCCTCCCATTCTGCCTTCTTCCCCCTTCCCAAGTCCACAGC CAAATCCTTCAAGAGGGCTCTGTTGAATTTGTTATTAAGAAAGGCTCTGGAAACTTTCTTAATAACATAC AGGCCCTCAGCTCCTGGAGGGCAGGGAGCAGATCTGTCCCGTCACCTGTGCTCCAGTCTCCTGCTTGGTA AATGCTTGATGACCGACTCACGGAAGGCCCCACCCTGCCAGTCAGATTGCTGGGTTACCCAAACCTGCTC TAAAAGTCTGCTGCTGATCCTGCTAGCATGAACCTCCCAGGAAGAAAGGAGAGGGGGATGCATTGAAGGG GCTGTGGGTGCCTTTGTCCTCTCCCCGCTAACATCATGGGAATAGGAGATTAAAGTTCTGGGCACCTGCT GGGGAAGGAGACGGAGGACACAGCCACAAGTCTGACACTGTCTCCTACCATCTGCCCAGTGTCTTTCCAT CGCAACCACACGGCAACCGTCCGATCCCATGCTGAGAACCCCACAGCCTCCCACGTGGATAACGAATACA GCCAACCTCCCAGGAACTCCCACCTGTCAGCTTATCCAGGTAAGCACCAGCGGGTGCTCCCAGGCACGCA GGCAGGGCTGCAGCAGGCCGCCAAGGCAGGTCACGTGATGCCCTTGAAGGTCGGGTGATGGCCCAAAGAC AGAATGGAGACCCCAGGGTGCAAGGGGATCAAACATCGCAGGATGCAGCAGGGGTACGGAAGGGATATAG CCTTTGCAGGCCCCAGCTGTCCCGTTTGAGACCACAAACTACTCATCTGGAGAGAAGAATGCACTCAGAA AGCCATCAACCCCAGCCTTGCTGTGACCTAATGGGCCTGTTTCCCTTCAGGGCCTTGAGGGGCCTGGATG GTAAATGAGGCCCCTTGGATTCCTCCGGGCTAGAAGAGAGCCATTAAGTGGGCTGCAGGAGCGCTGTACT AAAGGCTCCCAGACTGGCACTACCAATTAAGCCCCAGGGAGCCTGGAGTTCAGCATCATTAGTGAACGTT TGGCTGATTTGATCAAAACAGATAAGGGGTTGTCACCGGCTCATAAAGCCCCTGTGATCTTCCAACCCAC TGTGGGCAGCGGCTCTAGGATCCAAGGGGCTCTGTGGAGTCAAAAACCCTCTGCAAACAGCGTCCTTTCT TTCCCCAGCTCTGGAAGGGGCTCTGCATCGCTCCTCCATGCAGCCTGACAACTCCTCCGACAGTGACTAT GATCTGCATGGGGCTCAGAGGCTGTAAAGGTGAGCCCGTCTCCAGCCTGACCCCAGCACCCCAGGGTCCC CCACAGTCCTGGGGGTGAGCAGCAGCCACTCAATGCCTCCCCTCAGTCCCACCCTGCCCATACATGAATA GGGGACCCCCAGCATCTTGATTCTTGTAAAGGGAAAGACAAACGTCAGCTGTTTAGGTTATACGAGCTGA TTTGACAATTAGAGCAGAGCAATCATGAACTTTAACAATCAACTCTTAAACTCCATTCGGAAGGCCACAC AGGCAGAGTCCCTAAGAGCTCCGTGATGGAGTCACACCCCTCTAGAGCTGCCAACCTGCCAGCGGGCAGA AGGAGCCCCTGGCTCAGGACAGCAGCCGTCCCCAAGATGGAGCCCAGAGCATGGGTCCTGTCCCCGACCT CCCTGCATCACCTCTGCGGTGTCTTCCTCTAACACGTTTCCTTCTCACCCCACAGAACTGGGATCCATGA GCAAAAAGCCGAGAGCCAGACCTGTTTGTCCTGAGAAAACTGTCCGCTCTTCACTTGAAATCATGTCCCT ATTTCTACCCCGGCCAGAACATGGACAGAGGCCAGAAGCCTTCCGGACAGGCGCTGCTGCCCCGAGTGGC AGGCCAGCTCACACTCTGCTGCACAACAGCTCGGCCGCCCCTCCACTTGTGGAAGCTGTGGTGGGCAGAG CCCCAAAACAAGCAGCCTTCCAACTAGAGACTCGGGGGTGTCTGAAGGGGGCCCCCTTTCCCTGCCCGCT GGGGAGCGGCGTCTCAGTGAAATCGGCTTTCTCCTCAGACTCTGTCCCTGGTAAGGAGTGACAAGGAAGC TCACAGCTGGGCGAGTGCATTTTGAATAGTTTTTTGTAAGTAGTGCTTTTCCTCCTTCCTGACAAATCGA GCGCTTTGGCCTCTTCTGTGCAGCATCCACCCCTGCGGATCCCTCTGGGGAGGACAGGAAGGGGACTCCC GGAGACCTCTGCAGCCGTGGTGGTCAGAGGCTGCTCACCTGAGCACAAAGACAGCTCTGCACATTCACCG CAGCTGCCAGCCAGGGGTCTGGGTGGGCACCACCCTGACCCACAGCGTCACCCCACTCCCTCTGTCTTAT GACTCCCCTCCCCAACCCCCTCATCTAAAGACACCTTCCTTTCCACTGGCTGTCAAGCCCACAGGGCACC AGTGCCACCCAGGGCCCGGCACAAAGGGGCGCCTAGTAAACCTTAACCAACTTGGTTTTTTGCTTCACCC AGCAATTAAAAGTCCCAAGCTGAGGTAGTTTCAGTCCATCACAGTTCATCTTCTAACCCAAGAGTCAGAG ATGGGGCTGGTCATGTTCCTTTGGTTTGAATAACTCCCTTGACGAAAACAGACTCCTCTAGTACTTGGAG ATCTTGGACGTACACCTAATCCCATGGGGCCTCGGCTTCCTTAACTGCAAGTGAGAAGAGGAGGTCTACC CAGGAGCCTCGGGTCTGATCAAGGGAGAGGCCAGGCGCAGCTCACTGCGGCGGCTCCCTAAGAAGGTGAA GCAACATGGGAACACATCCTAAGACAGGTCCTTTCTCCACGCCATTTGATGCTGTATCTCCTGGGAGCAC AGGCATCAATGGTCCAAGCCGCATAATAAGTCTGGAAGAGCAAAAGGGAGTTACTAGGATATGGGGTGGG CTGCTCCCAGAATCTGCTCAGCTTTCTGCCCCCACCAACACCCTCCAACCAGGCCTTGCCTTCTGAGAGC CCCCGTGGCCAAGCCCAGGTCACAGATCTTCCCCCGACCATGCTGGGAATCCAGAAACAGGGACCCCATT TGTCTTCCCATATCTGGTGGAGGTGAGGGGGCTCCTCAAAAGGGAACTGAGAGGCTGCTCTTAGGGAGGG CAAAGGTTCGGGGGCAGCCAGTGTCTCCCATCAGTGCCTTTTTTAATAAAAGCTCTTTCATCTATAGTTT GGCCACCATACAGTGGCCTCAAAGCAACCATGGCCTACTTAAAAACCAAACCAAAAATAAAGAGTTTAGT TGAGGAGAAA (SEQ ID NO: 182)

[0194] A representative mRNA sequence for CD5 is provided by NCBI reference sequence number: NM_014207.4 and is shown below:

[0195] [ka]

[0196] A representative amino acid sequence of CD5 is provided by NCBI reference sequence number NP_055022.2 and is shown below: MPMGSLQPLATLYLLGMLVASCLGRLSWYDPDFQARLTRSNSKCQGQLEVYLKDGWHMVCSQSWGRSSKQ WEDPSQASKVCQRLNCGVPLSLGPFLVTYTPQSSIICYGQLGSFSNCSHSRNDMCHSLGLTCLEPQKTTP PTTRPPPTTTPEPTAPPRLQLVAQSGGQHCAGVVEFYSSGSLGGTISYEAQDKTQDLENFLCNNLQCGSFL KHLPETEAGRAQDPGEPREHQPLPIQWKIQNSSCTSLEHCFRKIKPQKSGRVLALLCSGFQPKVQSRLVG GSSICEGTVEVRQGAQWAALCDSSSARSSLRWEEVCREQQCGSVNSYRVLDAGDPTSRGLFCPHQKLSQC HELWERNSYCKKVFVTCQDPNPAGLAAGTVASIILALVLLVVLLVVCGPLAYKKLVKKFRQKKQRQWIGP TGMNQNMSFHRNHTATVRSHAENPTASHVDNEYSQPPRNSHLSAYPALEGALHRSSMQPDNSSDSDYDLH GAQRL (Accession No. 184)

[0197] A representative DNA sequence of the CD47 gene is provided by NCBI Gene ID: 961 and is shown below. GCAGCCTGGGCAGTGGGTCCTGCCTGTGACGCGCGGCGGCGGTCGGTCCTGCCTGTAACGGCGGCGGCGG CTGCTGCTCCGGACACCTGCGGCGGCGGCGGCGACCCCGCGGCGGGCGCGGAGATGTGGCCCCTGGTAGC GGCGCTGTTGCTGGGCTCGGCGTGCTGCGGTGAGTGGCTCCTCGCTCCCAGCCCTGCGGCTGCTGTCGCT TCGCCCCCGCGGGCGTGTGGGCTGCGCCCCAGCCAGCCCGGCGGCGCCCTGAAGAGGGTGGCCGGGGCGC AGAACACTCGGGCCCTGAGCGCCCGAAGTGCAGACGTGGGAGGGCCCCACGGGGAATCGGGCGCCCCCCT TCTTCCTCCCTTCCTTTCCCTGGTCGTCTTCTTCCCCCTGGGTGAGAGCGGGGCTCATCTCCTCCACCCG GTTCTCCATCTCCAAATGCACACACACAGGAAGACTCCAAACCGCGCACCTCGCGCAAAAATTAACTAGC AAGAAAGGGCGCGATCAGAGGCAAGGGGCTGCAGCTGCTAGGGATCCCCTTCTTTCGCACCCCTCTCCCT TCTCAGTGCTTAGACGCATTTGGGGTTGAGGGAGGGAGAGTGGGAGGGCCCGGGCCTCTTGCGATGGAAG TGCGCATTTTGGCGAAGTCGGTGAGAAGGGGTTTCTGCCGTTTGCCTCCCACATGAACATAGGAGCGAAG AGGACGTAAAGGACACAATTAAGTTTCATTTTCAACTCAGCATTCAATCAGAAGAATCTTCCGGCCGTGT AATTTTTGCTGTCGTTTTTAATCCTAAAAATAAGCTTGCTGGAAACTCTTCCTTTCTCGTGACCCTCACG CCCCATCAGCCACTTGCATACATTCTAAATTGTACGTTGAAGTTTTTCCCACTTTATTTGGGGGAACCGT TTTAAAAGTAATCTGGTTTTCGCCTGAAATAGGAAGACAGTAACCTCCAGTCAAAACATGTGCAGCAGAA TGGGATTTGGTGTTTTTCGACGCCAAAGAACCTCCACCCCCCCACCCCCACCCCGAGCTTTTGGAATCCA TTTCGCTTTTGTAAAACGTGTGCTTCGTCTGTAAAAACTGAGCAAGGAATAGAAACATTCGTAGCTCTAC AGTGAGTGCCTGTCACACTTCACATCCATAGAGCCTTTTGTGAACTTTTATAAGATTCAGAATTCAGCTT GAGCACCAGTGACAGCAATTGTTACATTTATTCTAGAATGCTAAATTAATTCGATTTTAAACTGATTATT AGCCTCGGTGTGCCGTTCCTAAAGGCTTGTGACTTTAGGTATTTCCAAGATGCCCTTAAGTCTCTGCTTA CCACTTTCCTCCCTCCCCGAGCATCCTGGATGTTGGGACTGTGAATCCAGGTCTCCGTTATCTAAATGGT TGATGTTAGTGTTTCCTGTCATCACGTTTAGTATGCTTGTTGCCTTTACTATCATTATAGCTAAAATAAT ACTGCTTTCAGAGATGTGTTGTATAATCGCATAATAATTTCAGAACGCCTTCATATTGAACCAGATATGA AGTGATACAGTATCATTTATTCAAACTGCCTAAAAAGTAAAAAGTATAAACCATATACTATTTTTAAAAC AGGTAGGTTAGATTTAAATCACGTAGTTTAGAACTGTTGGAATGGTACTTTGAGTGATAGGATTTATGTT AGGCCTTCTTTAGTAATAGTTAATATCATGTAATTAGCACTCGTTTACACACAATTTTAATGTGTTCGAA TCCCTAGAGTCATCGAATTCAGAATTTATAGTATTTTATTTTACTTAGTTAATATTAACCTAAAAAAAAA GCAAAATACAGGCATTATGCAGTTGAGTTGTGTTAAGTGTGGACTGTAAAACAGGATAGTATTTGTTATA AAATATGTTTTTGTATGTGTTTAATATATAGCTTCAAAAGGACATGTATGAAGAAAGATGTCTCAGCACA TAGGTAGTTATAAACCAAGGGTTTGAGAGACTGACTTTAGAATCTAGAATGAGTAAGAAAGTGATGGATC TGTACATTCATTTGTATTGAAGTTTGATTTTGCTTTCAGTTTCGTTAATTAAGCCCCGGGAATCAGGGAC CTTTCCTGGCACCCTCTACAGTGTTAGTGGCCTTTGTGGTAAAAGAAGTTATTCTCAGATACTCATTTCAT GCAATTACAGGCAAACTGGAAGGCACCTTAATGGTATGCAAGATGTGAAATGAATTACTATGTTGCATTC CACTCTGCCCCCACTCATGTACAATTATACTTTGTTTAAAAGTGCATAGTTTCAGTGGTATTTATTAGCT AGCAAATAAAACTTTAAATAAATAATGATGGTGTTGTGAACTGTCTTTTCAGTGGTTGAATGATTCCTCT TTGTTCAAAATGAGTTGTTTTTTTTTTTAAATCAGGGTACATATGATTAAATAAAATTTTTTTCCATCGT CTAAGTCTAGTAGTAATTTGGCTTGTTTAAATAAAAATGTTTTCTCTTAAAGAATATATTATTTTTAATT ACAAGTTGCCATTTTAAGGAAGTAAATGCCTATTTAAAAGTACGTATTTATGGGGCCGCCCCAGCAGTCT AGAGGCAGTGTTTTTTAAAGCATTACACTAAATGCCTACCATTAGGACACTTGCTCATGCTGTCATCTGT GTTTTGGCAGATGTTTTATCTTTGTCACTGAGTTGTTCATGGAGATTTGAAAGGCCAAGTTCTAAAGATG AGAGATAGATGACTTTCCCCCCCAAATTGATACATATTCTAAATCCAAAATAATCACGCATATTCGCAAAA CTAGATTTGCATTTCATAGTATGAATTCAGCTATAGCTAGTATATAGGCATTGCTTTTAATATAGTAATG TTCTTTTGTGGCATAATTTTCAATCACTTTCCCTGTCTTAGGTTTCTGCCTACCTTTTTATACAAGATA AACATTTCTATTTTCATATCTTATCTCTCAGTCTGATTTTAATAATATCGTCTTGGGTTATCATAAAGTT ACTTTCACTTTGTATACTAGTGTAGATATTTCTTCTGTGATAAAAAAGAACAAGAATAATAGTGTAGGATC AGTTTTTTAAACTCATTGTAGTACTAAAGTAGAAAATACAGAGCACCCAGGAAATTTGATTCTGATATA CCTAGACTAAGACAATAGAATCTGGGTTTTTCCTCTTTACGATTATACAGTGTTGAAACAGAAATCAAGC TACACCCACCTCTGAAATTTCATTCCAAATCCATGGAGCAGTTGCATAAGACTCAGAATCTGGTCAATC TCATGGTTTGAGATTCAGATTAGGAGAATTAATGTGCATATTTGAATATATATCACACTTGTAGGTTAAA AGGTAAAGCAAGGGGTTTGGCTAGTTCAAGGCCTGTCAAAACCACTTTTTTCCATTTCAAGCAAGTATGA GCCTATACTACACACCAAGCAGTTTGCAAAGCCCTCTTTGGCATATAGACAAATTTCAGACAGTATGTCA GTATGTCTCTTAATGACTTAAGATGTAGTATATAGACAGTAGGGATATCTTGTTTTTATTCCCTCATGGT ACTTAGGTGCTTGTTAAGGCCATAATTCTGGATGTTATTGACTTTAAACTGCCCTGCCCTTTTTTAATTCT AAGTTGGCCTCTTCCATTCTTTATAAGGCCACTTTTAAATTCATTAGGGGGATTGCGTTGGGTCAGAACTA AGTACAACTTGTGGACACCCCACTCTACCCCAGACCTTTTATTTTAGGTTCCATTGAGGAGAGATCTGACT TCATCCTTTTTGTACGTAACCCAGTGTAGCTAGAGACTCCAGTTCTTCAAGCCAGACACTAGTTCTAAGG GGCAAAAGTAGGTAGGATTGCTTTCTGGCATGTGTCTTTACTGATGATCTTTGCTTCTTTACCTAAGAGG CTGCTGAGACTTCTTACTCCATTCCTGAACATCCATTCCCAGCAGATAATTAACTTCCTACTCTACTGCA AAAAATAGTGGTGTGTGGTTTGGACTCCCTCTGCTTCCCTCCCACCATTACTAAACTGATCTATAACATC TTAGATAACCTTCGTCTCATTCTCTTCCGTCATCCAGCTTCAGAAGAAAACCTACTCACTTTTCCAACGA AAGGACAACCTGTCTACCACCTTTTCACTTCTACCATCTTGGAGACCTGCTCAGTCAGCATAATTCTTCA GTCTTGCATCTTAGATCTTTCCCTCTCCATTGACTCCTTCCCCGTAGCTTGCACACATGATCAAATTTTA CCCCATATTAAAAAATAAAACAAAAACCTCAAGAACCAAAATGTCCTTGCCCTTGTTCCTGCCTGCACA TTGTCAGTTGTTGCCTTCTCTGCTTTCTTACACTGCCAAATTTATTGTAAAAGAATCACTGCCACTTAAT ATATTTTAAGTGCTTGATAAAGCCATCTTGTTTCTCTCTCATTTCTCAAACATAGTGTGCATCAGATCTC ACTAGTCCATTGACAGATGGAAAGATATTCTCCCCAAATATTTAAGCCTCTTTTCTCTCTATAATTAGCCTA CCATCTAACTAGCTACACATACTATTTTGGTTCACAATTTTAATTTTCCCCTCTTTATCTCCCCTGCTTC CCACAACTAACTTCTGTTGATTACTGTAAGTTTTAGGTCTTCTCTCCAGTTGTTCTTTACCTGGACTATT GCAGTAGTCTGTTAACGGATCTCCCTTTCTCTTTTCCCTTGCTGCCCTCAGTCCTTCCTTTACACTGCTG TCAATGTTTTTTTTTTAATGAAATTAAATCTGAGTGGTTTACAGTTGTTTATGTTATCAAGTCTAAACTC AGCATGACATTCAAGGAAGCCCTTTAGAATTTTTCTTATTCATCTTTTCCAACCTCATTTCCTAGAGCTC ACCTAAATAATTCTCTGCTTTCAGTTCTTCACTTTCTTTATCTGGCTGTACACGTTACTCTGCCTAGAAA GCCCTTCTTTCCCATAACATTCTTACTCATCCTTTAAGACCCAGTTTAAACATCACCCTCTAGGAAGAT CATTCATAGACACATTCTTGTGATTTTTAAGGAACTTTTCTATCCAACCATACAAGTGACTTGAGATTTT CCATGAAAATATGCAGCTTCATGATTTCATAATCAAGTCTATACAAGTGAGAAGCAGTGGCAGGTTCTCT TGAATACAGAAGAAAAATCTATCTTCTCCCACATGATTTTTAGATTTTTCTTCTATGGAATTTATACTT TAAACTTTTTACATTCACAAGGAAGGTATTGAATCCTACTTTCTGGACCCTGTGTTAGATTCTTAGGATA CCAAATGAAGACAGTGTCTTACTTTCTGAGCCTGGAGAGATCAGTTAAGTAAAAATAGTAATTACACAGTA GTAGGTACAGTGACAACATTTGAACACAAGGCAATGGGAGCAATAAGGAGGGGAAGAATACAATTGAGTA ATGGTAGGGAAGGGGAACAAGATTGAACAACTCAACTGTGCTTTAGTAGGAAGAGCAAAAGTTAGCAAAC TGTAGCTGCCATTTATTGAGTACTTGCTGTATATGTGAAAGGTGATACAAAAACATGTTATCTTATTAAA TCTTAACAATATCTCTATGAGGTATATACCATCACTATGCACATTTTATAGATGAGGAAACTGAGGAGCA GAGGTAAGTAACTTGCCTAAGGTTATACAGCTAAGAAGTACATGAGGTGAATCTTAAACTAGGGTCGACC CCTTAATGTGTGCACTTAACTATTATACACCCTATGTACTCAGTGGGGTAATAGTGTATAACAGTTAAGA GGCTATTTAGGTTTGAGGAACAGTATGTGCTAAGGCATGGGGATGAGAGTGCATTTGCATAGTCATGGAA TTGCAATAACGTCATTATGATTGGAGCTTAGTGTGGAAATGAGGGAGTACAGGAGGTGGGGCTGGAAAGG ATCTTGAAGGACCTCGTTTGTATTCCATGCTAATGTACTTAGAATTTAGCCTGAAAAGGGTTGAGGGTGT TGTTAAAGTATCTTAGGCAAAGGAGTGCACATTTGCATTTAGAACAGTTATTTTGGGAACTGTAGAAAAT ACATCAGGGCCAGGATGGAGAAACCTTGAGGATGGAGGCAGGGAGCTGGTTAGAAGGGTATGGCAGTAGT TCGAGGGGGGAGGATGAGTGTCTAATTCATTCATTCAACAGACATTGGATTTAGGAACTATTAATTAGGT TGAACCTACTAAGAATTCTCCATGGCTAAGGAGAGGGAGTAATCTAATGTGATTCTTAGGTCCAGGTCTG GTTCACTGGTAGATGATAGAAAATACTTTAGCAAAGTGATTTAGGCTGAGGGTAAGAAGTGGGGGTACGT AAGTTTTTGGACATACTGACTTTGAAGTAACTGTATTACATCCAAGTGGAAGTGCACAGCAGACAGTTGG CTGTCCAACATCTGCAGTTTAGAAGAAAGGCAGTGCATGATTCAGTTTTTTATTTAAAGTTTTGCAGGTG AAGAAAACCATGGGAGTCAGGGAAAATACTAGGGGGCAGAGGGTACTGAATCTGAAGCCCTTAGGGACAC TAACATTTAGTGTGAGTCAAGGAATAGATTTCAACAAAGAAAGCAAACTAGGATGGGGTGGACAAAGCAG GAGGAAATTCGTAATCAGGAGATACCATGGAGGCCAAGAGAGAAGAGCAAGAAAGAGACTGCCCACTGGC TCAAGAACTGTGGAAAAGTCAAGACAGAAAGTGAACCATGCATATTTGATTTTTCACAAGGGATTGGTGG TGACTTTGGCAAGAGCTGTAGTAATGAAGAAGTAGGGGTGAATGGGTGGAGGCCACGTAACAGATTTAAG GAGTGAATTGGAAGTGAGGAAATGCTTATAGATTGTTAATTTGGGATGCTAGGGAGAGAGAAAGTATGAG GGATTTTTTTTAAGTTGGCAAATATTCAAGCAATTTTTAAAGACACACACAAAAACAACACCTTGTAAAT GGAATGGGGGATGTGGGAGGCTGATAATCAACTAAGGAAAGTCGTTGAGGAGACTGATGGGTATGGAGGC AGAAATGGAAACTAGCTTTCAGGTGATGGGATTCATTTCAGAAAAGGAAGCAAAAAAAAAAAAAAGGTGT GGATAGTTGGGGTTACAGGTAGGTTTATAGGGAAGGAGTATTGGAAGCTTCAGGATTCTCCTCTGAAGGT TTCAGTTTGTATTAAGAAATAGGGAGAGAGGACTGTTGGAGAGTATGGGATAAAGGGCAGAAGGGAGTAA TTTAAGAGTTGTTAAAAAGAAGCGAACATTTACATGAAATATGTAAAATGGTAAAGATTGAAGGCCCGGA GGAGCAGGGACTATGACATTCTTCTGTTTTTGCCTATTAGCAACATGAATTTTCTTAGAAATCTGTAAGA CCAATTACTCTTCTGCCCATCCATAAGGACGTTGTCCATGCAGAAAAGATAACACTTGTAACCTTGTATT ATATACTTATCATCGCCCATTTGAGAATTGTAAGCTCCTAAAAGATAATAACTATATTTTTTCATCACTA TATCCCCACATCCTATCACAATATTTCATCACAGGTAATTCTTGACAAATGTTGATTGCATTTTTAATT TTCTAACCTGAACTTGTGTGCTGTGACCACCATGGATTGAGTCTTCTCTGCCACTACAAAGCTCTTTTCT AGACTATGATATGAGATGGTTTGGGGCTGATAGTCTATATTCACCAATACTTGTACAGTTCCAATGAAGGT TTCAAGTCTAATACTTTTGGCATTTGATATAAAATCATTTTCCCATTTTATTTGCTAATTTATCTATAAC TCTGGCATTACTCTTGGTTACATTTGTCTGCTGCTATCTGATTTAGACTGCTATAAGCATATCTTGTTTA TTCAGAGTTTTCTTATTCATGTTAATTATCTGTGTTTCTTTATTTGCTCTCTACATTTTAAATAGTTTCT TCACTTTGCTGTTTTATGAGAAGGGAGTAGGCACAAGGAAAAAACCCCAAATCAGACAGTTTTACTACTT AATAGTTTTTTAATGCATCTTTATAGAGATTGAAGTTGTAGTTAACAGCTAGAGTGATATTTTTGGCCTG CCCTTATTTATTAATTACTAGTGCAAAGGTTATTCAAATTGTGGTTTATCCAGGTCAATTTTACTGTTAT TTTTACTAATAGCATTTATTCTACTTAAATTGCTTCAGCTATAAAATGTTTTTATTGTAACAAATAATGC AGTATAATTATTGCTTTCTGTATTCCTTTGAAAATTCATTCTCTAAACATATGTTATGAAATGGTGGATT GTTCACCAACTACTTCTTACTTACTTAATTAAAGCCTTTGCAAAAAGTTTCATAGGATGACTGAGTTCTT CATTCGTACCTCTTTTTTTTAAGAATAATCTCTTGAAAGTCAAACCATGATTCATACAAAATAAAATGA ACATGTGTCAAAGATTTTATTTCACTAATTAAATTAACAAGCAAACTAGCAAGAAGGCAAACCCTTTTAA AAGAAAATTGGAAAAACAGACACTTATAGGCAATCAAGAATGCCTAAATGAATTTGCTAATAAATGCA AAATTGGTCAATATCCTCAGGGCAGTAAAATGTAGTCTTTTGGAATCCTCTCTCCACCAGGTGGAAATC AATTGCAGAACAAGATTTATTTTTATCTGTATGGACAGCAATCCATTTGAATTACTCTCAGTTTTCTACA ACTTATGAAACTGTAAAACAGCCCAGTCAAAGTCAGTGAAAGGCACAGGCTTCTATAGAGTCAGATAATT CCAAAGCGTCTGTTAGACATTGCCCAGCACTTGACTGAAAGGATACCCAGTAGTCTTTGTATCCCTTCA AAATCTTCACCATATTTCTGATACTTTCCTTCCTTATAATTTGAACATCTTCACCCCTTTATTCTTCT CTGGTATACTATGTCCTCTCTCCCTCAGTGTTTCTCTCTCAGGGGAAATAATTCACAATTCCAAAGTTTT AAAGAATGCAACGGAAGTCCAGGTTTTGCCTTGGGCTTCCTAGTATTTGGGTGGCTAGAAATGTAGAAAA CTGGGAAGAGGTGAGCTGTGGATGCCCACAATATAGGTTCAGACTGCAATTTCCCAGAAATATAACAATT TGGACTAGTCAAAGAGGGCCCATATCATTACATTAAAATGCCAGATTATTATGTTTTTATAGTATCACCC TACATTTTTACAGCATAGTATTGTTTAGAGAGAGCACTTGCCGCTATGTTTTCCATGTTAATGCTCAACACA GTTCTGTGAAGTAGCCCGGTGTTTGTTTTTGATCCTCCTACTTAAGAGATCCTCCTATTTTAGAGAGTAA ACCAAGGCATAGAAAAGTGAGTTGCTTAAGGCAGCATATTAAAAGGGGCAGAAATAGTATTTTTACTCA GGTGTTTTGAACATTGTCCAGTACCTATATTCCAGTATGCATCCTTGCATTCAATTCATGGGGTATTTAT TGAGTAAACATAGTGTTCTTACAATAGAAACATTCTATGCCTCTGATTTTTATTCCAGTTTATGTAGAAG TAAATCTTAAGTGTGAACTATTAACAAAGTTGATATTTTATTTATATTTGTTAGTAATTTGTGTTTTGTT TTTGTTTATGTTTTGAGGGGAAGGCCAAGTAGCTACTTAGGTAAAAGAGTTGCTGAGTGGCTGAAGAATA TGGAAGACAACTACAATTCCTACACATTCTTGTACATTTTAGTTGAACAATGAGTGATTACATTTATTTA CCCAGTGCCTCTTCTATAAGGCAACAACTGGTAGTTTATCCTATTGAGAAGCGTAGATAGGATGCTTATT AGCAGTAACTGCTCTTGGTTTCAACTTGCATCTTACTTAGCTTTTTCACCGTTTTGTGGTTTCTTGGAGG AAGAATACCCATAATATACATTTGGAGACTGTTGTTCTGTAGTGTCAATGAAATGTGGGGGTGGGAAAAT GTCATTCAAGACTCCCATACAAAATGTCTATTGCTGCCTATATTTGCTATGGGAAAGTAGCCACAGATA ATGTTTTTTTTTTCCTCATTAGTATTTAAGATTTTCCATCCTAGTGGAAAGATATGATTTGATTCATCC TATTTACTTTGTATATTAAAGTACAGTAGAACCTGCCACTTTTTTTGGAAATGCAGCATAAGGATAAAGA TAAATTTCATATCAGTTCAGCAAGTTCTATTTAGCAGTGTGTTGAAGTTGAGACTGAATAAAATATTTTGG TTTGGTTTTCTGTTCAAAATTTTACCTTGATAAGGACAATATTTTTCTACATATATCAGTAGGCAGTAAT GATTACTTCAAAGCTTCCAAAGCCAGATACTACACCTGCATGTTCCAACATAGTTGCTGAATTTATTCCC AAGATGCATGTAATGTATAACTTTGTATTATTGAGAATGAATAAAGAAAAGTCATAATGATGCCTTCCAGC TGTGCAAGTTAATATTAAAAATAATTTGTTTGCATATTTCACCTAATAGGTCTTCTTCCATTGCTATACT GTTTACTTAAGTGAACAATGGAAATGTTGCTGTTTATCTTAAGGATTTGTAACATGCCTAAGATCTTACA GTACAGACTTCTATAATTAATGAAACATTTTTCTTTTTCCTTTCCAGGATCAGCTCAGCTACTATTTAAT AAAACAAAATCTGTAGAATTCACGTTTTGTAATGACACTGTCGTCATTCCATGCTTTGTTACTAATATGG AGGCACAAAACACTACTGAAGTATACGTAAAGTGGAAATTTAAAGGAAGAGATATTTACACCTTTGATGG AGCTCTAAACAAGTCCACTGTCCCCACTGACTTTAGTAGTGCAAAAATTGAAGTCTCACAATTACTAAAA GGAGATGCCTCTTTGAAGATGGATAAGAGTGATGCTGTCTCACACACAGGAAACTACACTTGTGAAGTAA CAGAATTAACCAGAGAAGGTGAAACGATCATCAGGAGCTAATATCGTGTTGGTAAGACTTCTATGAAAGC TTCTTTTTTTTATTTGTCCTGGTGCAACCTGATCCTCTTTCGAGAGGAGGCCAAATGGGGATAGGTACTCC TTTGAATCAAAAAGCAGGCTGTTATTAATAAATGTTGATGTAATGTTTAGCAAGTTTAAGATTGTGATTT CTATCCTATTTGTTAATCTACTCTTCATGGTAAAACACATTTACTATTTAATTTGTATTACTTGTTAATT TGTATATTGTGTGGGTTCTCAAACTTTTGGTCTAAGGACCATTTTATTTGGCTGATTGGTCCTGGGGTAAT GTACTCTTCATTTCTGTTTGCTGCCATCCAAGTGCTTTCTTTCCTGTGACTAATTTTCAAGTCTCTTTCC AATTATTCAGCCTTCTGTTTTTTGTGTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTAACCTTACACTTTGAC CAAGGGATGACAAGTCCCAGCCATGCTCCCTCTGGAGAGAATAGAGGAGAGTCAGGAGATTAAATCACGT TATCATGGATGATTTCTTCAGATTTTCTGTGCTTAGCCTTACTTTGGTCTTCTTTTTGTTGGCCAAAATA TGAGAGAACTAGAGATTCAAATTCATTTATTCAAATTCAGACTTGATGATAGCACCATGATTCTGCCCTT TTTATCACAAACTTGGGTTGCTGAGGTGTAGAAGGTAGTAAAATTAGCTGAAGTAAATAGTCTCCTCTCC TTGCTGTTTCCTGCTGTGCTAGCTGAGTTCTTCAACCCCATATGAGTATGTTCTTTCTGTTTTGAAATCT CTATTTAGAATTACCTGTCATATTACCGATCATGGCTTAACCACATTTTGAGAACCTTAACTCTAGATAG GTTCAGTTCTGTTCAGTTGGATCAACTATTTGCCTAGCTTTTATAAAATAATCTACTACCAGATTAGGAA TGGGTGTTTGGTTTGATGAAATGTTTGCACATTACTTAGGATTGAGAATCAGGAGGTCTGAGTTACAGC CCCTGTTCAGCAACTTACTATTCACCTGTGTGATCTTGAGAAGGTGCTTAACCGAGTCCAAGTTACCTCA TGTCCGAAATGGAGATGACAATAGATACTTGCTTTTATGTATTCTACAGTCTAATGAGCATAAACCAAAG TGTGTTGTAAATGTTAGTTGTTTTCCTGGGATGTTTATTTTATGAAGTAAGGTATCAAGTTGTTGAAGTT AGCCTAGAGCATTGTGAGAGGGATTTGTGTGCAGTGAGGAATGGGCAAGGGATGGCTCCGAGTAGGCAAG GAGAATTTGAAAGGTCTTTCTGCTCAGAAATCTCTCTTACATTGTAGCTTTTACTATTGATATTAATTTG GGCAAAGGAGCTTTTTTTGTTGTTGTTAAGTATACCTTTAATGAAGGGTGTAATCAGTCAGTCATATCCA CTGCAACTAACTGGTATTAGATCATCTAATACCTAGTTGAAATTTTTCTGAGGTAATGTACAATTTTGTG GAAATGAAAGAATAAGATATGGGGGAACAGTAACTTTTGGGGGGTAATGCAGGTCACTGAGAGGTAAACT GGAAAACTCAAAATATGAGATTTGAAAGGCCAGAAGATGAAATGAAGATGGAAGAGAAAGGAATGCCAGG ATCTGAGATAAAAACAGTTTATTATTTTGCGTGTGATTGAAAGACTGTTTTATCTTTGTTTGCAAAATAC CCTATAAAAATAAAAACTAGACTTATTTTGGGAGCCATTAAAAGGAGAAAGTATTGTTTTATTATAGAGA ATGTAGAAAATTCTACCAGAGGTTTGAGACATTTGGCTTTGCATCCATTTTTTAATTTTTTGAAATAATTAT TGATTTACAGGACGATTCAAAGATAGAGAGCTTCCATGTGCCCTTCACCCAGTCCCCCCAATGGTTTCCT CTTATGTGACACAATATGAAAATCAGGAAGTTCACAATGGTGCAGTGTGTGTTAATGGTTCGCTCTCATT TTATCACGTGTTTATTTGCGTAACTACCTACACAATCAGGACATAGACCTATCCATCATAACAAACATCT CCCTCATGCTTATTCTTACTAGTCATACCGCCACCTCCCACTGTCCCTAACCTGGCAAGCACCAATTTGT CCTCCATATCTATAATTTTGTCATTTCAACAATGCTATATAAATGGAATGATACAAGATGTGACCTTTTG AGAAGGGCTTTTTCACTAAGCACAATGCCCTTGAGATCTATCCAAGTTTTTGCATCTATCATAATAGTTCTT TTTTCCTTACTTTTTTTTTTGCTGAGTAGTAGTCCATAGTATGAATATATCACAGTTTGTTTAACCACTT ACCTATAGTAGGACATTTTGGTTGTTTCCAAGTTTGGGCTGTTACAAATGAAACTGCTCTGAACAATTGT GTACAGGTTCTTGTGTAGACACAGTTTTCATTTCTCTGGGATAAATGCTCAGAAGTATGGTTACTGGGCT GTATGGTAAGTATATGCTTAGTTTTTAAAGAAACTGCCAGACTTTTCCAGAGTGGCTATTTCATTTTACA TTTCCATCAACAATGTAAGAGTGATCTGGTTTCTCCACATCCTCACCAGTATCTGGTGCCACTATGTTGT AACCATTTTTAACTGAAAAAAACAACAAACAAACAAAAACCAGTTAAAAGTTATCTGCACAAAGGGTT AAATGGGGCAGGACTCTTACTATGGAGTATAAGTTCTTTTCTAATAAGAATACTTACTTGTCACGGACTT TGAGGATTTAAGCATTAGAAACCATTTTTACTATGTCGTGATTTTTGCAAATACAGGCATAAATGAGAAA CAGAATTTGCTCAATAGAGAAGTGAATTCTACTTAATAGAATGCAGATAATAGTGACCATTCTTGTGCC TTTTTAAATTTTTTGGAGGACTAGGACTGAATACATGGAAACACTATAAGATATAGTTTTATATACCTCT GTGCCTATGTATGATATAGTCCATTAGAAGGAGCACTGGACTTGAAATTAGAAAGCTGTTTTCTAGACTT CACTGTTTAATAGCTTAAGAAAAGTTGGATAAAAAACTCAACCTATCTGTCCCTCAAATTCTTCTTAGTA CAGTAAAGATGGCATCACCATCATGGAAATGTTTGAGATATAAATAATACCTCTCTTACTTAACAAAATT TTACTAAGTGCCTACCATGTGCCAGTCACCGTACTAGGCATCACAGATTCTGTGATGAATAAGACATTAT TGCTGATTTCAAGGAAGTGGTTGCAGTACAAATATGAAAGGTGAAGTGACCTAACATTTATTTGCACATG TGCCACATACTTTATGTAAGTCGTGTCCTGTTATCTCTCAAGAACTCTAGGAGATAGATGGAGTTCTTA CCATTTAACAAATAGGGAAGCAAAAGAGATTCAAGGTATTGTTCAAGGTCAGAGTAATAGCCAAGAATTA AACCTAGATCTCTTTGGCATCAAAACCCAGTATTTTTACCACCGTAATATGTGGTTCATGTGAAAACAC TTGTAAATAACTAATACTGAATGCAAATACTAGTAATCATAAAATTTATCATTAATATTTCTGTTCTAT TAGCAATACAAAAACTTGAAAACTTAAAGTTTATTTTTCTCTAAGCTATTAGAGTTTTGTTTAGAAAGGT CAGTTCAAGTTCCGCAGTGTGCCAATTCCATTATTATCAAACAGGACATATGGTCTGTTTATAATGAA CATGAAGGCATTGCAAGATCTTGTATTAGTTTTCTAGTGCTGCATAACACTACAAATATAGCAGCTTA AAACACTACCCATTTTTTAGCTCAAAGTTTTGTAGGTGGCAAGCCTGGTCCATGGCATGACTGGGTTCTCT GATCAAAGTCTTAAAAAGACTAAAAATCAAGGTGTTGCCCAGGGCATACTTTATTTGGAGTTTGGGGTCTTG TTCTAGGCTCACATAATTGTGACAGAATTCAATTCCTTGTGATTGTAAGACTGGGGTCCCTGTTTCTTTG CTAAGTATCTGCCAAGGAATTGTAGCAGCTCCTAGAGATTGCCCTCATTTCTTGCCCCATGGCCCCTTGC ATATTTAAAGCCAGCAAAGGAGAATCTTCCTCTTGTTGAATCCCTCTCACATATTGAGTTTATTTTGCCA GGAAGAACCCAGACCCTTTTAAGAGACCACTTGGTTAGGTCAGCTCTCTCTCCCTCAAAATATAATCTCTC TTTCTTAAAGTCAGCTGATTTGGGATCTTAATTACATCAGCAAACTCCCTTTTGCTGGGTGATATAATCA TGAAAATGAAATCCAATACATTCACAGTCCTAGTCTCCCACACACAAGGGGAGGGAATTATAAAGATGC ATGGCTAGGGCAGGGGTTTTGGGACCACCTTAGAATTCTGGGTTTGATTAAATAAATAATGGGACTGTAG CCTAGCAAGTCGACACATCAGAAAAGCCATCACAATCCTGAAACATCAAGTAAAAATTTGGTTTGCATTT TAGGATTGTAAATGTTACTGTGGTGTATGCGTGTGTGTGTGTGTGTAACCTGGTGTTCCCTATGTTTGTG GGAGTTTGAAGGAGACACTTTGTTGATAGGAATGGTGTCTTCACTTTTTTAGGTTGTCTCATTTTTGTAC AGTGATAAGACAATGAGGTCCTGGGTTTTAATAACACTTCAGCTTGAAAGCAAAAATTAAACACTTATT CATTGACTACACCCTTGTAATATCACTTCTTTGCCTTTCCACTAGCAAGAAGTTCATTTTCGTGGAAGCC ATTCTGCCTGTCCAGGAATTGGAGGAGAGTGATAGACACAGTTGTCAGCCATAGCTTGGGTAGAATAAGG ATGTGAATGTCCTTGGCTTATCTTTATTAATCTTGTGATGGAAAAATATCTGACATTGTTCTTAGTCCAT TTTAAGCTTAATTTATGTTCTTAGTGGCATAGAAATTCAGAGCTGAAAGAAGTATCATGTCTCACTCTCC CCTTGAGGAACAAGAGTTAACGTCATCTGACAGTACTACCATAACAAATCAATAGCTTAATAGGCATATA AGTGGCTTTATATAAAATGTTGGTTTTTTTCCCCCAGCATCTCAGTTGGTTCTTAAATATCTAATTCCAT GATCCTCAAACTTTTCCCACTGTAACAAATTAGAGAGAGGAGGAACATGTTCATCAGTGACTGTAGTTCA AATACAGCAGAAATGTGCAGCAGTGATTTCAAACTGAGAGAATCCTGGATGCCCTTCTCCATTGTGTGCC CCCCCCCCCCACCCGCCCCACATATCAGGGAACAATTTAAAATCCTGGCACAATAATGAGAAGGGAGAGT GACAAACTGATAAGTTCCAGTTAAGAATCACTTAGACAGGCCAGGTGTGGCAGCCCACGCCTGTAATACC AGCACTTTGGGAGGCTGAGGCAGGTGGATCACCTGAGGTCAGGAATTTGAGACCAGCCTGGCCAACATGA TAAAACCCTGTCTCTACCAAAATTACAAAATTTAGCTGGGCATGGTGGCGCCTGCCTATAATCCCAGCTA CTTGGGAGGCTGAGACAGGAGAATTGCTTGAACCCGGGAGGTGGAGGTTGCAGTGAGCTGAGACCACACC ACTGCACTCCAGCCTGGGCAACAGAGTGAGACTCCATCTCAAAAATAAAAAAAAAGAATCACTTAGACA GTATTTTTTGTTCTGTAATCTTCCTCTCTGTCATTGAAATGTCTCATATTTCTTGTTTCACATAGAATCT GGGAGTCATCAAGTAGTCATCTAGTCTATCTCATCATCTAATTTACGAATTATTTCTATCACACCCCAAG TAGCTGGCGATTCAGATTTTTTACTGAAAGCTTGACATGATGAGACATTATTACTTCCCAGAGGATCTTG GTCTGTTATCAGGCAATTTCAGTTCTTCCTTAAATCAAGCTAAAACTTGCTACTATTCTCACTCACTGGG AGGGCTGTAGTATTTCTCTCAAGTTTATACCTCTCTGTGATGAAAATCTCTGTTTCTTTTAGCCATTGCTC ATGGTTTCGGGATCCCCATTCTAGTCATCTTCCTTTGGAGCAATTCCCTGTTAGATCTTCAAATGTGATG AGTAAAACTGAGCTCAGGACTGCGTTTGTTGGTCACACCCTTCTCCCTTCCAAATGACTGCTGCTGTAAC TTTCTGACAGGTCCTGCTTCCCTGTAATCCATTCTCCATGTGGCTGCCAAAGAAGATTTGAAATAAGTCA GGTTATTTTTCTTCCCTATTTAAAGCCTTCCAGCCGCTTGTCATTGCTCTTAGAATAAAATCCAAACTCT CCCAACAAGCTTTTAAGACTCTACATGGTCTTGCCCCAGCAAACTCTTCCCATTTGATCTCATCACTGAA TTCCAGTCACAATGGCCATCCTTTGGTTTCTCAAAGTTGCCGTTCCTTTGGCATCTCTTCCTCCCTGTCT TCACAATGTAGGCTCCTTCCCATCCTAGGCTTAGTTTCGTTCC...

Claims

1. A genetically engineered hematopoietic cell, or progeny thereof, comprising a genomic modification of a gene encoding a lineage-specific cell surface antigen, said genomic modification altering the amino acid sequence of an epitope recognized by an agent that specifically binds to said lineage-specific cell surface antigen resulting in a modified lineage-specific cell surface antigen, said modified lineage-specific cell surface antigen being characterized by reduced or no binding of said agent.

2. The genetically engineered hematopoietic cell or progeny thereof of claim 1 , wherein the genomic modification alters 1, 2, 3, 4, or 5 amino acid residues in the lineage-specific cell surface antigen.

3. 2. The genetically engineered hematopoietic cell or progeny thereof of claim 1 , wherein the genomic modification alters no more than one, no more than two, no more than three, no more than four, or no more than five amino acid residues in the lineage-specific cell surface antigen.

4. The genetically engineered hematopoietic cell or its progeny according to any one of claims 1 to 3, wherein the genomic modification results in a deletion, substitution, insertion or inversion of one or more amino acid residues, or a combination thereof.

5. The genetically engineered hematopoietic cell or its progeny according to any one of claims 1 to 4, wherein said genomic modification results in the substitution of one or more amino acid residues.

6. The genetically engineered hematopoietic cell or its progeny according to any one of claims 1 to 5, wherein said epitope is characterized by an endogenous post-translational modification.

7. The genetically engineered hematopoietic cell or its progeny according to claim 6 , wherein the endogenous post-translational modification is glycosylation.

8. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 7, wherein the agent is an immunotherapeutic agent.

9. The genetically engineered hematopoietic cell or progeny thereof of claim 8 , wherein the immunotherapeutic agent comprises an antibody or an antigen-binding fragment thereof.

10. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 7, wherein said modified lineage-specific cell surface antigen is not recognized by said agent.

11. 9. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 8, wherein the modified lineage-specific cell surface antigen is recognized by a second agent that specifically binds to the lineage-specific cell surface antigen in a region that is distinct from the epitope recognized by the first agent.

12. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 11, wherein said genomic modification does not substantially alter the function of said lineage-specific cell surface antigen.

13. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 12, wherein said genomic modification does not substantially alter the expression of said lineage-specific cell surface antigens.

14. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 13, wherein said genomic modification does not substantially alter the viability or growth of the cell.

15. 15. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 14, wherein the hematopoietic cell or progeny thereof retains the ability to differentiate normally compared to a reference population of hematopoietic cells, optionally a population of hematopoietic cells that does not comprise the genomic modification.

16. The genetically engineered hematopoietic cell or its progeny according to any one of claims 1 to 15, wherein the hematopoietic cell is a hematopoietic stem cell (HSC).

17. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 16, wherein the hematopoietic cell is a CD34+ cell.

18. The genetically engineered hematopoietic cell or its progeny according to any one of claims 1 to 17, wherein the hematopoietic cell is obtained from bone marrow, blood, umbilical cord, or peripheral blood mononuclear cells (PBMC).

19. The genetically engineered hematopoietic cell or a progeny thereof according to any one of claims 1 to 18, wherein the hematopoietic cell is a human cell.

20. 18. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 17, wherein the lineage-specific cell surface antigen is selected from the group consisting of CD123, CD47, CD34, CD38, CD19, CD33, CLL-1, CD30, CD5, CD6, CD7, EMR2, and BCMA.

21. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 20, wherein the lineage-specific cell surface antigen is CD123.

22. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 20, wherein the lineage-specific cell surface antigen is CD38.

23. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 20, wherein the lineage-specific cell surface antigen is CD19.

24. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 20, wherein the lineage-specific cell surface antigen is EMR2.

25. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 20, wherein the lineage-specific cell surface antigen is CD5.

26. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 20, wherein the lineage-specific cell surface antigen is CD47.

27. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 1 to 20, wherein the lineage-specific cell surface antigen is CD34.

28. The genetically engineered hematopoietic cell or its progeny according to any one of claims 1 to 21, wherein said epitope is encoded by exon 3 and / or exon 4 of the gene encoding CD123.

29. 30. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 21, or 28, wherein the epitope is a region of CD123 bound by the murine anti-CD123 antibody 7G3, a humanized variant thereof (e.g., antibody CSL-362), or talacuzumab.

30. 30. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 21, 28, or 29, wherein the agent comprises the murine anti-CD123 antibody 7G3, a humanized variant thereof (e.g., antibody CSL-362), talacuzumab, or an antigen-binding fragment thereof.

31. 31. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 21 or 28 to 30, wherein the epitope comprises 1, 2, 3, 4, or 5 amino acids at positions 51, 59, 61, 82, or 84 of the wild-type gene encoding CD123.

32. 32. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 21 or 28 to 31, wherein the genomic modification results in a deletion, substitution, insertion or inversion of one or more amino acids at positions 51, 59, 61, 82 or 84 of the wild-type gene encoding CD123 or at the corresponding positions of a homologous CD123 gene.

33. 33. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 21 or 28 to 32, wherein the genomic modification results in the substitution of one or more (e.g. one, two, three, four or all) amino acids at positions 51, 59, 61, 82 or 84 of the wild-type gene encoding CD123, or at the corresponding positions of a homologous CD123 gene.

34. The genetically engineered hematopoietic cell or progeny thereof according to any one of claims 4 to 33, wherein said one or more substitutions are conservative substitutions.

35. 35. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 21 or 28 to 34, wherein the genomic modification results in an amino acid substitution at position 51 of the wild-type gene encoding CD123 or at the corresponding position of a homologous CD123 gene.

36. 35. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 21 or 28 to 34, wherein the genomic modification results in a substitution of lysine for glutamic acid at position 51 of the wild-type gene encoding CD123 or at the corresponding position of a homologous CD123 gene.

37. 23. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20 or 22, wherein said epitope is encoded by exon 7 of the gene encoding CD38.

38. 38. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 22, or 37, wherein the epitope is a region of CD38 bound by the murine anti-CD38 antibody HB7, a humanized variant thereof, or daratumumab.

39. 40. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 22, 37, or 38, wherein the agent comprises the murine anti-CD38 antibody HB7, a humanized variant thereof, daratumumab, or an antigen-binding fragment thereof.

40. 40. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 22, or 37 to 39, wherein the epitope comprises 1, 2, 3, 4, or 5 amino acids at positions 270 to 274 of the wild-type gene encoding CD38.

41. 41. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 22, or 37 to 40, wherein said genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 270 to 274 of the wild-type gene encoding CD38, or at the corresponding positions of a homologous CD38 gene.

42. 42. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 22, or 37-41, wherein the genomic modification results in the substitution of one or more (e.g., 1, 2, 3, 4, or all) amino acids at positions 270-274 of the wild-type gene encoding CD38, or at the corresponding positions of a homologous CD38 gene.

43. 43. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 4 to 20, 22, or 37 to 42, wherein said one or more substitutions are conservative substitutions.

44. 44. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 22, or 37 to 43, wherein said genomic modification results in an amino acid substitution at position 272 of the wild-type gene encoding CD38 or at the corresponding position of a homologous CD38 gene.

45. 45. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 22, or 37-44, wherein the genomic modification results in a substitution of arginine, histidine, or alanine for glutamine at position 272 of the wild-type gene encoding CD38, or at the corresponding position of a homologous CD38 gene.

46. 24. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20 or 23, wherein said epitope is encoded by exon 2 or exon 4 of CD19.

47. 47. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 23, or 46, wherein the epitope is a region of CD19 bound by anti-CD19 antibody B43, anti-CD19 antibody FMC63, or an antigen-binding fragment thereof.

48. 48. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 23, 46, or 47, wherein the agent comprises anti-CD19 antibody B43, anti-CD19 antibody FMC63, tafasitamab, roncatuximab, blinatumomab, or an antigen-binding fragment thereof.

49. 49. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 23, or 46 to 48, wherein the epitope comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at positions 216 to 224 or 218 to 238 of the wild-type gene encoding CD19.

50. 50. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 23, or 46 to 49, wherein said genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 163, 164, 216-224, or 218-238 of the wild-type gene encoding CD19, or at the corresponding positions of a homologous CD19 gene.

51. 51. The genetically engineered hematopoietic cell of any one of claims 1 to 20, 23, or 46 to 50, or a progeny thereof, wherein the genomic modifications result in the substitution of one or more (e.g. 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more, e.g. all) amino acids at positions 163, 164, 216-224, or 218-238 of the wild-type gene encoding CD19, or at the corresponding positions of a homologous CD19 gene.

52. 52. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 4 to 20, 23, or 46 to 51, wherein said one or more substitutions are conservative substitutions.

53. 53. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 23, or 46-52, wherein said genomic modification results in an amino acid substitution at position 163 of the wild-type gene encoding CD19 or at the corresponding position of a homologous CD19 gene.

54. 54. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 23, or 46 to 53, wherein said genomic modification results in amino acid substitutions at positions 163 and 220 of the wild-type gene encoding CD19 or at the corresponding positions of a homologous CD19 gene.

55. 54. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 23, or 46 to 53, wherein the genomic modification results in an amino acid substitution at positions 163 and 164 of the wild-type gene encoding CD19 or at the corresponding positions of a homologous CD19 gene.

56. 54. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 23, or 46 to 53, wherein the genomic modification results in a substitution of the amino acid cysteine ​​or leucine at position 163 of the wild-type gene encoding CD19 or at the corresponding position of a homologous CD19 gene.

57. 54. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 23, or 46-53, wherein the genomic modification results in a substitution of the amino acid phenylalanine at position 164 of the wild-type gene encoding CD19 or at the corresponding position of a homologous CD19 gene.

58. 54. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 23, or 46 to 53, wherein said genomic modification results in an amino acid substitution at positions 163 and 164 of a wild-type gene encoding CD19 or at the corresponding positions of a homologous CD19 gene, wherein said substitution of the amino acid at position 163 is a cysteine ​​or leucine and said substitution of the amino acid at position 164 is a phenylalanine.

59. 25. The genetically engineered hematopoietic cell of any one of claims 1 to 20 and 24, or a progeny thereof, wherein the epitope comprises 1, 2, 3, 4, 5, or 6 amino acids at positions 124, 132, 146, 292, 294, 295, 296, 298, 299, 303, 304, 305, 306, 307, 308, 312, 318, 320, 328, 329, 331, 332, 335, 340, 347, 527, or 708 of the wild-type gene encoding EMR2.

60. 60. The genetically engineered hematopoietic cell of any one of claims 1 to 20, 24 or 59, or a progeny thereof, wherein the genomic modification results in a deletion, substitution, insertion or inversion of one or more amino acids at positions 124, 132, 146, 292, 294, 295, 296, 298, 299, 303, 304, 305, 306, 307, 308, 312, 318, 328, 329, 331, 332, 335, 340, 347, 527 or 708 of the wild-type gene encoding EMR2, or at the corresponding positions of a homologous EMR2 gene.

61. 27. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20 or 26, wherein the epitope is a region of CD47 bound by anti-CD47 antibody B6H12, anti-CD47 antibody 2D3, or an antigen-binding fragment thereof.

62. 62. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 26, or 61, wherein the agent comprises anti-CD47 antibody B6H12, anti-CD47 antibody 2d3, ligfalimab, or an antigen-binding fragment thereof.

63. 63. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 26, 61, or 62, wherein the epitope comprises 1, 2, 3, 4, 5, or 6 amino acids at positions 117 to 122 of the wild-type gene encoding CD47.

64. 64. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 26, or 61-63, wherein the epitope comprises 1, 2, 3, or 4 amino acids at positions 47, 49, 52-55, or 117-122 of the wild-type gene encoding CD47.

65. 65. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 26, or 61-64, wherein the genomic modification results in a deletion, substitution, insertion, or inversion of one or more amino acids at positions 31, 47, 49, 52-55, 117-122, or 124 of the wild-type gene encoding CD47, or at the corresponding positions of a homologous CD47 gene.

66. 66. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 4-20, 26, or 61-65, wherein said one or more substitutions are conservative substitutions.

67. 67. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 26, or 61-66, wherein the genomic modification results in the substitution of one or more amino acids at positions 31, 47, 49, 52-55, 117-122, or 124 of the wild-type gene encoding CD47, or at the corresponding positions of a homologous CD47 gene.

68. 68. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 26, or 61-67, wherein said genomic modification results in an amino acid substitution at position 49 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene.

69. The genome modification (i) a substitution of the amino acid at position 4 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene for histidine; (ii) a substitution of the amino acid at position 49 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene with an arginine; (iii) a substitution of the amino acid at position 49 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene for proline; (iv) a substitution of the amino acid at position 52 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene with an alanine; (v) a substitution of the amino acid at position 53 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene with an alanine; (vi) a substitution of the amino acid at position 53 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene for proline; (v) a substitution of an alanine for the amino acid at position 120 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene; or (vi) a substitution of an amino acid at position 124 of the wild-type gene encoding CD47 or at the corresponding position of a homologous CD47 gene for a lysine; 69. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 26, or 61 to 68, which results in

70. 28. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20 or 27, wherein the epitope is a region of CD34 bound by anti-CD34 antibody QBend10, anti-CD34 antibody 561, or an antigen-binding fragment thereof.

71. 71. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 27 or 70, wherein the genomic modification results in a deletion, substitution, insertion or inversion of one or more amino acids at positions 42, 45, 46, 47, 49, 50, 51, 54 or 55 of the wild-type gene encoding CD34, or at the corresponding positions of a homologous CD34 gene.

72. 72. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 4 to 20, 27, 70, or 71, wherein said one or more substitutions are conservative substitutions.

73. 73. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 27, or 70-72, wherein the genomic modification results in the substitution of one or more amino acids at positions 42, 45, 46, 47, 49, 50, 51, 54, or 55 of the wild-type gene encoding CD34, or at the corresponding positions of a homologous CD34 gene.

74. 74. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1-20, 27, or 70-73, wherein the genomic modification results in an alanine substitution at the amino acid at any one or more of positions 45, 46, 50, 51, 54, 55 of the wild-type gene encoding CD34, or at the corresponding positions of a homologous CD34 gene.

75. The genome modification (i) a substitution of the amino acid at position 46 of the wild-type gene encoding CD34 or at the corresponding position of a homologous CD34 gene for a phenylalanine; (ii) a substitution of the amino acid lysine at position 47 of the wild-type gene encoding CD34 or at the corresponding position of a homologous CD34 gene; (iii) a substitution of the amino acid at position 47 of the wild-type gene encoding CD34 or at the corresponding position of a homologous CD34 gene for glutamic acid; (iv) a substitution of the amino acid at position 49 of the wild-type gene encoding CD34 or at the corresponding position of a homologous CD34 gene with a phenylalanine; or (v) a substitution of an amino acid serine at position 49 of the wild-type gene encoding CD34 or at the corresponding position of a homologous CD34 gene; 75. The genetically engineered hematopoietic cell or progeny thereof of any one of claims 1 to 20, 27, or 70 to 74,

76. A method for providing a method to a subject in need thereof, (i) administering a population of genetically engineered hematopoietic cells or progeny thereof according to any one of claims 1 to 75.

77. 77. The method of claim 76, further comprising: (ii) administering an effective amount of an agent that specifically binds to the lineage-specific cell surface antigen.

78. 78. The method of claim 76 or 77, wherein the subject has a hematopoietic malignancy.

79. 79. The method of claim 77 or 78, wherein the agent is a single chain antibody fragment (scFv).

80. 80. The method of any one of claims 77 to 79, wherein the agent is an antibody or an antibody-drug conjugate (ADC).

81. 80. The method of claim 77 or 78, wherein the agent is an immune cell expressing a chimeric antigen receptor comprising an antigen-binding fragment.

82. 82. The method of claim 81 , wherein the immune cell is a T cell.

83. 83. The method of claim 82, wherein the T cells express CD3, CD4, and / or CD8.

84. The chimeric antigen receptor (a) a hinge domain, (b) a transmembrane domain, (c) at least one costimulatory domain; (d) a cytoplasmic signaling domain, or The method of any one of claims 81 to 83, further comprising (e) a combination thereof.

85. 85. The method of claim 84, wherein the chimeric antigen receptor comprises at least one costimulatory signaling domain derived from a costimulatory receptor selected from the group consisting of CD27, CD28, 4-1BB, OX40, CD30, ICOS, CD2, CD7, LIGHT, NKG2C, B7-H3, GITR, HVEM, and combinations thereof.

86. 86. The method of claim 84 or claim 85, wherein the chimeric antigen receptor comprises a cytoplasmic signaling domain derived from CD3zeta.

87. 87. The method of any one of claims 84 to 86, wherein the chimeric antigen receptor comprises a hinge domain derived from CD8α or CD28.

88. 88. The method of any one of claims 77 to 87, wherein the agent comprises the murine anti-CD123 antibody 7G3, a humanized variant thereof (e.g., antibody CSL-362), or talacuzumab; the murine anti-CD38 antibody HB7, a humanized variant thereof, or daratumumab; B43; or the anti-CD19 antibodies blinatumomab, FMC63, or HIB19; or the anti-CD47 antibodies B6H12 or 2D3; or the anti-CD34 antibodies QBend10 or 561; or the anti-CD5 antibody H65.

89. 89. The method of any one of claims 78 to 88, wherein the hematopoietic malignancy is Hodgkin's lymphoma, non-Hodgkin's lymphoma, leukemia, multiple myeloma (MM), myelodysplastic syndrome (MDS), or blastic plasmacytoid dendritic cell neoplasm (BPDCN).

90. 90. The method of any one of claims 78 to 89, wherein the hematopoietic malignancy is acute myeloid leukemia, B-cell acute lymphoblastic leukemia (B-ALL), chronic myeloid leukemia, acute lymphoblastic leukemia, or chronic lymphoblastic leukemia.

91. The method of any one of claims 78 to 90, wherein the hematopoietic malignancy is B-cell acute lymphoblastic leukemia (B-ALL).

92. The method of any one of claims 78 to 90, wherein the hematopoietic malignancy is acute myeloid leukemia (AML).

93. The method of any one of claims 78 to 90, wherein the hematopoietic malignancy is multiple myeloma (MM).

94. The method of any one of claims 78 to 90, wherein the hematopoietic malignancy is myelodysplastic syndrome (MDS).

95. 1. A method comprising: genetically modifying hematopoietic cells to introduce a genomic modification in a gene encoding a lineage-specific cell surface antigen, said genomic modification altering the amino acid sequence of an epitope recognized by an agent that specifically binds to said lineage-specific cell surface antigen resulting in a modified lineage-specific cell surface antigen, said modified lineage-specific cell surface antigen being characterized by reduced or no binding of said agent; thereby producing engineered hematopoietic cells that have reduced or no binding to an agent that targets said lineage-specific cell surface antigen.

96. 96. The method of claim 95, further comprising providing hematopoietic cells.

97. The method of claim 95 or 96, wherein the genetically engineered hematopoietic cell is a genetically engineered hematopoietic cell according to any one of claims 1 to 75.

98. Genetically modifying the hematopoietic cells comprises: (a) a clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) (CRISPR / Cas) system comprising a Cas nuclease, and a guide RNA (gRNA) comprising a nucleotide sequence that hybridizes to a gene encoding a lineage-specific cell surface antigen in the genome of said hematopoietic cell (e.g., a sequence encoding an epitope bound by an agent that specifically binds to said lineage-specific cell surface antigen); The method of any one of claims 95 to 97, comprising (b) contacting with a template polynucleotide.

99. The contacting comprises: (c) (i) the expansion factor, (ii) Homologous recombination repair (HDR) promoting factor Either or both of 99. The method of claim 98, further comprising contacting the

100. 100. The method of any one of claims 98 or 99, wherein the CRISPR / Cas system generates a double stranded break (DSB) in the gene encoding the lineage specific cell surface antigen in the genome of the hematopoietic cell.

101. The method of any one of claims 98 to 100, wherein the template polynucleotide is a single-stranded donor oligonucleotide (ssODN) or a double-stranded donor oligonucleotide (dsODN).

102. 102. The method of any one of claims 98 to 101, wherein the template polynucleotide hybridizes to genomic sequences adjacent to the DSB in the gene encoding the lineage-specific cell surface antigen and is integrated into the gene encoding the lineage-specific cell surface antigen.

103. 103. The method of any one of claims 98-102, wherein the template polynucleotide comprises a donor sequence, a first flanking sequence that is homologous to genomic sequence upstream of the DSB in the gene encoding the lineage-specific cell surface antigen, and a second flanking sequence that is homologous to genomic sequence downstream of the DSB in the gene encoding the lineage-specific cell surface antigen.

104. The method of claim 103, wherein the donor sequence of the template polynucleotide is integrated into the genome of the hematopoietic cell by homology directed repair (HDR).

105. 105. The method of any one of claims 99 to 104, wherein the expansion factors include SR1 and UM171.

106. 106. The method of any one of claims 99 to 105, wherein the HDR promoting factor comprises at least one of SCR7, NU7441, rucaparib, and RS-1.

107. 107. The method of any one of claims 101 to 106, wherein the ssODN is 50 to 200 nucleotides in length.

108. The method of any one of claims 101 to 107, wherein the ssODN is 120 nucleotides in length.

109. 109. The method of any one of claims 98-108, wherein said contacting comprises contacting a population of hematopoietic cells.

110. 110. The method of claim 109, further comprising sorting the population of hematopoietic cells.

111. 111. The method of claim 110, wherein sorting comprises selecting viable hematopoietic cells.

112. 112. The method of claim 110 or 111, wherein sorting comprises selecting hematopoietic cells that have integrated the donor sequences into their genome.

113. The method of any one of claims 110 to 112, wherein sorting comprises fluorescence activated cell sorting (FACS).

114. 114. The method of any one of claims 110 to 113, wherein sorting comprises selecting viable, long-term engrafting HSCs.

115. 115. The method of any one of claims 110-114, wherein the editing efficiency in the population of hematopoietic cells is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%.

116. The method of any one of claims 110 to 115, wherein the percentage of viability in the population of hematopoietic cells is at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, or at least 99%.

117. 117. The method of any one of claims 110 to 116, wherein the efficiency of HDR is 50% or more.

118. 118. The method of any one of claims 110 to 117, wherein the efficiency of the HDR is 60% or more.

119. 119. The method of any one of claims 110 to 118, wherein the efficiency of the HDR is 80% or more.

120. 120. The method of any one of claims 95 to 119, wherein the lineage specific cell surface antigen is selected from the group consisting of CD33, CD123, CD19, CLL-1, CD30, CD5, EMR2, CD6, CD7, CD38, CD34, CD47, and BCMA.

121. 121. The method of any one of claims 95 to 120, wherein the lineage-specific cell surface antigen is CD123.

122. 122. The method of Claim 121, wherein the gRNA comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 3, 6, 9, and 12.

123. 123. The method of claim 121 or 122, wherein the first flanking sequence is homologous to a first portion of the CD123 gene and the second flanking sequence is homologous to a second portion of the CD123 gene.

124. The method of claim 123, wherein the first portion of the CD123 gene comprises a portion of exon 3 or a sequence proximal thereto.

125. The method of claim 123, wherein the first portion of the CD123 gene comprises a portion of exon 4 or a sequence proximal thereto.

126. 126. The method of any one of claims 123 to 125, wherein the second portion of the CD123 gene comprises a portion of exon 3 or a sequence proximal thereto.

127. 126. The method of any one of claims 123 to 125, wherein the second portion of the CD123 gene comprises a portion of exon 4 or a sequence proximal thereto.

128. 128. The method of any one of claims 123 to 127, wherein the first portion and the second portion are not identical.

129. 129. The method of any one of claims 121 to 128, wherein the donor sequence comprises a sequence corresponding to a codon encoding 1, 2, 3, 4, or 5 amino acids at positions 51, 59, 61, 82, or 84 of a wild-type gene encoding CD123.

130. 130. The method of any one of claims 121 to 129, wherein the first flanking sequence comprises a flanking sequence set forth in any one of SEQ ID NOs: 93 to 99.

131. The method of any one of claims 121 to 130, wherein the second flanking sequence comprises a flanking sequence as set forth in any one of SEQ ID NOs: 93 to 99.

132. The method of any one of claims 121 to 131, wherein the donor sequence comprises a donor sequence set forth in any one of SEQ ID NOs: 93 to 99.

133. The method of any one of claims 121 to 132, wherein the template polynucleotide comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 93 to 99.

134. 121. The method of any one of claims 95 to 120, wherein the lineage-specific cell surface antigen is CD38.

135. 135. The method of claim 134, wherein the gRNA comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 18, 21, 24, 27, 30, 33, 36, 39, 42, 45, 48, 51, 54, 57, and 60.

136. 136. The method of claim 134 or 135, wherein the first flanking sequence is homologous to a first portion of the CD38 gene and the second flanking sequence is homologous to a second portion of the CD38 gene.

137. 137. The method of claim 136, wherein the first portion of the CD38 gene comprises a portion of exon 7 or a sequence proximal thereto.

138. 138. The method of claim 136 or 137, wherein the second portion of the CD38 gene comprises a portion of exon 7 or a sequence proximal thereto.

139. The method of any one of claims 136 to 138, wherein the first portion and the second portion are not identical.

140. 140. The method of any one of claims 134 to 139, wherein the donor sequence comprises a sequence corresponding to codons encoding 1, 2, 3, 4, or 5 amino acids at positions 270 to 274 of a wild-type gene encoding CD38.

141. 121. The method of any one of claims 95 to 120, wherein the lineage-specific cell surface antigen is CD19.

142. 142. The method of claim 141, wherein the gRNA comprises a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 66, 69, 72, 75, 78, 81, and 84.

143. 143. The method of claim 141 or 142, wherein the first flanking sequence is homologous to a first portion of the CD19 gene and the second flanking sequence is homologous to a second portion of the CD19 gene.

144. 144. The method of claim 143, wherein the first portion of the CD19 gene comprises a portion of exon 2 or a sequence proximal thereto.

145. 144. The method of claim 143, wherein the first portion of the CD19 gene comprises a portion of exon 4 or a sequence proximal thereto.

146. 146. The method of any one of claims 143 to 145, wherein the second portion of the CD19 gene comprises a portion of exon 2 or a sequence proximal thereto.

147. 146. The method of any one of claims 143 to 145, wherein the second portion of the CD19 gene comprises a portion of exon 4 or a sequence proximal thereto.

148. 148. The method of any one of claims 143 to 147, wherein the first portion and the second portion are not identical.

149. 149. The method of any one of claims 141 to 148, wherein the donor sequence comprises a sequence corresponding to codons encoding 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids at positions 216 to 224 or 218 to 238 of a wild-type gene encoding CD19.

150. 150. The method of any one of claims 95 to 149, wherein said genomic modification results in expression of a variant form of said lineage specific cell surface antigen that is not recognized by said agent.

151. 151. The method of any one of claims 95-150, wherein the genomic modification results in expression of a variant form of the lineage-specific cell surface antigen that is recognized by a second agent that specifically binds to the lineage-specific cell surface antigen in a region that is distinct from the agent that binds the epitope.

152. The method of any one of claims 96 to 151, wherein the Cas nuclease is a Cas9 nuclease.

153. The method of any one of claims 96 to 152, wherein the Cas nuclease is a Streptococcus pyogenes Cas9 (spCas9) nuclease.

154. The method of any one of claims 96 to 152, wherein the Cas nuclease is a Staphylococcus aureus Cas9 (saCas9) nuclease.

155. The method of any one of claims 96 to 152, wherein the Cas nuclease is a Cas12a nuclease.

156. The method of any one of claims 96 to 152, wherein the Cas nuclease is a Cas12b nuclease.

157. 157. The method of any one of claims 96-156, wherein said contacting comprises introducing the CRISPR / Cas system into said cell in the form of a preformed ribonucleoprotein (RNP) complex.

158. The method of claim 157, wherein the ribonucleoprotein complex is introduced into the hematopoietic cells via electroporation.

159. The method of any one of claims 98 to 158, wherein the template polynucleotide and the CRISPR / Cas system are simultaneously electroporated into the cell.

160. A genetically engineered hematopoietic cell, wherein said cell is obtained or obtainable by a method according to any one of claims 95 to 159.

161. A population of genetically engineered hematopoietic cells comprising a plurality of the genetically engineered hematopoietic cells of any one of claims 1 to 75, or the genetically engineered hematopoietic cells of claim 160.

162. 162. A pharmaceutical composition comprising a genetically engineered hematopoietic cell according to any one of claims 1 to 75 or a progeny thereof, a genetically engineered hematopoietic cell according to claim 160, or a population of genetically engineered hematopoietic cells according to claim 161.

163. 1. A method for producing a genetically engineered hematopoietic stem or progenitor cell, or a plurality thereof, comprising at least one nucleotide substitution in a gene encoding a lineage-specific cell surface antigen, said method comprising: (i) a guide RNA (gRNA) comprising a targeting domain that targets a nucleotide sequence within the genome of the hematopoietic stem or progenitor cell; (ii) a base editor comprising a catalytically damaging Cas9 endonuclease fused to a cytosine (CBE) or an adenosine deaminase (CBE), thereby producing said engineered hematopoietic stem or progenitor cell, or a plurality thereof.

164. 164. The method of claim 163, wherein said at least one substitution produces a missense variant in the gene encoding said lineage-specific cell surface antigen.

165. 164. The method of claim 163, wherein said at least one substitution produces an alteration in the translation start site of said gene encoding said lineage-specific cell surface antigen.

166. 164. The method of claim 163, wherein said at least one substitution produces a splice region variant in said gene encoding said lineage-specific cell surface antigen.

167. 167. The method of any one of claims 163 to 166, wherein the gene encoding the lineage-specific cell surface antigen is selected from the group consisting of CD123, CD47, CD34, CD38, CD19, CD33, CLL-1, CD30, CD5, CD6, CD7, and BCMA.

168. 168. The method of any one of claims 163 to 167, wherein the gene encoding the lineage-specific cell surface antigen is selected from the group consisting of CD123, CD47, CD34, CD38, CD19, and CD5.

169. The method of any one of claims 163 to 168, wherein the gene encoding the lineage-specific cell surface antigen is CD123.

170. 170. The method of any one of claims 163 to 169, wherein the gene encoding the lineage-specific cell surface antigen is CD47.

171. 170. The method of any one of claims 163 to 169, wherein the gene encoding the lineage-specific cell surface antigen is CD34.

172. 170. The method of any one of claims 163 to 169, wherein the gene encoding the lineage-specific cell surface antigen is CD38.

173. 170. The method of any one of claims 163 to 169, wherein the gene encoding the lineage-specific cell surface antigen is CD19.

174. 170. The method of any one of claims 163 to 169, wherein the gene encoding the lineage-specific cell surface antigen is CD5.

175. The method of any one of claims 163 to 174, wherein the gRNA comprises a nucleotide sequence set forth in any one of SEQ ID NOs: 1-12, 16-60, 64-84, 100-181, 195, 196, and 204-423.

176. 176. The method of any one of claims 163-175, wherein the catalytically impaired Cas9 nuclease is SpRY Cas9.

177. 176. The method of any one of claims 163-175, wherein the catalytically impaired Cas9 nuclease is SpG Cas9.

178. 178. The method of any one of Claims 163-177, wherein the base editor is introduced into the cell as mRNA.

179. The method of any one of claims 163 to 178, wherein the base editor and gRNA are introduced into the cell via electroporation.

180. 180. The method of any one of claims 163-179, wherein the method further comprises sorting the genetically engineered hematopoietic stem or progenitor cells, or a plurality thereof, via fluorescence activated cell sorting (FACS).

181. 181. The method of any one of claims 163 to 180, wherein said substitution results in the reduction or elimination of expression of a gene encoding a wild-type form of said lineage specific cell surface antigen.

182. 182. A genetically engineered hematopoietic stem or progenitor cell produced by the method of any one of claims 163 to 181.

183. A cell population comprising a plurality of the genetically engineered hematopoietic stem or progenitor cells of claim 182.

184. 184. A pharmaceutical composition comprising the genetically engineered hematopoietic stem or progenitor cells of claim 182, or the cell population of claim 183.

185. A method for treating a hematopoietic disorder, comprising administering to a subject in need thereof an effective amount of a genetically engineered hematopoietic stem or progenitor cell described in claim 182, a cell population described in claim 183, or a pharmaceutical composition described in claim 184.

186. The method of claim 185, wherein the hematopoietic disease is a hematopoietic malignancy.

187. 187. The method of claim 185 or 186, wherein the method further comprises administering an effective amount of an agent that targets a wild-type form of the lineage-specific cell surface antigen.

188. 88. The method of claim 87, wherein the agent comprises an antibody or antigen-binding fragment that binds to the wild-type form of the lineage-specific cell surface antigen.

189. The cell of claim 188, wherein the agent is an immune cell.

190. 190. The method of claim 189, wherein the immune cell is a cytotoxic T cell.

191. 191. The method of claim 190, wherein the cytotoxic T cells express a chimeric antigen receptor (CAR) comprising the antibody or antigen-binding fragment that binds to the wild-type form of the lineage-specific cell surface antigen.

192. 192. The method of any one of claims 188 to 191, wherein the antibody is selected from the group consisting of anti-CD123 antibody 7G3, talatumumab, anti-CD38 antibody HB7, daratumumab, anti-CD38 antibody B43, blinatumomab, anti-CD19 antibody FMC63, anti-CD19 antibody HIB19, anti-CD47 antibody B6H12, anti-CD47 antibody 2D3, anti-CD34 antibody QBend10, anti-CD34 antibody 561, and anti-CD5 antibody H65.

193. 193. The method of any one of claims 185-192, wherein the genetically engineered hematopoietic stem or progenitor cells, the immune cells, or both, are allogeneic.

194. 194. The method of any one of claims 185-193, wherein the genetically engineered hematopoietic stem or progenitor cells, the immune cells, or both, are autologous.

195. 195. The method of any one of claims 185-194, wherein the subject is a human patient with Hodgkin's lymphoma, non-Hodgkin's lymphoma, leukemia, acute myeloid leukemia (AML), chronic myelogenous leukemia, acute lymphoblastic leukemia, or chronic lymphoblastic leukemia.

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