SCFV-and FAB-modified AAV capsids and uses thereof

Abstract

The present disclosure provides modified adeno-associated virus (AAV) virions with altered capsid proteins comprising single-chain fragment variable (scFv) or antigen-binding fragment (Fab), where the modified AAV virions exhibit enhanced infectivity of target cells with corresponding ligands on the cell surface. The present disclosure further provides methods of delivering a nucleic acid segment encoding various agents (e.g., chimeric antigen receptors, T cell receptors, antibodies, peptides, polypeptides, toxins, ribozymes, and catalytic RNA molecules) to a cell (e.g., in an individual), and methods of treating genetic defects, cancerous diseases, and disorders.

Description

Attorney Docket No.2014202-0027 SCFV- AND FAB-MODIFIED AAV CAPSIDS AND USES THEREOF CROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the benefit of U.S. Prov. Application No. 63 / 732,954 filed October 11, 2024, the entire contents of which are hereby incorporated by reference herein. FIELD OF THE INVENTION

[0002] Embodiments of the present disclosure relate to antibody-modified viral capsid proteins, including adeno-associated virus (AAV) capsid proteins modified with single-chain fragment variable (scFv) or antigen-binding fragment (Fab), as well as viruses and viral vectors comprising these modified capsid proteins. The disclosure further relates to methods of using such viruses and viral vectors for delivering nucleic acids encoding polypeptides, RNA molecules, or other therapeutic agents to cells for the treatment or prevention of diseases and disorders. BACKGROUND OF THE INVENTION

[0003] A promising approach to treating and preventing genetic and other diseases and disorders is delivery of therapeutic agents with a gene therapy vector such as a viral vector.

[0004] Certain challenges that remain with regard to the design of AAV vectors for use in gene and cell therapy include optimizing viral cell tropism and tissue-specific delivery. Antibody fragments, such as single-chain fragment variable (scFv) and antigen-binding fragment (Fab), which contain the antigen-binding domains of an antibody, offer advantages such as high specificity and affinity for antigens, low immunogenicity, and, in the case of scFv, the ability to penetrate tumor tissues and diffuse. The scFv fragment has been used to retarget lentivirus to specific lymphocyte subtypes for CAR-T cell therapies (Frank and Buchholz, Mol Ther Methods Clin Dev.2019 Mar 15; 12: 19–31).1However, attempts to modify AAV capsids with scFv have consistently resulted in significantly reduced viral titers, rendering such approaches impractical for AAV retargeting (Yang et al., Hum. Gene Ther.1998, 9, 1929 - 1937; Burning and Srivastava, Mol Ther Methods Clin Dev.2019 Mar 15; 12: 248–265).2,3- 1 - 12961406v1Attorney Docket No.2014202-0027

[0005] The present invention overcomes these limitations by providing novel AAV capsid proteins modified with scFv that, unexpectedly, can be packaged into AAV virions with titers comparable to unmodified AAV vectors. Additionally, the invention introduces a pioneering approach utilizing Fab fragments to modify AAV capsids, a strategy not previously reported for either AAV or lentiviral vectors. These modified capsids enhance the specificity and efficiency of viral vector delivery to target cells and tissues. By addressing the critical need for precise and effective targeting in gene therapy, the present invention represents a significant advancement in the field. SUMMARY OF THE INVENTION

[0006] The present disclosure relates generally to the fields of cancer therapy, autoimmune disease therapy, and gene therapy, and in particular, to viral vectors useful for the delivery of nucleic acid segments encoding various agents, e.g., therapeutic agents (e.g., chimeric antigen receptors, T cell receptors, antibodies, peptides, polypeptides, toxins, ribozymes, and catalytic RNA molecules), to selected cells and / or tissues of vertebrate animals.

[0007] In some aspects, the present disclosure provides antibody-modified capsid proteins, including those modified with single-chain fragment variable (scFv) or antigen-binding fragment (e.g., Fab), that are useful in cancer and gene therapy vectors (e.g., viral vectors), such as herpes simplex virus (HSV), alphavirus (AV), and adeno-associated virus (AAV) vectors. In some embodiments, such vectors that include antibody-modified capsid proteins can be used for the delivery of agents to desired cells or tissues, e.g., B-cells, T-cells, brain, liver, muscles, lung, kidney, heart, and other tissues, and for the treatment of mammalian diseases, disorders, and dysfunctions. In certain embodiments, viral vectors of the present invention are AAV6, which exhibits improved infectivity or specificity toward hematopoietic stem / progenitor cells, as well as other cells of the hematopoietic lineage, compared to many other AAV serotypes.4,5

[0008] The disclosed compositions may be utilized in a variety of investigative, diagnostic and therapeutic regimens, including the prevention and treatment of a variety of human diseases.

[0009] In certain embodiments, the present invention includes a non-naturally-occurring modified AAV capsid protein comprising one or more antibody fragments, such as scFv or Fab units, or extracellular domains such as CD80 and CD58. In certain embodiments, the modified - 2 - 12961406v1Attorney Docket No.2014202-0027 AAV capsid protein comprises a CD19-scFv or CD19-Fab fragment, e.g., a CD19-scFv fragment comprising or consisting of an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 093. In some embodiments, the modified AAV capsid protein comprises a CD3-scFv or CD3-Fab fragment, e.g., comprising or consisting of an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 091 or SEQ ID NO: 094. In some embodiments, the modified AAV capsid protein comprises a GD2-scFv or GD2-Fab fragment, e.g., comprising or consisting of an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to the amino acid sequence of SEQ ID NO: 095. In certain embodiments, the AAV is AAV6, and the scFv is located between amino acid residues 447 and 448, between 448 and 449, between 449 and 450, between 450 and 451, or between 451 and 452 in the Loop III region of the AAV6 capsid protein VP1 (SEQ ID NO: 096), VP2 (SEQ ID NO: 097), or VP3 (SEQ ID NO: 098), or a combination of two or three of them. In certain embodiments, the scFv is located between amino acid residues 585 and 586, between 586 and 587, between 587 and 588, between 588 and 589, or between 589 and 590 in the Loop IV region of the AAV6 capsid protein. In yet additional embodiments, the scFv or Fab fragment is located at the N-terminal of the VP2 protein of AAV5 or AAV6. Unless otherwise indicated, the capsid protein amino acid sequences referred to herein are VP1 sequences, and the AAV6 VP1 capsid protein comprises or consists of the amino acid sequence of SEQ ID NO: 096. A skilled artisan will understand that equivalent sequences exist in VP2 and VP3 capsid protein amino acid sequences, and the present disclosure also includes modified VP2 and VP3 capsid proteins having any of the modifications, e.g., insertions, described herein. The amino acid sequence for AAV6 VP2 capsid protein is provided as SEQ ID NO: 097, and the amino acid sequence for AAV6 VP3 capsid protein is provided as SEQ ID NO: 098. In certain embodiments, the AAV is a different AAV serotype, such as AAV1, AAV2, AAV5, AAV6, AAV8, AAV9, or AAV10, and the scFv is located in the capsid proteins of these other AAVs between amino acid residues corresponding to amino acid residues 447 and 448, between 448 and 449, between 449 and 450, between 450 and 451, or between 451 and 452 in the Loop III region, or between amino acid residues 585 and 586, between 586 and 587, between 587 and 588, between 588 and 589, or - 3 - 12961406v1Attorney Docket No.2014202-0027 between 589 and 590 in the Loop IV region of the capsid sequence. In certain embodiments, the scFv or Fab fragment is located at the N-terminal or C-terminal of the capsid protein VP1, VP2, or VP3. In some embodiments, the VP proteins can be from different AAV serotypes, for example, VP1 and VP3 from AAV2 but VP2 inserted with scFv or Fab from AAV6 serotype. It is understood that the corresponding residues in other AAVs may have different amino acid numbers. The skilled artisan could determine these residues based on sequence alignments, crystal structure, and the location of heparan sulfate proteoglycan (HSPG) binding sites. In particular embodiments, the insertion is close to but does not completely disrupt the HSPG binding site’s activity. In certain embodiments, it is adjacent to an amino acid residue identified as being important for HSPG binding.

[0010] In a related embodiment, the present disclosure includes a polynucleotide comprising a nucleic acid sequence encoding a modified AAV capsid protein described herein. In certain embodiments, the nucleic acid sequence encoding the modified AAV capsid protein is operably linked to a promoter sequence. In particular embodiments, the polynucleotide further comprises a nucleic acid sequence encoding a rep protein.

[0011] A further related embodiment of the present disclosure includes a cell comprising an expression vector described herein. In further embodiments, the cell comprises a polynucleotide that encodes a therapeutic protein.

[0012] Another embodiment is a recombinant virus or viral vector comprising a modified capsid protein described herein. The AAV virion of this disclosure may exhibit altered affinity for heparan sulfate binding, e.g., relative to AAV6. In certain embodiments, the recombinant virus or viral vector comprises a polynucleotide sequence that encodes a therapeutic protein. In particular embodiments, the therapeutic protein is a chimeric antigen receptor (CAR), a transgenic T cell receptor (TCR), a toxin protein, a bi-specific T-cell engager, or a tri-specific T- cell engager. In particular embodiments, the therapeutic protein is a CD19-specific CAR, a GD2- specific CAR, a TCR specific for a tumor-associated antigen (e.g., PRAME, NY-ESO-1, MAGE-A4), alpha-1 antitrypsin, factor IX, factor VIII, C1-esterase inhibitor, β-globin, or γ- globin. In certain embodiments, the therapeutic protein is one that exerts its therapeutic effect when expressed systemically, e.g., wherein the viral vector transduces the liver, which then produces the therapeutic protein, resulting in it being delivered systemically. - 4 - 12961406v1Attorney Docket No.2014202-0027

[0013] In certain embodiments, a recombinant virus or viral vector described herein has an altered cellular tropism as compared to a corresponding virus or viral vector having a wild- type capsid protein, i.e., the same capsid protein absent the scFv or Fab insertion. In some embodiments, the recombinant virus or viral vector has a greater tropism for T-cells, B-cells, NK cells, myeloid cells, dendritic cells, stem cells, epithelial cells, endothelial cells, tumor-specific cells, neurons, and pancreatic cells. In one embodiment, the recombinant virus or viral vector comprises an AAV6 capsid having a CD3-scFv inserted between amino acid residues 449 and 450 of VP2 protein. In one embodiment, the recombinant virus or viral vector comprises an AAV6 capsid having a CD19-scFv or CD19-Fab inserted at the N-terminal of VP2 protein. In one embodiment, the recombinant virus or viral vector comprises an AAV6 capsid having a CD3-scFv inserted between amino acid residues 587 and 588 of VP2 protein. In another embodiment, the recombinant virus or viral vector comprises an AAV5 capsid having a GD2- scFv or GD2-Fab inserted at the N-terminal of VP2 protein. In another embodiment, the recombinant virus or viral vector comprises an AAV8 capsid having a CD3-scFv inserted between amino acid residues 451 and 452 of VP1 and VP2 proteins.

[0014] In further related embodiments, the present disclosure includes a pharmaceutical composition comprising a recombinant virus or viral vector described herein.

[0015] The present disclosure also includes a method of providing a CAR or a transgenic TCR to a T-cell of a subject, comprising administering to the subject, a recombinant virus or viral vector or pharmaceutical composition described herein, wherein the recombinant virus or viral vector comprises a polynucleotide sequence that encodes the protein. In one embodiment, the recombinant virus or viral vector comprises an AAV6 capsid having a CD3-scFv inserted between amino acid residues 448 and 449, amino acid residues 449 and 450, or amino acid residues 450 and 451.

[0016] The present disclosure also includes a method of providing a protein to the liver of a subject, comprising administering to the subject, e.g., intravenously, a recombinant virus or viral vector or pharmaceutical composition described herein, wherein the recombinant virus or viral vector comprises a polynucleotide sequence that encodes the protein. In one embodiment, the recombinant virus or viral vector comprises an AAV6 capsid having a CD3-scFv inserted - 5 - 12961406v1Attorney Docket No.2014202-0027 between amino acid residues 448 and 449, amino acid residues 449 and 450, or amino acid residues 450 and 451.

[0017] The present disclosure further includes a method of providing a therapeutic gene product (e.g., a therapeutic protein) to a subject in need thereof, comprising administering to the subject, e.g., by peritoneal injection, a pharmaceutical composition comprising a recombinant virus or viral vector described herein, wherein the recombinant virus or viral vector comprises a polynucleotide encoding the therapeutic gene product. In particular embodiments, a subject has been diagnosed with peritoneal carcinomatosis disease or disorder. In particular embodiments, the subject has been diagnosed with one or more conditions selected from the group consisting of: B cell lymphoma or leukemia, ovarian cancer, colorectal cancer, stomach cancer, pancreatic cancer, appendiceal cancer, peritoneal mesothelioma, and endometrial cancer or B cell-mediated autoimmune disease. In one embodiment, the recombinant virus or viral vector comprises an AAV6 capsid having a scFv inserted between amino acid residues 448 and 449, amino acid residues 449 and 450, or amino acid residues 450 and 451.

[0018] The present disclosure further includes a method of providing a therapeutic gene product (e.g., a therapeutic protein) to the liver of a subject in need thereof, comprising administering to the subject, e.g., intravenously, a pharmaceutical composition comprising a recombinant virus or viral vector described herein, wherein the recombinant virus or viral vector comprises a polynucleotide encoding the therapeutic gene product. In particular embodiments, a subject has been diagnosed with a liver disease or disorder. In particular embodiments, the subject has been diagnosed with having one or more conditions selected from the group consisting of: alpha- 1 antitrypsin deficiency, hemophilia B, hemophilia A, hereditary angioedema, and β- thalassemia. In one embodiment, the recombinant virus or viral vector comprises an AAV6 capsid having a scFv insert between amino acid residues 448 and 449, amino acid residues 449 and 450, or amino acid residues 450 and 451.

[0019] The present disclosure further includes a method of providing a therapeutic gene product (e.g., a therapeutic protein) to the brain of a subject in need thereof, comprising administering to the subject, e.g., intraventricularly or intrathecally, a pharmaceutical composition comprising a recombinant virus or viral vector described herein, wherein the recombinant virus or viral vector comprises a polynucleotide encoding the therapeutic gene - 6 - 12961406v1Attorney Docket No.2014202-0027 product. In particular embodiments, the subject has been diagnosed with having one or more conditions selected from the group consisting of: gliomas, meningiomas, medulloblastomas, Schwannomas, pituitary adenomas, primary CNS lymphomas, chordomas, craniopharyngiomas, and hemangioblastomas.

[0020] The present disclosure further provides a method of altering the tropism of an AAV6, AAV5, AAV8, AAV1, or AAV2 virus or viral vector, comprising introducing a scFv or Fab into the capsid protein of the virus or viral vector, e.g., at the N-terminal of VP2, or a scFv between any of amino acid residues 448 and 449, amino acid residues 449 and 450, or amino acid residues 450 and 451, or 585 and 590 of VP1, VP2, or VP3. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The novel features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings.

[0022] FIGs.1A – 1L illustrate exemplary adeno-associated virus (AAV) capsid proteins VP1, VP2, and VP3 engineered with single-chain fragment variable (scFv) or Fab sequences. (A) A CD19-scFv sequence was inserted at the N-terminus of the AAV6 VP2 capsid protein, expressed under the control of the polyhedrin (polh) promoter (V775). (B) A CD3-scFv sequence was inserted at the N-terminus of the AAV6 VP2 capsid protein, driven by the polh promoter (V685). (C) A CD3-scFv sequence was inserted between amino acids 587 and 588 of the AAV6 VP2 capsid protein, under the control of the polh promoter (V689). (D) A CD3-scFv sequence was inserted between amino acids 587 and 588 of the AAV6 VP3 capsid protein, driven by the polh promoter (V697). (E) A CD3-scFv sequence was inserted between amino acids 449 and 450 of the AAV6 VP2 capsid protein, expressed under the polh promoter (V705). (F) A CD3-scFv sequence was inserted between amino acids 451 and 452 of the AAV8 VP1 / VP2 capsid protein, driven by the polh promoter (V735), with a separate expression cassette in the same construct encoding the AAV2 Rep sequence under the p10 promoter. (G) A GD2-scFv sequence was inserted at the N-terminus of the AAV5 VP2 capsid protein, driven by the polh promoter (V784), with two expression cassettes in the same construct: one driving AAV5 VP1 / VP3 expression - 7 - 12961406v1Attorney Docket No.2014202-0027 under the polh promoter and the other driving AAV2 Rep expression under the p10 promoter. (H) A CD3-scFv sequence was inserted at the N-terminus of the AAV6 VP2 capsid protein, expressed under the polh promoter (V797). (I) The AAV6 VP2 start codon ACG was mutated to ACC, resulting in expression of only VP1 and VP3 under the polh promoter (V334), with a separate AAV2 Rep expression cassette driven by the p10 promoter. (J) The AAV2 VP2 start codon ACG was mutated to ACC, resulting in expression of only VP1 and VP3 under the polh promoter (V076), with a separate AAV2 Rep expression cassette driven by the p10 promoter. (K) The AAV8 VP1 and VP2 sequences were removed, leaving only the VP3 sequence expressed under the polh promoter (V734). (L) A CD3-Fab sequence was inserted at the N- terminus of the AAV6 VP2 capsid protein, driven by the polh promoter (V794).

[0023] FIGs.2A – 2G illustrate exemplary adeno-associated virus (AAV) genomes encoding therapeutic genes. (A) A CD19-CAR sequence was cloned between two AAV inverted terminal repeats (ITRs), with expression driven by the cytomegalovirus (CMV) promoter (V694). (B) A GD2-CAR sequence was cloned between two AAV ITRs, with expression driven by the CMV promoter (V695). (C) A DT-A-P2A-TRAIL fusion sequence was cloned between two AAV ITRs, with expression of the fusion protein driven by the survivin promoter (V767). (D) A DT-A-P2A-TRAIL fusion sequence was cloned between two AAV ITRs, with expression of the fusion protein driven by a synthetic glioblastoma (GBM) promoter (V769). (E) A CD19- CAR sequence was cloned between two AAV ITRs, with expression driven by the CMV promoter, and five copies of miR-122 binding sites were inserted downstream of the coding sequence and upstream of the human growth hormone polyadenylation (hGHpA) signal (V786). (F) A tri-specific T-cell engager sequence was cloned between two AAV ITRs, with expression of the fusion protein driven by the CMV promoter (V800). (G) An eGFP sequence was cloned between two AAV ITRs, with expression of the protein driven by the CMV promoter (V545).

[0024] FIGs.3A - 3G present exemplary SDS-PAGE analyses of purified adeno- associated virus (AAV) capsid proteins, demonstrating the incorporation of various single-chain fragment variable (scFv) sequences into AAV particles. (A) Lane M: protein markers; Lane 1: AAV8-CMV-GFP control virus; Lane 2: AAV6-SURV-DTA-P2A-TRAIL (25-255) with CD19- scFv inserted at the N-terminus of VP2; Lane 3: mosaic AAV-CMV-CD19-CAR (25-256) comprising AAV2 VP1 / VP3 and AAV6 VP2 with CD3-scFv inserted between amino acids 449 and 450. (B) Lane M: protein markers; Lane 1: AAV8-CMV-GFP control virus; Lane 2: AAV5- - 8 - 12961406v1Attorney Docket No.2014202-0027 SURV-DTA-P2A-TRAIL (25-273) with GD2-scFv inserted at the N-terminus of VP2; Lane 3: AAV6-CMV-CD19-CAR (25-274) with CD3-scFv inserted at the N-terminus of VP2. (C) Lane M: protein markers; Lane 1: AAV8-CMV-GFP control virus; Lanes 2–4: AAV6-CMV-GFP (24- 365), AAV6-CMV-CD19-CAR (24-366), and AAV6-CMV-GD2-CAR (24-367), each with CD3-scFv inserted between amino acids 587 and 588 of VP2. (D) Lane M: protein markers; Lane 1: AAV8-CMV-GFP control virus; Lanes 2–4: AAV6-CMV-GFP (24-374), AAV6-CMV- CD19-CAR (24-375), and AAV6-CMV-GD2-CAR (24-376), each with CD3-scFv inserted between amino acids 449 and 450 of VP2. (E) Lane M: protein markers; Lane 1: AAV8-CMV- GFP control virus; Lane 2: AAV8-CMV-GFP (25-182) with VP1, VP2, and partial VP3, where CD3-scFv was inserted between amino acids 451 and 452, and the majority of non-modified VP3 was provided by a separate construct V734. (F) Lane M: protein markers; Lane 1: AAV8- CMV-GFP control virus; Lanes 2–4: AAV6-CMV-GFP (24-341), AAV6-CMV-CD19-CAR (24- 342), and AAV6-CMV-GD2-CAR (24-343), each with CD3-scFv inserted between amino acids 587 and 588 of VP3, with the majority of VP3 supplied by an additional construct V290. The VP3-CD3-scFv band overlapped with VP1 due to their similar molecular weights. (G) Lane M: protein markers; Lane 1: AAV8-CMV-GFP control virus; Lane 2: AAV6CD3Fab-GFP (25-298) with CD3Fab inserted at the N-terminus of VP2 showing a molecular weight of ~ 125 kDa.

[0025] FIG.4 illustrates a comparative analysis of transduction efficiencies of activated human T cells using AAV6, AAV8, and AAV-DJ vectors, each encoding eGFP, across various dosages. T cells were transduced with 2x104, 2x105, 5x105, 1x106, or 2x106vector genomes (vg) of AAV vectors, and the percentages of eGFP-expressing T cells were measured using the Cytek Aurora Flow Cytometry System (Cytek Biosciences).

[0026] FIG.5 presents a schematic representation of the transduction protocol and planned assessment of CAR-T cells when non-activated human T cells are transduced with either wild-type (wt) AAV6 (top row) or CD3-scFv-modified AAV6 (bottom row).

[0027] FIGs.6A – 6C depict exemplary comparisons of transduction efficiencies of non- activated human T cells from five healthy donors by either wild type AAV6 or CD3scFv- modified AAV6, both of which encoded eGFP. Non-transduced (NT) were used as a negative control. The percentages of eGFP-expressing cells were acquired by the Cytek Aurora Flow Cytometry System. Top row in FIG.6A shows the non-transduced peripheral blood mononuclear - 9 - 12961406v1Attorney Docket No.2014202-0027 cell (PBMC) data for 5 healthy donors, while the middle and bottom rows in FIG.6A show the fraction of eGFP+ T cells (per donor) upon transduction by wt AAV6 or AAV6-CD3scFv, respectively. FIG.6B summarizes the data and evaluates statistically significant differences using ordinary one-way Anova and Tuckey’s multiple comparisons post-test. FIG.6C shows the fold difference in transduction efficiency for the individual donors.

[0028] FIGs.7A – 7C depict exemplary comparisons of transduction efficiencies of non- activated human T cells from five healthy donors by either wild type AAV6 or CD3scFv- modified AAV6, both of which encoded a CD19-specific CAR (FMC63). Non-transduced (NT) were used as a negative control. Five days post-transduction, the cells were harvested, stained with antibodies specific for CD3 and the CAR, and the percentages of CAR-expressing cells were acquired by the Cytek Aurora Flow Cytometry System. Top row in FIG.7A shows the non- transduced peripheral blood mononuclear cell (PBMC) data for 5 healthy donors, while the middle and bottom rows in FIG.7A show the fraction of CAR+ T cells (per donor) upon transduction by wt AAV6 or AAV6-CD3scFv, respectively. FIG.7B summarizes the data and evaluates statistically significant differences using ordinary one-way Anova and Tuckey’s multiple comparisons post-test. FIG.7C shows the fold difference in transduction efficiency for the individual donors.

[0029] FIGs.8A – 8B depict exemplary comparisons of transduction efficiencies of Daudi cells (a human B cell line) by either wild type AAV6 or CD19scFv-modified AAV6, both of which encoded eGFP, at different dosages. Each cell was transduced with 1x103, 1x104, 1x105vg of AAV vectors and the percentages of eGFP-expressing cells were acquired by the Cytek Aurora Flow Cytometry System. FIG.8A shows flow cytometry graphs and FIG.8B shows the percentages of eGFP-expressing cells (left panel) and mean fluorescence intensity (right panel) by either wild type AAV6 (unfilled bar) or CD19scFv-modified AAV6 (filled bar), respectively.

[0030] FIG.9 depicts an exemplary comparison of transduction efficiencies of non- activated human T cells from two healthy donors by CD3scFv-modified AAV6, one of which had the CD3scFv inserted in VP2, while the other one had it inserted in VP3, and encoded either a CD19-specific CAR (FMC63) or a GD2-specific CAR (14G2A). The percentages of CAR- positive T cells were acquired by the Cytek Aurora Flow Cytometry System. - 10 - 12961406v1Attorney Docket No.2014202-0027

[0031] FIG.10 depicts an exemplary comparison of transduction efficiencies of non- activated human T cells from three healthy donors by CD3scFv-modified AAV6, one of which had the CD3scFv inserted in loop IV of VP2, while the other one had it inserted in loop III of VP2, and encoded eGFP. The percentages of eGFP-expressing T cells were acquired by the Cytek Aurora Flow Cytometry System.

[0032] FIG.11 depicts an exemplary comparison of transduction efficiencies of non- activated human T cells from three healthy donors by CD3scFv-modified AAV6, one of which had the CD3scFv inserted in loop IV of VP2, while the other one had it inserted in loop III of VP2, and encoded either a CD19-specific CAR (FMC63) or a GD2-specific CAR (14G2A). The percentages of CAR-positive T cells were acquired by the Cytek Aurora Flow Cytometry System.

[0033] FIGs.12A – 12B depict analysis of anti-tumor cytotoxicity of in vitro-generated human CAR-T cells. The CAR-T cells were generated from three healthy donors using CD3scFv-modified AAV6, one of which had the CD3scFv inserted in loop IV (aa587) of VP2, while the other one had it inserted in loop III (aa449) of VP2, and encoded either a CD19- specific CAR (FMC63) or a GD2-specific CAR (14G2A). The killing assay set-up and analysis is shown in FIG.12A, while the % specific lysis of antigen-positive tumor targets (CD19+ Raji cells for the FMC63 CAR or GD2-positive KM-H2 cells for the 14G2A CAR) is shown in FIG. 12B.

[0034] FIGs.13A-C depict an exemplary comparison of the anti-tumor efficacy of in vivo-generated human CAR-T cells. Luciferase-modified Raji cells were implanted intraperitoneally (IP) in NSG mice on Day -3 (three days prior to treatment). On Day 0, one group of 5 mice received human PBMCs and AAV6-CD3scFv-FMC63CAR IP, another group of 5 received human PBMCs and AAV6-CD3scFv-EGFP IP and the last group of 5 mice was left untreated. Tumor growth was monitored twice a week via IVIS imaging for 31 days. The experimental scheme is shown in FIG.13A and the IVIS imaging data are shown in FIG.13B. Measurement of the average radiance (tumor burden) is shown in FIG.13C (Panel A) and mouse survival is shown in FIG.13C (Panel B).

[0035] FIGs.14A – 14B depict an exemplary analysis of the persistence of in vivo- generated CAR-T cells using flow cytometry. FIG.14A shows the percentages of human T cells - 11 - 12961406v1Attorney Docket No.2014202-0027 in mice that had received human PBMCs and either AAV6-CD3scFv-FMC63CAR (top) or AAV6-CD3scFv-EGFP (bottom) out of viable human hematopoietic cells, as determined using a viability dye and anti-human CD45 and anti-human CD3 antibodies. FIG.14B shows the percentages of human T cells that were CAR-positive (top) or EGFP-positive (bottom) in mice that had received human PBMCs and AAV6-CD3scFv-FMC63CAR or AAV6-CD3scFv-EGFP, respectively.

[0036] FIGs.15A-C depict another exemplary comparison of the anti-tumor efficacy of in vivo-generated human CAR-T cells. Luciferase-modified Raji cells administered intravenously (IV) via the tail vain to NSG mice on Day -3 (three days prior to treatment). On Day 0, one group of 10 mice received human PBMCs and AAV6-CD3scFv-FMC63 CAR IV, while the other group of 10 was left untreated. Tumor growth was monitored twice a week via IVIS imaging for 24 days. The experimental scheme is shown in FIG.15A and the IVIS imaging data are shown in FIG.15B. Measurement of the average radiance (tumor burden) is shown in FIG.15C (Panel A) and mouse survival are shown in FIG.15C (Panel B).

[0037] FIGs.16A – 16B depict another exemplary analysis of the persistence of in vivo- generated CAR-T cells using flow cytometry. FIG.16A shows the percentages of human T cells in mice that had received human PBMCs and AAV6-CD3scFv-FMC63CAR out of viable human hematopoietic cells, as determined using a viability dye and anti-human CD45 and anti-human CD3 antibodies. FIG.16B shows the percentages of human T cells that were CAR-positive in the mice that had received human PBMCs and AAV6-CD3scFv-FMC63 CAR.

[0038] FIG.17 depicts an exemplary analysis of various mouse organs for CD19-specific CAR DNA by quantitative PCR (qPCR) using primers and a probe specific for the CAR. Following mouse euthanasia, the following organs: liver, kidney, heart, spleen, brain, lung, ovaries and bone marrow were collected from 6 mice that had received human PBMCs and AAV-CD3scFv-FMC63CAR and 3 control animals. DNA was isolated from the organs and the copy number of the CAR per mg of tissue was determined by qPCR. DETAILED DESCRIPTION OF THE INVENTION

[0039] The present disclosure provides modified capsid proteins and virions and viral vectors having one or more modified or altered capsid proteins, wherein the capsid proteins are modified with antibody fragments, such as single-chain fragment variable (scFv) or antigen- - 12 - 12961406v1Attorney Docket No.2014202-0027 binding fragment (Fab), to enhance targeting and / or infectivity. In various embodiments, the virions exhibit: 1) increased infectivity or specificity for a T cell or a B cell; 2) altered tropism; 3) increased cell or tissue specificity as compared to one or more other cells or tissues; 4) increased binding to target cells displaying corresponding ligands; and / or 5) an increased ability to infect and / or deliver a therapeutic gene product to a target cell when administered peritoneally, intravenously, intraventricularly, or intrathecally, as compared to a corresponding virion comprising its native or wild-type capsid protein instead of a modified capsid protein disclosed herein. In certain embodiments, the modified capsid proteins are adeno-associated virus (AAV) capsid proteins modified with scFv or Fab fragments specific for ligands such as CD19, CD3, or GD2, enabling targeted delivery to cells expressing these ligands, such as hematopoietic cells or tumor cells. Also provided are pharmaceutical compositions and methods for the use of any of the compositions disclosed herein for promoting the expression of a gene in cells, e.g., T and B cells, in an individual, e.g., for the treatment of a disease or disorder. These and other objects, advantages, and features of the invention will become apparent to those persons skilled in the art upon reading the details of the compositions and methods as more fully described below. DEFINITIONS

[0040] A "vector" as used herein refers to a macromolecule or association of macromolecules that comprises or associates with a polynucleotide and which can be used to mediate delivery of the polynucleotide to a cell. Illustrative vectors include, for example, plasmids, viral vectors, lipid nanoparticles, liposomes, and other gene delivery vehicles.

[0041] The term "AAV" is an abbreviation for adeno-associated virus, and may be used to refer to the virus itself or derivatives thereof. The term covers all subtypes and both naturally occurring and recombinant forms, except where required otherwise. The term "AAV" includes AAV type 1 (AAV1), AAV type 2 (AAV2), AAV type 3 (AAV3), AAV type 4 (AAV4), AAV type 5 (AAV5), AAV type 6 (AAV6), AAV type 7 (AAV7), AAV type 8 (AAV8), avian AAV, bovine AAV, canine AAV, equine AAV, primate AAV, non-primate AAV, and ovine AAV. "Primate AAV" refers to AAV that infect primates, "non-primate AAV" refers to AAV that infect non-primate mammals, "bovine AAV" refers to AAV that infect bovine mammals, etc. - 13 - 12961406v1Attorney Docket No.2014202-0027

[0042] The genomic sequences of various serotypes of AAV, as well as the sequences of the native terminal repeats (TRs), Rep proteins, and capsid subunits are known in the art. Such sequences may be found in the literature or in public databases such as GenBank. See, e.g., GenBank Accession Numbers NC_002077 (AAV1), AF063497 (AAV1), NC_001401 (AAV2), AF043303 (AAV2), NC_001729 (AAV3), NC_001829 (AAV4), U89790 (AAV4), NC_006152 (AAV5), AF028704 and AAB95450 (AAV6), AF513851 (AAV7), AF513852 (AAV8), and NC_006261 (AAV8), AY530579.1 (AAV9). The disclosures of which are incorporated by reference herein for teaching AAV nucleic acid and amino acid sequences. See also, e.g., Srivistava et al. (1983) J. Virology 45:555;6Chiorini et al. (1997) J. Virology 71 :6823;7Chiorini et al. (1999) J. Virology 73: 1309;8Bantel-Schaal et al. (1999) J. Virology 73:939;9Xiao et al. (1998) J. Virology 73:3994;10Muramatsu et al. (1996) Virology 221 :208;11Shade et al. (1986) J. Virol.58:921;12Gao et al. (2002) Proc. Nat. Acad. Sci. USA 99: 11854;13Moris et al. (2004) Virology 33:375-383;14international patent publications WO 00 / 28061, WO 99 / 61601, WO 98 / 11244; and U.S. Pat. No.6,156,303. In addition, polynucleotide sequences encoding any of the capsid proteins may be readily generated based on the amino acid sequence and the known genetic code, including codon-optimized sequences.

[0043] An "AAV virus" or "AAV viral particle" or "rAAV vector particle" refers to a viral particle composed of at least one AAV capsid protein (typically by all of the capsid proteins of a wild-type AAV) and an encapsidated polynucleotide rAAV vector. If the particle comprises a heterologous polynucleotide (i.e. a polynucleotide other than a wild-type AAV genome such as a transgene to be delivered to a mammalian cell), it is typically referred to as a "rAAV vector particle" or simply a "rAAV vector". Thus, production of a rAAV particle necessarily includes production of a rAAV vector, as such a vector is contained within a rAAV particle.

[0044] The term "replication defective" as used herein relative to an AAV viral vector of the invention means the AAV vector cannot independently replicate and package its genome. For example, when a cell of a subject is infected with rAAV virions, the heterologous gene is expressed in the infected cells, however, due to the fact that the infected cells lack AAV rep and cap genes and accessory function genes, the rAAV is not able to replicate further.

[0045] An "AAV variant" or "AAV mutant" as used herein refers to a viral particle composed of: a) a variant AAV capsid protein, where the variant AAV capsid protein comprises - 14 - 12961406v1Attorney Docket No.2014202-0027 at least one amino acid difference (e.g., amino acid substitution, amino acid insertion, amino acid deletion) relative to a corresponding parental AAV capsid protein, where the AAV capsid protein does not correspond to the amino acid sequence present of a naturally occurring AAV capsid protein; and, optionally, b) a heterologous nucleic acid comprising a nucleotide sequence encoding a heterologous gene product, wherein the variant AAV capsid protein confers increased binding to heparan or a heparan sulfate proteoglycan as compared to the binding by an AAV virion comprising the corresponding parental AAV capsid protein. In certain embodiments, an scFv-modified capsid protein confers: a) increased infectivity of a specific cell type compared to the infectivity of the cell type by an AAV virion comprising the corresponding parental AAV capsid protein; and / or b) altered cellular tropism as compared to the tropism of an AAV virion comprising the corresponding parental AAV capsid protein.

[0046] The abbreviation "rAAV" refers to recombinant adeno-associated virus, also referred to as a recombinant AAV vector (or "rAAV vector"). A "rAAV vector" as used herein refers to an AAV vector comprising a polynucleotide sequence not of AAV origin (i.e., a polynucleotide heterologous to AAV), typically a sequence of interest for the genetic transformation of a cell. In general, the heterologous polynucleotide is flanked by at least one, and generally by two AAV inverted terminal repeat sequences (ITRs). The term rAAV vector encompasses both rAAV vector particles and rAAV vector plasmids.

[0047] "Packaging" refers to a series of intracellular events that result in the assembly and encapsidation of an AAV particle.

[0048] "Mosaic packaging" refers to a method of producing recombinant adeno- associated virus (AAV) particles wherein the viral capsid is assembled from a combination of capsid proteins derived from two or more distinct AAV serotypes or variants. Specifically, mosaic packaging involves the incorporation of at least one capsid protein (e.g., VP1, VP2, or VP3) from a first AAV serotype or variant and at least one capsid protein from a second, different AAV serotype or variant into a single capsid structure. Such capsids, referred to as mosaic capsids, exhibit a heterogeneous composition of capsid proteins, which may confer altered functional properties, including but not limited to modified tropism, transduction efficiency, or immunogenicity, compared to capsids composed of proteins from a single AAV serotype or variant. Mosaic packaging encompasses the production of AAV particles using any - 15 - 12961406v1Attorney Docket No.2014202-0027 combination of capsid proteins from multiple serotypes or variants, whether naturally occurring, engineered, or synthetic, to encapsidate a viral genome.

[0049] AAV "rep" and "cap" genes refer to polynucleotide sequences encoding replication and encapsidation proteins of adeno-associated virus. AAV rep and cap are referred to herein as AAV "packaging genes."

[0050] A "helper virus" for AAV refers to a virus that allows AAV (e.g. wild-type AAV) to be replicated and packaged by a mammalian cell. A variety of such helper viruses for AAV are known in the art, including adenoviruses, herpesviruses and poxviruses such as vaccinia. The adenoviruses encompass a number of different subgroups, although Adenovirus type 5 of subgroup C is most commonly used. Numerous adenoviruses of human, non-human mammalian and avian origin are known and available from depositories such as the ATCC. Viruses of the herpes family include, for example, herpes simplex viruses (HSV) and Epstein- Barr viruses (EBV), as well as cytomegaloviruses (CMV) and pseudorabies viruses (PRV); which are also available from depositories such as ATCC.

[0051] "Helper virus function(s)" refers to function(s) encoded in a helper virus genome which allow AAV replication and packaging (in conjunction with other requirements for replication and packaging described herein). As described herein, "helper virus function" may be provided in a number of ways, including by providing helper virus or providing, for example, polynucleotide sequences encoding the requisite function(s) to a producer cell in trans. For example, a plasmid or other expression vector comprising nucleotide sequences encoding one or more adenoviral proteins is transfected into a producer cell along with an rAAV vector.

[0052] An "infectious" virus or viral particle is one that comprises a competently assembled viral capsid and is capable of delivering a polynucleotide component into a cell for which the viral species is tropic. The term does not necessarily imply any replication capacity of the virus. Assays for counting infectious viral particles are described elsewhere in this disclosure and in the art. Viral infectivity can be expressed as the ratio of infectious viral particles to total viral particles. Methods of determining the ratio of infectious viral particles to total viral particles are known in the art. See, e.g., Francois et al. (2018) Mol. Ther. Methods Clin. Dev., 10:223– 236;15and Zolotukhin et al. (1999) Gene Ther.6:973.16- 16 - 12961406v1Attorney Docket No.2014202-0027

[0053] A "replication-competent" virus (e.g. a replication-competent AAV) refers to a phenotypically wild-type virus that is infectious, and is also capable of being replicated in an infected cell (i.e. in the presence of a helper virus or helper virus functions). In the case of AAV, replication competence generally requires the presence of functional AAV packaging genes. In general, rAAV vectors as described herein are replication-incompetent in mammalian cells (especially in human cells) even in the presence of helper functions, by virtue of the lack of one or more AAV packaging genes. Typically, such rAAV vectors lack any AAV packaging gene sequences in order to minimize the possibility that replication competent AAV are generated by recombination between AAV packaging genes and an incoming rAAV vector. In many embodiments, rAAV vector preparations as described herein are those which contain few if any replication competent AAV (rcAAV, also referred to as RCA) (e.g., less than about 1 rcAAV per 102rAAV particles, less than about 1 rcAAV per 104rAAV particles, less than about 1 rcAAV per 108rAAV particles, less than about 1 rcAAV per 1012rAAV particles, or no rcAAV).

[0054] The term "polynucleotide" refers to a polymeric form of nucleotides of any length, including deoxyribonucleotides or ribonucleotides, or analogs thereof. A polynucleotide may comprise modified nucleotides, such as methylated nucleotides and nucleotide analogs, and may be interrupted by non-nucleotide components. If present, modifications to the nucleotide structure may be imparted before or after assembly of the polymer. The term polynucleotide, as used herein, refers interchangeably to double- and single-stranded molecules. Unless otherwise specified or required, any embodiment of the invention described herein that is a polynucleotide encompasses both the double-stranded form and each of two complementary single-stranded forms known or predicted to make up the double-stranded form.

[0055] A polynucleotide or polypeptide has a certain percent "sequence identity" to another polynucleotide or polypeptide, meaning that, when aligned, that percentage of bases or amino acids are the same when comparing the two sequences. Sequence similarity can be determined in a number of different manners. To determine sequence identity, sequences can be aligned using the methods and computer programs, including BLAST, available over the world wide web at ncbi.nlm.nih.gov / BLAST / . Another alignment algorithm is FASTA, available in the Genetics Computing Group (GCG) package, from Madison, Wis., USA, a wholly owned subsidiary of Oxford Molecular Group, Inc. Other techniques for alignment are described in Methods in Enzymology, vol.266: Computer Methods for Macromolecular Sequence Analysis - 17 - 12961406v1Attorney Docket No.2014202-0027 (1996), ed. Doolittle, Academic Press, Inc., a division of Harcourt Brace & Co., San Diego, Calif, USA. Of particular interest are alignment programs that permit gaps in the sequence. The Smith- Waterman is one type of algorithm that permits gaps in sequence alignments. See Meth. Mol. Biol.70: 173-187 (1997). Also, the GAP program using the Needleman and Wunsch alignment method can be utilized to align sequences. See J. Mol. Biol.48: 443-453 (1970).

[0056] Of interest is the BestFit program using the local homology algorithm of Smith and Waterman (Advances in Applied Mathematics 2: 482-489 (1981) to determine sequence identity. The gap generation penalty will generally range from 1 to 5, usually 2 to 4 and in many embodiments will be 3. The gap extension penalty will generally range from about 0.01 to 0.20 and in many instances will be 0.10. The program has default parameters determined by the sequences inputted to be compared. Preferably, the sequence identity is determined using the default parameters determined by the program. This program is available also from the Genetics Computing Group (GCG) package, from Madison, Wis., USA.

[0057] Another program of interest is the FastDB algorithm. FastDB is described in Current Methods in Sequence Comparison and Analysis, Macromolecule Sequencing and Synthesis, Selected Methods and Applications, pp.127-149, 1988, Alan R. Liss, Inc. Percent sequence identity is calculated by FastDB based upon the following parameters: Mismatch Penalty: 1.00; Gap Penalty: 1.00; Gap Size Penalty: 0.33; and Joining Penalty: 30.0.

[0058] A "gene" refers to a polynucleotide containing at least one open reading frame that is capable of encoding a particular gene product after being transcribed, and sometimes also translated. The term "gene" or "coding sequence" refers to a nucleotide sequence in vitro or in vivo that encodes a gene product. In some instances, the gene consists or consists essentially of coding sequence, that is, sequence that encodes the gene product. In other instances, the gene comprises additional, non-coding, sequences. For example, the gene may or may not include regions preceding and following the coding region, e.g.5' untranslated (5' UTR) or "leader" sequences and 3' UTR or "trailer" sequences, as well as intervening sequences (introns) between individual coding segments (exons).

[0059] A "gene product" is a molecule resulting from expression of a particular gene. Gene products include, e.g., a polypeptide, an aptamer, an interfering RNA, an mRNA, and the like. In particular embodiments, a "gene product" is a polypeptide, peptide, protein or interfering - 18 - 12961406v1Attorney Docket No.2014202-0027 RNA including short interfering RNA (siRNA), miRNA or small hairpin RNA (shRNA). In particular embodiments, a gene product is a therapeutic gene product, e.g., a therapeutic protein.

[0060] As used herein, a "therapeutic gene" refers to a gene that, when expressed, produces a therapeutic gene product that confers a beneficial effect on the cell or tissue in which it is present, or on a mammal in which the gene is expressed. Examples of beneficial effects include amelioration of a sign or symptom of a condition or disease, prevention or inhibition of a condition or disease, or conferral of a desired characteristic. Therapeutic genes include, but are not limited to, genes that correct a genetic deficiency in a cell or mammal.

[0061] As used herein, a "transgene" is a gene that is delivered to a cell by a vector.

[0062] "Recombinant," as applied to a polynucleotide, means that the polynucleotide is the product of various combinations of cloning, restriction or ligation steps, and other procedures that result in a construct that is distinct from a polynucleotide found in nature. A recombinant virus is a viral particle comprising a recombinant polynucleotide. The terms respectively include replicates of the original polynucleotide construct and progeny of the original virus construct.

[0063] A "control element" or "control sequence" is a nucleotide sequence involved in an interaction of molecules that contributes to the functional regulation of a polynucleotide, including replication, duplication, transcription, splicing, translation, or degradation of the polynucleotide. The regulation may affect the frequency, speed, or specificity of the process, and may be enhancing or inhibitory in nature. Control elements known in the art include, for example, transcriptional regulatory sequences such as promoters and enhancers. A promoter is a DNA region capable under certain conditions of binding RNA polymerase and initiating transcription of a coding region usually located downstream (in the 3' direction) from the promoter.

[0064] "Operatively linked" or "operably linked" refers to a juxtaposition of genetic elements, wherein the elements are in a relationship permitting them to operate in the expected manner. For instance, a promoter is operatively linked to a coding region if the promoter helps initiate transcription of the coding sequence. There may be intervening residues between the promoter and coding region so long as this functional relationship is maintained. - 19 - 12961406v1Attorney Docket No.2014202-0027

[0065] "Heterologous" means derived from a genotypically distinct entity from that of the rest of the entity to which it is being compared. For example, a polynucleotide introduced by genetic engineering techniques into a plasmid or vector derived from a different species is a heterologous polynucleotide. A promoter removed from its native coding sequence and operatively linked to a coding sequence with which it is not naturally found linked is a heterologous promoter. Thus, for example, an rAAV that includes a heterologous nucleic acid encoding a heterologous gene product is an rAAV that includes a nucleic acid not normally included in a naturally-occurring, wild-type AAV, and the encoded heterologous gene product is a gene product not normally encoded by a naturally-occurring, wild-type AAV.

[0066] As used herein, the terms "polypeptide," "peptide," and "protein" refer to polymers of amino acids of any length. The terms also encompass an amino acid polymer that has been modified; for example, disulfide bond formation, glycosylation, lipidation, phosphorylation, or conjugation with a labeling component.

[0067] By "comprising" it is meant that the recited elements are required in, for example, the composition, method, kit, etc., but other elements may be included to form the, for example, composition, method, kit etc. within the scope of the claim. For example, an expression cassette "comprising" a gene encoding a therapeutic polypeptide operably linked to a promoter is an expression cassette that may include other elements in addition to the gene and promoter, e.g. poly-adenylation sequence, enhancer elements, other genes, linker domains, etc.

[0068] By "consisting essentially of”, it is meant a limitation of the scope of the, for example, composition, method, kit, etc., described to the specified materials or steps that do not materially affect the basic and novel characteristic(s) of the, for example, composition, method, kit, etc. For example, an expression cassette "consisting essentially of a gene encoding a therapeutic polypeptide operably linked to a promoter and a polyadenylation sequence” may include additional sequences, e.g. linker sequences, so long as they do not materially affect the transcription or translation of the gene.

[0069] By "consisting of”, it means the exclusion from the composition, method, or kit of any element, step, or ingredient not specified in the claim. For example, an expression cassette "consisting of a gene encoding a therapeutic polypeptide operably linked to a promoter, and a polyadenylation sequence” consists only of the promoter, polynucleotide sequence encoding the - 20 - 12961406v1Attorney Docket No.2014202-0027 therapeutic polypeptide, and polyadenlyation sequence. As another example, a polypeptide "consisting of a recited sequence” contains only the recited sequence.

[0070] An "expression vector" as used herein encompasses a vector, e.g. plasmid, minicircle, viral vector, liposome, and the like as discussed above or as known in the art, comprising a polynucleotide which encodes a gene product of interest, and is used for effecting the expression of a gene product in an intended target cell. An expression vector also comprises control elements operatively linked to the encoding region to facilitate expression of the gene product in the target. The combination of control elements, e.g. promoters, enhancers, UTRs, miRNA targeting sequences, etc., and a gene or genes to which they are operably linked for expression is sometimes referred to as an "expression cassette." Many such control elements are known and available in the art or can be readily constructed from components that are available in the art.

[0071] A "promoter" as used herein encompasses a DNA sequence that directs the binding of RNA polymerase and thereby promotes RNA synthesis, i.e., a minimal sequence sufficient to direct transcription. Promoters and corresponding protein or polypeptide expression may be ubiquitous, meaning strongly active in a wide range of cells, tissues and species or cell- type specific, tissue-specific, or species specific. Promoters may be "constitutive," meaning continually active, or "inducible," meaning the promoter can be activated or deactivated by the presence or absence of biotic or abiotic factors. Also included in the nucleic acid constructs or vectors of the invention are enhancer sequences that may or may not be contiguous with the promoter sequence. Enhancer sequences influence promoter-dependent gene expression and may be located in the 5' or 3' regions of the native gene.

[0072] An "enhancer" as used herein encompasses a cis-acting element that stimulates or inhibits transcription of adjacent genes. An enhancer that inhibits transcription also is termed a "silencer". Enhancers can function (i.e., can be associated with a coding sequence) in either orientation, over distances of up to several kilobase pairs (kb) from the coding sequence and from a position downstream of a transcribed region.

[0073] A "termination signal sequence" as used herein encompasses any genetic element that causes RNA polymerase to terminate transcription, such as for example a polyadenylation signal sequence. - 21 - 12961406v1Attorney Docket No.2014202-0027

[0074] A "polyadenylation signal sequence" as used herein encompasses a recognition region necessary for endonuclease cleavage of an RNA transcript that is followed by the polyadenylation consensus sequence AATAAA. A polyadenylation signal sequence provides a "poly A site", i.e. a site on an RNA transcript to which adenine residues will be added by post- transcriptional polyadenylation.

[0075] The term "endogenous" as used herein with reference to a nucleotide molecule or gene product refers to a nucleic acid sequence, e.g. gene or genetic element, or gene product, e.g. RNA, protein, that is naturally occurring in or associated with a host virus or cell.

[0076] The term "native" as used herein refers to a nucleotide sequence, e.g. gene, or gene product, e.g. RNA, protein, that is present in a wildtype virus or cell. The term "variant" as used herein refers to a mutant of a reference polynucleotide or polypeptide sequence, for example a native polynucleotide or polypeptide sequence, i.e. having less than 100% sequence identity with the reference polynucleotide or polypeptide sequence. Put another way, a variant comprises at least one amino acid difference (e.g., amino acid substitution, amino acid insertion, amino acid deletion) relative to a reference polynucleotide or polypeptide sequence, e.g. a native polynucleotide or polypeptide sequence. For example, a variant may be a polynucleotide having a sequence identity of 70% or more with a full-length native polynucleotide sequence, e.g. an identity of 75% or 80% or more, such as 85%, 90%, or 95% or more, for example, 98% or 99% identity with the full-length native polynucleotide sequence. As another example, a variant may be a polypeptide having a sequence identity of 70% or more with a full-length native polypeptide sequence, e.g. an identity of 75% or 80% or more, such as 85%, 90%, or 95% or more, for example, 98% or 99% identity with the full-length native polypeptide sequence. Variants may also include variant fragments of a reference, e.g. native, sequence sharing a sequence identity of 70% or more with a fragment of the reference, e.g. native, sequence, e.g. an identity of 75% or 80% or more, such as 85%, 90%, or 95% or more, for example, 98% or 99% identity with the native sequence.

[0077] A “single-chain fragment variable” (scFv) is a fusion protein composed of the variable regions of the heavy (VH) and light (VL) chains of antibodies, connected by a short linker peptide. This structure enables the scFv to retain the antigen-binding specificity of the parent antibody, but in a smaller and more versatile format. scFvs are commonly used in - 22 - 12961406v1Attorney Docket No.2014202-0027 research, diagnostics, and therapeutic applications due to their ease of genetic manipulation and ability to penetrate tissues more effectively than full-length antibodies.

[0078] As used herein, a "Fab antibody" or "Fab fragment" refers to an antibody fragment that comprises a complete light chain and the variable and first constant domain (CH1) of a heavy chain, connected via a disulfide bond. The Fab fragment includes one antigen-binding site and lacks the Fc (crystallizable) region of the full-length antibody. It may be generated through proteolytic digestion of an immunoglobulin (e.g., papain cleavage) or produced recombinantly. The Fab fragment retains the ability to specifically bind to an antigen and may be human, humanized, murine, chimeric, or synthetic in origin.

[0079] CD molecules, or “Cluster of Differentiation” molecules, are cell surface proteins that serve as markers to differentiate and identify immune cells and other hematopoietic cells. They play key roles in cell signaling, communication, and immune responses. Each CD molecule is designated by a number (e.g., CD1, CD3, CD19, and CD28) and is associated with specific functions: For example, CD3 is part of the T-cell receptor complex, crucial for T-cell activation. CD28 is a co-stimulatory molecule found on the surface of T cells. It plays a crucial role in the activation and survival of T cells during the immune response.

[0080] As used herein, the terms "biological activity" and "biologically active" refer to the activity attributed to a particular biological element in a cell. For example, the "biological activity" of an "immunoglobulin", "antibody" or fragment or variant thereof refers to the ability to bind an antigenic determinant and thereby facilitate immunological function. As another example, the biological activity of a polypeptide or functional fragment or variant thereof refers to the ability of the polypeptide or functional fragment or variant thereof to carry out its native functions of, e.g., binding, enzymatic activity, etc. As a third example, the biological activity of a gene regulatory element, e.g. promoter, enhancer, Kozak sequence, and the like, refers to the ability of the regulatory element or functional fragment or variant thereof to regulate, i.e. promote, enhance, or activate the translation of, respectively, the expression of the gene to which it is operably linked.

[0081] The terms "administering" or "introducing", as used herein, refer to delivery of a vector for recombinant gene or protein expression to a cell, to cells and / or organs of a subject, or to a subject. Such administering or introducing may take place in vivo, in vitro or ex vivo. A - 23 - 12961406v1Attorney Docket No.2014202-0027 vector for expression of a gene product may be introduced into a cell by transfection, which typically means insertion of heterologous DNA into a cell by physical means (e.g., calcium phosphate transfection, electroporation, microinjection or lipofection); infection, which typically refers to introduction by way of an infectious agent, i.e. a virus; or transduction, which typically means stable infection of a cell with a virus or the transfer of genetic material from one microorganism to another by way of a viral agent (e.g., a bacteriophage).

[0082] "Transformation" is typically used to refer to bacteria comprising heterologous DNA or cells which express an oncogene and have therefore been converted into a continuous growth mode such as tumor cells. A vector used to "transform" a cell may be a plasmid, virus or other vehicle.

[0083] Typically, a cell is referred to as "transduced", "infected"; "transfected" or "transformed" dependent on the means used for administration, introduction or insertion of heterologous DNA (i.e., the vector) into the cell. The terms "transduced", "transfected" and "transformed" may be used interchangeably herein regardless of the method of introduction of heterologous DNA.

[0084] The term "host cell", as used herein refers to a cell which has been transduced, infected, transfected or transformed with a vector. The vector may be a plasmid, a viral particle, a phage, etc. The culture conditions, such as temperature, pH and the like, are those previously used with the host cell selected for expression, and will be apparent to those skilled in the art. It will be appreciated that the term "host cell" refers to the original transduced, infected, transfected or transformed cell and progeny thereof.

[0085] The terms "treatment", "treating" and the like are used herein to generally mean obtaining a desired pharmacologic and / or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof, e.g. reducing the likelihood that the disease or symptom thereof occurs in the subject, and / or may be therapeutic in terms of a partial or complete cure for a disease and / or adverse effect attributable to the disease. "Treatment" as used herein covers any treatment of a disease in a mammal, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; or (c) relieving the disease, i.e., causing regression of the disease. The therapeutic agent may be - 24 - 12961406v1Attorney Docket No.2014202-0027 administered before, during or after the onset of disease or injury. The treatment of ongoing disease, where the treatment stabilizes or reduces the undesirable clinical symptoms of the patient, is of particular interest. Such treatment is desirably performed prior to complete loss of function in the affected tissues. The subject therapy will desirably be administered during the symptomatic stage of the disease, and in some cases after the symptomatic stage of the disease.

[0086] The terms "individual," "host," "subject," and "patient" are used interchangeably herein, and refer to a mammal, including, but not limited to, human and non-human primates, including simians and humans; mammalian sport animals (e.g., horses); mammalian farm animals (e.g., sheep, goats, etc.); mammalian pets (dogs, cats, etc.); and rodents (e.g., mice, rats, etc.).

[0087] The various compositions and methods of the invention are described below. Although particular compositions and methods are exemplified herein, it is understood that any of a number of alternative compositions and methods are applicable and suitable for use in practicing the invention. It will also be understood that an evaluation of the expression constructs and methods of the invention may be carried out using procedures standard in the art.

[0088] The practice of the present invention will employ, unless otherwise indicated, conventional techniques of cell biology, molecular biology (including recombinant techniques), microbiology, biochemistry and immunology, which are within the scope of those of skill in the art. Such techniques are explained fully in the literature, such as, "Molecular Cloning: A Laboratory Manual", second edition (Sambrook et al, 1989); "Oligonucleotide Synthesis" (M. J. Gait, ed., 1984); "Animal Cell Culture" (R. I. Freshney, ed., 1987); "Methods in Enzymology" (Academic Press, Inc.); "Handbook of Experimental Immunology" (D. M. Weir & C. C. Blackwell, eds.); "Gene Transfer Vectors for Mammalian Cells" (J. M. Miller & M. P. Calos, eds., 1987); "Current Protocols in Molecular Biology" (F. M. Ausubel et al, eds., 1987); "PCR: The Polymerase Chain Reaction", (Mullis et al, eds., 1994); and "Current Protocols in Immunology" (J. E. Coligan et al, eds., 1991), each of which is expressly incorporated by reference herein.

[0089] Several aspects of the invention are described below with reference to example applications for illustration. It should be understood that numerous specific details, relationships, and methods are set forth to provide a full understanding of the invention. One having ordinary - 25 - 12961406v1Attorney Docket No.2014202-0027 skill in the relevant art, however, will readily recognize that the invention can be practiced without one or more of the specific details or with other methods. The present invention is not limited by the illustrated ordering of acts or events, as some acts may occur in different orders and / or concurrently with other acts or events. Furthermore, not all illustrated acts or events are required to implement a methodology in accordance with the present invention.

[0090] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. Furthermore, to the extent that the terms "including", "includes", "having", "has", "with", or variants thereof are used in either the detailed description and / or the claims, such terms are intended to be inclusive in a manner similar to the term "comprising".

[0091] The term "about" or "approximately" means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, "about" can mean within 1 or more than 1 standard deviation, per the practice in the art. Alternatively, "about" can mean a range of up to 20%, preferably up to 10%, more preferably up to 5%, and more preferably still up to 1% of a given value. Alternatively, particularly with respect to biological systems or processes, the term can mean within an order of magnitude, preferably within 5 -fold, and more preferably within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term "about" meaning within an acceptable error range for the particular value should be assumed.

[0092] All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited. It is understood that the present disclosure supersedes any disclosure of an incorporated publication to the extent there is a contradiction.

[0093] It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as "solely", "only" and the like in connection with the recitation of claim elements, or the use of a "negative" limitation. - 26 - 12961406v1Attorney Docket No.2014202-0027

[0094] The publications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing-herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed.

[0095] Unless otherwise indicated, all terms used herein have the same meaning as they would to one skilled in the art and the practice of the present invention will employ, conventional techniques of microbiology and recombinant DNA technology, which are within the knowledge of those of skill of the art. SCFV- AND FAB-MODIFIED AAV CAPSID POLYPEPTIDES

[0096] The present disclosure provides antibody-modified AAV capsid proteins, e.g., VP1, VP2, or VP3 proteins, where the antibody-modified AAV capsid protein comprises one or more antibody fragments, such as single-chain fragment variable (scFv) or a Fab fragment, as compared to the corresponding wild-type AAV or parental AAV. In particular embodiments, the antibody-modified AAV capsid proteins comprise an scFv or Fab inserted at the AAV capsid as a fusion protein. In particular embodiments, the antibody-modified AAV capsid protein comprises an insertion within a capsid protein, e.g., VP1, VP2, or VP3 of AAV type 1 (AAV-1), AAV type 2 (AAV-2), AAV type 3 (AAV-3), AAV type 4 (AAV-4), AAV type 5 (AAV-5), AAV type 6 (AAV-6), AAV type 7 (AAV-7), AAV type 8 (AAV-8), AAV type 9 (AAV-9), AAV type 10 (AAV-10), AAV rh.10, avian AAV, bovine AAV, canine AAV, equine AAV, primate AAV, non-primate AAV, bovine AAV, AAVShH10, or AAV2.5T.

[0097] In certain embodiments, the parental AAV capsid protein is an AAV6, AAV5, AAV8, AAV1, or AAV2 capsid protein. The amino acid sequence of the AAV1 capsid protein can be found under GENBANK Accession No. NP_049542. The amino acid sequence of AAV2 can be found under GENBANK Accession No. YP_680426.1. The amino acid sequence of AAV5 capsid protein can be found under UniProt ID No. Q9YIJ1. The amino acid sequence of the AAV6 capsid protein can be found under GENBANK Accession No. AAB95450. The amino acid sequence of AAV8 can be found under UniProt accession No. YP_077180. While references are made herein to amino acid modifications of capsid proteins (including specific amino acid insertions) using the amino acid numbering corresponding to the AAV6, AAV2, - 27 - 12961406v1Attorney Docket No.2014202-0027 AAV5, and AAV8 VP1 capsid protein, it is understood that any of these amino acid modifications may also be introduced in the capsid proteins of AAVs of other serotypes, e.g., at positions corresponding to those of these AAV serotypes. Additional exemplary sequences are included in the List of Sequences described herein.

[0098] In particular embodiments, the antibody-modified capsid protein, when present in an AAV virion, confers increased infectivity of a lymphocyte as compared to the infectivity of the hematopoietic cell by an AAV virion comprising the corresponding parental AAV capsid protein. In some cases, the hematopoietic cell is a T lymphocyte or a B lymphocyte (e.g., T cell or B cell). In other cases, the lymphocyte is a natural killer (NK) or natural killer T (NKT) cell. In other cases, the T lymphocyte is a CD4+ Helper T cell or Regulatory T cell. In other cases, the T lymphocyte is a CD8+ Cytotoxic T cell. In other cases, the B lymphocyte is a naïve B cell, an activated B cell, a plasma cell, or a memory B cell. In other cases, the B lymphocyte is a follicular B cell, a marginal zone B cell, B1 cell, or germinal center B cell. In particular embodiments, the antibody-modified capsid protein, when present in an AAV virion, confers altered tropism as compared to the tropism of the lymphocyte by an AAV virion comprising the corresponding parental AAV capsid protein.

[0099] In some embodiments, an antibody-modified capsid protein comprises an antibody or antibody fragment that binds to CD3, CD19, or GD2. Exemplary antibody or antibody fragments comprise or consist of an amino acid sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 048, 091, 093, 094, or 095. Exemplary antibody or antibody fragments are encoded by nucleic acid comprising or consisting of a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 012, 026, 041, or 048.

[0100] In certain embodiments, the antibody-modified capsid protein, e.g., VP2, includes more than one unit of scFv or Fab. For example, the antibody-modified AAV6 VP2 capsid protein includes a CD3-scFv, CD28-scFv, CD19-scFv, CD3-Fab, CD28-Fab, or CD19-Fab.

[0101] In certain embodiments, the antibody-modified capsid protein, e.g., VP2, includes an insertion of a scFv or Fab polypeptide linked to an extracellular binding domain of a CD molecule. In particular embodiments, the extracellular binding domain is selected from a group - 28 - 12961406v1Attorney Docket No.2014202-0027 of CD4, CD8, CD19, CD40, CD28, CD152, CD279, CD137, CD2, CD11a / CD18, CD31, CD21, CD14, CD95, CD47, CD55, CD59, CD58, CD80, and CD86.

[0102] In particular embodiments, any of these scFvs or Fabs, or scFv or Fab linked to an extracellular domain of a CD molecule, are present at the N-terminal of AAV6, AAV5, AAV8, or AAV2.

[0103] In some embodiments, a subject antibody-modified AAV capsid does not include any other amino acid substitutions, insertions, or deletions, other than the scFv or Fab insertion relative to a corresponding parental AAV capsid protein. In other embodiments, a subject antibody-modified AAV capsid includes from 1 to about 25 amino acid insertions, deletions, or substitutions, compared to the parental AAV capsid protein, in addition to the scFv or Fab insertion relative to a corresponding parental AAV capsid protein.

[0104] In some embodiments, a subject antibody-modified AAV capsid polypeptide is a chimeric capsid, e.g., the capsid comprises a portion of an AAV capsid of a first AAV serotype and a portion of an AAV capsid of a second serotype; and comprises a scFv or Fab insertion relative to a corresponding parental AAV capsid protein.

[0105] In some embodiments, a subject antibody-modified capsid protein is isolated, e.g., purified. In some cases, a subject antibody-modified capsid protein is included in an AAV vector, which is also provided. As described in detail below, a subject antibody-modified capsid protein can be included in a recombinant AAV virion.

[0106] The scFv sequence may be inserted at various sites within the AAV capsid protein. In particular embodiments, the antibody-modified capsid protein is an AAV6 capsid protein, e.g., VP1 (SEQ ID NO: 096), and the scFv insertion site is in the Loop III region of the AAV6 capsid protein. The wild-type AAV1 virus is closely related to AAV6, with an identical capsid amino acid sequence, but for 5 amino acid substitutions. In certain other embodiments, the antibody-modified capsid is AAV6, AAV5, AAV8, AAV1, AAV2, or any other AAV serotype or derivative, including but not limited to those described herein, and the scFv insertion is present in a region of the AAV that corresponds to the Loop III region of AAV6. In certain embodiments, the scFv insertion is present at or near the tip of the 3-fold protrusion, at or near the basic amino acid residue 449, or at or near the heparan sulfate proteoglycan binding site of AAV6 capsid protein, or a corresponding site of another AAV serotype or derivative. In - 29 - 12961406v1Attorney Docket No.2014202-0027 particular embodiments, "near" indicates within two, within three, within four, or within five amino acid residues of the indicated amino acid or site. For example, the scFv insertion site can be within amino acids 447–460 or amino acids 448–464 of AAV6 capsid protein, e.g., between two adjacent amino acids selected from 447 and 448, 448 and 449, 449 and 450, 450 and 451, 451 and 452, 452 and 453, 453 and 454, 454 and 455, 455 and 456, 456 and 457, 457 and 458, 458 and 459, 459 and 460, 460 and 461, 461 and 462, 462 and 463, or 463 and 464 of AAV6 capsid protein, e.g., VP1 (SEQ ID NO: 096), or the corresponding amino acid residues in any other AAV capsid protein, e.g., VP1, VP2, or VP3.

[0107] In particular embodiments, the antibody-modified capsid protein is an AAV6 capsid protein, e.g., VP1 (SEQ ID NO: 096), and the scFv insertion site is in the Loop IV region of the AAV6 capsid protein. In certain other embodiments, the antibody-modified capsid is AAV6, AAV5, AAV8, AAV1, AAV2, or any other AAV serotype or derivative, including but not limited to those described herein, and the scFv insertion is present in a region of the AAV that corresponds to the Loop IV region of AAV6.

[0108] In particular embodiments, the scFv insertion is present at or near amino acid residue 587 of the AAV6 capsid protein, or at a corresponding site in another AAV serotype or derivative. As used herein, “near” refers to within two, within three, within four, or within five amino acid residues of the indicated amino acid or site. For example, the scFv insertion site can be within amino acids 585–590 of the AAV6 capsid protein, e.g., between two adjacent amino acids selected from 585 and 586, 586 and 587, 587 and 588, 588 and 589, or 589 and 590 of the AAV6 capsid protein, e.g., VP1 (SEQ ID NO: 096), or the corresponding amino acid residues in any other AAV capsid protein, e.g., VP1, VP2, or VP3.

[0109] In particular embodiments, the antibody-modified capsid protein is an AAV6 capsid protein, and the scFv or Fab insertion site is at the N-terminus of the VP2 capsid protein (SEQ ID NO: 097). In certain other embodiments, the antibody-modified capsid is AAV6, AAV5, AAV8, AAV1, AAV2, or any other AAV serotype or derivative, including but not limited to those described herein, and the scFv or Fab insertion is present at the N-terminal region of the VP2 protein of such AAV capsids.

[0110] In particular embodiments, the scFv or Fab sequence is fused in-frame at the amino terminus of the VP2 protein, such that the scFv or Fab is displayed on the exterior of the - 30 - 12961406v1Attorney Docket No.2014202-0027 AAV particle following capsid assembly. The insertion may include a flexible linker between the scFv or Fab and VP2 to facilitate proper folding and surface presentation. This approach may allow for receptor targeting, antigen binding, or cell-specific tropism modulation, while maintaining overall capsid integrity and packaging efficiency. The corresponding N-terminal site in other AAV serotypes (e.g., AAV1, AAV2, AAV5, AAV8) may also be used for such insertions, with appropriate adjustment based on sequence alignment and capsid architecture.

[0111] In particular embodiments, the antibody-modified capsid protein is an AAV6 capsid protein, and the scFv or Fab insertion site is at the C-terminus of one of the capsid proteins, such as VP1 (SEQ ID NO: 096), VP2 (SEQ ID NO: 097), or VP3 (SEQ ID NO: 098). In certain other embodiments, the antibody-modified capsid is AAV6, AAV5, AAV8, AAV1, AAV2, or any other AAV serotype or derivative, including but not limited to those described herein, and the scFv or Fab insertion is present at the C-terminal region of one or more of the capsid proteins of such AAV variants.

[0112] In particular embodiments, the scFv or Fab sequence is fused in-frame at the carboxyl terminus of VP1, VP2, or VP3, such that the scFv or Fab is displayed on the exterior of the AAV particle following capsid assembly. A flexible peptide linker may be included between the capsid protein and the scFv or Fab to promote proper folding, surface exposure, and functional presentation. This strategy allows for antigen binding, receptor targeting, or cell-type specific transduction, while maintaining capsid stability and genome packaging efficiency. The corresponding C-terminal site in other AAV serotypes (e.g., AAV1, AAV2, AAV5, AAV8) may also be used for such insertions, with appropriate adjustments based on structural homology and capsid architecture.

[0113] In some embodiments, a modified capsid protein is encoded by a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 014, 039, 046, or 053.

[0114] AAV capsids are known to have icosahedral structures in which about 60 viral capsid proteins are associated, at a ratio (e.g., ratio of copy number or molar ratio) of VP1:VP2:VP3 of about 1:1:10 (Matsuzaka et al., Curr. Issues Mol. Biol.46:8464-8498 (2024)). In some embodiments, a modified capsid of the disclosure includes antibody-modified VP1 capsid proteins described herein and unmodified VP2 and VP3 capsid proteins. For example, in - 31 - 12961406v1Attorney Docket No.2014202-0027 some embodiments, a modified capsid of the disclosure includes (i) about 5 copies of VP1 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, or all copies) of VP1 capsid proteins are antibody-modified VP1 capsid proteins; and (ii) unmodified VP2 and VP3 capsid proteins. In some embodiments, a modified capsid of the disclosure includes (i) a plurality of VP1 capsid proteins where about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the plurality are antibody-modified VP1 capsid proteins; and (ii) a plurality of VP2 and VP3 capsid proteins where 100% are unmodified VP2 and VP3 capsid proteins.

[0115] In some embodiments, a modified capsid of the disclosure includes antibody- modified VP2 capsid proteins described herein and unmodified VP1 and VP3 capsid proteins. For example, in some embodiments, a modified capsid of the disclosure includes (i) about 5 copies of VP2 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, or all copies) of VP2 capsid proteins are antibody-modified VP2 capsid proteins; and (ii) unmodified VP1 and VP3 capsid proteins. In some embodiments, a modified capsid of the disclosure includes (i) a plurality of VP2 capsid proteins where about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the plurality are antibody-modified VP2 capsid proteins; and (ii) a plurality of VP1 and VP3 capsid proteins where 100% are unmodified VP1 and VP3 capsid proteins.

[0116] In some embodiments, a modified capsid of the disclosure includes antibody- modified VP3 capsid proteins described herein and unmodified VP1 and VP2 capsid proteins. For example, in some embodiments, a modified capsid of the disclosure includes (i) about 50 copies of VP3 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 20 copies) of VP3 capsid proteins are antibody-modified VP3 capsid proteins; and (ii) unmodified VP1 and VP2 capsid proteins. In some embodiments, a modified capsid of the disclosure includes (i) a plurality of VP3 capsid proteins where about 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 30%, 40%, or 50% of the plurality are antibody-modified VP3 capsid proteins; and (ii) a plurality of VP1 and VP2 capsid proteins where 100% are unmodified VP1 and VP2 capsid proteins.

[0117] In some embodiments, a modified capsid of the disclosure includes antibody- modified VP1 and antibody-modified VP2 capsid proteins described herein and unmodified VP3 capsid proteins. For example, in some embodiments, a modified capsid of the disclosure includes (i) about 5 copies of VP1 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, or all - 32 - 12961406v1Attorney Docket No.2014202-0027 copies) of VP1 capsid proteins are antibody-modified VP1 capsid proteins; (ii) about 5 copies of VP2 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, or all copies) of VP2 capsid proteins are antibody-modified VP2 capsid proteins; and (iii) unmodified VP3 capsid proteins. In some embodiments, a modified capsid of the disclosure includes (i) a plurality of VP1 capsid proteins where about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the plurality are antibody-modified VP1 capsid proteins; (ii) a plurality of VP2 capsid proteins where about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the plurality are antibody-modified VP2 capsid proteins; and (iii) a plurality VP3 capsid proteins where 100% are unmodified VP3 capsid proteins.

[0118] In some embodiments, a modified capsid of the disclosure includes antibody- modified VP1 and antibody-modified VP3 capsid proteins described herein and unmodified VP2 capsid proteins. For example, in some embodiments, a modified capsid of the disclosure includes (i) about 5 copies of VP1 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, or all copies) of VP1 capsid proteins are antibody-modified VP1 capsid proteins; (ii) about 50 copies of VP3 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 20 copies) of VP3 capsid proteins are antibody-modified VP3 capsid proteins; and (iii) unmodified VP2 capsid proteins. In some embodiments, a modified capsid of the disclosure includes (i) a plurality of VP1 capsid proteins where about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100% of the plurality are antibody-modified VP1 capsid proteins; (ii) a plurality of VP3 capsid proteins where about 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 30%, 40%, or 50% of the plurality are antibody-modified VP3 capsid proteins; and (iii) a plurality of VP2 capsid proteins where 100% are unmodified VP2 capsid proteins.

[0119] In some embodiments, a modified capsid of the disclosure includes antibody- modified VP2 and antibody-modified VP3 capsid proteins described herein and unmodified VP1 capsid proteins. For example, in some embodiments, a modified capsid of the disclosure includes (i) about 5 copies of VP2 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, or all copies) of VP2 capsid proteins are antibody-modified VP2 capsid proteins; (ii) about 50 copies of VP3 capsid protein, and at least 1 copy (e.g., at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 20 copies) of VP3 capsid proteins are antibody-modified VP3 capsid proteins; and (iii) unmodified VP1 capsid proteins. In some embodiments, a modified capsid of the disclosure includes (i) a plurality of VP2 capsid proteins where about 10%, 20%, 30%, 40%, 50%, 60%, - 33 - 12961406v1Attorney Docket No.2014202-0027 70%, 80%, 90%, or 100% of the plurality are antibody-modified VP2 capsid proteins; (ii) a plurality of VP3 capsid proteins where about 2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 30%, 40%, or 50% of the plurality are antibody-modified VP3 capsid proteins; and (iii) a plurality of VP1 capsid proteins where 100% are unmodified VP1 capsid proteins.

[0120] The present disclosure also includes polynucleotides that encode one or more antibody-modified capsids described herein. In particular embodiments, the polynucleotide is an expression vector, and the expression vector comprises a polynucleotide sequence encoding a scFv- or Fab-modified capsid described herein operably linked to a promoter sequence, e.g., a promoter sequence that drives expression of the polynucleotide in a cell. In particular embodiments, the promoter sequence is a tissue-specific promoter that preferentially drives expression in one or more tissue or cell types, e.g., the B lymphocyte, the T lymphocyte, or hepatocytes. In some embodiments, one or more modified capsid proteins are encoded by a nucleotide sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to SEQ ID NO: 013, 018, 022, 032, 040, 044, 047, 052, or 057, or a portion thereof.

[0121] The present disclosure also includes cells comprising a polynucleotide or vector that encodes a scFv- or Fab-modified capsid described herein. In particular embodiments, the polynucleotide is an expression vector, and the expression vector comprises a polynucleotide sequence encoding a scFv- or Fab-modified capsid described herein operably linked to a promoter sequence, e.g., a promoter sequence that drives expression of the polynucleotide in the cell. In certain embodiments, the polynucleotide or vector further comprises a sequence that encodes a rep protein, e.g., an AAV2 rep protein. In certain embodiments, the cell is a helper cell or host cell, such as, e.g., an HEK293 cell that may be used to produce virions comprising the scFv- or Fab-modified capsid protein. In preparing the subject rAAV compositions, any host cells for producing rAAV virions may be employed, including, for example, mammalian cells (e.g., 293 cells), insect cells (e.g., Sf9 cells), and microorganisms such as yeast and bacteria. Host cells can also be packaging cells in which the AAV rep and cap genes are stably maintained in the host cell or producer cells in which the AAV vector genome is stably maintained and packaged. Exemplary packaging and producer cells are derived from Sf-9, 293, A549, or HeLa cells. AAV vectors are purified and formulated using standard techniques known in the art. - 34 - 12961406v1Attorney Docket No.2014202-0027 RECOMBINANT VIRIONS AND VIRAL VECTORS

[0122] In some embodiments, the recombinant virion or virus is, e.g., an AAV, further comprising a polynucleotide cassette comprising a sequence that encodes a gene product, e.g., a therapeutic gene product.

[0123] Illustrative examples of viral vectors suitable for gene therapy include but are not limited to retroviral vectors, lentiviral vectors, adenovirus vectors, herpes virus vectors, alphavirus vectors, baculovirus vectors and adeno-associated virus (AAV) vectors.

[0124] AAV is a 4.7 kb, single-stranded DNA virus widely utilized in gene therapy due to its favorable safety profile and robust gene delivery capabilities. Wild-type AAV is nonpathogenic, with no known etiologic association with human diseases, making recombinant AAV vectors highly attractive for clinical applications. These vectors demonstrate efficient gene delivery and sustained transgene expression across diverse tissues, including the retina, skeletal muscle, lung, and central nervous system. Notably, several AAV-based gene therapy drugs have received regulatory approval for human use, including Luxturna (voretigene neparvovec) for retinal dystrophy, Zolgensma (onasemnogene abeparvovec) for spinal muscular atrophy, Glybera (alipogene tiparvovec) for lipoprotein lipase deficiency, Hemgenix (etranacogene dezaparvovec) for hemophilia B, and Roctavian (valoctocogene roxaparvovec) for hemophilia A.

[0125] In certain embodiments, the sequence that encodes the gene product is operably linked to a promoter sequence. In certain embodiments, the polynucleotide cassette is flanked on the 5' and 3' ends by functional AAV inverted terminal repeat (ITR) sequences. By "functional AAV ITR sequences" is meant that the ITR sequences function as intended for the rescue, replication, and packaging of the AAV virion. Hence, AAV ITRs for use in the gene delivery vectors of the invention need not have a wild-type nucleotide sequence, and may be altered by the insertion, deletion, or substitution of nucleotides, or the AAV ITRs may be derived from any of several AAV serotypes, e.g., AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10. Certain AAV vectors have the wild-type REP and CAP genes deleted in whole or part, but retain functional flanking ITR sequences.

[0126] In certain embodiments, recombinant viruses or virions described herein comprise a heterologous nucleic acid comprising a nucleotide sequence encoding a gene product, e.g., a therapeutic gene product. In some embodiments, the gene product is an interfering RNA. In - 35 - 12961406v1Attorney Docket No.2014202-0027 some embodiments, the gene product is an aptamer. In some embodiments, the gene product is a polypeptide. In some embodiments, the gene product is a site-specific nuclease that provides for site-specific knock-down of gene function.

[0127] Where the gene product is an interfering RNA (RNAi), suitable RNAi include RNAi that decrease the level of an apoptotic or angiogenic factor in a cell. For example, an RNAi can be an shRNA or siRNA that reduces the level of a gene product that induces or promotes apoptosis in a cell. Genes whose gene products induce or promote apoptosis are referred to herein as "pro-apoptotic genes" and the products of those genes (mRNA; protein) are referred to as "pro-apoptotic gene products." Pro-apoptotic gene products include, e.g., Bax, Bid, Bak, and Bad gene products. See, e.g., U.S. Pat. No.7,846,730.

[0128] Interfering RNAs could also be against an angiogenic product, for example VEGF (e.g., Cand5; see, e.g., U.S. Patent Publication No.2011 / 0143400; U.S. Patent Publication No. 2008 / 0188437; and Reich et al. (2003) Mol. Vis.9:21017), VEGFR1 (e.g., Sirna-027; see, e.g., Kaiser et al. (2010) Am. J. Ophthalmol.150:33;18and Shen et al. (2002) Lab Invest 82:16719). See also, U.S. Pat. Nos.6,649,596, 6,399,586, 5,661,135, 5,639,872, and 5,639,736; and U.S. Pat. Nos.7,947,659 and 7,919,473.

[0129] Where the gene product is an aptamer, exemplary aptamers of interest include an aptamer against vascular endothelial growth factor (VEGF). See, e.g., Ng et al. (2006) Nat. Rev. Drug Discovery 5: 123;20and Lee et al. (2005) Proc. Natl. Acad. Sci. USA 102: 18902.21For example, a VEGF aptamer can comprise the nucleotide sequence 5'- cgcaaucagugaaugcuuauacauccg-3'. Also suitable for use is a PDGF-specific aptamer, e.g., E10030; see, e.g., Ni and Hui (2009) Ophthalmologica 223:401;22and Akiyama et al. (2006) J. Cell Physiol.207:407).23

[0130] Where the gene product is a polypeptide, in certain embodiments, the polypeptide is a chimeric antigen receptor (CAR) or a transgenic T cell receptor (TCR). For example, a CD19-specific CAR is designed to target the CD19 antigen, which is predominantly expressed on the surface of B cells. It is used in CAR-T cell therapies to treat various B-cell malignancies, such as Acute Lymphoblastic Leukemia (ALL), Diffuse Large B Cell Lymphoma (DLBCL), Mantle Cell Lymphoma (MCL), Follicular Lymphoma (FL), Non-Hodgkin Lymphoma (NHL), and Chronic Lymphocytic Leukemia (CLL). A GD2-specific CAR is designed to target the GD2 - 36 - 12961406v1Attorney Docket No.2014202-0027 antigen, which is highly expressed on the surface of certain solid tumors, particularly neuroblastoma, as well as some other cancers like osteosarcoma and melanoma. GD2-specific CAR T cell therapy is used to recognize and kill cancer cells expressing this antigen, making it a promising approach for treating these types of cancers. A transgenic TCR can target tumor- associated antigens (e.g., PRAME, NY-ESO-1, MAGE-A4), expressed by multiple solid tumors (e.g., melanoma, sarcoma, ovarian carcinoma, and breast cancer, among others). Nucleotide sequences for exemplary CARs are included as SEQ ID NOs: 063 and 064. In some embodiments, recombinant viruses or virions described herein comprise a heterologous nucleic acid comprising a nucleotide sequence having at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO: 063 or SEQ ID NO: 064.

[0131] In some cases, a gene product of interest is a toxin, e.g., Pseudomonas exotoxin, diphtheria toxin, and ricin are used to target cancer cells because they are the most potent cell- killing agents. One molecule of a toxin delivered to the cytoplasm of a cancer cell will be lethal for that cell. For example, a CD19-scFv- or CD19-Fab-modified AAV capsid is designed to target CD19-positive cancer cells specifically and destroy the cancer cells upon delivering and expressing the toxin inside the cells.

[0132] In some cases, a gene product of interest is a site-specific endonuclease that provide for site-specific knock-down of gene function, e.g., where the CRISPR-based gene editing nucleases knockout or decrease the expression of the PD-1 protein on T cells to improve their ability to target and kill cancer cells (Su et al., 2017. Scientific reports).24

[0133] In addition to knocking out a gene function, a site-specific nuclease can also be used to stimulate homologous recombination with a donor DNA that encodes a functional copy of the protein encoded by a defective allele. Thus, e.g., a subject rAAV virion can be used to deliver both a site-specific endonuclease that knocks out a defective allele, and can be used to deliver a functional copy of the defective allele, resulting in repair of the defective allele, thereby providing for production of a functional protein.

[0134] Site-specific endonucleases that are suitable for use include, e.g., zinc finger nucleases (ZFNs); and transcription activator-like effector nucleases (TALENs), where such site- specific endonucleases are non-naturally occurring and are modified to target a specific gene. Such site- specific nucleases can be engineered to cut specific locations within a genome, and - 37 - 12961406v1Attorney Docket No.2014202-0027 nonhomologous end joining can then repair the break while inserting or deleting several nucleotides. Such site-specific endonucleases (also referred to as "INDELs") then throw the protein out of frame and effectively knock out the gene. See, e.g., U.S. Patent Publication No. 2011 / 0301073.

[0135] In some embodiments, a nucleotide sequence encoding a gene product is operably linked to a constitutive promoter. In other embodiments, a nucleotide sequence encoding a gene product of interest is operably linked to an inducible promoter. In some instances, a nucleotide sequence encoding a gene product of interest is operably linked to a tissue-specific or cell type- specific regulatory element. In certain embodiments, the promoter selected from cytomegalovirus (CMV) promoter, Rous sarcoma virus (RSV) promoter, MMT promoter, EF- 1 alpha promoter, UB6 promoter, chicken beta-actin promoter, CAG promoter, RPE65 promoter and opsin promoter.

[0136] For example, in some instances, a nucleotide sequence encoding a gene product of interest is operably linked to a photoreceptor-specific regulatory element (e.g., a photoreceptor- specific promoter), e.g., a regulatory element that confers selective expression of the operably linked gene in a photoreceptor cell. Suitable photoreceptor-specific regulatory elements include, e.g., a rhodopsin promoter; a rhodopsin kinase promoter (Young et al. (2003) Ophthalmol. Vis. Sci.44:4076);25a beta phosphodiesterase gene promoter (Nicoud et al. (2007) J. Gene Med.9: 1015);26a retinitis pigmentosa gene promoter (Nicoud et al. (2007) supra); an interphotoreceptor retinoid-binding protein (IRBP) gene enhancer (Nicoud et al. (2007) supra); an IRBP gene promoter (Yokoyama et al. (1992) Exp Eye Res.55:225).27

[0137] For example, in some instances, a nucleotide sequence encoding a gene product of interest is operably linked to a liver or hepatocyte specific regulatory element (e.g., a liver- specific promoter), e.g., a regulatory element that confers selective expression of the operably linked gene in a liver cell, e.g., a hepatocyte. Suitable liver-specific regulatory elements include, e.g., the Apolipoprotein E / C-I hepatic control region, alone or combined with the human alpha- 1-antitrypsin core promoter; one or two copies of alpha 1 microglobulin / bikunin enhancer coupled to the core promoter of human thyroxine-binding globulin (TBG); or the promoter region referred to as "ET," and described as randomly assembled hepatocyte-specific transcription factor binding sites linked to the murine transthyretin promoter, as summarized in - 38 - 12961406v1Attorney Docket No.2014202-0027 Kattenhorn, L.M. et al, Human Gene Therapy, 2016 Dec 1; 27(12): 947-961.28Expression may be further stabilized by the inclusion of a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE).

[0138] Recombinant AAV vectors comprising scFv- or Fab-modified capsid proteins described herein, and optionally encapsulating polynucleotide cassettes of the present disclosure, may be produced using standard methodology. For example, an AAV expression vector comprising a polynucleotide cassette may be introduced into a producer cell, followed by introduction of an AAV helper construct comprising a polynucleotide sequence encoding a scFv- or Fab-modified capsid protein disclosed herein, and where the helper construct includes AAV coding regions capable of being expressed in the producer cell and which complement AAV helper functions absent in the AAV vector. This is followed by introduction of helper virus and / or additional vectors into the producer cell, wherein the helper virus and / or additional vectors provide accessory functions capable of supporting efficient rAAV virus production. The producer cells are then cultured to produce rAAV. These steps are carried out using standard methodology. Replication-defective AAV virions comprising scFv- or Fab-modified capsid proteins described herein are made by standard techniques known in the art using AAV packaging cells and packaging technology. Examples of these methods may be found, for example, in U.S. Pat. Nos.5,436,146; 5,753,500, 6,040,183, 6,093,570, 6,548,286, expressly incorporated by reference herein in their entirety. Further compositions and methods for packaging of AAV in Sf9 cells are described in Chen (Patent No: US 8,945,918 B2), also incorporated by reference herein in its entirety.

[0139] As disclosed in the accompanying Examples, scFv- or Fab-modified capsid proteins described herein confer enhanced or altered cellular tropism to virions comprising the scFv- or Fab-modified capsid proteins. Accordingly, the scFv- or Fab-modified capsids may be used to enhance or alter the tropism of a virus or virion in order to enhance its tropism for a desired cell type.

[0140] In particular embodiments, the virions or viral vectors comprising a scFv- or Fab- modified capsid protein described herein are capable of delivering a gene product to lymphocytes when delivered via intravenous injection, e.g., wherein they result in the expression of at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 20-fold, at - 39 - 12961406v1Attorney Docket No.2014202-0027 least 50-fold, 100-fold, or at least 200-fold gene product as compared to a corresponding virion or viral vector that does not include a scFv- or Fab-modified capsid protein disclosed herein. In certain embodiments, these virions or viral vectors comprising a scFv- or Fab-modified capsid protein described herein are capable of selectively transducing lymphocytes at a higher level than they transduce one or more other cell types.

[0141] In particular embodiments, the virions or viral vectors comprising a scFv- or Fab- modified capsid protein described herein are capable of delivering a gene product to cancer cells such as neuroblastoma and melanoma when delivered via intravenous injection or infusion, e.g., wherein they result in the expression of at least 2-fold, at least 3-fold, at least 4-fold, at least 5- fold, at least 10-fold, at least 20-fold, at least 50-fold, 100-fold, or at least 200-fold gene product as compared to a corresponding virion or viral vector that does not include a scFv- or Fab- modified capsid protein disclosed herein. In certain embodiments, these virions or viral vectors comprising a scFv- or Fab-modified capsid protein described herein are capable of selectively transducing cancer cells at a higher level than they transduce one or more other normal cells. PHARMACEUTICAL COMPOSITIONS

[0142] Pharmaceutical compositions comprising a virion or viral vector comprising a scFv- or Fab-modified capsid protein described herein and one or more pharmaceutically acceptable diluent, carrier, or excipient are also disclosed. The subject virions or vectors can be combined with pharmaceutically acceptable carriers, diluents, and reagents useful in preparing a formulation that is generally safe, non-toxic, and desirable, and includes excipients that are acceptable for primate use. Such excipients can be solid, liquid, semisolid, or, in the case of an aerosol composition, gaseous. Examples of such carriers or diluents include, but are not limited to, water, saline, Ringer’s solutions, dextrose solution, and 5% human serum albumin. Supplementary active compounds can also be incorporated into the formulations. Solutions or suspensions used for the formulations can include a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol, or other synthetic solvents; antibacterial compounds such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating compounds such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates, or phosphates; detergents such as Tween 20 to prevent aggregation; and compounds for the adjustment of tonicity such as sodium chloride or - 40 - 12961406v1Attorney Docket No.2014202-0027 dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. In particular embodiments, the pharmaceutical compositions are sterile.

[0143] Pharmaceutical compositions may further include sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, or phosphate buffered saline (PBS). In some cases, the composition is sterile and should be fluid to the extent that easy syringeability exists. In certain embodiments, it is stable under the conditions of manufacture and storage and is preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be, e.g., a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion, and by the use of surfactants. Prevention of the action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Prolonged absorption of the internal compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

[0144] Sterile solutions can be prepared by incorporating the vector or virion in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

[0145] In one embodiment, the pharmaceutical compositions are prepared with carriers that will protect the virion or vector against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, - 41 - 12961406v1Attorney Docket No.2014202-0027 biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

[0146] It may be advantageous to formulate oral, ocular or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specifications for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

[0147] Any concentration of viral particles suitable to effectively transduce mammalian cells can be prepared. For example, the viral particles may be formulated at a concentration of 1 x 108vector genomes per mL or more, for example, 5 x 108vector genomes per mL; 1 x 109vector genomes per mL; 5 x 109vector genomes per mL, 1 x 1010vector genomes per mL, 5 x l010vector genomes per mL; 1 x 1011vector genomes per mL; 5 x l0nvector genomes per mL; 1 x 1012vector genomes per mL; 5 x 1012vector genomes per mL; 1 x 1013vector genomes per mL; 1.5 x l013vector genomes per mL; 3 x 1013vector genomes per mL; 5 x 1013vector genomes per mL; 7.5 x l013vector genomes per mL; 9 x 1013vector genomes per mL; 1 x 1014vector genomes per mL, 5 x 1014vector genomes per mL or more, but typically not more than 1 x 1015vector genomes per mL.

[0148] The subject viral vector may be formulated into any suitable unit dosage, including, without limitation, 1 x 108vector genomes or more, for example, 1 x 109, 1 x 1010, 1 x 1011, 1 x 1012, or 1 x 1013vector genomes or more, in certain instances, 1 x 1014vector genomes, but usually no more than 1 x 1015vector genomes. In some cases, the unit dosage is at most - 42 - 12961406v1Attorney Docket No.2014202-0027 about 5 x 1015vector genomes, e.g.1 x 1014vector genomes or less, for example 1 x 1013, 1 x 1012, 1 x 1011, 1 x 1010, or 1 x 109vector genomes or less, in certain instances 1 x 108vector genomes or less, and typically no less than 1 x 108vector genomes. In some cases, the unit dosage is 1 x 1010to 1 x 1011vector genomes. In some cases, the unit dosage is 1 x 1010to 3 x 1012vector genomes. In some cases, the unit dosage is 1 x 109to 3 x 1013vector genomes. In some cases, the unit dosage is 1 x 108to 3 x 1014vector genomes.

[0149] In some cases, the unit dosage of a pharmaceutical composition may be measured using multiplicity of infection (MOI). By MOI it is meant the ratio, or multiple, of vector or viral genomes to the cells to which the nucleic acid may be delivered. In some cases, the MOI may be 1 x 106. In some cases, the MOI may be 1 x 105– 1 x 107. In some cases, the MOI may be 1 x 104– 1 x 108. In some cases, recombinant viruses of the disclosure are at least about 1 x 101, 1 x 102, 1 x 103, 1 x 104, 1 x 105, 1 x 106, 1 x 107, 1 x 108, 1 x 109, 1 x 1010, 1 x 1011, 1 x 1012, 1 x 1013, 1 x 1014MOI.

[0150] In some aspects, the amount of pharmaceutical composition comprises about 1 x 108to about 1 x 1015recombinant viruses, about 1 x 109to about 1 x 1014recombinant viruses, about 1 x 1010to about 1 x 1013recombinant viruses, or about 1 x 1011to about 3 x 1012recombinant viruses.

[0151] The pharmaceutical compositions can be included in a container, pack, or dispenser, e.g. syringe, e.g. a prefilled syringe, together with instructions for administration.

[0152] The pharmaceutical compositions of the invention encompass any pharmaceutically acceptable salts, esters, or salts of such esters, or any other compound which, upon administration to an animal comprising a human, is capable of providing (directly or indirectly) the biologically active metabolite or residue thereof.

[0153] The term "pharmaceutically acceptable salt" refers to physiologically and pharmaceutically acceptable salts of the compounds of the invention: i.e., salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects thereto. A variety of pharmaceutically acceptable salts are known in the art and described, e.g., in in "Remington's Pharmaceutical Sciences", 17th edition, Alfonso R. Gennaro (Ed.), Mark Publishing Company, Easton, PA, USA, 1985 (and more recent editions thereof), in the "Encyclopaedia of Pharmaceutical Technology", 3rd edition, James Swarbrick (Ed.), Informa - 43 - 12961406v1Attorney Docket No.2014202-0027 Healthcare USA (Inc.), NY, USA, 2007, and in J. Pharm. Sci.66: 2 (1977). Also, for a review on suitable salts, see Handbook of Pharmaceutical Salts: Properties, Selection, and Use by Stahl and Wermuth (Wiley -VCH, 2002).

[0154] Pharmaceutically acceptable base addition salts are formed with metals or amines, such as alkali and alkaline earth metals or organic amines. Metals used as cations comprise sodium, potassium, magnesium, calcium, and the like. Amines comprise N-N'- dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, dicyclohexylamine, ethylenediamine, N-methylglucamine, and procaine (see, for example, Berge et al., "Pharmaceutical Salts," J. Pharma Sci., 1977, 66, 119). The base addition salts of said acidic compounds are prepared by contacting the free acid form with a sufficient amount of the desired base to produce the salt in the conventional manner. The free acid form may be regenerated by contacting the salt form with an acid and isolating the free acid in the conventional manner. The free acid forms differ from their respective salt forms somewhat in certain physical properties such as solubility in polar solvents, but otherwise the salts are equivalent to their respective free acid for purposes of the present invention.

[0155] The subject polynucleotide cassette or gene delivery vector, e.g., recombinant virus (virions), can be incorporated into pharmaceutical compositions for administration to mammalian patients, particularly primates and more particularly humans. The subject polynucleotide cassette or gene delivery vector, e.g. virions can be formulated in nontoxic, inert, pharmaceutically acceptable aqueous carriers, preferably at a pH ranging from 3 to 8, more preferably ranging from 6 to 8. Such sterile compositions will comprise the vector or virion containing the nucleic acid encoding the therapeutic molecule dissolved in an aqueous buffer having an acceptable pH upon reconstitution.

[0156] In some embodiments, the pharmaceutical composition provided herein comprise a therapeutically effective amount of a vector or virion in admixture with a pharmaceutically acceptable carrier and / or excipient, for example saline, phosphate buffered saline, phosphate and amino acids, polymers, polyols, sugar, buffers, preservatives and other proteins. Exemplary amino acids, polymers and sugars and the like are octylphenoxy polyethoxy ethanol compounds, polyethylene glycol monostearate compounds, polyoxyethylene sorbitan fatty acid esters, sucrose, fructose, dextrose, maltose, glucose, mannitol, dextran, sorbitol, inositol, galactitol, - 44 - 12961406v1Attorney Docket No.2014202-0027 xylitol, lactose, trehalose, bovine or human serum albumin, citrate, acetate, Ringer's and Hank's solutions, cysteine, arginine, carnitine, alanine, glycine, lysine, valine, leucine, polyvinylpyrrolidone, polyethylene and glycol. Preferably, this formulation is stable for at least six months at 4° C.

[0157] In some embodiments, the pharmaceutical composition provided herein comprises a buffer, such as phosphate buffered saline (PBS) or sodium phosphate / sodium sulfate, tris buffer, glycine buffer, sterile water and other buffers known to the ordinarily skilled artisan such as those described by Good et al. (1966) Biochemistry 5:467. In particular embodiments, the pH of the buffer may be in the range of 6.5 to 7.75, preferably 7 to 7.5, and most preferably 7.2 to 7.4. The pharmaceutical composition may be formulated for various types of deliveries, including, e.g., parenteral administration, intravenous injection or infusion, and injection into the liver. METHOD OF ENHANCING OR ALTERING VIRAL TROPISM

[0158] As disclosed in the accompanying Examples, scFv- or Fab-modified capsid proteins described herein confer enhanced or altered cellular tropism or tissue specificity to virions comprising the scFv- or Fab-modified capsid proteins. For example, certain scFv- or Fab- modified capsid proteins described herein are associated with increased infectivity of hematopoietic cells or cancer cells, and increased expression levels of gene products in hematopoietic cells or cancer cells.

[0159] The scFv- or Fab-modified capsids disclosed herein may be used to enhance or alter the tropism of a virus or virion in order to enhance its tropism for a desired cell type.

[0160] In particular embodiments, the virions or viral vectors comprising a scFv- or Fab- modified capsid protein described herein are used to deliver a gene product to T cells. In particular embodiments, the virion has increased tropism for T cells.

[0161] In particular embodiments, the virions or viral vectors comprising a scFv- or Fab- modified capsid protein described herein are used to deliver a gene product to cancer cells, e.g., by intravenous injection or infusion, or by direct injection into the tumor.

[0162] In certain embodiments, the disclosure provides a method of altering the tropism of a virus, e.g., an AAV6, AAV5, AAV8, AAV1, or AAV2 virus, comprising introducing one or - 45 - 12961406v1Attorney Docket No.2014202-0027 more scFv or Fab sequences into a capsid protein of the virus at a position described herein. In related embodiments, the method comprises a method of altering the tropism of a virus by incorporating a scFv- or Fab-modified capsid protein disclosed herein into the virus. METHODS OF EXPRESSING GENE PRODUCTS AND TREATING DISEASES AND DISORDERS

[0163] Virions and viral vectors described herein, comprising a scFv- or Fab-modified capsid protein described herein, may be used for delivering a transgene to a cell, e.g., cells of an animal. For example, they may be used in research, e.g., to determine the effect that the gene has on cell viability and / or function. As another example, they may be used in medicine, e.g., to treat a disorder, for example, by delivering a therapeutic gene product to a cell or tissue. Thus, in some aspects of the invention, methods are provided for the expression of a gene in cells, the method comprising contacting cells with a composition of the present disclosure. In some embodiments, contacting cells with a composition of the present disclosure occurs in vitro. In some embodiments, contacting cells with a composition of the present disclosure occurs in vivo, i.e., the subject composition is administered to a subject. In particular embodiments, a viral vector is administered parenterally, e.g., intravenously, orally, or by injection. In certain embodiments, it is administered to the subject by intravenous injection. In certain embodiments, it is administered by peritoneal injection. In certain embodiments, it is administered parenterally, e.g., via intravenous injection or infusion. In certain embodiments, it is administered locally or directly to a tissue or organ of interest, e.g., via injection into the tumor.

[0164] For instances in particular embodiments in which mammalian cells are to be contacted in vitro with a subject virion or vector comprising a scFv- or Fab-modified capsid protein disclosed herein, the cells may be from any mammalian species, e.g., rodent (e.g., mice, rats, gerbils, squirrels), rabbit, feline, canine, goat, ovine, pig, equine, bovine, primate, human. Cells may be from established cell lines, e.g., HEK293 cells, HeLa cells, C2C12 cells, and CHO cells, or they may be primary cells, where "primary cells", "primary cell lines", and "primary cultures" are used interchangeably herein to refer to cells and cell cultures that have been derived from a subject and allowed to grow in vitro for a limited number of passages, i.e., splittings, of the culture. For example, primary cultures are cultures that may have been passaged 0 times, 1 time, 2 times, 4 times, 5 times, 10 times, or 15 times, but not enough times to go through the - 46 - 12961406v1Attorney Docket No.2014202-0027 crisis stage. Typically, the primary cell lines of the present disclosure are maintained for fewer than 10 passages in vitro.

[0165] If the cells are primary cells, they may be harvested from a mammal by any convenient method, e.g., whole explant, biopsy, leukapheresis, blood collection, etc. An appropriate solution may be used for dispersion or suspension of the harvested cells. Such solution will generally be a balanced salt solution, e.g., normal saline, PBS, Hank’s balanced salt solution, etc., conveniently supplemented with fetal calf serum or other naturally occurring factors, in conjunction with an acceptable buffer at low concentration, generally from 5-25 mM. Convenient buffers include HEPES, phosphate buffers, lactate buffers, etc. The cells may be used immediately, or they may be stored, frozen, for long periods of time, being thawed and capable of being reused. In such cases, the cells will usually be frozen in 10% DMSO, 50% serum, 40% buffered medium, or some other such solution as is commonly used in the art to preserve cells at such freezing temperatures, and thawed in a manner as commonly known in the art for thawing frozen cultured cells.

[0166] In certain embodiments, to promote expression of the transgene, the subject virion or gene delivery vector comprising a scFv- or Fab-modified capsid protein is contacted with the cells for about 30 minutes to 24 hours or more, e.g., 1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 3.5 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 16 hours, 18 hours, 20 hours, 24 hours, etc. The subject virion or gene delivery vector comprising a scFv- or Fab- modified capsid protein may be provided to the subject cells one or more times, e.g., one time, twice, three times, or more than three times, and the cells allowed to incubate with the agent(s) for some amount of time following each contacting event, e.g., 16-24 hours, after which time the media is replaced with fresh media and the cells are cultured further. Contacting the cells may occur in any culture media and under any culture conditions that promote the survival of the cells. For example, cells may be suspended in any appropriate nutrient medium that is convenient, such as Iscove’s modified DMEM or RPMI 1640, supplemented with fetal calf serum, human AB serum, or heat-inactivated goat serum (about 5-10%), L-glutamine, a thiol, particularly 2-mercaptoethanol, and antibiotics, e.g., penicillin and streptomycin. The culture may contain growth factors to which the cells are responsive (e.g., cytokines). Growth factors, as defined herein, are molecules capable of promoting survival, growth, and / or differentiation of - 47 - 12961406v1Attorney Docket No.2014202-0027 cells, either in culture or in the intact tissue, through specific effects on a transmembrane receptor. Growth factors include polypeptides and non-polypeptide factors.

[0167] In certain embodiments, an effective amount of a virion or gene delivery vector comprising a scFv- or Fab-modified capsid protein is provided to produce the expression of the transgene in cells. In particular embodiments, the effective amount may be readily determined empirically, e.g., by detecting the presence or levels of transgene gene product, by detecting an effect on the viability or function of the cells, etc. In certain embodiments, an effective amount of subject virion or gene delivery vector comprising a scFv- or Fab-modified capsid protein will promote equal or greater expression of the transgene in cells than the same amount of a reference virion or viral vector known in the art, e.g., AAV1, AAV2, or AAV6. In certain embodiments, expression is enhanced 2-fold or more relative to the expression from a reference, or control virion or viral vector, for example, 3-fold, 4-fold, or 5-fold or more, in some instances 10-fold, 20-fold, or 50-fold or more, e.g., 200-fold. In certain embodiments, the enhanced expression occurs in a particular cell type, e.g., any of the hematopoietic cells described herein.

[0168] For instances in which cells are contacted in vivo with a subject virion or gene delivery vector comprising a scFv- or Fab-modified capsid protein described herein, the subject may be any mammal, e.g., rodent (e.g., mice, rats, gerbils), rabbit, feline, canine, goat, ovine, pig, equine, bovine, or primate. The methods and compositions of the present disclosure find use in the treatment of any condition that can be addressed, at least in part, by gene therapy of cells. Thus, the compositions and methods of the present disclosure find use in the treatment of individuals in need of a cell therapy. Cells include but are not limited to blood, eye, liver, kidney, heart, muscle, stomach, intestine, pancreas, and skin.

[0169] In certain embodiments, the disclosure provides a method of providing a gene product to a hematopoietic cell of a subject, comprising administering to the subject by intravenous injection a pharmaceutical composition comprising a recombinant virion or vector described herein, wherein the recombinant virus comprises a scFv- or Fab-modified capsid protein disclosed herein and a polynucleotide sequence that encodes the gene product. In certain embodiments, the hematopoietic cell can be a T cell, a B cell, an NK cell, an NK-T cell, a monocyte, a macrophage, or a dendritic cell. - 48 - 12961406v1Attorney Docket No.2014202-0027

[0170] In certain embodiments, the disclosure provides a method of treating or preventing a cancer disease or disorder in a subject in need thereof, comprising administering to the subject by intravenous injection, a pharmaceutical composition comprising a recombinant virion or vector described herein, wherein the recombinant virus comprises a scFv- or Fab- modified capsid protein disclosed herein and a polynucleotide sequence that encodes a therapeutic gene product. The therapeutic gene product may be any therapeutic gene product, including but not limited to any of those described herein.

[0171] The cancers that can be treated using a subject method include, but are not limited to, adenocarcinoma, squamous cell carcinoma, basal cell carcinoma, osteocarcinoma, chondrosarcoma, liposarcoma, acute lymphoblastic leukemia, acute myeloid leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia, diffuse large B cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma, multiple myeloma, cutaneous melanoma, gliomas, meningiomas, germ cell tumors, neuroendocrine tumors, melanoma, mesothelioma, blastomas, breast cancer, pancreatic cancer, and lung cancer.

[0172] In particular embodiments, the subject has been diagnosed with one or more diseases or disorders selected from the group consisting of: acute lymphoblastic leukemia (ALL), diffuse large B cell lymphoma (DLBCL), non-Hodgkin lymphoma (NHL), chronic lymphocytic leukemia (CLL), small lymphocytic lymphoma (SLL), hairy cell leukemia (HCL), neuroblastoma, osteosarcoma, melanoma, soft tissue sarcomas, and diffuse intrinsic pontine glioma (DIPG).

[0173] In particular embodiments, the gene product is a CD19-specific CAR for treating acute lymphoblastic leukemia (ALL), refractory large B-cell lymphoma (LBCL), high-grade B- cell lymphoma, and primary mediastinal large B-cell lymphoma. In particular embodiments, the gene product is a CD19-specific CAR for treating B cell-mediated autoimmune disease (e.g., systemic lupus erythematosus (SLE), idiopathic inflammatory myositis, and systemic sclerosis). In particular embodiments, the gene product is a GD2-specific CAR for treating neuroblastoma, melanoma, osteosarcoma, glioblastoma, glioma, and other solid tumors. In particular embodiments, the gene product is a transgenic TCR that targets tumor-associated antigens (e.g., PRAME, NY-ESO-1, MAGE-A4), expressed by multiple solid tumors (e.g., melanoma, sarcoma, ovarian carcinoma, and breast cancer, among others). - 49 - 12961406v1Attorney Docket No.2014202-0027

[0174] In certain embodiments, virions and viral vectors comprising a scFv- or Fab- modified capsid protein that confers altered tropism to the virion or viral vector are used to treat a disease or disorder of the cells for which tropism of the virion or viral vector is increased, e.g., T lymphocytes or B lymphocytes.

[0175] In certain embodiments, the disclosure provides a method of providing a gene product to the liver of a subject, comprising parenteral administration to the subject, e.g., by intravenous injection, a pharmaceutical composition comprising a recombinant virion or vector described herein, wherein the recombinant virus comprises a scFv- or Fab-modified capsid protein disclosed herein and a polynucleotide sequence that encodes the gene product.

[0176] In certain embodiments, the disclosure provides a method of treating or preventing a liver disease or disorder in a subject in need thereof, comprising administering to the subject, e.g., parenterally or intravenously, a pharmaceutical composition comprising a recombinant virion or vector described herein, wherein the recombinant virus comprises a scFv- or Fab-modified capsid protein disclosed herein and a polynucleotide sequence that encodes a therapeutic gene product.

[0177] In particular embodiments, the subject has been diagnosed with one or more diseases or disorders selected from the group consisting of: inherited metabolic defects, chronic viral hepatitis, liver cirrhosis, primary and metastatic liver cancer, alpha-1 antitrypsin deficiency, hemophilia B, hemophilia A, hereditary angioedema, or β-thalassemia. Gene transfer to the liver can also be used to convert this organ into a factory of secreted proteins needed to treat conditions that do not affect the liver itself.

[0178] In some aspects, the disclosure provides for administering the pharmaceutical composition comprising the viral vector or virion described herein at a frequency of at least once per 3, 6, 9, 12, 18, 24, or 36 months in a human subject in need of treatment. In some aspects, the disclosure provides for administering the pharmaceutical composition comprising the viral vector or virion described herein at a frequency of at most once per 3, 6, 9, 12, 18, 24, or 36 months in a human subject in need of treatment. In some aspects, the disclosure provides for administering the pharmaceutical composition comprising the viral vector or virion described herein at a frequency less than 3 times, less than twice a year, less than once a year, less than once every two years, or less than once every three years in the human subject. - 50 - 12961406v1Attorney Docket No.2014202-0027

[0179] In certain embodiments, the viral vector of the present invention is administered concurrently or during an overlapping time period with the additional therapeutic agent, while in other embodiments, either the viral vector is administered first or the additional therapeutic agent is delivered first, a time period is allowed to pass, and then the other of the viral vector or the additional agent is administered. In particular embodiments, the time period is at least one day, at least one week, at least two weeks, at least one month, at least two months, at least four months, at least six months, at least one year, at least eighteen months, at least two years, or at least three years.

[0180] In certain embodiments, the additional therapeutic agent is a viral vector or virion comprising a polynucleotide sequence that encodes the additional therapeutic agent, i.e., an additional viral vector or virion. In certain embodiments, the additional viral vector or virion comprises a capsid protein, e.g., VP1, that is not the same as the modified capsid protein, e.g., VP1, present in the viral vector or virion of the present invention. Such methods may be used to reduce the possibility of an undesirable immune response when a subject has developed an innate or adaptive immune response against viral particles or capsid proteins therein following administration of the first viral vector or virion (whether that is a viral vector or virion of the present invention or the additional viral vector or virion), such that a second administration of the same viral vector or capsid protein would result in an undesired immune response in the subject. In certain embodiments, a subject is administered an AAV6-scFv- or AAV6-Fab-modified viral vector described herein in combination with a different viral vector, e.g., AAV type 1 (AAV1), AAV type 2 (AAV2), AAV type 3 (AAV3), AAV type 4 (AAV4), AAV type 5 (AAV5), AAV type 6 (AAV6), AAV type 7 (AAV7), AAV type 8 (AAV8), AAV type 9 (AAV9), AAV type 10 (AAV10), AAV rh.10, avian AAV, bovine AAV, canine AAV, equine AAV, primate AAV, non- primate AAV, bovine AAV, AAV.7m8, AAVShH10, or AAV2.5T.

[0181] In certain embodiments, the present invention includes a method of treating a disease or disorder in a subject in need thereof, comprising administering to the subject a first pharmaceutical composition comprising a pharmaceutically acceptable excipient and a first recombinant virus or viral vector, said first virus or vector comprising: (a) a first scFv- or Fab- modified capsid protein, wherein the first scFv- or Fab-modified capsid protein is a modified AAV6, AAV1, or AAV2 capsid protein comprising a peptide insertion relative to a corresponding parental AAV6, AAV1, or AAV2 capsid protein, wherein the peptide insertion - 51 - 12961406v1Attorney Docket No.2014202-0027 comprises the amino acid sequence of a scFv or Fab, and wherein the insertion site is located in Loop III, Loop IV, or at the C-terminal of VP1, VP2, or VP3 capsid proteins, or at the N- terminal of VP2 or VP3 proteins of the AAV6 capsid protein or the corresponding residues of the AAV1, AAV2, AAV5, or AAV8 capsid protein, and (b) a first polynucleotide sequence that encodes a first therapeutic gene product; in combination with a second pharmaceutical composition comprising a pharmaceutically acceptable excipient and a second recombinant virus or viral vector, said second virus or vector comprising: (a) a second scFv- or Fab-modified capsid protein, wherein the scFv- or Fab-modified capsid protein is not the modified AAV6, AAV5, AAV8, AAV1, or AAV2 capsid protein, and (b) a second polynucleotide sequence that encodes a second therapeutic gene product. In certain embodiments, the second modified capsid protein is an AAV2 capsid protein, or a modified AAV2 capsid protein, optionally an AAV2.7m8 capsid protein. The first and second therapeutic gene products are the same or different. In certain embodiments, the disease or disorder is a cancer, and the first and second pharmaceutical compositions are administered to the subject, e.g., intravenously. In particular embodiments, one or both of the first and second therapeutic gene products is a CD19 CAR. In particular embodiments, the disease or disorder is selected from the group consisting of: acute lymphoblastic leukemia (ALL), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), follicular lymphoma (FL), primary mediastinal large B-cell lymphoma (PMBCL), high- grade B-cell lymphoma (HGBCL), chronic lymphocytic leukemia (CLL), or B cell-mediated autoimmune disease.

[0182] In some embodiments, the disease or disorder is a liver disease or disorder, and the first and second pharmaceutical compositions are administered parenterally, optionally intravenously. In various embodiments, the first pharmaceutical composition and the second pharmaceutical composition are administered sequentially in either order, wherein a period of time passes between the sequential administrations. For example, the period of time may be at least one month, at least 3 months, at least 6 months, at least one year, at least 18 months, at least two years, or at least three years. In particular embodiments, one or both of the first and second therapeutic gene products is alpha-1 antitrypsin, factor IX, factor VIII, C1-esterase inhibitor, β- globin, or γ-globin. In certain embodiments, the disease or disorder is selected from the group consisting of: alpha-1 antitrypsin deficiency, hemophilia B, hemophilia A, hereditary angioedema, or β-thalassemia. - 52 - 12961406v1Attorney Docket No.2014202-0027

[0183] In some embodiments, the subject methods result in a therapeutic benefit, e.g., preventing the development of a disorder, halting the progression of a disorder, reversing the progression of a disorder, etc. In some embodiments, the subject method comprises the step of detecting that a therapeutic benefit has been achieved. The ordinarily skilled artisan will appreciate that such measures of therapeutic efficacy will be applicable to the particular disease being modified, and will recognize the appropriate detection methods to use to measure therapeutic efficacy.

[0184] In some instances, the expression of the transgene, e.g., as detected by measuring levels of gene product, by measuring therapeutic efficacy, etc., may be observed two months or less after administration, e.g., 4, 3, or 2 weeks or less after administration, for example, 1 week after administration of the subject composition. Expression of the transgene is also expected to persist over time. Accordingly, in some instances, the expression of the transgene, e.g., as detected by measuring levels of gene product, by measuring therapeutic efficacy, etc., may be observed 2 months or more after administration of the subject composition, e.g., 4, 6, 8, or 10 months or more, in some instances 1 year or more, for example 2, 3, 4, or 5 years, in certain instances, more than 5 years.

[0185] In particular embodiments, a subject is administered with about 1 x 108vector genomes or more, in some cases 1 x 109, 1 x 1010, 1 x 1011, 1 x 1012, or 1 x 1013vector genomes or more, in certain instances, 1 x 1014vector genomes or more. In some cases, the amount of vector genomes that is delivered is at most about 1 x 1015vector genomes, e.g.1 x 1014vector genomes or less, for example 1 x 1013, 1 x 1012, 1 x 1011, 1 x 1010, or 1 x 109vector genomes or less, in certain instances 1 x 108vector genomes, and sometimes no less than 1 x 108vector genomes. In some cases, the amount of vector genomes that is delivered is 1 x 1010to 1 x 1011vector genomes. In some cases, the amount of vector genomes that is delivered is 1 x 1010to 3 x 1012vector genomes. In some cases, the amount of vector genomes that is delivered is 1 x 109to 3 x 1013vector genomes. In some cases, the amount of vector genomes that is delivered is 1 x 108to 3 x 1014vector genomes.

[0186] In some cases, the amount of pharmaceutical composition to be administered may be measured using multiplicity of infection (MOI). In some cases, MOI may refer to the ratio, or multiple of vector or viral genomes to the cells to which the viral genomes may be delivered. In - 53 - 12961406v1Attorney Docket No.2014202-0027 some cases, the MOI may be 1 x 106. In some cases, the MOI may be 1 x 105– 1 x 107. In some cases, the MOI may be 1 x 104– 1 x 108. In some cases, recombinant viruses of the disclosure are at least about 1 x 101, 1 x 102, 1 x 103, 1 x 104, 1 x 105, 1 x 106, 1 x 107, 1 x 108, 1 x 109, 1 x 1010, 1 x 1011, 1 x 1012, 1 x 1013, 1 x 1014MOI.

[0187] In some aspects, the amount of pharmaceutical composition comprises about 1 x 108to about 1 x 1015particles of recombinant virions or viruses, about 1 x 109to about 1 x 1014particles of recombinant virions or viruses, about 1 x 1010to about 1 x 1013particles of recombinant virions or viruses, or about 1 x 1011to about 3 x 1012particles of recombinant virions or viruses.

[0188] In some aspects, no virion or vector is detected in the human subject's tear, blood, saliva or urine samples 7, 14, 21 or 30 days after administering said pharmaceutical composition. In some aspects, the presence of the viral vector is detected by qPCR or ELBA as known in the art.

[0189] All of the above U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications referred to in this specification are incorporated herein by reference, in their entirety.

[0190] From the foregoing it will be appreciated that, although specific embodiments of the invention have been described herein for purposes of illustration, various modifications may be made without deviating from the spirit and scope of the invention. Accordingly, the invention is not limited except as by the appended claims. EXAMPLES

[0191] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Centigrade, and pressure is at or near atmospheric. - 54 - 12961406v1Attorney Docket No.2014202-0027

[0192] General methods in molecular and cellular biochemistry can be found in such standard textbooks as Molecular Cloning: A Laboratory Manual, 3rd Ed. (Sambrook et al, Cold Spring Harbor Laboratory Press 2001); Short Protocols in Molecular Biology, 4th Ed. (Ausubel et al. eds., John Wiley & Sons 1999); Protein Methods (Bollag et al, John Wiley & Sons 1996); Nonviral Vectors for Gene Therapy (Wagner et al. eds., Academic Press 1999); Viral Vectors (Kaplift & Loewy eds., Academic Press 1995); Immunology Methods Manual (I. Lefkovits ed., Academic Press 1997); and Cell and Tissue Culture: Laboratory Procedures in Biotechnology (Doyle & Griffiths, John Wiley & Sons 1998), the disclosures of which are incorporated herein by reference. Reagents, cloning vectors, and kits for genetic manipulation referred to in this disclosure are available from commercial vendors such as BioRad, Stratagene, Invitrogen, Sigma-Aldrich, and TaKaRa. Example 1: Construction of scFv- and Fab-Modified Capsid Mutants

[0193] Standard molecular biology techniques were utilized to construct scFv-modified capsid mutants. Specifically, the plasmid V076-pFBD-inRepOpt-hr2-inCap2-VP1VP3 was generated as follows: Plasmid V062 (SEQ ID NO: 001) was digested with EcoNI and BsiWI to isolate the backbone fragment. Concurrently, a PCR fragment was amplified using primer 0297 (SEQ ID NO: 002), which introduces a point mutation converting the VP2 start codon from ACG to ACC, and primer 0298 (SEQ ID NO: 003). The resulting PCR fragment was digested with EcoNI and BsiWI and ligated to the backbone fragment to produce plasmid V076 (SEQ ID NO: 004). This plasmid encodes the AAV2 VP1, VP3, and Rep sequences.

[0194] The plasmid V334-pFB-inCap6VP1VP3-hr2-inRep was constructed as follows: Plasmid V290 (SEQ ID NO: 005) was digested with AvrII and Bsu36I to isolate the backbone fragment. A first PCR fragment was amplified using primers 3821 (SEQ ID NO: 006) and 3822 (SEQ ID NO: 007), introducing a point mutation that alters the VP2 start codon from ACG to ACA. A second PCR fragment was amplified using primers 3823 (SEQ ID NO: 008), which also introduces a point mutation converting the VP2 start codon from ACG to ACA, and primer 2671 (SEQ ID NO: 009). These two PCR fragments were fused by a second round of PCR amplification using primers 3821 and 2671. The resulting fused PCR fragment was digested with Bsu36I and AvrII and ligated to the backbone to generate plasmid V334 (SEQ ID NO: 010). This plasmid contains the AAV6 VP1, VP3, and AAV2 Rep coding sequences. - 55 - 12961406v1Attorney Docket No.2014202-0027

[0195] The plasmid V685-pFB-polh-CD3-scFv-3xGGGGS-Cap6-VP2 was constructed as follows: Plasmid V683 (SEQ ID NO: 011) was digested with BamHI and BstEII to isolate the backbone fragment. The CD3-scFv fragment (SEQ ID NO: 012) was synthesized de novo and assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit (New England Biolabs) to generate plasmid V685 (SEQ ID NO: 013). This construct contains the CD3-scFv sequence inserted in-frame at the N-terminus of the AAV6 VP2 capsid.

[0196] The plasmid V689-pFB-polh-Cap6VP2-3xGGGGS-CD3scFv-587 was constructed as follows: Plasmid V290 (SEQ ID NO: 005) was digested with NcoI and SnaBI to isolate the backbone fragment. The CD3scFv-3’-partial Cap6VP2-HSVtkpA fragment (SEQ ID NO: 014) was synthesized de novo. The 5’-partial Cap6VP2 fragment (SEQ ID NO: 015) was amplified by PCR using primers 9466 (SEQ ID NO: 016) and 9467 (SEQ ID NO: 017), with plasmid V290 as the template. The backbone fragment was assembled with the two insert fragments using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V689 (SEQ ID NO: 018). This construct encodes the CD3-scFv sequence inserted between amino acids 587 and 588 of the AAV6 VP2 capsid.

[0197] The plasmid V697-pFB-polh-Cap6VP3-3xGGGGS-CD3scFv-587 was constructed as follows: Plasmid V689 (SEQ ID NO: 018) was digested with BamHI and BsiWI to isolate the backbone fragment. The 5’-Cap6VP3 fragment (SEQ ID NO: 019) was amplified by PCR using primers 9482 (SEQ ID NO: 020) and 5388 (SEQ ID NO: 021), with plasmid V689 as the template. The backbone and insert fragments were assembled using the NEBuilder HiFi Assembly Cloning Kit (New England Biolabs) to generate plasmid V697 (SEQ ID NO: 022). This construct encodes the CD3-scFv sequence inserted between amino acids 587 and 588 of the AAV6 VP3 capsid.

[0198] The plasmid V705-pFB-polh-Cap6VP2-3xGGGGS-CD3scFv-449 was constructed as follows: Plasmid V689 (SEQ ID NO: 018) was digested with AvrII and SnaBI to isolate the backbone fragment. The polh-VP2 fragment (SEQ ID NO: 023) was amplified by PCR using primers 6711 (SEQ ID NO: 024) and 9554 (SEQ ID NO: 025), with plasmid V689 as the template. The CD3-scFv fragment (SEQ ID NO: 026) was amplified by PCR using primers 9555 (SEQ ID NO: 027) and 9556 (SEQ ID NO: 028), with plasmid V689 as the template. The Cap6VP2-HSVtkpA fragment (SEQ ID NO: 029) was amplified by PCR using primers 9557 - 56 - 12961406v1Attorney Docket No.2014202-0027 (SEQ ID NO: 030) and 9468 (SEQ ID NO: 031), with a de novo synthesized fragment as the template. These three PCR fragments were assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V705 (SEQ ID NO: 032). This construct encodes the CD3-scFv sequence inserted between amino acids 449 and 450 of the AAV6 VP2 capsid.

[0199] The plasmid V734 was constructed as follows: Plasmid V288 (SEQ ID NO: 033) was digested with NcoI and SphI to isolate the backbone fragment. The AAV8 VP3 coding sequence (SEQ ID NO: 034) was amplified by PCR using primers 9624 (SEQ ID NO: 035) and 9625 (SEQ ID NO: 036), with plasmid V288 as the template. The PCR fragment was assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V734 (SEQ ID NO: 037). This construct contains the AAV8 VP3 capsid coding sequence.

[0200] The plasmid V735-pFB-inCap8VP1VP2-CD3scFv-451-inRep-hr2 was constructed as follows: Plasmid V733 (SEQ ID NO: 038) was digested with BsiWI-HF and MluI to isolate the backbone fragment. The partial Cap8VP1VP2-CD3scFv-451 fragment (SEQ ID NO: 039) was synthesized de novo and assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit (New England Biolabs) to generate plasmid V735 (SEQ ID NO: 040). This construct encodes the CD3-scFv sequence inserted between amino acids 451 and 452 of the AAV8 VP1 and VP2 capsids.

[0201] The plasmid V775-pFB-polh-sigCD19scFv-3xGGGGS-Cap6VP2-NT was constructed as follows: Plasmid V683 (SEQ ID NO: 011) was digested with NcoI and XhoI to isolate the backbone fragment. The 5’-CD19scFv fragment (SEQ ID NO: 041), lacking the secretion signal peptide, was amplified by PCR using primers 9797 (SEQ ID NO: 042) and 9798 (SEQ ID NO: 043), with plasmid V683 as the template. The PCR fragment was assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V775 (SEQ ID NO: 044). This construct encodes the CD19-scFv sequence, without the secretion signal peptide, inserted at the N-terminus of the AAV6 VP2 capsid.

[0202] The plasmid V784-pFB-inCap5GD2scFv-NT-inRep was constructed as follows: Plasmid V758 (SEQ ID NO: 045) was digested with SnaBI and EagI to isolate the backbone fragment. The GD2scFv-Cap5VP2-hGHpA fragment (SEQ ID NO: 046) was synthesized de novo and assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to - 57 - 12961406v1Attorney Docket No.2014202-0027 generate plasmid V784 (SEQ ID NO: 047). This construct contains coding sequences for the AAV5 VP1, VP3, GD2scFv at the N-terminus of VP2, and AAV2 Rep coding sequences.

[0203] The plasmid V794-pFB-polh-CD3Fab-3xGGGGS-Cap6VP2-NT was constructed as follows: Plasmid V775 (SEQ ID NO: 044) was digested with NcoI and BstEII to isolate the backbone fragment. The CD3Fab fragment (SEQ ID NO: 048) was amplified by PCR using primers 9841 (SEQ ID NO: 049) and 9842 (SEQ ID NO: 050), with plasmid V756 (SEQ ID NO: 051) as the template. The PCR fragment was assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V794 (SEQ ID NO: 052). This construct encodes the CD3Fab sequence inserted at the N-terminus of the AAV6 VP2 capsid.

[0204] The plasmid V797-pFB-inCap6CD3scFv-NT-hr2-inRep was constructed as follows: Plasmid V334 (SEQ ID NO: 010) was digested with EcoRV and NdeI to isolate the backbone fragment. The CD3-scFv-Cap6VP2 fragment (SEQ ID NO: 053) was amplified by PCR using primers 9835 (SEQ ID NO: 054) and 9836 (SEQ ID NO: 055), with plasmid V787 (SEQ ID NO: 056) as the template. The PCR fragment and backbone were assembled using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V797 (SEQ ID NO: 057). This construct contains coding sequences for the AAV6 VP1, VP3, CD3scFv at the N-terminus of VP2, and AAV2 Rep coding sequences. Example 2: Construction of Gene of Interests into an AAV Genome

[0205] The plasmid V545-pFB-CMV-GFP-hGHpA was constructed as follows: Plasmid ADV05 (SEQ ID NO: 058) was digested with PmeI and BsiWI to isolate the backbone fragment. The optimized eGFP-hGHpA fragment (SEQ ID NO: 059) was synthesized de novo and amplified by PCR using primers 7733 (SEQ ID NO: 060) and 7779 (SEQ ID NO: 061). The PCR fragment was assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V545 (SEQ ID NO: 062).

[0206] The plasmids V694-pFB-CMV-CD19-CAR and V695-pFB-CMV-GD2-CAR were constructed as follows: Plasmid V545 (SEQ ID NO: 062) was digested with MluI and PmeI to isolate the backbone fragment. The CD19-CAR fragment (SEQ ID NO: 063) and the GD2- CAR fragment (SEQ ID NO: 064) were amplified by PCR using primers 9496 (SEQ ID NO: 065) and 9497 (SEQ ID NO: 066), with plasmids V681 (SEQ ID NO: 099) and V686 (SEQ ID NO: 100) as templates, respectively. Each PCR fragment was assembled with the backbone - 58 - 12961406v1Attorney Docket No.2014202-0027 using the NEBuilder HiFi Assembly Cloning Kit to generate plasmids V694 (SEQ ID NO: 067) and V695 (SEQ ID NO: 068), respectively. These constructs contain AAV genomes with the CMV promoter driving expression of CD19-CAR and GD2-CAR, respectively.

[0207] The plasmid V767-pFB-SURV-inDTA-P2A-Trail-WPRE was constructed as follows: Plasmid V766 (SEQ ID NO: 069) was digested with SpeI and BamHI to isolate the backbone fragment. The P2A-Trail fragment (SEQ ID NO: 070) was amplified by PCR using primers 9731 (SEQ ID NO: 071) and 5418 (SEQ ID NO: 072), with plasmid V259 (SEQ ID NO: 073) as the template. The WPRE element (SEQ ID NO: 074) was amplified by PCR using primers 9732 (SEQ ID NO: 075) and 9733 (SEQ ID NO: 076), with plasmid V729 (SEQ ID NO: 077) as the template. The two PCR fragments were assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V767 (SEQ ID NO: 078).

[0208] The plasmid V769-pFB-synGBM-inDTA-P2A-Trail-WPRE was constructed as follows: Plasmid V767 (SEQ ID NO: 078) was digested with SpeI and AgeI to isolate the backbone fragment. The synthetic GBM promoter fragment (SEQ ID NO: 079) was synthesized de novo and assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit (New England Biolabs) to generate plasmid V769 (SEQ ID NO: 080).

[0209] The plasmid V786-pFB-CMV-CD19-CAR-5xmiR-122-hGHpA was constructed as follows: Plasmid V694 (SEQ ID NO: 067) was digested with MluI to isolate the backbone fragment. A fragment containing five copies of the miR-122 binding sites (SEQ ID NO: 081) was synthesized de novo and assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V786 (SEQ ID NO: 082).

[0210] The plasmid V800-pFB-CMV-Tri-Specific-T-Cell-Engager-hGHpA was constructed as follows: Plasmid V694 (SEQ ID NO: 067) was digested with MluI and PmeI to isolate the backbone fragment. The TriTE fragment (SEQ ID NO: 101) was synthesized de novo and assembled with the backbone using the NEBuilder HiFi Assembly Cloning Kit to generate plasmid V800 (SEQ ID NO: 083).

[0211] All plasmids were verified by restriction enzyme digestion and confirmed by whole-plasmid sequencing to ensure sequence accuracy. - 59 - 12961406v1Attorney Docket No.2014202-0027 Example 3: Insect and Mammalian Cell Culture for Production of scFv- and Fab-Modified AAVs

[0212] Sf9 cells (Expression Systems, Davis, CA) were cultured in Corning storage bottles at 28°C in ESF AF media (Expression Systems) supplemented with 100 units / mL penicillin and 100 µg / mL streptomycin (HyClone, Logan, UT). The cells were split 1:4 once the cell density reached about 8 x 106cells / mL for maintenance. HEK293T cells (ATCC, Manassas, VA) were cultured in T-75 flasks at 37°C in DMEM medium (ATCC) supplemented with 100 units / mL penicillin and 100 µg / mL streptomycin, and 10% FBS (ATCC) in a CO2incubator with 5% CO2. Once the cells reached confluency, they were split 1:10 every twice a week for maintenance. These cell culture methods were used to produce recombinant AAV virions comprising scFv- or Fab-modified capsid proteins, as described herein. For example, Sf9 cells were employed for baculovirus-based production of AAV6 virions modified with CD19-scFv or CD19-Fab, while HEK293T cells were used for triple transfection to generate scFv- or Fab- modified AAV6 virions, achieving titers of 1 x 1012vector genomes per mL or higher. Example 4: T-Cell Culture for Testing scFv- and Fab-Modified AAVs

[0213] Peripheral blood mononuclear cells (PBMC) were isolated from leukapheresis material of healthy donors using manual Ficoll® Paque Plus or a closed system instrument (Lovo) and cryopreserved in Cryostor® CS10. The cryopreserved PBMCs were thawed and washed with CTL medium, containing 47% Advanced RPMI medium, 47% Click’s medium, 5% Human AB serum, GlutaMAX, and Antibiotic / Antimycotic. Supernatants were removed, and cell pellets were resuspended at 5 x 107cells / mL in the presence of Benzonase®. Cells were incubated with Benzonase® at 37°C for 30 minutes, and then spun at 400 x g for 5 minutes at RT. The supernatants were removed, and the cell pellets were resuspended at approximately 2 x 107cells / mL in CTL medium. Cells were counted and plated in a G-Rex®100M or a T-75 flask at 1 x 107cells / mL in the presence of interleukin (IL)-7. T cells were either activated with CD3 and CD28 antibodies or left unstimulated. These T-cell cultures were used to evaluate the transduction efficiency and tropism of scFv- or Fab-modified AAV virions, such as CD3-scFv- modified AAV6 virions, demonstrating over 200-fold increased transduction efficiency in non- activated T cells compared to unmodified AAV6. - 60 - 12961406v1Attorney Docket No.2014202-0027 Example 5: Generation of Recombinant Baculovirus (rBV) and Determination of rBV Titer

[0214] Recombinant baculoviruses (rBVs) were generated using the Bac-to-Bac Baculovirus Expression System (Invitrogen, Carlsbad, CA) according to the manufacturer’s protocol. Briefly, shuttle plasmids containing the genes of interest, including those encoding scFv- or Fab-modified AAV capsid proteins (e.g., CD3-scFv or CD3-Fab fused to AAV6 VP2), were respectively transformed into DH10Bac competent cells. After 2 days of incubation on LB- agar plates containing appropriate concentrations of antibiotics, X-gal, and IPTG at 37°C, white colonies were picked, and miniprep bacmid DNAs were prepared. The miniprep bacmid DNAs were then used to transfect Sf9 cells to generate rBVs. The rBVs were collected and stored at 4°C in the dark if not used immediately. To amplify the rBV, Sf9 cells at 2 x 106cells / mL were infected with the rBVs at 0.01–0.1 MOI for 3 days, and supernatants were harvested and stored in the dark before use. These rBVs were used to produce scFv- or Fab-modified AAV virions.

[0215] The rBV titers were measured by quantitative PCR (QPCR) method. Briefly, 50 µL of rBV samples were respectively mixed with 50 µL of 0.2% SDS solution and heated at 95°C for 30 minutes. The heat-treated samples were diluted in ultrapure water and used for QPCR reaction. One MOI was empirically defined to contain an average of 20 rBV genome copies. This titration method was applied to rBVs encoding both scFv- and Fab-modified AAV capsid proteins, ensuring accurate quantification for downstream AAV production. Example 6: Production and Purification of scFv- and Fab-Modified AAV Particles

[0216] Recombinant baculoviruses (rBVs) were used to infect Sf9 insect cells for the production of recombinant adeno-associated virus (AAV) particles, including those with mosaic capsids comprising scFv- or Fab-modified capsid proteins. For mosaic packaging, rBVs were engineered to express capsid proteins from two or more distinct AAV serotypes or variants, including scFv- or Fab-modified variants, to form a heterogeneous capsid structure. In one embodiment, rBVs were designed to express AAV2 capsid proteins VP1 and VP3 and AAV6 capsid protein VP2 modified with either a CD3-scFv or CD19-Fab. Additional rBVs containing AAV Rep genes and rBVs carrying the AAV genome (e.g., flanked by inverted terminal repeats, ITRs, from a specified serotype such as AAV2) were used. The mosaic packaging approach described herein is applicable to any combination of capsid proteins (VP1, VP2, or VP3) from - 61 - 12961406v1Attorney Docket No.2014202-0027 any AAV serotypes or variants, including but not limited to AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, or engineered variants thereof, modified with scFv or Fab fragments, to produce mosaic capsids with tailored properties. Sf9 cells were co-infected with the appropriate rBVs at suitable multiplicities of infection and incubated for 3 days at 28°C in a shaker incubator. Cell pellets were collected by centrifugation and lysed in SF9 lysis buffer (50 mM Tris-HCl, pH 7.8, 50 mM NaCl, 2 mM MgCl2, 1% Sarkosyl, 1% Triton X-100, and 140 units / mL Benzonase nuclease (Sigma-Aldrich)) by sonication. Cell debris was removed by centrifugation, and the cleared lysates were transferred to ultraclear centrifuge tubes for an SW28 rotor (Beckman Coulter, Brea, CA). CsCl solutions with densities of 1.32 g / cc and 1.55 g / cc were under-layered, and the samples were centrifuged at 28,000 rpm at 15°C for approximately 20 hours to separate the AAV or mosaic particles from cellular contaminants. The separated AAV vector band, containing the AAV or mosaic capsids (e.g., composed of AAV2 VP1 / VP3 + AAV6 CD3-scFv-VP2 or AAV6 CD19-Fab-VP2), was collected using a syringe attached to an 18G needle. The collected AAV vectors were transferred into ultraclear centrifuge tubes for a 70.1 Ti rotor (Beckman Coulter), filled with 1.38 g / cc CsCl solution, and sealed. After centrifugation at 65,000 rpm for approximately 20 hours, the AAV vector band was collected with a syringe needle. The AAV vectors were buffer-exchanged using PD-10 desalting columns (GE Healthcare Bio-Sciences, Pittsburgh, PA). After filter sterilization, the scFv- or Fab- modified AAV vectors were diluted to appropriate concentrations for further experiments, achieving titers of >1 x 1013vector genomes per mL for CD3-Fab-modified AAV6. If not used immediately, they were stored at -80°C.

[0217] The titers of the AAV or mosaic samples were determined using a quantitative PCR (qPCR) method. Briefly, the samples were diluted in ultrapure water and treated with DNase I to eliminate potential contaminating DNA molecules. The treated samples were then heated at 95°C in EDTA buffer for 30 minutes to inactivate DNase I. Subsequently, the heat- treated samples were further diluted and used for qPCR reactions. The copy numbers of the scFv- or Fab-modified AAV samples were calculated based on a standard curve. This titration method was applied to both scFv- and Fab-modified AAVs, ensuring accurate quantification for downstream applications. - 62 - 12961406v1Attorney Docket No.2014202-0027 Example 7: SDS-Polyacrylamide Gel Electrophoresis (SDS-PAGE) of scFv- and Fab- Modified AAV Capsids

[0218] Purified AAV samples, including those with scFv- or Fab-modified capsid proteins, were subjected to SDS-PAGE to separate the capsid proteins according to their molecular weights. Unmodified AAV capsid proteins VP1, VP2, and VP3 have approximate molecular weights of 87 kDa, 73 kDa, and 62 kDa, respectively, while scFv- or Fab-modified capsid proteins (e.g., CD3-scFv-VP2 or CD19-Fab-VP2) exhibit higher molecular weights due to the addition of the antibody fragment. For example, CD3-Fab-modified VP2 was observed at approximately 123 kDa due to the larger size of the Fab fragment compared to scFv. The AAV samples were mixed with a loading buffer and heated at 95°C for 5 minutes to denature the capsid proteins. Approximately 1 x 1011vector genomes per lane of denatured samples were loaded and electrophoresed at about 100 V until the loading dye reached the bottom of the gel. After disassembly, the gels were stained and destained with the SimplyBlue SafeStain kit according to the manufacturer’s instructions (Thermo Fisher). The gel images were recorded with the iBright CL750 Image System (Thermo Fisher). This SDS-PAGE analysis confirmed the presence and integrity of scFv- or Fab-modified capsid proteins in AAV vectors. Example 8: AAV Transduction of HEK293 Cells

[0219] HEK293 cells were seeded on 24-well plates and were transduced with the scFv- modified AAV vectors and compared with non-modified AAV vectors. Briefly, the cells were trypsinized and resuspended in serum-containing media. Each well was seeded with 1.5 x 105cells in 0.5 mL medium overnight. The cells were transduced with different quantities of AAV vectors for 3 days and gene expression was detected with fluorescent microscope. Example 9: AAV Transduction of B cell lymphoma cell line (Daudi)

[0220] Daudi cells were rinsed with culture medium without serum and concentrated to 1 x 107cells / mL in culture medium without serum. One hundred microliters of the cells were mixed with different quantities of AAV vectors (1x103- 2x105vg / cell) and incubated for 2 hours. The cells were then transferred to a 24-well plate and 0.4 mL of culture medium containing 10% FBS were added per well. After 24 hours of incubation, 7.5 mL of culture medium containing 10% FBS were added per well. After 48-72 hours of incubation, the gene - 63 - 12961406v1Attorney Docket No.2014202-0027 expression was detected using the Cytek Aurora flow cytometry system (Cytek Biosciences, Fremont, CA). Example 10: AAV Transduction of Primary Lymphocytes

[0221] Antibody-activated or resting T cells were mixed with rAAVs at viral titers ranging from 2x104to 2x106vg / cell for 24 - 96 hours in vitro, following the Daudi transduction protocol above. T cell transduction was measured using flow cytometry, using either eGFP fluorescence or fluorescently-labeled antibodies specific for the CARs. Example 11: Killing of Cancer Cells by AAV transduced T-Cells

[0222] CAR-T cells generated using rAAV encoding a CAR specific for CD19 or GD2 were mixed with tumor targets expressing the relevant antigens at 1:1 effector-to-target ratio. The CD19-positive Raji tumor cells were used as targets for the CD19-specific CAR-T cells, while the GD2-positive KM-H2 tumor cells were used as targets for the GD2-specific CAR-T cells. Both tumor cell lines were modified to express eGFP-luciferase, so that their disappearance from the culture due to CAR-T cell-mediated cytotoxicity could be monitored via the loss of eGFP fluorescence using flow cytometry. Non-transduced or T cells transduced with rAAV encoding eGFP were used as negative controls. Specific killing was determined 48 hours after the initiation of the co-culture using the following formula: %Specific lysis = 100*(#tumor cells in control well - #tumor cells in CAR well) / #tumor cells in control well. Example 12: rBVs Carrying scFv-or Fab-Modified Capsid Protein Exhibited normal Baculovirus Titers

[0223] The recombinant baculoviruses (rBVs) carrying scFv- or Fab-modified AAV capsid proteins were generated, amplified, and compared with regular rBVs without modified capsid proteins for their ability to replicate and amplify. The results are shown in Table 1. rBV- V290 was the rBV carrying the wild-type AAV6 capsid gene, and rBV-V545 was the rBV carrying the GFP marker gene, whereas the other rBVs carried either scFv-modified AAV6 capsid genes (e.g., CD19-scFv-VP2) or Fab-modified AAV6 capsid genes (e.g., CD3-Fab-VP2). The rBV titers from all these rBVs were comparable, with titers of 9 x 108to 1 x 109genome copies / mL as measured by QPCR (as described in Example 5). These results demonstrate that - 64 - 12961406v1Attorney Docket No.2014202-0027 neither scFv- nor Fab-modified capsid proteins have a negative impact on rBV generation and amplification. Example 13: scFv- or Fab-Modified Capsid Proteins Maintain Comparable AAV Production Yields Regardless of Insertion Site

[0224] To evaluate the impact of insertion site on AAV production, scFv or Fab sequences were inserted at various positions within the VP1, VP2, or VP3 capsid proteins of AAV6. The insertion sites included the N-terminus, C-terminus, Loop III, and Loop IV of the respective capsid proteins. AAV vectors were produced in Sf9 insect cells using triple or double baculovirus infection (as described in Example 6), and the resulting yields were quantified. The data are summarized in Table 2. - 65 - 12961406v1Attorney Docket No.2014202-0027

[0225] Constructs with scFv- or Fab-modified AAV6 capsids were produced in 200 mL or 400 mL cultures using the triple or double baculovirus infection of Sf9 cells (as described in Example 6), and yields of purified AAV vectors were calculated per liter of cell culture.

[0226] The results demonstrate that insertion of scFv or Fab sequences at different capsid locations did not significantly affect AAV production yields. For instance, construct V705, which harbors CD3-scFv inserted between amino acids 449 and 450 of AAV6 VP2 within Loop III (SEQ ID NO.085), yielded 8.35 × 1013vg of purified AAV particles from a 200 mL culture (lot 24-374), equivalent to 4.18 × 10^14 vg / L.

[0227] In comparison, construct V683, with CD19-scFv fused to the N-terminus of AAV6 VP2, yielded 4.07 × 1013vg from a 200 mL culture (lot 24-299), or 2.03 × 10^14 vg / L of purified AAV vectors. A new construct, V794, with CD3-Fab fused to the N-terminus of AAV6 VP2, yielded 4.40 × 1013vg from a 200 mL culture (lot 25-298), or 2.20 × 1014vg / L of purified AAV vectors. Construct V689, in which CD3-scFv was inserted at the Loop IV position (between amino acids 587 and 588) of VP2, and construct V697, with the same insertion site on VP3, also produced high titers in multiple production lots (24-365 to 367, and 24-341 to 343, respectively). - 66 - 12961406v1Attorney Docket No.2014202-0027

[0228] Notably, V705 (Loop III insertion of CD3-scFv in VP2; lots 24-374 to 376) and V794 (N-terminal insertion of CD3-Fab in VP2; lot 25-298) consistently yielded titers comparable to those of constructs with Loop IV or C-terminal insertions.

[0229] Additionally, production yields of scFv- or Fab-modified AAV6 capsids were comparable to those of the wild-type AAV6 capsid. For example, V290 (lot 24-362), which contains unmodified AAV6 capsid sequences, produced AAV particles at levels similar to those of scFv- or Fab-modified constructs.

[0230] These results indicate that the location of scFv or Fab insertion within the AAV6 capsid does not significantly influence vector production efficiency. Example 14: Efficient Assembly of scFv-Modified Capsid Proteins into Virus Particles Regardless of Insertion Site

[0231] SDS-PAGE analysis demonstrated that incorporation of scFv-modified capsid proteins into AAV virus particles was not significantly affected by the position of the scFv insertion within the capsid protein.

[0232] As shown in FIG.3A, lane 2, a strong band corresponding to CD19scFv (SEQ ID NO: 093) fused to the N-terminus of AAV6 VP2 (SEQ ID NO: 084) was detected, indicating successful expression and incorporation. Lane 3 illustrates a mosaic AAV capsid composed of AAV2 VP1 and VP3, and AAV6 VP2 containing CD3scFv inserted between amino acids 449 and 450 (SEQ ID NO: 085). The molecular weight of the VP2-scFv fusion protein exceeded that of VP1, as expected.

[0233] In FIG.3B, lane 2 shows GD2scFv fused to the N-terminus of AAV5 VP2 (SEQ ID NO: 086) and successfully packaged into AAV5 virus particles. Similarly, lane 3 displays CD3scFv inserted at the N-terminus of AAV6 VP2 (SEQ ID NO: 087), which was efficiently packaged into AAV6 particles.

[0234] FIG.3C (lanes 2–4) shows CD3scFv inserted between amino acids 587 and 588 of AAV6 VP2 (SEQ ID NO: 088), which was effectively assembled into AAV6 virus particles. Likewise, FIG.3D (lanes 2–4) illustrates that insertion of CD3scFv between amino acids 449 and 450 of AAV6 VP2 (SEQ ID NO: 085) resulted in successful packaging into virus particles. - 67 - 12961406v1Attorney Docket No.2014202-0027

[0235] In FIG.3E, lane 2 demonstrates that CD3scFv inserted simultaneously between amino acids 451 and 452 of both AAV8 VP1 and VP2 (SEQ ID NO: 089) was successfully assembled into AAV8 virus particles. Interestingly, a partial CD3scFv-VP3 fusion protein was also detected. Despite mutation of the canonical double ATG start codons of VP3 to ATT, an alternative GTG codon located 54 bp downstream was utilized as a cryptic start site, resulting in unexpected expression and incorporation of the CD3scFv-VP3 fusion. The majority of VP3 protein in this configuration was supplied in trans from a separate plasmid construct (V734).

[0236] FIG.3F (lanes 2–4) shows that CD3scFv inserted between amino acids 587 and 588 of AAV6 VP3 (SEQ ID NO: 090) was successfully incorporated into AAV6 virus particles. The resulting CD3scFv-VP3 fusion protein exhibited a molecular weight similar to that of VP1, and the two bands overlapped, forming a strong composite band at the VP1 position. The majority of AAV6 VP3 in these particles was supplied by a separate construct (V290).

[0237] FIG.3G, lane 2, demonstrates successful incorporation of a CD3-Fab fragment fused to the N-terminus of the AAV6 VP2 capsid protein (SEQ ID NO: 092) into assembled AAV6 virions. The CD3-Fab-VP2 fusion protein exhibited the expected molecular weight of approximately 125 kDa, as confirmed by SDS-PAGE analysis. Example 15: Superior Lymphocyte Transduction Efficiency of Wild-Type AAV6 Capsid Compared to AAV-DJ and AAV8 Justifies Its Selection for scFv- and Fab-Modifications

[0238] To evaluate the transduction efficiency of recombinant adeno-associated virus (rAAV) vectors for lymphocyte gene delivery, activated T cells were transduced with rAAVs encoding enhanced green fluorescent protein (eGFP) using capsids derived from AAV6, AAV- DJ, or AAV8. As shown in FIG.4, AAV6 exhibited significantly higher transduction efficiency, achieving approximately 40% transduction at a multiplicity of infection (MOI) of 1 × 10^6 vector genomes (vg) per cell, compared to only ~2% for both AAV-DJ and AAV8 under identical conditions. This superior performance of the wild-type AAV6 capsid in lymphocyte transduction justified its selection as the primary vector for further modifications, including the development of scFv- and Fab-modified AAV6 capsids (e.g., CD3-scFv-VP2 or CD3-Fab-VP2), to enhance targeted gene delivery to lymphocytes. - 68 - 12961406v1Attorney Docket No.2014202-0027 Example 16: scFv-Modified AAV6 Capsids Enhance Transduction Efficiency in Non- Activated T Cells

[0239] To compare transduction efficiencies of recombinant adeno-associated virus (rAAV) vectors with wild-type (WT) AAV6 capsids, or CD3-scFv-modified AAV6 capsids, non-activated T cells from four healthy donors were transduced with rAAVs encoding enhanced green fluorescent protein (eGFP). The experimental design is depicted in FIG.5. Transduction results are shown in FIG.6. The top row of FIG.6A displays non-transduced peripheral blood mononuclear cell (PBMC) data, while the middle and bottom rows show the fraction of eGFP- positive T cells per donor for WT AAV6 and CD3-scFv-modified AAV6 capsids, respectively. FIG.6B demonstrates that the CD3-scFv-modified VP2 capsid significantly enhanced transduction efficiency compared to WT AAV6. FIG.6C indicates that the fold increase in transduction efficiency, after subtracting the non-transduced baseline, reached up to 280-fold for individual donors.

[0240] To further evaluate transduction efficiencies, non-activated T cells from four healthy donors were transduced with rAAVs encoding a CD19-specific chimeric antigen receptor (CAR, FMC63) using WT AAV6 capsids, or CD3-scFv-modified AAV6 capsids. The experimental design is depicted in FIG.5. Transduction results are shown in FIG.7. The top row of FIG.7A displays non-transduced PBMC data, while the middle and bottom rows show the fraction of CAR-positive T cells per donor for WT AAV6 and CD3-scFv-modified AAV6 capsids, respectively. FIG.7B demonstrates that the CD3-scFv-modified VP2 capsid significantly enhanced transduction efficiency compared to WT AAV6. FIG.7C indicates that the fold increase in transduction efficiency reached up to 200-fold for individual donors. This antibody-mediated enhancement of transduction efficiency underscores the novelty of antibody- modified AAV capsids for targeted gene delivery to non-activated T cells. Example 17: CD19-scFv-Modified AAV6 Capsids Enhance Transduction Efficiency in Daudi Cells

[0241] To compare transduction efficiencies of recombinant adeno-associated virus (rAAV) vectors with wild-type (WT) AAV6 capsids, and CD19-scFv-modified AAV6 capsids, Daudi cells (a CD19-positive B-cell line) were transduced with rAAVs encoding enhanced green fluorescent protein (eGFP) at multiplicities of infection (MOI) ranging from 1 × 103to 2 × 105- 69 - 12961406v1Attorney Docket No.2014202-0027 vector genomes (vg) per cell. As shown in FIGs.8A and 8B, CD19-scFv-modified VP2 capsids enhanced transduction efficiency at low MOI compared to WT AAV6 capsids (94.0% vs.65.4% at 1 × 103vg / cell), while at higher MOIs, efficiencies converged due to transduction saturation (98.3% vs.94.9% at 1 × 104vg / cell; 99.2% vs.98.3% at 1 × 105vg / cell). Example 18: Comparison of CD3-scFv-Modified VP2 vs. VP3 Capsid Proteins in Transduction Efficiency of Non-Activated T Cells

[0242] To compare transduction efficiencies of recombinant adeno-associated virus (rAAV) vectors with CD3-scFv-modified capsid proteins inserted into either VP2 or VP3 of AAV6, non-activated T cells from two healthy donors were transduced with rAAVs encoding either a CD19-specific chimeric antigen receptor (CAR, FMC63) or a GD2-specific CAR (14G2A). The rAAVs included constructs with CD3-scFv fused to VP2 or VP3. As shown in FIG.9, CD3-scFv-modified VP3 capsids exhibited higher transduction efficiencies than CD3- scFv-modified VP2 capsids: 13.2% vs.9.05% and 23.9% vs.19.3% for the CD19-specific CAR, and 20.3% vs.7.52% and 31.3% vs.12.2% for the GD2-specific CAR. Due to donor-to-donor variability, these differences did not reach statistical significance. Example 19: CD3-scFv-Modified VP2 Capsid Proteins with Loop IV or Loop III Insertions Exhibit Comparable Transduction Efficiencies in Non-Activated T Cells

[0243] To compare transduction efficiencies of recombinant adeno-associated virus (rAAV) vectors with CD3-scFv-modified VP2 capsid proteins of AAV6, with insertions in Loop IV or Loop III, non-activated T cells from three healthy donors were transduced with rAAVs encoding enhanced green fluorescent protein (eGFP). As shown in FIG.10, CD3-scFv-modified VP2 capsids with Loop IV versus Loop III insertions exhibited comparable transduction efficiencies: 39.2% vs.46.7% for Donor 735, 13.6% vs.12.9% for Donor 088, and 42.6% vs. 45.1% for Donor 992. Due to donor-to-donor variability, these differences were not statistically significant. Example 20: CD3-scFv-Modified VP2 Capsid Proteins with Loop IV or Loop III Insertions Exhibit Comparable CAR-T Cell Transduction Efficiencies

[0244] To compare transduction efficiencies of recombinant adeno-associated virus (rAAV) vectors with CD3-scFv-modified VP2 capsid proteins of AAV6, with insertions in Loop - 70 - 12961406v1Attorney Docket No.2014202-0027 IV or Loop III, non-activated T cells from three healthy donors were transduced with rAAVs encoding either a CD19-specific chimeric antigen receptor (CAR, FMC63) or a GD2-specific CAR (14G2A). As shown in FIG.11, CD3-scFv-modified VP2 capsids with Loop IV versus Loop III insertions exhibited comparable transduction efficiencies: 47.2% vs.49.9% (Donor 735, FMC63 CAR), 9.92% vs.15.2% (Donor 088, FMC63 CAR), 36.9% vs.43.8% (Donor 992, FMC63 CAR), 33.3% vs.44.9% (Donor 735, 14G2A CAR), 9.50% vs.9.62% (Donor 088, 14G2A CAR), and 47.4% vs.52.6% (Donor 992, 14G2A CAR). Due to donor-to-donor variability, these differences were not statistically significant. Example 21: CAR-T Cells Generated by rAAV6 Transduction Using CD3-scFv-Modified VP2 Capsids with Loop IV or Loop III Insertions Exhibit Comparable Tumor-Killing Efficiency

[0245] To compare the anti-tumor activity of CAR-T cells generated by transduction with recombinant adeno-associated virus (rAAV) vectors containing CD3-scFv-modified VP2 capsid proteins of AAV6, with insertions in Loop IV or Loop III, non-activated T cells from three healthy donors were transduced with rAAVs encoding a CD19-specific chimeric antigen receptor (CAR, FMC63) or a GD2-specific CAR (14G2A). Seventy-two hours post-transduction, CAR-T cells were co-cultured with tumor cells expressing the relevant antigens at a 1:1 effector- to-target ratio for 48 hours, and tumor killing was assessed by flow cytometry, measuring the reduction in eGFP-positive tumor cells. The experimental design is shown in FIG.12A, and tumor-killing results are summarized in FIG.12B. CD3-scFv-modified VP2 capsids with Loop IV versus Loop III insertions yielded comparable specific lysis: 68.0% vs.34.0% (Donor 735, FMC63 CAR), 24.0% vs.38.0% (Donor 088, FMC63 CAR), 62.0% vs.64.0% (Donor 992, FMC63 CAR), 48.0% vs.45.0% (Donor 735, 14G2A CAR), 14.0% vs.11.0% (Donor 088, 14G2A CAR), and 60.0% vs.55.0% (Donor 992, 14G2A CAR). Due to donor-to-donor variability, these differences were not statistically significant. These data demonstrate successful use of CD3-scFv-modified capsids with either Loop IV or Loop III insertions to generate CAR-T cells with comparable tumor-killing efficiency. - 71 - 12961406v1Attorney Docket No.2014202-0027 Example 22: CD19-Specific CAR-T Cells Generated In Vivo Using Human PBMCs and CD3-scFv-Modified AAV6-FMC63CAR Administered Intraperitoneally Control Raji Tumor Growth and Improve Mouse Survival

[0246] To evaluate the in vivo efficacy of CD19-specific CAR-T cells against B-cell lymphoma, luciferase-modified Raji cells were implanted intraperitoneally (IP) in NSG mice on Day -3. On Day 0, one group received human peripheral blood mononuclear cells (PBMCs) with CD3-scFv-modified AAV6 encoding a CD19-specific CAR (FMC63; CD3scFv-AAV- FMC63CAR) IP, another received human PBMCs with CD3-scFv-modified AAV6 encoding eGFP (CD3scFv-AAV-EGFP) IP, and a third was untreated. Tumor growth was monitored twice weekly via In Vivo Imaging System (IVIS) for 31 days. The experimental design is shown in FIG.13A. IVIS images (FIG.13B) showed progressive tumor growth in untreated mice and those receiving CD3scFv-AAV-EGFP, with no untreated mice surviving past Day 28 and CD3scFv-AAV-EGFP-treated mice beginning to die by Day 18 (left-most and right-most columns, respectively). In contrast, mice treated with CD3-scFv-modified AAV6-FMC63CAR effectively controlled tumor growth through Day 31 (middle columns). FIG.13C, Panel A, quantifies average radiance (tumor burden), showing comparable tumor growth in untreated and CD3scFv-AAV-EGFP-treated groups, while CD3-scFv-modified AAV6-FMC63CAR group suppressed Raji tumor growth until Day 31. FIG.13C, Panel B, shows survival data: untreated mice died by Day 28, CD3scFv-AAV-EGFP-treated mice began dying after Day 17, while all mice treated with CD3-scFv-modified AAV6-FMC63CAR survived until Day 31. Example 23: In Vivo Generated CAR-T Cells and EGFP-Positive T Cells Can Be Found in Mouse Blood on Day 31.

[0247] To assess the persistence of the in vivo generated CAR-T and EGFP+ T cells (described in Example 22), all surviving mice were terminally bled on day 31 via cardiac puncture. The blood was analyzed for the presence of CD19-specific CAR-T cells and EGFP- positive T cells using flow cytometry. The data, shown in FIG.14, revealed that 70%-100% of the viable human hematopoietic cells, as determined using a viability dye and anti-human CD45 and CD3 antibodies, were CD3-positive T cells (FIG.14, panel A). Analysis of the CAR and EGFP expression showed that 0.16%-30.3% of the human T cells expressed the CAR and 0.23%-1.33% expressed EGFP (FIG.14, panel B). These results suggest that in vivo generated - 72 - 12961406v1Attorney Docket No.2014202-0027 CAR-T cells had undergone preferential CD19-specific expansion in response to tumor recognition, unlike the EGFP-positive T cells generated by CD3scFv-AAV-EGFP. Example 24: In Vivo Generated CD19-Specific CAR-T Cells Using Human PBMCs and CD3scFv-AAV-FMC63CAR Control Raji Lymphoma Growth upon Intravenous (IV) Administration and Improve Mouse Survival.

[0248] Having established that CD19-specific CAR-T cells can be generated in vivo following IP administration of human PBMCs and CD3scFv-AAV-FMC63CAR, and they are effective at controlling Raji tumors that are also growing IP, we evaluated whether similar results could be obtained using systemic IV administration of Raji cells, PBMCs and CD3scFv-AAV- FMC63CAR. The treatment schema is shown in FIG.15A. The IVIS images (FIG.15B) demonstrated that without treatment, Raji tumor cells grew over time and the mice did not survive past day 16 (four left-most image columns), while mice that received Raji, PBMCs and CD3scFv-AAV-FMC63CAR successfully controlled tumor growth (four right-most image columns) through day 24. Measurements of average radiance, which correlates with tumor burden (shown in FIG.15C, panel A), demonstrated that the tumors in the untreated animals were statistically significantly larger than the tumors in the mice treated with human PBMCs and CD3scFv-AAV-FMC63CAR starting on Day 3 post treatment initiation. Survival analysis (shown in panel B of FIG.15C) revealed that the untreated animals did not live past day 16, while all animals treated with human PBMCs and CD3scFv-AAV-FMC63CAR remained alive until day 24. Example 25: In Vivo Generated CAR-T Cells Using IV Administration of Human PBMCs and CD3scFv-AAV-FMC63CAR Can Be Found in Mouse Blood

[0249] To assess the persistence of the in vivo generated CAR-T cells (described in Example 24), the blood samples were analyzed for the presence of CD19-specific CAR-T cells using flow cytometry. The data revealed that more than 99% of the viable human hematopoietic cells, as determined using a viability dye and an anti-human CD45 antibody, were CD3-positive T cells (FIG.16 panel A). Analysis of the CAR expression showed that 0.3%-69.9% of the human T cells expressed the CAR (FIG.16 panel B). - 73 - 12961406v1Attorney Docket No.2014202-0027 Example 26: FMC63CAR DNA was detected in multiple mouse tissues following intravenous administration of human PBMCs and CD3scFv-AAV6-FMC63CAR in Raji- tumor-bearing animals, with biodistribution consistent with published data for wild-type AAV

[0250] To evaluate the biodistribution of FMC63CAR, liver, kidney, heart, spleen, brain, lung, ovaries, and bone marrow were collected from six mice administered human PBMCs and CD3scFv-AAV6-FMC63CAR, and three control mice, following euthanasia. Organ DNA was extracted using the DNeasy Blood and Tissue Kit, and the FMC63CAR copy number per mg of tissue was quantified by qPCR using CAR-specific primers and probe. The liver exhibited the highest CAR copy numbers, followed by the kidney, heart, and spleen, while the brain, lung, ovaries, and bone marrow showed copy numbers comparable to controls (FIG.17). EQUIVALENTS

[0251] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. - 74 - 12961406v1Attorney Docket No.2014202-0027 LIST OF SEQUENCES SEQ ID NO: 001 V062 DNA sequence TTCTCTGTCACAGAATGAAAATTTTTCTGTCATCTCTTCGTTATTAATGTTTGTAATTG ACTGAATATCAACGCTTATTTGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCG GCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCC AGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCG GCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTT ACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATC GCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGA CTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTAT AAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAAT TTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCAGGTGGCACTTTTCGGG GAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTA TGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCT GTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGT GCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTT CGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCG GTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCT CAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCAT GACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCA ACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACA TGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATA CCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAA ACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGAT GGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTT TATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAG CATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTC ATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAA TCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAG GATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACC ACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAA GGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTA GTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAAT CCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTC AAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCA CACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAG - 75 - 12961406v1Attorney Docket No.2014202-0027 CATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAG CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGG TATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGAT GCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGG TTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTC TGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAAC GACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATT TTCTCCTTACGCATCTGTGCGGTATTTCACACCGCAGACCAGCCGCGTAACCTGGCA AAATCGGTTACGGTTGAGTAATAAATGGATGCCCTGCGTAAGCGGGTGTGGGCGGA CAATAAAGTCTTAAACTGAACAAAATAGATCTAAACTATGACAATAAAGTCTTAAA CTAGACAGAATAGTTGTAAACTGAAATCAGTCCAGTTATGCTGTGAAAAAGCATACT GGACTTTTGTTATGGCTAAAGCAAACTCTTCATTTTCTGAAGTGCAAATTGCCCGTCG TATTAAAGAGGGGCGTGGCCAAGGGCATGGTAAAGACTATATTCGCGGCGTTGTGA CAATTTACCGAACAACTCCGCGGCCGGGAAGCCGATCTCGGCTTGAACGAATTGTTA GGTGGCGGTACTTGGGTCGATATCAAAGTGCATCACTTCTTCCCGTATGCCCAACTT TGTATAGAGAGCCACTGCGGGATCGTCACCGTAATCTGCTTGCACGTAGATCACATA AGCACCAAGCGCGTTGGCCTCATGCTTGAGGAGATTGATGAGCGCGGTGGCAATGC CCTGCCTCCGGTGCTCGCCGGAGACTGCGAGATCATAGATATAGATCTCACTACGCG GCTGCTCAAACCTGGGCAGAACGTAAGCCGCGAGAGCGCCAACAACCGCTTCTTGG TCGAAGGCAGCAAGCGCGATGAATGTCTTACTACGGAGCAAGTTCCCGAGGTAATC GGAGTCCGGCTGATGTTGGGAGTAGGTGGCTACGTCTCCGAACTCACGACCGAAAA GATCAAGAGCAGCCCGCATGGATTTGACTTGGTCAGGGCCGAGCCTACATGTGCGA ATGATGCCCATACTTGAGCCACCTAACTTTGTTTTAGGGCGACTGCCCTGCTGCGTA ACATCGTTGCTGCTGCGTAACATCGTTGCTGCTCCATAACATCAAACATCGACCCAC GGCGTAACGCGCTTGCTGCTTGGATGCCCGAGGCATAGACTGTACAAAAAAACAGT CATAACAAGCCATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAG GTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACAGTTTACGAACCGAA CAGGCTTATGTCAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGG CAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGC AAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGC AAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGTAGACCTCGGCCGTCG CGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTG GAAGGCGAGCATCGTTTGTTCGCCCAGGACTCTAGCTATAGTTCTAGTGGTTGGCCT ACGTACCCGTAGTGGCTATGGCAGGGCTTGCCGCCCCGACGTTGGCTGCGAGCCCTG GGCCTTCACCCGAACTTGGGGGTTGGGGTGGGGAAAAGGAAGAAACGCGGGCGTAT TGGTCCCAATGGGGTCTCGGTGGGGTATCGACAGAGTGCCAGCCCTGGGACCGAAC CCCGCGTTTATGAACAAACGACCCAACACCCGTGCGTTTTATTCTGTCTTTTTATTGC CGTCATAGCGCGGGTTCCTTCCGGTATTGTCTCCTTCCGTGTTTCAGTTAGCCTCCCC CATCTCCCGGTACCGCATGCTCCTTCAGAGAGAGTGTCCTCGAGCCAATCTGAAACA ATACCATCGGCAGCCATACCTGATTTAAATCATTTATTGTTCAAAGATGCAGTCATC - 76 - 12961406v1Attorney Docket No.2014202-0027 CAAATCCACATTGACCAGATCGCAGGCAGTGCAAGCGTCTGGCACCTTTCCCATGAT ATGATGAATGTAGCACAGTTTCTGATACGCCTTTTTGACGACAGAAACGGGTTGAGA TTCTGACACGGGAAAGCACTCTAAACAGTCTTTCTGTCCGTGAGTGAAGCAGATATT TGAATTCTGATTCATTCTCTCGCATTGTCTGCAGGGAAACAGCATCAGATTCATGCC CACGTGACGAGAACATTTGTTTTGGTACCTGTCTGCGTAGTTGATCGAAGCTTCCGC GTCTGACGTCGATGGCTGCGCAACTGACTCGCGCACCCGTTTGGGCTCACTTATATC TGCGTCACTGGGGGCGGGTCTTTTCTTGGCTCCACCCTTTTTGACGTAGAATTCATGC TCCACCTCAACCACGTGATCCTTTGCCCACCGGAAAAAGTCTTTGACTTCCTGCTTG GTGACCTTCCCAAAGTCATGATCCAGACGGCGGGTGAGTTCAAATTTGAACATCCGG TCTTGCAACGGCTGCTGGTGTTCGAAGGTCGTTGAGTTCCCGTCAATCACGGCGCAC ATGTTGGTGTTGGAGGTGACGATCACGGGAGTCGGGTCTATCTGGGCCGAGGACTTG CATTTCTGGTCCACGCGCACCTTGCTTCCTCCGAGAATGGCTTTGGCCGACTCCACG ACCTTGGCGGTCATCTTCCCCTCCTCCCACCAGATCACCATCTTGTCGACACAGTCGT TGAAGGGAAAGTTCTCATTGGTCCAGTTTACGCACCCGTAGAAGGGCACAGTGTGG GCTATGGCCTCCGCGATGTTGGTCTTCCCGGTAGTTGCAGGCCCAAACAGCCAGATG GTGTTCCTCTTGCCGAACTTTTTCGTGGCCCATCCCAGAAAGACGGAAGCCGCATAT TGGGGATCGTACCCGTTTAGTTCCAAAATTTTATAAATCCGATTGCTGGAAATGTCC TCCACGGGCTGCTGGCCCACCAGGTAGTCGGGGGCGGTTTTAGTCAGGCTCATAATC TTTCCCGCATTGTCCAAGGCAGCCTTGATTTGGGACCGCGAGTTGGAGGCCGCATTG AAGGAGATGTATGAGGCCTGGTCCTCCTGGATCCACTGCTTCTCCGAGGTAATCCCC TTGTCCACGAGCCACCCGACCAGCTCCATGTACCTGGCTGAAGTTTTTGATCTGATC ACCGGCGCATCAGAATTGGGATTCTGATTCTCTTTGTTCTGCTCCTGCGTCTGCGACA CGTGCGTCAGATGCTGCGCCACCAACCGTTTACGCTCCGTGAGATTCAAACAGGCGC TGAAACAATAGGAAGGGAGTGGATGTCAGTGTGTGCTGCCCGGGGGCTCTGACTAC AGGTCTCCCCCTTCGCGCCCGATGGTGGGACGGTATGAATAATCCGGAATATTTATA GGTTTTTTTATTACAAAACTGTTACGAAAACAGTAAAATACTTATTTATTTGCGAGAT GGTTATCATTTTAATTATCTCCATGATAGATCTCTATCACTGATAGGGAGTACTTACC TTAAATACTGTTCCATATTAGTCCACGCCCACTGGAGCTCAGGCTGGGTTTTGGGGA GCAAGTAATTGGGGATGTAGCACTCATCCACCACCTTGTTCCCGCCTCCGGCGCCAT TTCTGGTCTTTGTGACCGCGAACCAGTTTGGCAAAGTCGGCTCGATCCCGCGGTAAA TTCTCTGAATCAGTTTTTCGCGAATCTGACTCAGGAAACGTCCCAAAACCATGGATT TCACCCCGGTGGTTTCCACGAGCACGTGCATGTGGAAGTAGCTCTCTCCCTTCTCAA ATTGCACAAAGAAAAGGGCCTCCGGGGCCTTACTCACACGGCGCCATTCCGTCAGA AAGTCGCGCTGCAGCTTCTCGGCCACGGTCAGGGGTGCCTGCTCAATCAGATTCAGA TCCATGTCAGAATCTGGCGGCAACTCCCATTCCTTCTCGGCCACCCAGTTCACAAAG CTGTCAGAAATGCCGGGCAGATGCTCGTCAAGGTCGCTGGGGACCTTAATCACAATC TCGTAAAACCCCGGCATGGCGGGTAGGGTGATCAAGTCTTCGTCGAGTGATTGTAAA TAAAATGTAATTTACAGTATAGTATTTTAATTAATATACAAATGATTTGATAATAATT CTTATTTAACTATAATATATTGTGTTGGGTTGAATTAAAGGTCCGTAGCGGAATAAT TGCCATATGTAAATGATGTCATCGTTCTAACTCGCTTTACGAGTAGAATTCTACGTGT - 77 - 12961406v1Attorney Docket No.2014202-0027 AAAACATAATCAAGAGATGATGTCATTTGTTTTTCAAAACTGAACTCAAGAAATGAT GTCATTTGTTTTTCAAAACTGAACTGGCTTTACGAGTAGAATTCTACTTGTAACGCAT GATCAAGGGATGATGTCATTGGATGAGTCATTTGTTTTTCAAAACTAAACTCGCTTT ACGAGTAGAATTCTACTTGTAAAACACAATCAAGGGATGATGTCATTATACAAATG ATGTCATTTGTTTTTCAAAACTAAACTCGCTTTACGGGTAGAATTCTACTTGTAAAAC ACAATCGAGGGATGATGTCATCCTTTACACATGATTATAAACGTGTTTATGTATGAC TCATTTGTTTTTCAAAACTAAACTCGCTTTACGAGTAGAATTCTACTTGTAACGCACG ATCAAGGGATGATGTCATTTATTTGTGCAAAGCTGATGTCATCTTTTGCACACGATT ATAAACACAATCAAATAATGACTCATTTGTTTTCAAAACTGAACTCGCTTTACGAGT AGAATTCTACTTGTAAAACACAATCAAGGGATGATGTCATTTTAAAAATGATGTCAT TTGTTTTTCAAAACTAAACTCGCTTTACGAGTAGAATTCTACGTGTAAAACACAATC AAGGGATGATGTCATTTACTAAAATAAAATAATTATTTAAATAAAAATGTTTTTATT GTAAAATACACATTGATTACACGTGACGTATACTCCGGAATATTAATAGATCATGGA GATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGTTTTCGTAAC AGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTCCCACCATC GGGCGCGGATCCTGTTAAGATGGCTGCCGATGGTTATCTTCCAGGTAAGTACTCCCT ATCAGTGATAGAGATCTATCATGGAGATAATTAAAATGATAACCATCTCGCAAATA AATAAGTATTTTACTGTTTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCC GGATTATTCATACCGTCCCACCATCGGGCGCGAAGGGGGAGACCTGTAGTCAGAGC CCCCGGGCAGCACACACTGACATCCACTCCCTTCCTATTGTTTCAGATTGGCTCGAG GACACTCTCTCTGAAGGAATAAGACAGTGGTGGAAGCTCAAACCTGGCCCACCACC ACCAAAGCCCGCAGAGCGGCATAAGGACGACAGCAGGGGTCTTGTGCTTCCTGGGT ACAAGTACCTCGGACCCTTCAACGGACTCGACAAGGGAGAGCCGGTCAACGAGGCA GACGCCGCGGCCCTCGAGCACGACAAAGCCTACGACCGGCAGCTCGACAGCGGAGA CAACCCGTACCTCAAGTACAACCACGCCGACGCGGAGTTTCAGGAGCGCCTTAAAG AAGATACGTCTTTTGGGGGCAACCTCGGACGAGCAGTCTTCCAGGCGAAAAAGAGG GTTCTTGAACCTCTGGGCCTGGTTGAGGAACCTGTTAAGACGGCTCCGGGAAAAAA GAGGCCGGTAGAGCACTCTCCTGTGGAGCCAGACTCCTCCTCGGGAACCGGAAAGG CGGGCCAGCAGCCTGCAAGAAAAAGATTGAATTTTGGTCAGACTGGAGACGCAGAC TCAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCTCTGGTCTGGGA ACTAATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACGAGGGCGC CGACGGAGTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGGGCG ACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCAC CTCTACAAACAAATTTCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGC TACAGCACCCCTTGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCAC GTGACTGGCAAAGACTCATCAACAACAACTGGGGATTCCGACCCAAGAGACTCAAC TTCAAGCTCTTTAACATTCAAGTCAAAGAGGTCACGCAGAATGACGGTACGACGAC GATTGCCAATAACCTTACCAGCACGGTTCAGGTGTTTACTGACTCGGAGTACCAGCT CCCGTACGTCCTCGGCTCGGCGCATCAAGGATGCCTCCCGCCGTTCCCAGCAGACGT CTTCATGGTGCCACAGTATGGATACCTCACCCTGAACAACGGGAGTCAGGCAGTAG - 78 - 12961406v1Attorney Docket No.2014202-0027 GACGCTCTTCATTTTACTGCCTGGAGTACTTTCCTTCTCAGATGCTGCGTACCGGAAA CAACTTTACCTTCAGCTACACTTTTGAGGACGTTCCTTTCCACAGCAGCTACGCTCAC AGCCAGAGTCTGGACCGTCTCATGAATCCTCTCATCGACCAGTACCTGTATTACTTG AGCAGAACAAACACTCCAAGTGGAACCACCACGCAGTCAAGGCTTCAGTTTTCTCA GGCCGGAGCGAGTGACATTCGGGACCAGTCTAGGAACTGGCTTCCTGGACCCTGTTA CCGCCAGCAGCGAGTATCAAAGACATCTGCGGATAACAACAACAGTGAATACTCGT GGACTGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCGGGC CCGGCCATGGCAAGCCACAAGGACGATGAAGAAAAGTTTTTTCCTCAGAGCGGGGT TCTCATCTTTGGGAAGCAAGGCTCAGAGAAAACAAATGTGGACATTGAAAAGGTCA TGATTACAGACGAAGAGGAAATCAGGACAACCAATCCCGTGGCTACGGAGCAGTAT GGTTCTGTATCTACCAACCTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGT CAACACACAAGGCGTTCTTCCAGGCATGGTCTGGCAGGACAGAGATGTGTACCTTCA GGGGCCCATCTGGGCAAAGATTCCACACACGGACGGACATTTTCACCCCTCTCCCCT CATGGGTGGATTCGGACTTAAACACCCTCCTCCACAGATTCTCATCAAGAACACCCC GGTACCTGCGAATCCTTCGACCACCTTCAGTGCGGCAAAGTTTGCTTCCTTCATCAC ACAGTACTCCACGGGACAGGTCAGCGTGGAGATCGAGTGGGAGCTGCAGAAGGAA AACAGCAAACGCTGGAATCCCGAAATTCAGTACACTTCCAACTACAACAAGTCTGTT AATGTGGACTTTACTGTGGACACTAATGGCGTGTATTCAGAGCCTCGCCCCATTGGC ACCAGATACCTGACTCGTAATCTGTAATTGCTTGTTAATCAATAAACCGTTTAATTCG TTTCAGTTGAACTTTGGTCTCTGCGTATTTCTTTCTTATCTAGTTTCCATGGCTACAGC TTGTCGAGAAGTACTAGAGGATCATAATCAGCCATACCACATTTGTAGAGGTTTTAC TTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAA TTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCAT CACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAA CTCATCAATGTATCTTATCATGTCTGGATCTGATCACTGCTTGAGCCTAGGAGATCCG AACCAGATAAGTGAAATCTAGTTCCAAACTATTTTGTCATTTTTAATTTTCGTATTAG CTTACGACGCTACACCCAGTTCCCATCTATTTTGTCACTCTTCCCTAAATAATCCTTA AAAACTCCATTTCCACCCCTCCCAGTTCCCAACTATTTTGTCCGCCCACAGCGGGGC ATTTTTCTTCCTGTTATGTTTTTAATCAAACATCCTGCCAACTCCATGTGACAAACCG TCATCTTCGGCTACTTT SEQ ID NO: 002 Primer 0297 GGGCCTGGTTGAGGAACCTGTTAAGACCGCTCCGGGAAAAAAGAGGCCGG SEQ ID NO: 003 Primer 0298 AGGACGTACGGGAGCTGGTA SEQ ID NO: 004 V076 DNA sequence TTCTCTGTCACAGAATGAAAATTTTTCTGTCATCTCTTCGTTATTAATGTTTGTAATTG ACTGAATATCAACGCTTATTTGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCG - 79 - 12961406v1Attorney Docket No.2014202-0027 GCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCC AGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCG GCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTT ACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATC GCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGA CTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTAT AAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAAT TTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCAGGTGGCACTTTTCGGG GAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTA TGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCT GTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGT GCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTT CGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCG GTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCT CAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCAT GACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCA ACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACA TGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATA CCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAA ACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGAT GGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTT TATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAG CATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTC ATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAA TCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAG GATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACC ACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAA GGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTA GTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAAT CCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTC AAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCA CACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAG CATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAG CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGG TATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGAT GCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGG TTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTC - 80 - 12961406v1Attorney Docket No.2014202-0027 TGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAAC GACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATT TTCTCCTTACGCATCTGTGCGGTATTTCACACCGCAGACCAGCCGCGTAACCTGGCA AAATCGGTTACGGTTGAGTAATAAATGGATGCCCTGCGTAAGCGGGTGTGGGCGGA CAATAAAGTCTTAAACTGAACAAAATAGATCTAAACTATGACAATAAAGTCTTAAA CTAGACAGAATAGTTGTAAACTGAAATCAGTCCAGTTATGCTGTGAAAAAGCATACT GGACTTTTGTTATGGCTAAAGCAAACTCTTCATTTTCTGAAGTGCAAATTGCCCGTCG TATTAAAGAGGGGCGTGGCCAAGGGCATGGTAAAGACTATATTCGCGGCGTTGTGA CAATTTACCGAACAACTCCGCGGCCGGGAAGCCGATCTCGGCTTGAACGAATTGTTA GGTGGCGGTACTTGGGTCGATATCAAAGTGCATCACTTCTTCCCGTATGCCCAACTT TGTATAGAGAGCCACTGCGGGATCGTCACCGTAATCTGCTTGCACGTAGATCACATA AGCACCAAGCGCGTTGGCCTCATGCTTGAGGAGATTGATGAGCGCGGTGGCAATGC CCTGCCTCCGGTGCTCGCCGGAGACTGCGAGATCATAGATATAGATCTCACTACGCG GCTGCTCAAACCTGGGCAGAACGTAAGCCGCGAGAGCGCCAACAACCGCTTCTTGG TCGAAGGCAGCAAGCGCGATGAATGTCTTACTACGGAGCAAGTTCCCGAGGTAATC GGAGTCCGGCTGATGTTGGGAGTAGGTGGCTACGTCTCCGAACTCACGACCGAAAA GATCAAGAGCAGCCCGCATGGATTTGACTTGGTCAGGGCCGAGCCTACATGTGCGA ATGATGCCCATACTTGAGCCACCTAACTTTGTTTTAGGGCGACTGCCCTGCTGCGTA ACATCGTTGCTGCTGCGTAACATCGTTGCTGCTCCATAACATCAAACATCGACCCAC GGCGTAACGCGCTTGCTGCTTGGATGCCCGAGGCATAGACTGTACAAAAAAACAGT CATAACAAGCCATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAG GTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACAGTTTACGAACCGAA CAGGCTTATGTCAACTGGGTTCGTGCCTTCATCCGTTTCCACGGTGTGCGTCACCCGG CAACCTTGGGCAGCAGCGAAGTCGAGGCATTTCTGTCCTGGCTGGCGAACGAGCGC AAGGTTTCGGTCTCCACGCATCGTCAGGCATTGGCGGCCTTGCTGTTCTTCTACGGC AAGGTGCTGTGCACGGATCTGCCCTGGCTTCAGGAGATCGGTAGACCTCGGCCGTCG CGGCGCTTGCCGGTGGTGCTGACCCCGGATGAAGTGGTTCGCATCCTCGGTTTTCTG GAAGGCGAGCATCGTTTGTTCGCCCAGGACTCTAGCTATAGTTCTAGTGGTTGGCCT ACGTACCCGTAGTGGCTATGGCAGGGCTTGCCGCCCCGACGTTGGCTGCGAGCCCTG GGCCTTCACCCGAACTTGGGGGTTGGGGTGGGGAAAAGGAAGAAACGCGGGCGTAT TGGTCCCAATGGGGTCTCGGTGGGGTATCGACAGAGTGCCAGCCCTGGGACCGAAC CCCGCGTTTATGAACAAACGACCCAACACCCGTGCGTTTTATTCTGTCTTTTTATTGC CGTCATAGCGCGGGTTCCTTCCGGTATTGTCTCCTTCCGTGTTTCAGTTAGCCTCCCC CATCTCCCGGTACCGCATGCTCCTTCAGAGAGAGTGTCCTCGAGCCAATCTGAAACA ATACCATCGGCAGCCATACCTGATTTAAATCATTTATTGTTCAAAGATGCAGTCATC CAAATCCACATTGACCAGATCGCAGGCAGTGCAAGCGTCTGGCACCTTTCCCATGAT ATGATGAATGTAGCACAGTTTCTGATACGCCTTTTTGACGACAGAAACGGGTTGAGA TTCTGACACGGGAAAGCACTCTAAACAGTCTTTCTGTCCGTGAGTGAAGCAGATATT TGAATTCTGATTCATTCTCTCGCATTGTCTGCAGGGAAACAGCATCAGATTCATGCC CACGTGACGAGAACATTTGTTTTGGTACCTGTCTGCGTAGTTGATCGAAGCTTCCGC - 81 - 12961406v1Attorney Docket No.2014202-0027 GTCTGACGTCGATGGCTGCGCAACTGACTCGCGCACCCGTTTGGGCTCACTTATATC TGCGTCACTGGGGGCGGGTCTTTTCTTGGCTCCACCCTTTTTGACGTAGAATTCATGC TCCACCTCAACCACGTGATCCTTTGCCCACCGGAAAAAGTCTTTGACTTCCTGCTTG GTGACCTTCCCAAAGTCATGATCCAGACGGCGGGTGAGTTCAAATTTGAACATCCGG TCTTGCAACGGCTGCTGGTGTTCGAAGGTCGTTGAGTTCCCGTCAATCACGGCGCAC ATGTTGGTGTTGGAGGTGACGATCACGGGAGTCGGGTCTATCTGGGCCGAGGACTTG CATTTCTGGTCCACGCGCACCTTGCTTCCTCCGAGAATGGCTTTGGCCGACTCCACG ACCTTGGCGGTCATCTTCCCCTCCTCCCACCAGATCACCATCTTGTCGACACAGTCGT TGAAGGGAAAGTTCTCATTGGTCCAGTTTACGCACCCGTAGAAGGGCACAGTGTGG GCTATGGCCTCCGCGATGTTGGTCTTCCCGGTAGTTGCAGGCCCAAACAGCCAGATG GTGTTCCTCTTGCCGAACTTTTTCGTGGCCCATCCCAGAAAGACGGAAGCCGCATAT TGGGGATCGTACCCGTTTAGTTCCAAAATTTTATAAATCCGATTGCTGGAAATGTCC TCCACGGGCTGCTGGCCCACCAGGTAGTCGGGGGCGGTTTTAGTCAGGCTCATAATC TTTCCCGCATTGTCCAAGGCAGCCTTGATTTGGGACCGCGAGTTGGAGGCCGCATTG AAGGAGATGTATGAGGCCTGGTCCTCCTGGATCCACTGCTTCTCCGAGGTAATCCCC TTGTCCACGAGCCACCCGACCAGCTCCATGTACCTGGCTGAAGTTTTTGATCTGATC ACCGGCGCATCAGAATTGGGATTCTGATTCTCTTTGTTCTGCTCCTGCGTCTGCGACA CGTGCGTCAGATGCTGCGCCACCAACCGTTTACGCTCCGTGAGATTCAAACAGGCGC TGAAACAATAGGAAGGGAGTGGATGTCAGTGTGTGCTGCCCGGGGGCTCTGACTAC AGGTCTCCCCCTTCGCGCCCGATGGTGGGACGGTATGAATAATCCGGAATATTTATA GGTTTTTTTATTACAAAACTGTTACGAAAACAGTAAAATACTTATTTATTTGCGAGAT GGTTATCATTTTAATTATCTCCATGATAGATCTCTATCACTGATAGGGAGTACTTACC TTAAATACTGTTCCATATTAGTCCACGCCCACTGGAGCTCAGGCTGGGTTTTGGGGA GCAAGTAATTGGGGATGTAGCACTCATCCACCACCTTGTTCCCGCCTCCGGCGCCAT TTCTGGTCTTTGTGACCGCGAACCAGTTTGGCAAAGTCGGCTCGATCCCGCGGTAAA TTCTCTGAATCAGTTTTTCGCGAATCTGACTCAGGAAACGTCCCAAAACCATGGATT TCACCCCGGTGGTTTCCACGAGCACGTGCATGTGGAAGTAGCTCTCTCCCTTCTCAA ATTGCACAAAGAAAAGGGCCTCCGGGGCCTTACTCACACGGCGCCATTCCGTCAGA AAGTCGCGCTGCAGCTTCTCGGCCACGGTCAGGGGTGCCTGCTCAATCAGATTCAGA TCCATGTCAGAATCTGGCGGCAACTCCCATTCCTTCTCGGCCACCCAGTTCACAAAG CTGTCAGAAATGCCGGGCAGATGCTCGTCAAGGTCGCTGGGGACCTTAATCACAATC TCGTAAAACCCCGGCATGGCGGGTAGGGTGATCAAGTCTTCGTCGAGTGATTGTAAA TAAAATGTAATTTACAGTATAGTATTTTAATTAATATACAAATGATTTGATAATAATT CTTATTTAACTATAATATATTGTGTTGGGTTGAATTAAAGGTCCGTAGCGGAATAAT TGCCATATGTAAATGATGTCATCGTTCTAACTCGCTTTACGAGTAGAATTCTACGTGT AAAACATAATCAAGAGATGATGTCATTTGTTTTTCAAAACTGAACTCAAGAAATGAT GTCATTTGTTTTTCAAAACTGAACTGGCTTTACGAGTAGAATTCTACTTGTAACGCAT GATCAAGGGATGATGTCATTGGATGAGTCATTTGTTTTTCAAAACTAAACTCGCTTT ACGAGTAGAATTCTACTTGTAAAACACAATCAAGGGATGATGTCATTATACAAATG ATGTCATTTGTTTTTCAAAACTAAACTCGCTTTACGGGTAGAATTCTACTTGTAAAAC - 82 - 12961406v1Attorney Docket No.2014202-0027 ACAATCGAGGGATGATGTCATCCTTTACACATGATTATAAACGTGTTTATGTATGAC TCATTTGTTTTTCAAAACTAAACTCGCTTTACGAGTAGAATTCTACTTGTAACGCACG ATCAAGGGATGATGTCATTTATTTGTGCAAAGCTGATGTCATCTTTTGCACACGATT ATAAACACAATCAAATAATGACTCATTTGTTTTCAAAACTGAACTCGCTTTACGAGT AGAATTCTACTTGTAAAACACAATCAAGGGATGATGTCATTTTAAAAATGATGTCAT TTGTTTTTCAAAACTAAACTCGCTTTACGAGTAGAATTCTACGTGTAAAACACAATC AAGGGATGATGTCATTTACTAAAATAAAATAATTATTTAAATAAAAATGTTTTTATT GTAAAATACACATTGATTACACGTGACGTATACTCCGGAATATTAATAGATCATGGA GATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGTTTTCGTAAC AGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTCCCACCATC GGGCGCGGATCCTGTTAAGATGGCTGCCGATGGTTATCTTCCAGGTAAGTACTCCCT ATCAGTGATAGAGATCTATCATGGAGATAATTAAAATGATAACCATCTCGCAAATA AATAAGTATTTTACTGTTTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCC GGATTATTCATACCGTCCCACCATCGGGCGCGAAGGGGGAGACCTGTAGTCAGAGC CCCCGGGCAGCACACACTGACATCCACTCCCTTCCTATTGTTTCAGATTGGCTCGAG GACACTCTCTCTGAAGGAATAAGACAGTGGTGGAAGCTCAAACCTGGCCCACCACC ACCAAAGCCCGCAGAGCGGCATAAGGACGACAGCAGGGGTCTTGTGCTTCCTGGGT ACAAGTACCTCGGACCCTTCAACGGACTCGACAAGGGAGAGCCGGTCAACGAGGCA GACGCCGCGGCCCTCGAGCACGACAAAGCCTACGACCGGCAGCTCGACAGCGGAGA CAACCCGTACCTCAAGTACAACCACGCCGACGCGGAGTTTCAGGAGCGCCTTAAAG AAGATACGTCTTTTGGGGGCAACCTCGGACGAGCAGTCTTCCAGGCGAAAAAGAGG GTTCTTGAACCTCTGGGCCTGGTTGAGGAACCTGTTAAGACCGCTCCGGGAAAAAAG AGGCCGGTAGAGCACTCTCCTGTGGAGCCAGACTCCTCCTCGGGAACCGGAAAGGC GGGCCAGCAGCCTGCAAGAAAAAGATTGAATTTTGGTCAGACTGGAGACGCAGACT CAGTACCTGACCCCCAGCCTCTCGGACAGCCACCAGCAGCCCCCTCTGGTCTGGGAA CTAATACGATGGCTACAGGCAGTGGCGCACCAATGGCAGACAATAACGAGGGCGCC GACGGAGTGGGTAATTCCTCGGGAAATTGGCATTGCGATTCCACATGGATGGGCGA CAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTACAACAACCACC TCTACAAACAAATTTCCAGCCAATCAGGAGCCTCGAACGACAATCACTACTTTGGCT ACAGCACCCCTTGGGGGTATTTTGACTTCAACAGATTCCACTGCCACTTTTCACCAC GTGACTGGCAAAGACTCATCAACAACAACTGGGGATTCCGACCCAAGAGACTCAAC TTCAAGCTCTTTAACATTCAAGTCAAAGAGGTCACGCAGAATGACGGTACGACGAC GATTGCCAATAACCTTACCAGCACGGTTCAGGTGTTTACTGACTCGGAGTACCAGCT CCCGTACGTCCTCGGCTCGGCGCATCAAGGATGCCTCCCGCCGTTCCCAGCAGACGT CTTCATGGTGCCACAGTATGGATACCTCACCCTGAACAACGGGAGTCAGGCAGTAG GACGCTCTTCATTTTACTGCCTGGAGTACTTTCCTTCTCAGATGCTGCGTACCGGAAA CAACTTTACCTTCAGCTACACTTTTGAGGACGTTCCTTTCCACAGCAGCTACGCTCAC AGCCAGAGTCTGGACCGTCTCATGAATCCTCTCATCGACCAGTACCTGTATTACTTG AGCAGAACAAACACTCCAAGTGGAACCACCACGCAGTCAAGGCTTCAGTTTTCTCA GGCCGGAGCGAGTGACATTCGGGACCAGTCTAGGAACTGGCTTCCTGGACCCTGTTA - 83 - 12961406v1Attorney Docket No.2014202-0027 CCGCCAGCAGCGAGTATCAAAGACATCTGCGGATAACAACAACAGTGAATACTCGT GGACTGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCGGGC CCGGCCATGGCAAGCCACAAGGACGATGAAGAAAAGTTTTTTCCTCAGAGCGGGGT TCTCATCTTTGGGAAGCAAGGCTCAGAGAAAACAAATGTGGACATTGAAAAGGTCA TGATTACAGACGAAGAGGAAATCAGGACAACCAATCCCGTGGCTACGGAGCAGTAT GGTTCTGTATCTACCAACCTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGT CAACACACAAGGCGTTCTTCCAGGCATGGTCTGGCAGGACAGAGATGTGTACCTTCA GGGGCCCATCTGGGCAAAGATTCCACACACGGACGGACATTTTCACCCCTCTCCCCT CATGGGTGGATTCGGACTTAAACACCCTCCTCCACAGATTCTCATCAAGAACACCCC GGTACCTGCGAATCCTTCGACCACCTTCAGTGCGGCAAAGTTTGCTTCCTTCATCAC ACAGTACTCCACGGGACAGGTCAGCGTGGAGATCGAGTGGGAGCTGCAGAAGGAA AACAGCAAACGCTGGAATCCCGAAATTCAGTACACTTCCAACTACAACAAGTCTGTT AATGTGGACTTTACTGTGGACACTAATGGCGTGTATTCAGAGCCTCGCCCCATTGGC ACCAGATACCTGACTCGTAATCTGTAATTGCTTGTTAATCAATAAACCGTTTAATTCG TTTCAGTTGAACTTTGGTCTCTGCGTATTTCTTTCTTATCTAGTTTCCATGGCTACAGC TTGTCGAGAAGTACTAGAGGATCATAATCAGCCATACCACATTTGTAGAGGTTTTAC TTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACATAAAATGAATGCAA TTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCAT CACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAA CTCATCAATGTATCTTATCATGTCTGGATCTGATCACTGCTTGAGCCTAGGAGATCCG AACCAGATAAGTGAAATCTAGTTCCAAACTATTTTGTCATTTTTAATTTTCGTATTAG CTTACGACGCTACACCCAGTTCCCATCTATTTTGTCACTCTTCCCTAAATAATCCTTA AAAACTCCATTTCCACCCCTCCCAGTTCCCAACTATTTTGTCCGCCCACAGCGGGGC ATTTTTCTTCCTGTTATGTTTTTAATCAAACATCCTGCCAACTCCATGTGACAAACCG TCATCTTCGGCTACTTT SEQ ID NO: 005 V290 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCGCCACCATGGCTGCCGATGGTTATCTTCCAGGTAAGT ACTCCCTATCAGTGATAGAGATCTATCATGGAGATAATTAAAATGATAACCATCTCG CAAATAAATAAGTATTTTACTGTTTTCGTAACAGTTTTGTAATAAAAAAACCTATAA ATATTCCGGATTATTCATACCGTCCCACCATCGGGCGCGAAGGGGGAGACCTGTAGT CAGAGCCCCCGGGCAGCACACACTGACATCCACTCCCTTCCTATTGTTTCAGATTGG CTCGAGGACAACCTCTCTGAGGGCATTCGCGAGTGGTGGGACTTGAAACCTGGAGC - 84 - 12961406v1Attorney Docket No.2014202-0027 CCCGAAACCCAAAGCCAACCAGCAAAAGCAGGACGACGGCCGGGGTCTGGTGCTTC CTGGCTACAAGTACCTCGGACCCTTCAACGGACTCGACAAGGGGGAGCCCGTCAAC GCGGCGGATGCAGCGGCCCTCGAGCACGACAAGGCCTACGACCAGCAGCTCAAAGC GGGTGACAATCCGTACCTGCGGTATAACCACGCCGACGCCGAGTTTCAGGAGCGTC TGCAAGAAGATACGTCTTTTGGGGGCAACCTCGGGCGAGCAGTCTTCCAGGCCAAG AAGAGGGTTCTCGAACCTTTTGGTCTGGTTGAGGAAGGTGCTAAGACGGCTCCTGGA AAGAAACGTCCGGTAGAGCAGTCGCCACAAGAGCCAGACTCCTCCTCGGGCATTGG CAAGACAGGCCAGCAGCCCGCTAAAAAGAGACTCAATTTTGGTCAGACTGGCGACT CAGAGTCAGTCCCCGACCCACAACCTCTCGGAGAACCTCCAGCAACCCCCGCTGCTG TGGGACCTACTACAATGGCTTCAGGCGGTGGCGCACCAATGGCAGACAATAACGAA GGCGCCGACGGAGTGGGTAATGCCTCAGGAAATTGGCATTGCGATTCCACATGGCT GGGCGACAGAGTCATCACCACCAGCACCCGAACATGGGCCTTGCCCACCTATAACA ACCACCTCTACAAGCAAATCTCCAGTGCTTCAACGGGGGCCAGCAACGACAACCAC TACTTCGGCTACAGCACCCCCTGGGGGTATTTTGATTTCAACAGATTCCACTGCCATT TCTCACCACGTGACTGGCAGCGACTCATCAACAACAATTGGGGATTCCGGCCCAAG AGACTCAACTTCAAGCTCTTCAACATCCAAGTCAAGGAGGTCACGACGAATGATGG CGTCACGACCATCGCTAATAACCTTACCAGCACGGTTCAAGTCTTCTCGGACTCGGA GTACCAGTTGCCGTACGTCCTCGGCTCTGCGCACCAGGGCTGCCTCCCTCCGTTCCC GGCGGACGTGTTCATGATTCCGCAGTACGGCTACCTAACGCTCAACAATGGCAGCC AGGCAGTGGGACGGTCATCCTTTTACTGCCTGGAATATTTCCCATCGCAGATGCTGA GAACGGGCAATAACTTTACCTTCAGCTACACCTTCGAGGACGTGCCTTTCCACAGCA GCTACGCGCACAGCCAGAGCCTGGACCGGCTGATGAATCCTCTCATCGACCAGTAC CTGTATTACCTGAACAGAACTCAGAATCAGTCCGGAAGTGCCCAAAACAAGGACTT GCTGTTTAGCCGGGGGTCTCCAGCTGGCATGTCTGTTCAGCCCAAAAACTGGCTACC TGGACCCTGTTACCGGCAGCAGCGCGTTTCTAAAACAAAAACAGACAACAACAACA GCAACTTTACCTGGACTGGTGCTTCAAAATATAACCTTAATGGGCGTGAATCTATAA TCAACCCTGGCACTGCTATGGCCTCACACAAAGACGACAAAGACAAGTTCTTTCCCA TGAGCGGTGTCATGATTTTTGGAAAGGAGAGCGCCGGAGCTTCAAACACTGCATTG GACAATGTCATGATCACAGACGAAGAGGAAATCAAAGCCACTAACCCCGTGGCCAC CGAAAGATTTGGGACTGTGGCAGTCAATCTCCAGAGCAGCAGCACAGACCCTGCGA CCGGAGATGTGCATGTTATGGGAGCCTTACCTGGAATGGTGTGGCAAGACAGAGAC GTATACCTGCAGGGTCCTATTTGGGCCAAAATTCCTCACACGGATGGACACTTTCAC CCGTCTCCTCTCATGGGCGGCTTTGGACTTAAGCACCCGCCTCCTCAGATCCTCATCA AAAACACGCCTGTTCCTGCGAATCCTCCGGCAGAGTTTTCGGCTACAAAGTTTGCTT CATTCATCACCCAGTATTCCACAGGACAAGTGAGCGTGGAGATTGAATGGGAGCTG CAGAAAGAAAACAGCAAACGCTGGAATCCCGAAGTGCAGTATACATCTAACTATGC AAAATCTGCCAACGTTGATTTCACTGTGGACAACAATGGACTTTATACTGAGCCTCG CCCCATTGGCACCCGTTACCTCACCCGTCCCCTGTAAGCGGTCGACTCTAGAGCCTG CAGTCTCGACAAGCTTGTCGAGAAGTACTAGAGGATCATAATCAGCCATACCACATT TGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTGAACCTGAAACA - 85 - 12961406v1Attorney Docket No.2014202-0027 TAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTATAATGGTTACAAA TAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGT TGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATCTGATCACTGCTTG AGCCTAGATTCGAAAGCTACGGACCTTTAATTCAACCCAACACAATATATTATAGTT AAATAAGAATTATTATCAAATCATTTGTATATTAATTAAAATACTATACTGTAAATT ACATTTTATTTACAATCACTCGACGAAGACTTGATCACCCTACCCGCCATGCCGGGG TTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGAGCATCTGCCCGGCATT TCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGCCGCCAGATTCT GACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCGAGAAGCTGCA GCGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGGAGGCCCTTTTCTT TGTGCAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTCGTGGAAACCA CCGGGGTGAAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGCGAAAAACTGA TTCAGAGAATTTACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTCGCGGTCACAA AGACCAGAAATGGCGCCGGAGGCGGGAACAAGGTGGTGGATGAGTGCTACATCCCC AATTACTTGCTCCCCAAAACCCAGCCTGAGCTCCAGTGGGCGTGGACTAATATGGAA CAGTATTTAAGGTAAGTACTCCCTATCAGTGATAGAGATCTATCATGGAGATAATTA AAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGTTTTCGTAACAGTTTTGTA ATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTCCCACCATCGGGCGCGAA GGGGGAGACCTGTAGTCAGAGCCCCCGGGCAGCACACACTGACATCCACTCCCTTC CTATTGTTTCAGCGCCTGTTTGAATCTCACGGAGCGTAAACGGTTGGTGGCGCAGCA TCTGACGCACGTGTCGCAGACGCAGGAGCAGAACAAAGAGAATCAGAATCCCAATT CTGATGCGCCGGTGATCAGATCAAAAACTTCAGCCAGGTACATGGAGCTGGTCGGG TGGCTCGTGGACAAGGGGATTACCTCGGAGAAGCAGTGGATCCAGGAGGACCAGGC CTCATACATCTCCTTCAATGCGGCCTCCAACTCGCGGTCCCAAATCAAGGCTGCCTT GGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCCCCCGACTACCTGGTGG GCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAAATTTTGGAACTAA ACGGGTACGATCCCCAATATGCGGCTTCCGTCTTTCTGGGATGGGCCACGAAAAAGT TCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTACCGGGAAGACCAAC ATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCGTAAACTGGACCAAT GAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCTGGTGGGAGGAGGG GAAGATGACCGCCAAGGTCGTGGAGTCGGCCAAAGCCATTCTCGGAGGAAGCAAGG TGCGCGTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCCGACTCCCGTGATC GTCACCTCCAACACCAACATGTGCGCCGTGATTGACGGGAACTCAACGACCTTCGA ACACCAGCAGCCGTTGCAAGACCGGATGTTCAAATTTGAACTCACCCGCCGTCTGGA TCATGACTTTGGGAAGGTCACCAAGCAGGAAGTCAAAGACTTTTTCCGGTGGGCAA AGGATCACGTGGTTGAGGTGGAGCATGAATTCTACGTCAAAAAGGGTGGAGCCAAG AAAAGACCCGCCCCCAGTGACGCAGATATAAGTGAGCCCAAACGGGTGCGCGAGTC AGTTGCGCAGCCATCGACGTCAGACGCGGAAGCTTCGATCAACTACGCAGACAGGT ACCAAAACAAATGTTCTCGTCACGTGGGCATGAATCTGATGCTGTTTCCCTGCAGAC AATGCGAGAGAATGAATCAGAATTCAAATATCTGCTTCACTCACGGACAGAAAGAC - 86 - 12961406v1Attorney Docket No.2014202-0027 TGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTTTCTGTCGTCAAAAAGGCGT ATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTGCCAGACGCTTGCACTG CCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGAACAATAAATGATTT AAATCAGGTATGGCTGCCGATGGTATTGTTTCAGATTGGCTCGAGGACACTCTCTCT GAAGGAGCATGCGGTACCGGGAGATGGGGGAGGCTAACTGAAACACGGAAGGAGA CAATACCGGAAGGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAACGCA CGGGTGTTGGGTCGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCACTCTG TCGATACCCCACCGAGACCCCATTGGGACCAATACGCCCGCGTTTCTTCCTTTTCCCC ACCCCAACCCCCAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGGGCGG CAAGCCCTGCCATAGCCACTACGGGTACGTACTATTAATATTCCGGAGTATACGTCA CGTGTAATCAATGTGTATTTTACAATAAAAACATTTTTATTTAAATAATTATTTTATT TTAGTAAATGACATCATCCCTTGATTGTGTTTTACACGTAGAATTCTACTCGTAAAGC GAGTTTAGTTTTGAAAAACAAATGACATCATTTTTAAAATGACATCATCCCTTGATT GTGTTTTACAAGTAGAATTCTACTCGTAAAGCGAGTTCAGTTTTGAAAACAAATGAG TCATTATTTGATTGTGTTTATAATCGTGTGCAAAAGATGACATCAGCTTTGCACAAAT AAATGACATCATCCCTTGATCGTGCGTTACAAGTAGAATTCTACTCGTAAAGCGAGT TTAGTTTTGAAAAACAAATGAGTCATACATAAACACGTTTATAATCATGTGTAAAGG ATGACATCATCCCTCGATTGTGTTTTACAAGTAGAATTCTACCCGTAAAGCGAGTTT AGTTTTGAAAAACAAATGACATCATTTGTATAATGACATCATCCCTTGATTGTGTTTT ACAAGTAGAATTCTACTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACTCATCC AATGACATCATCCCTTGATCATGCGTTACAAGTAGAATTCTACTCGTAAAGCCAGTT CAGTTTTGAAAAACAAATGACATCATTTCTTGAGTTCAGTTTTGAAAAACAAATGAC ATCATCTCTTGATTATGTTTTACACGTAGAATTCTACTCGTAAAGCGAGTTAGAACG ATGACATCATTTACATATGGCAATTATTCCGCCCAGTTGACATAAGCCTGTTCGGTTC GTAAACTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCG AACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTT TTGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGAT GTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGCAACGATGTTACGCAGCA GGGCAGTCGCCCTAAAACAAAGTTAGGTGGCTCAAGTATGGGCATCATTCGCACAT GTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTC GTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCG GGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGG TTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGA TCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCG CGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACG TGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATAC GGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAACAATTCGTTC AAGCCGAGATCGGCTTCCCGGCCGCGGAGTTGTTCGGTAAATTGTCACAACGCCGC GAATATAGTCTTTACCATGCCCTTGGCCACGCCCCTCTTTAATACGACGGGCAATTT GCACTTCAGAAAATGAAGAGTTTGCTTTAGCCATAACAAAAGTCCAGTATGCTTTTT - 87 - 12961406v1Attorney Docket No.2014202-0027 CACAGCATAACTGGACTGATTTCAGTTTACAACTATTCTGTCTAGTTTAAGACTTTAT TGTCATAGTTTAGATCTATTTTGTTCAGTTTAAGACTTTATTGTCCGCCCACACCCGC TTACGCAGGGCATCCATTTATTACTCAACCGTAACCGATTTTGCCAGGTTACGCGGC TGGTCTGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGC GCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGC GGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACG CAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGC CGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCG ACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTC CCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCT GTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTAT CTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTT CAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGA CACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTA TGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAG GACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGG TAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAA GCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTAC GGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATT ATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAAT CTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGC ACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTG TAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCG CGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAG GGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTG TTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGC CATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCC GGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTT AGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTC ATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTT CTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGA GTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAA AAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGC TGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTT TACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAA AGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATT ATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTA GAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGAAA TTGTAAACGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATT TTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCG - 88 - 12961406v1Attorney Docket No.2014202-0027 AGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGG ACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAA CCATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAAC CCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAG AAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCG GTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGC GTCCCATTCGCCATTCAGGCTGCAAATAAGCGTTGATATTCAGTCAATTACAAACAT TAATAACGAAGAGATGACAGAAAAATTTTCATTCTGTGACAGAGAA SEQ ID NO: 006 Primer 3821 CAATTTCCTGAGGCATTACCCACTC SEQ ID NO: 007 Primer 3822 GCTAAGACAGCTCCTGGAAAGAAACGTCC SEQ ID NO: 008 Primer 3823 GTTTCTTTCCAGGAGCTGTCTTAGCACCTTCCTCAACCAG SEQ ID NO: 009 Primer 2671 CTGGGAGGGGTGGAAATGGAGTTTT SEQ ID NO: 010 V334 DNA sequence TTCTCTGTCACAGAATGAAAATTTTTCTGTCATCTCTTCGTTATTAATGTTTGTAATTG ACTGAATATCAACGCTTATTTGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCG GCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCC AGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCG GCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTT ACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATC GCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGA CTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTAT AAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAAT TTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTCAGGTGGCACTTTTCGGG GAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTA TGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCT GTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGT GCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTT CGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCG GTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCT - 89 - 12961406v1Attorney Docket No.2014202-0027 CAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCAT GACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCA ACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACA TGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATA CCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAA ACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGAT GGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTT TATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGG CAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAG CATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTC ATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAA TCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAG GATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACC ACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAA GGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTA GTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAAT CCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTC AAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCA CACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAG CATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAG CGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGG TATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGAT GCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGG TTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTC TGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAAC GACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCTGATGCGGTATT TTCTCCTTACGCATCTGTGCGGTATTTCACACCGCAGACCAGCCGCGTAACCTGGCA AAATCGGTTACGGTTGAGTAATAAATGGATGCCCTGCGTAAGCGGGTGTGGGCGGA CAATAAAGTCTTAAACTGAACAAAATAGATCTAAACTATGACAATAAAGTCTTAAA CTAGACAGAATAGTTGTAAACTGAAATCAGTCCAGTTATGCTGTGAAAAAGCATACT GGACTTTTGTTATGGCTAAAGCAAACTCTTCATTTTCTGAAGTGCAAATTGCCCGTCG TATTAAAGAGGGGCGTGGCCAAGGGCATGGTAAAGACTATATTCGCGGCGTTGTGA CAATTTACCGAACAACTCCGCGGCCGGGAAGCCGATCTCGGCTTGAACGAATTGTTA GGTGGCGGTACTTGGGTCGATATCAAAGTGCATCACTTCTTCCCGTATGCCCAACTT TGTATAGAGAGCCACTGCGGGATCGTCACCGTAATCTGCTTGCACGTAGATCACATA AGCACCAAGCGCGTTGGCCTCATGCTTGAGGAGATTGATGAGCGCGGTGGCAATGC CCTGCCTCCGGTGCTCGCCGGAGACTGCGAGATCATAGATATAGATCTCACTACGCG GCTGCTCAAACCTGGGCAGAACGTAAGCCGCGAGAGCGCCAACAACCGCTTCTTGG TCGAAGGCAGCAAGCGCGATGAATGTCTTACTACGGAGCAAGTTCCCGAGGTAATC - 90 - 12961406v1Attorney Docket No.2014202-0027 GGAGTCCGGCTGATGTTGGGAGTAGGTGGCTACGTCTCCGAACTCACGACCGAAAA GATCAAGAGCAGCCCGCATGGATTTGACTTGGTCAGGGCCGAGCCTACATGTGCGA ATGATGCCCATACTTGAGCCACCTAACTTTGTTTTAGGGCGACTGCCCTGCTGCGTA ACATCGTTGCTGCTGCGTAACATCGTTGCTGCTCCATAACATCAAACATCGACCCAC GGCGTAACGCGCTTGCTGCTTGGATGCCCGAGGCATAGACTGTACAAAAAAACAGT CATAACAAGCCATGAAAACCGCCACTGCGCCGTTACCACCGCTGCGTTCGGTCAAG GTTCTGGACCAGTTGCGTGAGCGCATACGCTACTTGCATTACAGTTTACGAACCGAA CAGGCTTATGTCAACTGGGCGGAATAATTGCCATATGTAAATGATGTCATCGTTCTA ACTCGCTTTACGAGTAGAATTCTACGTGTAAAACATAATCAAGAGATGATGTCATTT GTTTTTCAAAACTGAACTCAAGAAATGATGTCATTTGTTTTTCAAAACTGAACTGGC TTTACGAGTAGAATTCTACTTGTAACGCATGATCAAGGGATGATGTCATTGGATGAG TCATTTGTTTTTCAAAACTAAACTCGCTTTACGAGTAGAATTCTACTTGTAAAACACA ATCAAGGGATGATGTCATTATACAAATGATGTCATTTGTTTTTCAAAACTAAACTCG CTTTACGGGTAGAATTCTACTTGTAAAACACAATCGAGGGATGATGTCATCCTTTAC ACATGATTATAAACGTGTTTATGTATGACTCATTTGTTTTTCAAAACTAAACTCGCTT TACGAGTAGAATTCTACTTGTAACGCACGATCAAGGGATGATGTCATTTATTTGTGC AAAGCTGATGTCATCTTTTGCACACGATTATAAACACAATCAAATAATGACTCATTT GTTTTCAAAACTGAACTCGCTTTACGAGTAGAATTCTACTTGTAAAACACAATCAAG GGATGATGTCATTTTAAAAATGATGTCATTTGTTTTTCAAAACTAAACTCGCTTTACG AGTAGAATTCTACGTGTAAAACACAATCAAGGGATGATGTCATTTACTAAAATAAA ATAATTATTTAAATAAAAATGTTTTTATTGTAAAATACACATTGATTACACGTGACG TATACTCCGGAATATTAATAGTACGTACCCGTAGTGGCTATGGCAGGGCTTGCCGCC CCGACGTTGGCTGCGAGCCCTGGGCCTTCACCCGAACTTGGGGGTTGGGGTGGGGA AAAGGAAGAAACGCGGGCGTATTGGTCCCAATGGGGTCTCGGTGGGGTATCGACAG AGTGCCAGCCCTGGGACCGAACCCCGCGTTTATGAACAAACGACCCAACACCCGTG CGTTTTATTCTGTCTTTTTATTGCCGTCATAGCGCGGGTTCCTTCCGGTATTGTCTCCT TCCGTGTTTCAGTTAGCCTCCCCCATCTCCCGGTACCGCATGCTCCTTCAGAGAGAGT GTCCTCGAGCCAATCTGAAACAATACCATCGGCAGCCATACCTGATTTAAATCATTT ATTGTTCAAAGATGCAGTCATCCAAATCCACATTGACCAGATCGCAGGCAGTGCAA GCGTCTGGCACCTTTCCCATGATATGATGAATGTAGCACAGTTTCTGATACGCCTTTT TGACGACAGAAACGGGTTGAGATTCTGACACGGGAAAGCACTCTAAACAGTCTTTC TGTCCGTGAGTGAAGCAGATATTTGAATTCTGATTCATTCTCTCGCATTGTCTGCAGG GAAACAGCATCAGATTCATGCCCACGTGACGAGAACATTTGTTTTGGTACCTGTCTG CGTAGTTGATCGAAGCTTCCGCGTCTGACGTCGATGGCTGCGCAACTGACTCGCGCA CCCGTTTGGGCTCACTTATATCTGCGTCACTGGGGGCGGGTCTTTTCTTGGCTCCACC CTTTTTGACGTAGAATTCATGCTCCACCTCAACCACGTGATCCTTTGCCCACCGGAA AAAGTCTTTGACTTCCTGCTTGGTGACCTTCCCAAAGTCATGATCCAGACGGCGGGT GAGTTCAAATTTGAACATCCGGTCTTGCAACGGCTGCTGGTGTTCGAAGGTCGTTGA GTTCCCGTCAATCACGGCGCACATGTTGGTGTTGGAGGTGACGATCACGGGAGTCGG GTCTATCTGGGCCGAGGACTTGCATTTCTGGTCCACGCGCACCTTGCTTCCTCCGAG - 91 - 12961406v1Attorney Docket No.2014202-0027 AATGGCTTTGGCCGACTCCACGACCTTGGCGGTCATCTTCCCCTCCTCCCACCAGAT CACCATCTTGTCGACACAGTCGTTGAAGGGAAAGTTCTCATTGGTCCAGTTTACGCA CCCGTAGAAGGGCACAGTGTGGGCTATGGCCTCCGCGATGTTGGTCTTCCCGGTAGT TGCAGGCCCAAACAGCCAGATGGTGTTCCTCTTGCCGAACTTTTTCGTGGCCCATCC CAGAAAGACGGAAGCCGCATATTGGGGATCGTACCCGTTTAGTTCCAAAATTTTATA AATCCGATTGCTGGAAATGTCCTCCACGGGCTGCTGGCCCACCAGGTAGTCGGGGG CGGTTTTAGTCAGGCTCATAATCTTTCCCGCATTGTCCAAGGCAGCCTTGATTTGGGA CCGCGAGTTGGAGGCCGCATTGAAGGAGATGTATGAGGCCTGGTCCTCCTGGATCC ACTGCTTCTCCGAGGTAATCCCCTTGTCCACGAGCCACCCGACCAGCTCCATGTACC TGGCTGAAGTTTTTGATCTGATCACCGGCGCATCAGAATTGGGATTCTGATTCTCTTT GTTCTGCTCCTGCGTCTGCGACACGTGCGTCAGATGCTGCGCCACCAACCGTTTACG CTCCGTGAGATTCAAACAGGCGCTGAAACAATAGGAAGGGAGTGGATGTCAGTGTG TGCTGCCCGGGGGCTCTGACTACAGGTCTCCCCCTTCGCGCCCGATGGTGGGACGGT ATGAATAATCCGGAATATTTATAGGTTTTTTTATTACAAAACTGTTACGAAAACAGT AAAATACTTATTTATTTGCGAGATGGTTATCATTTTAATTATCTCCATGATAGATCTC TATCACTGATAGGGAGTACTTACCTTAAATACTGTTCCATATTAGTCCACGCCCACT GGAGCTCAGGCTGGGTTTTGGGGAGCAAGTAATTGGGGATGTAGCACTCATCCACC ACCTTGTTCCCGCCTCCGGCGCCATTTCTGGTCTTTGTGACCGCGAACCAGTTTGGCA AAGTCGGCTCGATCCCGCGGTAAATTCTCTGAATCAGTTTTTCGCGAATCTGACTCA GGAAACGTCCCAAAACCATGGATTTCACCCCGGTGGTTTCCACGAGCACGTGCATGT GGAAGTAGCTCTCTCCCTTCTCAAATTGCACAAAGAAAAGGGCCTCCGGGGCCTTAC TCACACGGCGCCATTCCGTCAGAAAGTCGCGCTGCAGCTTCTCGGCCACGGTCAGGG GTGCCTGCTCAATCAGATTCAGATCCATGTCAGAATCTGGCGGCAACTCCCATTCCT TCTCGGCCACCCAGTTCACAAAGCTGTCAGAAATGCCGGGCAGATGCTCGTCAAGG TCGCTGGGGACCTTAATCACAATCTCGTAAAACCCCGGCATGGCGGGTAGGGTGATC AAGTCTTCGTCGAGTGATTGTAAATAAAATGTAATTTACAGTATAGTATTTTAATTA ATATACAAATGATTTGATAATAATTCTTATTTAACTATAATATATTGTGTTGGGTTGA ATTAAAGGTCCGTAGCTTTCGAATCTAGGCTCAAGCAGTGATCAGATCCAGACATGA TAAGATACATTGATGAGTTTGGACAAACCACAACTAGAATGCAGTGAAAAAAATGC TTTATTTGTGAAATTTGTGATGCTATTGCTTTATTTGTAACCATTATAAGCTGCAATA AACAAGTTAACAACAACAATTGCATTCATTTTATGTTTCAGGTTCAGGGGGAGGTGT GGGAGGTTTTTTAAAGCAAGTAAAACCTCTACAAATGTGGTATGGCTGATTATGATC CTCTAGTACTTCTCGACAAGCTTGTCGAGACTGCAGGCTCTAGAGTCGACCGCTTAC AGGGGACGGGTGAGGTAACGGGTGCCAATGGGGCGAGGCTCAGTATAAAGTCCATT GTTGTCCACAGTGAAATCAACGTTGGCAGATTTTGCATAGTTAGATGTATACTGCAC TTCGGGATTCCAGCGTTTGCTGTTTTCTTTCTGCAGCTCCCATTCAATCTCCACGCTC ACTTGTCCTGTGGAATACTGGGTGATGAATGAAGCAAACTTTGTAGCCGAAAACTCT GCCGGAGGATTCGCAGGAACAGGCGTGTTTTTGATGAGGATCTGAGGAGGCGGGTG CTTAAGTCCAAAGCCGCCCATGAGAGGAGACGGGTGAAAGTGTCCATCCGTGTGAG GAATTTTGGCCCAAATAGGACCCTGCAGGTATACGTCTCTGTCTTGCCACACCATTC - 92 - 12961406v1Attorney Docket No.2014202-0027 CAGGTAAGGCTCCCATAACATGCACATCTCCGGTCGCAGGGTCTGTGCTGCTGCTCT GGAGATTGACTGCCACAGTCCCAAATCTTTCGGTGGCCACGGGGTTAGTGGCTTTGA TTTCCTCTTCGTCTGTGATCATGACATTGTCCAATGCAGTGTTTGAAGCTCCGGCGCT CTCCTTTCCAAAAATCATGACACCGCTCATGGGAAAGAACTTGTCTTTGTCGTCTTTG TGTGAGGCCATAGCAGTGCCAGGGTTGATTATAGATTCACGCCCATTAAGGTTATAT TTTGAAGCACCAGTCCAGGTAAAGTTGCTGTTGTTGTTGTCTGTTTTTGTTTTAGAAA CGCGCTGCTGCCGGTAACAGGGTCCAGGTAGCCAGTTTTTGGGCTGAACAGACATGC CAGCTGGAGACCCCCGGCTAAACAGCAAGTCCTTGTTTTGGGCACTTCCGGACTGAT TCTGAGTTCTGTTCAGGTAATACAGGTACTGGTCGATGAGAGGATTCATCAGCCGGT CCAGGCTCTGGCTGTGCGCGTAGCTGCTGTGGAAAGGCACGTCCTCGAAGGTGTAGC TGAAGGTAAAGTTATTGCCCGTTCTCAGCATCTGCGATGGGAAATATTCCAGGCAGT AAAAGGATGACCGTCCCACTGCCTGGCTGCCATTGTTGAGCGTTAGGTAGCCGTACT GCGGAATCATGAACACGTCCGCCGGGAACGGAGGGAGGCAGCCCTGGTGCGCAGA GCCGAGGACGTACGGCAACTGGTACTCCGAGTCCGAGAAGACTTGAACCGTGCTGG TAAGGTTATTAGCGATGGTCGTGACGCCATCATTCGTCGTGACCTCCTTGACTTGGA TGTTGAAGAGCTTGAAGTTGAGTCTCTTGGGCCGGAATCCCCAATTGTTGTTGATGA GTCGCTGCCAGTCACGTGGTGAGAAATGGCAGTGGAATCTGTTGAAATCAAAATAC CCCCAGGGGGTGCTGTAGCCGAAGTAGTGGTTGTCGTTGCTGGCCCCCGTTGAAGCA CTGGAGATTTGCTTGTAGAGGTGGTTGTTATAGGTGGGCAAGGCCCATGTTCGGGTG CTGGTGGTGATGACTCTGTCGCCCAGCCATGTGGAATCGCAATGCCAATTTCCTGAG GCATTACCCACTCCGTCGGCGCCTTCGTTATTGTCTGCCATTGGTGCGCCACCGCCTG AAGCCATTGTAGTAGGTCCCACAGCAGCGGGGGTTGCTGGAGGTTCTCCGAGAGGT TGTGGGTCGGGGACTGACTCTGAGTCGCCAGTCTGACCAAAATTGAGTCTCTTTTTA GCGGGCTGCTGGCCTGTCTTGCCAATGCCCGAGGAGGAGTCTGGCTCTTGTGGCGAC TGCTCTACCGGACGTTTCTTTCCAGGAGCTGTCTTAGCACCTTCCTCAACCAGACCA AAAGGTTCGAGAACCCTCTTCTTGGCCTGGAAGACTGCTCGCCCGAGGTTGCCCCCA AAAGACGTATCTTCTTGCAGACGCTCCTGAAACTCGGCGTCGGCGTGGTTATACCGC AGGTACGGATTGTCACCCGCTTTGAGCTGCTGGTCGTAGGCCTTGTCGTGCTCGAGG GCCGCTGCATCCGCCGCGTTGACGGGCTCCCCCTTGTCGAGTCCGTTGAAGGGTCCG AGGTACTTGTAGCCAGGAAGCACCAGACCCCGGCCGTCGTCCTGCTTTTGCTGGTTG GCTTTGGGTTTCGGGGCTCCAGGTTTCAAGTCCCACCACTCGCGAATGCCCTCAGAG AGGTTGTCCTCGAGCCAATCTGAAACAATAGGAAGGGAGTGGATGTCAGTGTGTGC TGCCCGGGGGCTCTGACTACAGGTCTCCCCCTTCGCGCCCGATGGTGGGACGGTATG AATAATCCGGAATATTTATAGGTTTTTTTATTACAAAACTGTTACGAAAACAGTAAA ATACTTATTTATTTGCGAGATGGTTATCATTTTAATTATCTCCATGATAGATCTCTAT CACTGATAGGGAGTACTTACCTGGAAGATAACCATCGGCAGCCATGGTGGCGGATC CGCGCCCGATGGTGGGACGGTATGAATAATCCGGAATATTTATAGGTTTTTTTATTA CAAAACTGTTACGAAAACAGTAAAATACTTATTTATTTGCGAGATGGTTATCATTTT AATTATCTCCATGATCTATTAATATTCCGGAGTATACCTAGGAGATCCGAACCAGAT AAGTGAAATCTAGTTCCAAACTATTTTGTCATTTTTAATTTTCGTATTAGCTTACGAC - 93 - 12961406v1Attorney Docket No.2014202-0027 GCTACACCCAGTTCCCATCTATTTTGTCACTCTTCCCTAAATAATCCTTAAAAACTCC ATTTCCACCCCTCCCAGTTCCCAACTATTTTGTCCGCCCACAGCGGGGCATTTTTCTT CCTGTTATGTTTTTAATCAAACATCCTGCCAACTCCATGTGACAAACCGTCATCTTCG GCTACTTT SEQ ID NO: 011 V683 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCGCCACCATGGCCCTCCCTGTCACCGCCCTGCTGCTTC CGCTGGCTCTTCTGCTCCACGCCGCTCGGCCCGAAATTGTGATGACCCAGTCACCCG CCACTCTTAGCCTTTCACCCGGTGAGCGCGCAACCCTGTCTTGCAGAGCCTCCCAAG ACATCTCAAAATACCTTAATTGGTATCAACAGAAGCCCGGACAGGCTCCTCGCCTTC TGATCTACCACACCAGCCGGCTCCATTCTGGAATCCCTGCCAGGTTCAGCGGTAGCG GATCTGGGACCGACTACACCCTCACTATCAGCTCACTGCAGCCAGAGGACTTCGCTG TCTATTTCTGTCAGCAAGGGAACACCCTGCCCTACACCTTTGGACAGGGCACCAAGC TCGAGATTAAAGGTGGAGGTGGCAGCGGAGGAGGTGGGTCCGGCGGTGGAGGAAG CCAGGTCCAACTCCAAGAAAGCGGACCGGGTCTTGTGAAGCCATCAGAAACTCTTT CACTGACTTGTACTGTGAGCGGAGTGTCTCTCCCCGATTACGGGGTGTCTTGGATCA GACAGCCACCGGGGAAGGGTCTGGAATGGATTGGAGTGATTTGGGGCTCTGAGACT ACTTACTACTCTTCATCCCTCAAGTCACGCGTCACCATCTCAAAGGACAACTCTAAG AATCAGGTGTCACTGAAACTGTCATCTGTGACCGCAGCCGACACCGCCGTGTACTAT TGCGCTAAGCATTACTATTATGGCGGGAGCTACGCAATGGATTACTGGGGACAGGG TACTCTGGTCACCGTGTCCAGCGGCGGTGGAGGAAGCGGTGGAGGTGGCAGCGGTG GAGGTGGCAGCACGGCTCCTGGAAAGAAACGTCCGGTAGAGCAGTCGCCACAAGA GCCAGACTCCTCCTCGGGCATTGGCAAGACAGGCCAGCAGCCCGCTAAAAAGAGAC TCAATTTTGGTCAGACTGGCGACTCAGAGTCAGTCCCCGACCCACAACCTCTCGGAG AACCTCCAGCAACCCCCGCTGCTGTGGGACCTACTACAATGGCTTCAGGCGGTGGCG CACCAATGGCAGACAATAACGAAGGCGCCGACGGAGTGGGTAATGCCTCAGGAAAT TGGCATTGCGATTCCACATGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAAC ATGGGCCTTGCCCACCTATAACAACCACCTCTACAAGCAAATCTCCAGTGCTTCAAC GGGGGCCAGCAACGACAACCACTACTTCGGCTACAGCACCCCCTGGGGGTATTTTG ATTTCAACAGATTCCACTGCCATTTCTCACCACGTGACTGGCAGCGACTCATCAACA ACAATTGGGGATTCCGGCCCAAGAGACTCAACTTCAAGCTCTTCAACATCCAAGTCA AGGAGGTCACGACGAATGATGGCGTCACGACCATCGCTAATAACCTTACCAGCACG GTTCAAGTCTTCTCGGACTCGGAGTACCAGTTGCCGTACGTCCTCGGCTCTGCGCAC - 94 - 12961406v1Attorney Docket No.2014202-0027 CAGGGCTGCCTCCCTCCGTTCCCGGCGGACGTGTTCATGATTCCGCAGTACGGCTAC CTAACGCTCAACAATGGCAGCCAGGCAGTGGGACGGTCATCCTTTTACTGCCTGGAA TATTTCCCATCGCAGATGCTGAGAACGGGCAATAACTTTACCTTCAGCTACACCTTC GAGGACGTGCCTTTCCACAGCAGCTACGCGCACAGCCAGAGCCTGGACCGGCTGAT GAATCCTCTCATCGACCAGTACCTGTATTACCTGAACAGAACTCAGAATCAGTCCGG AAGTGCCCAAAACAAGGACTTGCTGTTTAGCCGGGGGTCTCCAGCTGGCATGTCTGT TCAGCCCAAAAACTGGCTACCTGGACCCTGTTACCGGCAGCAGCGCGTTTCTAAAAC AAAAACAGACAACAACAACAGCAACTTTACCTGGACTGGTGCTTCAAAATATAACC TTAATGGGCGTGAATCTATAATCAACCCTGGCACTGCTATGGCCTCACACAAAGACG ACAAAGACAAGTTCTTTCCCATGAGCGGTGTCATGATTTTTGGAAAGGAGAGCGCCG GAGCTTCAAACACTGCATTGGACAATGTCATGATCACAGACGAAGAGGAAATCAAA GCCACTAACCCCGTGGCCACCGAAAGATTTGGGACTGTGGCAGTCAATCTCCAGAG CAGCAGCACAGACCCTGCGACCGGAGATGTGCATGTTATGGGAGCCTTACCTGGAA TGGTGTGGCAAGACAGAGACGTATACCTGCAGGGTCCTATTTGGGCCAAAATTCCTC ACACGGATGGACACTTTCACCCGTCTCCTCTCATGGGCGGCTTTGGACTTAAGCACC CGCCTCCTCAGATCCTCATCAAAAACACGCCTGTTCCTGCGAATCCTCCGGCAGAGT TTTCGGCTACAAAGTTTGCTTCATTCATCACCCAGTATTCCACAGGACAAGTGAGCG TGGAGATTGAATGGGAGCTGCAGAAAGAAAACAGCAAACGCTGGAATCCCGAAGT GCAGTATACATCTAACTATGCAAAATCTGCCAACGTTGATTTCACTGTGGACAACAA TGGACTTTATACTGAGCCTCGCCCCATTGGCACCCGTTACCTCACCCGTCCCCTGTAA GCATGCGGTACCGGGAGATGGGGGAGGCTAACTGAAACACGGAAGGAGACAATAC CGGAAGGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAACGCACGGGTG TTGGGTCGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCACTCTGTCGATA CCCCACCGAGACCCCATTGGGACCAATACGCCCGCGTTTCTTCCTTTTCCCCACCCC AACCCCCAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGGGCGGCAAGC CCTGCCATAGCCACTACGGGTACGTAGGCCAACCACTAGAACTATAGCTAGAGTCCT GGGCGAACAAACGATGCTCGCCTTCCAGAAAACCGAGGATGCGAACCACTTCATCC GGGGTCAGCACCACCGGCAAGCGCCGCGACGGCCGAGGTCTACCGATCTCCTGAAG CCAGGGCAGATCCGTGCACAGCACCTTGCCGTAGAAGAACAGCAAGGCCGCCAATG CCTGACGATGCGTGGAGACCGAAACCTTGCGCTCGTTCGCCAGCCAGGACAGAAAT GCCTCGACTTCGCTGCTGCCCAAGGTTGCCGGGTGACGCACACCGTGGAAACGGAT GAAGGCACGAACCCAGTTGACATAAGCCTGTTCGGTTCGTAAACTGTAATGCAAGT AGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAAC GGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGTACAGTCTATGCCTC GGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGC AGCAACGATGTTACGCAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAA CAAAGTTAGGTGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACC AAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAG CCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTA AGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGG - 95 - 12961406v1Attorney Docket No.2014202-0027 CTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCG CAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCA AGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTG ACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCAC TTTGATATCGACCCAAGTACCGCCACCTAACAATTCGTTCAAGCCGAGATCGGCTTC CCGGCCGCGGAGTTGTTCGGTAAATTGTCACAACGCCGCGAATATAGTCTTTACCAT GCCCTTGGCCACGCCCCTCTTTAATACGACGGGCAATTTGCACTTCAGAAAATGAAG AGTTTGCTTTAGCCATAACAAAAGTCCAGTATGCTTTTTCACAGCATAACTGGACTG ATTTCAGTTTACAACTATTCTGTCTAGTTTAAGACTTTATTGTCATAGTTTAGATCTA TTTTGTTCAGTTTAAGACTTTATTGTCCGCCCACACCCGCTTACGCAGGGCATCCATT TATTACTCAACCGTAACCGATTTTGCCAGGTTACGCGGCTGGTCTGCGGTGTGAAAT ACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCT CACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAA GGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAG CAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTC CATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTG GCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCG TGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTC GGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGT CGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGC CTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACT GGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAG AGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCT GCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCA AACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCA GAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGT GGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTC ACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAG TAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATC TGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATAC GGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCA CCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAG TGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGA GTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATC GTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAA GGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTC CGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCAC TGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTA CTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGC GTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGG - 96 - 12961406v1Attorney Docket No.2014202-0027 AAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTC GATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTT TCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGA CACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCA GGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAAT AGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGAAATTGTAAACGTTAATAT TTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCC GAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGT TGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAG GGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCA AGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCC CCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAG AAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGT AACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATT CAGGCTGCAAATAAGCGTTGATATTCAGTCAATTACAAACATTAATAACGAAGAGA TGACAGAAAAATTTTCATTCTGTGACAGAGAA SEQ ID NO: 012 CD3-scFv DNA sequence CCGTCCCACCATCGGGCGCGGATCCGCCACCATGCAGGTCCAGCTGCAGCAGTCTG GTGCTGAATTGGCTCGTCCTGGTGCTTCCGTGAAGATGTCCTGCAAGACCTCCGGTT ACACCTTCACTCGTTACACCATGCACTGGGTCAAGCAGCGTCCTGGACAAGGCCTGG AATGGATCGGTTACATCAACCCCTCCAGGGGTTACACCAACTACAACCAGAAGTTC AAGGACAAGGCTACCCTGACTACCGACAAGTCCTCCTCCACCGCTTACATGCAGCTG TCCTCACTGACCTCCGAGGACTCCGCTGTGTACTACTGCGCTCGTTACTACGACGAC CACTACTCCCTGGACTACTGGGGCCAGGGTACTACTCTGACCGTGTCCTCCGGTTCT ACCTCTGGTTCCGGAAAGCCTGGTTCTGGCGAGGGTTCCACCAAGGGACAGATCGTG CTGACCCAGTCTCCTGCTATCATGTCCGCTTCTCCCGGCGAGAAAGTGACTATGACC TGCCGTGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGTCCGGCACC TCTCCTAAGCGTTGGATCTACGACACCTCCAAGGTGGCCTCTGGTGTCCCCTACCGT TTCTCCGGATCAGGTTCCGGTACTTCCTACTCTCTGACCATCTCCTCTATGGAAGCTG AGGACGCCGCTACCTACTACTGCCAGCAGTGGTCTAGCAACCCTCTGACCTTCGGTG CTGGCACCAAGCTGGAACTGAAGGGCGGTGGAGGAAGCGGTGG SEQ ID NO: 013 V685 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT - 97 - 12961406v1Attorney Docket No.2014202-0027 TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCGCCACCATGCAGGTCCAGCTGCAGCAGTCTGGTGCT GAATTGGCTCGTCCTGGTGCTTCCGTGAAGATGTCCTGCAAGACCTCCGGTTACACC TTCACTCGTTACACCATGCACTGGGTCAAGCAGCGTCCTGGACAAGGCCTGGAATGG ATCGGTTACATCAACCCCTCCAGGGGTTACACCAACTACAACCAGAAGTTCAAGGA CAAGGCTACCCTGACTACCGACAAGTCCTCCTCCACCGCTTACATGCAGCTGTCCTC ACTGACCTCCGAGGACTCCGCTGTGTACTACTGCGCTCGTTACTACGACGACCACTA CTCCCTGGACTACTGGGGCCAGGGTACTACTCTGACCGTGTCCTCCGGTTCTACCTCT GGTTCCGGAAAGCCTGGTTCTGGCGAGGGTTCCACCAAGGGACAGATCGTGCTGAC CCAGTCTCCTGCTATCATGTCCGCTTCTCCCGGCGAGAAAGTGACTATGACCTGCCG TGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGTCCGGCACCTCTCCT AAGCGTTGGATCTACGACACCTCCAAGGTGGCCTCTGGTGTCCCCTACCGTTTCTCC GGATCAGGTTCCGGTACTTCCTACTCTCTGACCATCTCCTCTATGGAAGCTGAGGAC GCCGCTACCTACTACTGCCAGCAGTGGTCTAGCAACCCTCTGACCTTCGGTGCTGGC ACCAAGCTGGAACTGAAGGGCGGTGGAGGAAGCGGTGGAGGTGGCAGCGGTGGAG GTGGCAGCACGGCTCCTGGAAAGAAACGTCCGGTAGAGCAGTCGCCACAAGAGCCA GACTCCTCCTCGGGCATTGGCAAGACAGGCCAGCAGCCCGCTAAAAAGAGACTCAA TTTTGGTCAGACTGGCGACTCAGAGTCAGTCCCCGACCCACAACCTCTCGGAGAACC TCCAGCAACCCCCGCTGCTGTGGGACCTACTACAATGGCTTCAGGCGGTGGCGCACC AATGGCAGACAATAACGAAGGCGCCGACGGAGTGGGTAATGCCTCAGGAAATTGGC ATTGCGATTCCACATGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAACATGG GCCTTGCCCACCTATAACAACCACCTCTACAAGCAAATCTCCAGTGCTTCAACGGGG GCCAGCAACGACAACCACTACTTCGGCTACAGCACCCCCTGGGGGTATTTTGATTTC AACAGATTCCACTGCCATTTCTCACCACGTGACTGGCAGCGACTCATCAACAACAAT TGGGGATTCCGGCCCAAGAGACTCAACTTCAAGCTCTTCAACATCCAAGTCAAGGA GGTCACGACGAATGATGGCGTCACGACCATCGCTAATAACCTTACCAGCACGGTTC AAGTCTTCTCGGACTCGGAGTACCAGTTGCCGTACGTCCTCGGCTCTGCGCACCAGG GCTGCCTCCCTCCGTTCCCGGCGGACGTGTTCATGATTCCGCAGTACGGCTACCTAA CGCTCAACAATGGCAGCCAGGCAGTGGGACGGTCATCCTTTTACTGCCTGGAATATT TCCCATCGCAGATGCTGAGAACGGGCAATAACTTTACCTTCAGCTACACCTTCGAGG ACGTGCCTTTCCACAGCAGCTACGCGCACAGCCAGAGCCTGGACCGGCTGATGAAT CCTCTCATCGACCAGTACCTGTATTACCTGAACAGAACTCAGAATCAGTCCGGAAGT GCCCAAAACAAGGACTTGCTGTTTAGCCGGGGGTCTCCAGCTGGCATGTCTGTTCAG CCCAAAAACTGGCTACCTGGACCCTGTTACCGGCAGCAGCGCGTTTCTAAAACAAA AACAGACAACAACAACAGCAACTTTACCTGGACTGGTGCTTCAAAATATAACCTTA ATGGGCGTGAATCTATAATCAACCCTGGCACTGCTATGGCCTCACACAAAGACGAC AAAGACAAGTTCTTTCCCATGAGCGGTGTCATGATTTTTGGAAAGGAGAGCGCCGG AGCTTCAAACACTGCATTGGACAATGTCATGATCACAGACGAAGAGGAAATCAAAG CCACTAACCCCGTGGCCACCGAAAGATTTGGGACTGTGGCAGTCAATCTCCAGAGC AGCAGCACAGACCCTGCGACCGGAGATGTGCATGTTATGGGAGCCTTACCTGGAAT - 98 - 12961406v1Attorney Docket No.2014202-0027 GGTGTGGCAAGACAGAGACGTATACCTGCAGGGTCCTATTTGGGCCAAAATTCCTCA CACGGATGGACACTTTCACCCGTCTCCTCTCATGGGCGGCTTTGGACTTAAGCACCC GCCTCCTCAGATCCTCATCAAAAACACGCCTGTTCCTGCGAATCCTCCGGCAGAGTT TTCGGCTACAAAGTTTGCTTCATTCATCACCCAGTATTCCACAGGACAAGTGAGCGT GGAGATTGAATGGGAGCTGCAGAAAGAAAACAGCAAACGCTGGAATCCCGAAGTG CAGTATACATCTAACTATGCAAAATCTGCCAACGTTGATTTCACTGTGGACAACAAT GGACTTTATACTGAGCCTCGCCCCATTGGCACCCGTTACCTCACCCGTCCCCTGTAA GCATGCGGTACCGGGAGATGGGGGAGGCTAACTGAAACACGGAAGGAGACAATAC CGGAAGGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAACGCACGGGTG TTGGGTCGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCACTCTGTCGATA CCCCACCGAGACCCCATTGGGACCAATACGCCCGCGTTTCTTCCTTTTCCCCACCCC AACCCCCAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGGGCGGCAAGC CCTGCCATAGCCACTACGGGTACGTAGGCCAACCACTAGAACTATAGCTAGAGTCCT GGGCGAACAAACGATGCTCGCCTTCCAGAAAACCGAGGATGCGAACCACTTCATCC GGGGTCAGCACCACCGGCAAGCGCCGCGACGGCCGAGGTCTACCGATCTCCTGAAG CCAGGGCAGATCCGTGCACAGCACCTTGCCGTAGAAGAACAGCAAGGCCGCCAATG CCTGACGATGCGTGGAGACCGAAACCTTGCGCTCGTTCGCCAGCCAGGACAGAAAT GCCTCGACTTCGCTGCTGCCCAAGGTTGCCGGGTGACGCACACCGTGGAAACGGAT GAAGGCACGAACCCAGTTGACATAAGCCTGTTCGGTTCGTAAACTGTAATGCAAGT AGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAAC GGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGTACAGTCTATGCCTC GGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGC AGCAACGATGTTACGCAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAA CAAAGTTAGGTGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACC AAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAG CCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTA AGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGG CTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCG CAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCA AGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTG ACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCAC TTTGATATCGACCCAAGTACCGCCACCTAACAATTCGTTCAAGCCGAGATCGGCTTC CCGGCCGCGGAGTTGTTCGGTAAATTGTCACAACGCCGCGAATATAGTCTTTACCAT GCCCTTGGCCACGCCCCTCTTTAATACGACGGGCAATTTGCACTTCAGAAAATGAAG AGTTTGCTTTAGCCATAACAAAAGTCCAGTATGCTTTTTCACAGCATAACTGGACTG ATTTCAGTTTACAACTATTCTGTCTAGTTTAAGACTTTATTGTCATAGTTTAGATCTA TTTTGTTCAGTTTAAGACTTTATTGTCCGCCCACACCCGCTTACGCAGGGCATCCATT TATTACTCAACCGTAACCGATTTTGCCAGGTTACGCGGCTGGTCTGCGGTGTGAAAT ACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCT CACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAA - 99 - 12961406v1Attorney Docket No.2014202-0027 GGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAG CAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTC CATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTG GCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCG TGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTC GGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGT CGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGC CTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACT GGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAG AGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCT GCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCA AACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCA GAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGT GGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTC ACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAG TAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATC TGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATAC GGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCA CCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAG TGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGA GTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATC GTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAA GGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTC CGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCAC TGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTA CTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGC GTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGG AAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTC GATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTT TCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGA CACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCA GGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAAT AGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGAAATTGTAAACGTTAATAT TTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCC GAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGT TGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAG GGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCA AGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCC CCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAG AAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGT - 100 - 12961406v1Attorney Docket No.2014202-0027 AACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATT CAGGCTGCAAATAAGCGTTGATATTCAGTCAATTACAAACATTAATAACGAAGAGA TGACAGAAAAATTTTCATTCTGTGACAGAGAA SEQ ID NO: 014 CD3scFv-3’-partial Cap6VP2-HSVtkpA AGTCAATCTCCAGAGCAGCAGCGGCGGTGGAGGAAGCGGTGGAGGTGGCAGCGGT GGAGGTGGCAGCCAGGTCCAGCTGCAGCAGTCTGGTGCTGAATTGGCTCGTCCTGGT GCTTCCGTGAAGATGTCCTGCAAGACCTCCGGTTACACCTTCACTCGTTACACCATG CACTGGGTCAAGCAGCGTCCTGGACAAGGCCTGGAATGGATCGGTTACATCAACCC CTCCAGGGGTTACACCAACTACAACCAGAAGTTCAAGGACAAGGCTACCCTGACTA CCGACAAGTCCTCCTCCACCGCTTACATGCAGCTGTCCTCACTGACCTCCGAGGACT CCGCTGTGTACTACTGCGCTCGTTACTACGACGACCACTACTCCCTGGACTACTGGG GCCAGGGTACTACTCTGACCGTGTCCTCCGGTTCTACCTCTGGTTCCGGAAAGCCTG GTTCTGGCGAGGGTTCCACCAAGGGACAGATCGTGCTGACCCAGTCTCCTGCTATCA TGTCCGCTTCTCCCGGCGAGAAAGTGACTATGACCTGCCGTGCTTCCTCCTCCGTGTC CTACATGAACTGGTATCAGCAGAAGTCCGGCACCTCTCCTAAGCGTTGGATCTACGA CACCTCCAAGGTGGCCTCTGGTGTCCCCTACCGTTTCTCCGGATCAGGTTCCGGTACT TCCTACTCTCTGACCATCTCCTCTATGGAAGCTGAGGACGCCGCTACCTACTACTGC CAGCAGTGGTCTAGCAACCCTCTGACCTTCGGTGCTGGCACCAAGCTGGAACTGAA GACAGACCCTGCGACCGGAGATGTGCATGTTATGGGAGCCTTACCTGGAATGGTGT GGCAAGACAGAGACGTATACCTGCAGGGTCCTATTTGGGCCAAAATTCCTCACACG GATGGACACTTTCACCCGTCTCCTCTCATGGGCGGCTTTGGACTTAAGCACCCGCCT CCTCAGATCCTCATCAAAAACACGCCTGTTCCTGCGAATCCTCCGGCAGAGTTTTCG GCTACAAAGTTTGCTTCATTCATCACCCAGTATTCCACAGGACAAGTGAGCGTGGAG ATTGAATGGGAGCTGCAGAAAGAAAACAGCAAACGCTGGAATCCCGAAGTGCAGTA TACATCTAACTATGCAAAATCTGCCAACGTTGATTTCACTGTGGACAACAATGGACT TTATACTGAGCCTCGCCCCATTGGCACCCGTTACCTCACCCGTCCCCTGTAAGCATG CGGTACCGGGAGATGGGGGAGGCTAACTGAAACACGGAAGGAGACAATACCGGAA GGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAACGCACGGGTGTTGGGT CGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCACTCTGTCGATACCCCAC CGAGACCCCATTGGGACCAATACGCCCGCGTTTCTTCCTTTTCCCCACCCCAACCCC CAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGGGCGGCAAGCCCTGCC ATAGCCACTACGGGTACGTACTATTAATATTCCGGAG SEQ ID NO: 015 5’-partial Cap6VP2 fragment ATCGGGCGCGGATCCGCCACCATGACGGCTCCTGGAAAGAAACGTCCGGTAGAGCA GTCGCCACAAGAGCCAGACTCCTCCTCGGGCATTGGCAAGACAGGCCAGCAGCCCG CTAAAAAGAGACTCAATTTTGGTCAGACTGGCGACTCAGAGTCAGTCCCCGACCCA CAACCTCTCGGAGAACCTCCAGCAACCCCCGCTGCTGTGGGACCTACTACAATGGCT TCAGGCGGTGGCGCACCAATGGCAGACAATAACGAAGGCGCCGACGGAGTGGGTA - 101 - 12961406v1Attorney Docket No.2014202-0027 ATGCCTCAGGAAATTGGCATTGCGATTCCACATGGCTGGGCGACAGAGTCATCACCA CCAGCACCCGAACATGGGCCTTGCCCACCTATAACAACCACCTCTACAAGCAAATCT CCAGTGCTTCAACGGGGGCCAGCAACGACAACCACTACTTCGGCTACAGCACCCCC TGGGGGTATTTTGATTTCAACAGATTCCACTGCCATTTCTCACCACGTGACTGGCAG CGACTCATCAACAACAATTGGGGATTCCGGCCCAAGAGACTCAACTTCAAGCTCTTC AACATCCAAGTCAAGGAGGTCACGACGAATGATGGCGTCACGACCATCGCTAATAA CCTTACCAGCACGGTTCAAGTCTTCTCGGACTCGGAGTACCAGTTGCCGTACGTCCT CGGCTCTGCGCACCAGGGCTGCCTCCCTCCGTTCCCGGCGGACGTGTTCATGATTCC GCAGTACGGCTACCTAACGCTCAACAATGGCAGCCAGGCAGTGGGACGGTCATCCT TTTACTGCCTGGAATATTTCCCATCGCAGATGCTGAGAACGGGCAATAACTTTACCT TCAGCTACACCTTCGAGGACGTGCCTTTCCACAGCAGCTACGCGCACAGCCAGAGCC TGGACCGGCTGATGAATCCTCTCATCGACCAGTACCTGTATTACCTGAACAGAACTC AGAATCAGTCCGGAAGTGCCCAAAACAAGGACTTGCTGTTTAGCCGGGGGTCTCCA GCTGGCATGTCTGTTCAGCCCAAAAACTGGCTACCTGGACCCTGTTACCGGCAGCAG CGCGTTTCTAAAACAAAAACAGACAACAACAACAGCAACTTTACCTGGACTGGTGC TTCAAAATATAACCTTAATGGGCGTGAATCTATAATCAACCCTGGCACTGCTATGGC CTCACACAAAGACGACAAAGACAAGTTCTTTCCCATGAGCGGTGTCATGATTTTTGG AAAGGAGAGCGCCGGAGCTTCAAACACTGCATTGGACAATGTCATGATCACAGACG AAGAGGAAATCAAAGCCACTAACCCCGTGGCCACCGAAAGATTTGGGACTGTGGCA GTCAATCTCCAGAGCAGCA SEQ ID NO: 016 Primer 9466 ATCGGGCGCGGATCCGCCACCATGACGGCTCCTGGAAAGAAACG SEQ ID NO: 017 Primer 9467 TGCTGCTCTGGAGATTGACT SEQ ID NO: 018 V689 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCGCCACCATGACGGCTCCTGGAAAGAAACGTCCGGTA GAGCAGTCGCCACAAGAGCCAGACTCCTCCTCGGGCATTGGCAAGACAGGCCAGCA GCCCGCTAAAAAGAGACTCAATTTTGGTCAGACTGGCGACTCAGAGTCAGTCCCCG ACCCACAACCTCTCGGAGAACCTCCAGCAACCCCCGCTGCTGTGGGACCTACTACAA TGGCTTCAGGCGGTGGCGCACCAATGGCAGACAATAACGAAGGCGCCGACGGAGTG - 102 - 12961406v1Attorney Docket No.2014202-0027 GGTAATGCCTCAGGAAATTGGCATTGCGATTCCACATGGCTGGGCGACAGAGTCATC ACCACCAGCACCCGAACATGGGCCTTGCCCACCTATAACAACCACCTCTACAAGCA AATCTCCAGTGCTTCAACGGGGGCCAGCAACGACAACCACTACTTCGGCTACAGCA CCCCCTGGGGGTATTTTGATTTCAACAGATTCCACTGCCATTTCTCACCACGTGACTG GCAGCGACTCATCAACAACAATTGGGGATTCCGGCCCAAGAGACTCAACTTCAAGC TCTTCAACATCCAAGTCAAGGAGGTCACGACGAATGATGGCGTCACGACCATCGCT AATAACCTTACCAGCACGGTTCAAGTCTTCTCGGACTCGGAGTACCAGTTGCCGTAC GTCCTCGGCTCTGCGCACCAGGGCTGCCTCCCTCCGTTCCCGGCGGACGTGTTCATG ATTCCGCAGTACGGCTACCTAACGCTCAACAATGGCAGCCAGGCAGTGGGACGGTC ATCCTTTTACTGCCTGGAATATTTCCCATCGCAGATGCTGAGAACGGGCAATAACTT TACCTTCAGCTACACCTTCGAGGACGTGCCTTTCCACAGCAGCTACGCGCACAGCCA GAGCCTGGACCGGCTGATGAATCCTCTCATCGACCAGTACCTGTATTACCTGAACAG AACTCAGAATCAGTCCGGAAGTGCCCAAAACAAGGACTTGCTGTTTAGCCGGGGGT CTCCAGCTGGCATGTCTGTTCAGCCCAAAAACTGGCTACCTGGACCCTGTTACCGGC AGCAGCGCGTTTCTAAAACAAAAACAGACAACAACAACAGCAACTTTACCTGGACT GGTGCTTCAAAATATAACCTTAATGGGCGTGAATCTATAATCAACCCTGGCACTGCT ATGGCCTCACACAAAGACGACAAAGACAAGTTCTTTCCCATGAGCGGTGTCATGATT TTTGGAAAGGAGAGCGCCGGAGCTTCAAACACTGCATTGGACAATGTCATGATCAC AGACGAAGAGGAAATCAAAGCCACTAACCCCGTGGCCACCGAAAGATTTGGGACTG TGGCAGTCAATCTCCAGAGCAGCAGCGGCGGTGGAGGAAGCGGTGGAGGTGGCAGC GGTGGAGGTGGCAGCCAGGTCCAGCTGCAGCAGTCTGGTGCTGAATTGGCTCGTCCT GGTGCTTCCGTGAAGATGTCCTGCAAGACCTCCGGTTACACCTTCACTCGTTACACC ATGCACTGGGTCAAGCAGCGTCCTGGACAAGGCCTGGAATGGATCGGTTACATCAA CCCCTCCAGGGGTTACACCAACTACAACCAGAAGTTCAAGGACAAGGCTACCCTGA CTACCGACAAGTCCTCCTCCACCGCTTACATGCAGCTGTCCTCACTGACCTCCGAGG ACTCCGCTGTGTACTACTGCGCTCGTTACTACGACGACCACTACTCCCTGGACTACT GGGGCCAGGGTACTACTCTGACCGTGTCCTCCGGTTCTACCTCTGGTTCCGGAAAGC CTGGTTCTGGCGAGGGTTCCACCAAGGGACAGATCGTGCTGACCCAGTCTCCTGCTA TCATGTCCGCTTCTCCCGGCGAGAAAGTGACTATGACCTGCCGTGCTTCCTCCTCCGT GTCCTACATGAACTGGTATCAGCAGAAGTCCGGCACCTCTCCTAAGCGTTGGATCTA CGACACCTCCAAGGTGGCCTCTGGTGTCCCCTACCGTTTCTCCGGATCAGGTTCCGG TACTTCCTACTCTCTGACCATCTCCTCTATGGAAGCTGAGGACGCCGCTACCTACTAC TGCCAGCAGTGGTCTAGCAACCCTCTGACCTTCGGTGCTGGCACCAAGCTGGAACTG AAGACAGACCCTGCGACCGGAGATGTGCATGTTATGGGAGCCTTACCTGGAATGGT GTGGCAAGACAGAGACGTATACCTGCAGGGTCCTATTTGGGCCAAAATTCCTCACAC GGATGGACACTTTCACCCGTCTCCTCTCATGGGCGGCTTTGGACTTAAGCACCCGCC TCCTCAGATCCTCATCAAAAACACGCCTGTTCCTGCGAATCCTCCGGCAGAGTTTTC GGCTACAAAGTTTGCTTCATTCATCACCCAGTATTCCACAGGACAAGTGAGCGTGGA GATTGAATGGGAGCTGCAGAAAGAAAACAGCAAACGCTGGAATCCCGAAGTGCAGT ATACATCTAACTATGCAAAATCTGCCAACGTTGATTTCACTGTGGACAACAATGGAC - 103 - 12961406v1Attorney Docket No.2014202-0027 TTTATACTGAGCCTCGCCCCATTGGCACCCGTTACCTCACCCGTCCCCTGTAAGCATG CGGTACCGGGAGATGGGGGAGGCTAACTGAAACACGGAAGGAGACAATACCGGAA GGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAACGCACGGGTGTTGGGT CGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCACTCTGTCGATACCCCAC CGAGACCCCATTGGGACCAATACGCCCGCGTTTCTTCCTTTTCCCCACCCCAACCCC CAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGGGCGGCAAGCCCTGCC ATAGCCACTACGGGTACGTACTATTAATATTCCGGAGTATACGTCACGTGTAATCAA TGTGTATTTTACAATAAAAACATTTTTATTTAAATAATTATTTTATTTTAGTAAATGA CATCATCCCTTGATTGTGTTTTACACGTAGAATTCTACTCGTAAAGCGAGTTTAGTTT TGAAAAACAAATGACATCATTTTTAAAATGACATCATCCCTTGATTGTGTTTTACAA GTAGAATTCTACTCGTAAAGCGAGTTCAGTTTTGAAAACAAATGAGTCATTATTTGA TTGTGTTTATAATCGTGTGCAAAAGATGACATCAGCTTTGCACAAATAAATGACATC ATCCCTTGATCGTGCGTTACAAGTAGAATTCTACTCGTAAAGCGAGTTTAGTTTTGA AAAACAAATGAGTCATACATAAACACGTTTATAATCATGTGTAAAGGATGACATCA TCCCTCGATTGTGTTTTACAAGTAGAATTCTACCCGTAAAGCGAGTTTAGTTTTGAAA AACAAATGACATCATTTGTATAATGACATCATCCCTTGATTGTGTTTTACAAGTAGA ATTCTACTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACTCATCCAATGACATC ATCCCTTGATCATGCGTTACAAGTAGAATTCTACTCGTAAAGCCAGTTCAGTTTTGA AAAACAAATGACATCATTTCTTGAGTTCAGTTTTGAAAAACAAATGACATCATCTCT TGATTATGTTTTACACGTAGAATTCTACTCGTAAAGCGAGTTAGAACGATGACATCA TTTACATATGGCAATTATTCCGCCCAGTTGACATAAGCCTGTTCGGTTCGTAAACTGT AATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAACGCAGC GGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGTACAGT CTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTTGATGT TATGGAGCAGCAACGATGTTACGCAGCAGCAACGATGTTACGCAGCAGGGCAGTCG CCCTAAAACAAAGTTAGGTGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGG CCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGG AGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCT CCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGC TCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTATATCTA TGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCA ATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAG ATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAA GTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAACAATTCGTTCAAGCCGAG ATCGGCTTCCCGGCCGCGGAGTTGTTCGGTAAATTGTCACAACGCCGCGAATATAGT CTTTACCATGCCCTTGGCCACGCCCCTCTTTAATACGACGGGCAATTTGCACTTCAG AAAATGAAGAGTTTGCTTTAGCCATAACAAAAGTCCAGTATGCTTTTTCACAGCATA ACTGGACTGATTTCAGTTTACAACTATTCTGTCTAGTTTAAGACTTTATTGTCATAGT TTAGATCTATTTTGTTCAGTTTAAGACTTTATTGTCCGCCCACACCCGCTTACGCAGG GCATCCATTTATTACTCAACCGTAACCGATTTTGCCAGGTTACGCGGCTGGTCTGCG - 104 - 12961406v1Attorney Docket No.2014202-0027 GTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCTTCCG CTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAG CTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAG AACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGC TGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAA GTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGA AGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCT TTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCAGTTC GGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGA CCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTT ATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCG GTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTAT TTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTT GATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGA TTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTG ACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAA AGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGT ATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATC TCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAA CTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGAC CCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGA GCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCG GGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGC TACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCC CAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCC TTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTT ATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGA CTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCT CTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTG CTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTG AGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTT TCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGG AATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTG AAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAA AAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGAAATTG TAAACGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTTTT TAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGAGA TAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGACT CCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAACCA TCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACCCT - 105 - 12961406v1Attorney Docket No.2014202-0027 AAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAA AGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGT CACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGT CCCATTCGCCATTCAGGCTGCAAATAAGCGTTGATATTCAGTCAATTACAAACATTA ATAACGAAGAGATGACAGAAAAATTTTCATTCTGTGACAGAGAA SEQ ID NO: 019 5’-Cap6VP3 DNA sequence CCGTCCCACCATCGGGCGCGGATCCGCCACCATGGCTTCAGGCGGTGGCGCACCAA TGGCAGACAATAACGAAGGCGCCGACGGAGTGGGTAATGCCTCAGGAAATTGGCAT TGCGATTCCACATGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAACATGGGC CTTGCCCACCTATAACAACCACCTCTACAAGCAAATCTCCAGTGCTTCAACGGGGGC CAGCAACGACAACCACTACTTCGGCTACAGCACCCCCTGGGGGTATTTTGATTTCAA CAGATTCCACTGCCATTTCTCACCACGTGACTGGCAGCGACTCATCAACAACAATTG GGGATTCCGGCCCAAGAGACTCAACTTCAAGCTCTTCAACATCCAAGTCAAGGAGG TCACGACGAATGATGGCGTCACGACCATCGCTAATAACCTTACCAGCACGGTTCAA GTCTTCTCGGACTCGGAGTACCAGTTGCCGTACGTCCTCGGCTCTGCGCACCA SEQ ID NO: 020 Primer 9482 CCGTCCCACCATCGGGCGCGGATCCGCCACCATGGCTTCAGGCGGTGGCG SEQ ID NO: 021 Primer 5388 TGGTGCGCAGAGCCGAGG SEQ ID NO: 022 V697 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCGCCACCATGGCTTCAGGCGGTGGCGCACCAATGGCA GACAATAACGAAGGCGCCGACGGAGTGGGTAATGCCTCAGGAAATTGGCATTGCGA TTCCACATGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAACATGGGCCTTGC CCACCTATAACAACCACCTCTACAAGCAAATCTCCAGTGCTTCAACGGGGGCCAGC AACGACAACCACTACTTCGGCTACAGCACCCCCTGGGGGTATTTTGATTTCAACAGA TTCCACTGCCATTTCTCACCACGTGACTGGCAGCGACTCATCAACAACAATTGGGGA TTCCGGCCCAAGAGACTCAACTTCAAGCTCTTCAACATCCAAGTCAAGGAGGTCACG ACGAATGATGGCGTCACGACCATCGCTAATAACCTTACCAGCACGGTTCAAGTCTTC - 106 - 12961406v1Attorney Docket No.2014202-0027 TCGGACTCGGAGTACCAGTTGCCGTACGTCCTCGGCTCTGCGCACCAGGGCTGCCTC CCTCCGTTCCCGGCGGACGTGTTCATGATTCCGCAGTACGGCTACCTAACGCTCAAC AATGGCAGCCAGGCAGTGGGACGGTCATCCTTTTACTGCCTGGAATATTTCCCATCG CAGATGCTGAGAACGGGCAATAACTTTACCTTCAGCTACACCTTCGAGGACGTGCCT TTCCACAGCAGCTACGCGCACAGCCAGAGCCTGGACCGGCTGATGAATCCTCTCATC GACCAGTACCTGTATTACCTGAACAGAACTCAGAATCAGTCCGGAAGTGCCCAAAA CAAGGACTTGCTGTTTAGCCGGGGGTCTCCAGCTGGCATGTCTGTTCAGCCCAAAAA CTGGCTACCTGGACCCTGTTACCGGCAGCAGCGCGTTTCTAAAACAAAAACAGACA ACAACAACAGCAACTTTACCTGGACTGGTGCTTCAAAATATAACCTTAATGGGCGTG AATCTATAATCAACCCTGGCACTGCTATGGCCTCACACAAAGACGACAAAGACAAG TTCTTTCCCATGAGCGGTGTCATGATTTTTGGAAAGGAGAGCGCCGGAGCTTCAAAC ACTGCATTGGACAATGTCATGATCACAGACGAAGAGGAAATCAAAGCCACTAACCC CGTGGCCACCGAAAGATTTGGGACTGTGGCAGTCAATCTCCAGAGCAGCAGCGGCG GTGGAGGAAGCGGTGGAGGTGGCAGCGGTGGAGGTGGCAGCCAGGTCCAGCTGCA GCAGTCTGGTGCTGAATTGGCTCGTCCTGGTGCTTCCGTGAAGATGTCCTGCAAGAC CTCCGGTTACACCTTCACTCGTTACACCATGCACTGGGTCAAGCAGCGTCCTGGACA AGGCCTGGAATGGATCGGTTACATCAACCCCTCCAGGGGTTACACCAACTACAACC AGAAGTTCAAGGACAAGGCTACCCTGACTACCGACAAGTCCTCCTCCACCGCTTACA TGCAGCTGTCCTCACTGACCTCCGAGGACTCCGCTGTGTACTACTGCGCTCGTTACT ACGACGACCACTACTCCCTGGACTACTGGGGCCAGGGTACTACTCTGACCGTGTCCT CCGGTTCTACCTCTGGTTCCGGAAAGCCTGGTTCTGGCGAGGGTTCCACCAAGGGAC AGATCGTGCTGACCCAGTCTCCTGCTATCATGTCCGCTTCTCCCGGCGAGAAAGTGA CTATGACCTGCCGTGCTTCCTCCTCCGTGTCCTACATGAACTGGTATCAGCAGAAGT CCGGCACCTCTCCTAAGCGTTGGATCTACGACACCTCCAAGGTGGCCTCTGGTGTCC CCTACCGTTTCTCCGGATCAGGTTCCGGTACTTCCTACTCTCTGACCATCTCCTCTAT GGAAGCTGAGGACGCCGCTACCTACTACTGCCAGCAGTGGTCTAGCAACCCTCTGA CCTTCGGTGCTGGCACCAAGCTGGAACTGAAGACAGACCCTGCGACCGGAGATGTG CATGTTATGGGAGCCTTACCTGGAATGGTGTGGCAAGACAGAGACGTATACCTGCA GGGTCCTATTTGGGCCAAAATTCCTCACACGGATGGACACTTTCACCCGTCTCCTCT CATGGGCGGCTTTGGACTTAAGCACCCGCCTCCTCAGATCCTCATCAAAAACACGCC TGTTCCTGCGAATCCTCCGGCAGAGTTTTCGGCTACAAAGTTTGCTTCATTCATCACC CAGTATTCCACAGGACAAGTGAGCGTGGAGATTGAATGGGAGCTGCAGAAAGAAAA CAGCAAACGCTGGAATCCCGAAGTGCAGTATACATCTAACTATGCAAAATCTGCCA ACGTTGATTTCACTGTGGACAACAATGGACTTTATACTGAGCCTCGCCCCATTGGCA CCCGTTACCTCACCCGTCCCCTGTAAGCATGCGGTACCGGGAGATGGGGGAGGCTA ACTGAAACACGGAAGGAGACAATACCGGAAGGAACCCGCGCTATGACGGCAATAA AAAGACAGAATAAAACGCACGGGTGTTGGGTCGTTTGTTCATAAACGCGGGGTTCG GTCCCAGGGCTGGCACTCTGTCGATACCCCACCGAGACCCCATTGGGACCAATACGC CCGCGTTTCTTCCTTTTCCCCACCCCAACCCCCAAGTTCGGGTGAAGGCCCAGGGCT CGCAGCCAACGTCGGGGCGGCAAGCCCTGCCATAGCCACTACGGGTACGTACTATT - 107 - 12961406v1Attorney Docket No.2014202-0027 AATATTCCGGAGTATACGTCACGTGTAATCAATGTGTATTTTACAATAAAAACATTT TTATTTAAATAATTATTTTATTTTAGTAAATGACATCATCCCTTGATTGTGTTTTACAC GTAGAATTCTACTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACATCATTTTTA AAATGACATCATCCCTTGATTGTGTTTTACAAGTAGAATTCTACTCGTAAAGCGAGT TCAGTTTTGAAAACAAATGAGTCATTATTTGATTGTGTTTATAATCGTGTGCAAAAG ATGACATCAGCTTTGCACAAATAAATGACATCATCCCTTGATCGTGCGTTACAAGTA GAATTCTACTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGAGTCATACATAAACA CGTTTATAATCATGTGTAAAGGATGACATCATCCCTCGATTGTGTTTTACAAGTAGA ATTCTACCCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACATCATTTGTATAATG ACATCATCCCTTGATTGTGTTTTACAAGTAGAATTCTACTCGTAAAGCGAGTTTAGTT TTGAAAAACAAATGACTCATCCAATGACATCATCCCTTGATCATGCGTTACAAGTAG AATTCTACTCGTAAAGCCAGTTCAGTTTTGAAAAACAAATGACATCATTTCTTGAGT TCAGTTTTGAAAAACAAATGACATCATCTCTTGATTATGTTTTACACGTAGAATTCTA CTCGTAAAGCGAGTTAGAACGATGACATCATTTACATATGGCAATTATTCCGCCCAG TTGACATAAGCCTGTTCGGTTCGTAAACTGTAATGCAAGTAGCGTATGCGCTCACGC AACTGGTCCAGAACCTTGACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTT CATGGCTTGTTATGACTGTTTTTTTGTACAGTCTATGCCTCGGGCATCCAAGCAGCAA GCGCGTTACGCCGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAG CAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGTGGCTCAA GTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGG CTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCA GCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGC TGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTT TGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCG GAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGC TTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTC TCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGT ACCGCCACCTAACAATTCGTTCAAGCCGAGATCGGCTTCCCGGCCGCGGAGTTGTTC GGTAAATTGTCACAACGCCGCGAATATAGTCTTTACCATGCCCTTGGCCACGCCCCT CTTTAATACGACGGGCAATTTGCACTTCAGAAAATGAAGAGTTTGCTTTAGCCATAA CAAAAGTCCAGTATGCTTTTTCACAGCATAACTGGACTGATTTCAGTTTACAACTATT CTGTCTAGTTTAAGACTTTATTGTCATAGTTTAGATCTATTTTGTTCAGTTTAAGACTT TATTGTCCGCCCACACCCGCTTACGCAGGGCATCCATTTATTACTCAACCGTAACCG ATTTTGCCAGGTTACGCGGCTGGTCTGCGGTGTGAAATACCGCACAGATGCGTAAGG AGAAAATACCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCG GTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCC ACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGG CCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTG ACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTA TAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACC - 108 - 12961406v1Attorney Docket No.2014202-0027 CTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTC AATGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCT GTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTC TTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAAC AGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCC TAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGT TACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTA GCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAG AAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTT AAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATT AAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTT ACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCAT AGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGG CCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGC AATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCG CCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTA ATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTT TGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCC CATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAA GTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTC ATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGA GAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACC GCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGA AAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCA CCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACA GGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATAC TCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAG CGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATT TCCCCGAAAAGTGCCACCTGAAATTGTAAACGTTAATATTTTGTTAAAATTCGCGTT AAATTTTTGTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCC TTATAAATCAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACA AGAGTCCACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTAT CAGGGCGATGGCCCACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGG TGCCGTAAAGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACG GGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGG CGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCACCACACCCGCCG CGCTTAATGCGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCAAATAAGCG TTGATATTCAGTCAATTACAAACATTAATAACGAAGAGATGACAGAAAAATTTTCAT TCTGTGACAGAGAA SEQ ID NO: 023 polh-VP2 fragment - 109 - 12961406v1Attorney Docket No.2014202-0027 CTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATAGATCATGGAGATA ATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGTTTTCGTAACAGTTT TGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTCCCACCATCGGGCG CGGATCCGCCACCATGACGGCTCCTGGAAAGAAACGTCCGGTAGAGCAGTCGCCAC AAGAGCCAGACTCCTCCTCGGGCATTGGCAAGACAGGCCAGCAGCCCGCTAAAAAG AGACTCAATTTTGGTCAGACTGGCGACTCAGAGTCAGTCCCCGACCCACAACCTCTC GGAGAACCTCCAGCAACCCCCGCTGCTGTGGGACCTACTACAATGGCTTCAGGCGG TGGCGCACCAATGGCAGACAATAACGAAGGCGCCGACGGAGTGGGTAATGCCTCAG GAAATTGGCATTGCGATTCCACATGGCTGGGCGACAGAGTCATCACCACCAGCACC CGAACATGGGCCTTGCCCACCTATAACAACCACCTCTACAAGCAAATCTCCAGTGCT TCAACGGGGGCCAGCAACGACAACCACTACTTCGGCTACAGCACCCCCTGGGGGTA TTTTGATTTCAACAGATTCCACTGCCATTTCTCACCACGTGACTGGCAGCGACTCATC AACAACAATTGGGGATTCCGGCCCAAGAGACTCAACTTCAAGCTCTTCAACATCCA AGTCAAGGAGGTCACGACGAATGATGGCGTCACGACCATCGCTAATAACCTTACCA GCACGGTTCAAGTCTTCTCGGACTCGGAGTACCAGTTGCCGTACGTCCTCGGCTCTG CGCACCAGGGCTGCCTCCCTCCGTTCCCGGCGGACGTGTTCATGATTCCGCAGTACG GCTACCTAACGCTCAACAATGGCAGCCAGGCAGTGGGACGGTCATCCTTTTACTGCC TGGAATATTTCCCATCGCAGATGCTGAGAACGGGCAATAACTTTACCTTCAGCTACA CCTTCGAGGACGTGCCTTTCCACAGCAGCTACGCGCACAGCCAGAGCCTGGACCGG CTGATGAATCCTCTCATCGACCAGTACCTGTATTACCTGAACAGAACTGGCGGTGGA G SEQ ID NO: 024 Primer 6711 CTTATCTGGTTCGGATCTCCTAGGTATACT SEQ ID NO: 025 Primer 9554 CTCCACCGCCAGTTCTGTTCAGGTAATACA SEQ ID NO: 026 CD3-scFv fragment GAACAGAACTGGCGGTGGAGGAAGCGGTGGAGGTGGCAGCGGTGGAGGTGGCAGC CAGGTCCAGCTGCAGCAGTCTGGTGCTGAATTGGCTCGTCCTGGTGCTTCCGTGAAG ATGTCCTGCAAGACCTCCGGTTACACCTTCACTCGTTACACCATGCACTGGGTCAAG CAGCGTCCTGGACAAGGCCTGGAATGGATCGGTTACATCAACCCCTCCAGGGGTTA CACCAACTACAACCAGAAGTTCAAGGACAAGGCTACCCTGACTACCGACAAGTCCT CCTCCACCGCTTACATGCAGCTGTCCTCACTGACCTCCGAGGACTCCGCTGTGTACT ACTGCGCTCGTTACTACGACGACCACTACTCCCTGGACTACTGGGGCCAGGGTACTA CTCTGACCGTGTCCTCCGGTTCTACCTCTGGTTCCGGAAAGCCTGGTTCTGGCGAGG GTTCCACCAAGGGACAGATCGTGCTGACCCAGTCTCCTGCTATCATGTCCGCTTCTC CCGGCGAGAAAGTGACTATGACCTGCCGTGCTTCCTCCTCCGTGTCCTACATGAACT GGTATCAGCAGAAGTCCGGCACCTCTCCTAAGCGTTGGATCTACGACACCTCCAAGG - 110 - 12961406v1Attorney Docket No.2014202-0027 TGGCCTCTGGTGTCCCCTACCGTTTCTCCGGATCAGGTTCCGGTACTTCCTACTCTCT GACCATCTCCTCTATGGAAGCTGAGGACGCCGCTACCTACTACTGCCAGCAGTGGTC TAGCAACCCTCTGACCTTCGGTGCTGGCACCAAGCTGGAACTGAAGCAGAATCAGT SEQ ID NO: 027 Primer 9555 GAACAGAACTGGCGGTGGAGGAAGCGGTGG SEQ ID NO: 028 Primer 9556 ACTGATTCTGCTTCAGTTCCAGCTTGGTGC SEQ ID NO: 029 Cap6VP2-HSVtkpA fragment GGAACTGAAGCAGAATCAGTCCGGAAGTGCCCAAAACAAGGACTTGCTGTTTAGCC GGGGGTCTCCAGCTGGCATGTCTGTTCAGCCCAAAAACTGGCTACCTGGACCCTGTT ACCGGCAGCAGCGCGTTTCTAAAACAAAAACAGACAACAACAACAGCAACTTTACC TGGACTGGTGCTTCAAAATATAACCTTAATGGGCGTGAATCTATAATCAACCCTGGC ACTGCTATGGCCTCACACAAAGACGACAAAGACAAGTTCTTTCCCATGAGCGGTGTC ATGATTTTTGGAAAGGAGAGCGCCGGAGCTTCAAACACTGCATTGGACAATGTCAT GATCACAGACGAAGAGGAAATCAAAGCCACTAACCCCGTGGCCACCGAAAGATTTG GGACTGTGGCAGTCAATCTCCAGAGCAGCAGCACAGACCCTGCGACCGGAGATGTG CATGTTATGGGAGCCTTACCTGGAATGGTGTGGCAAGACAGAGACGTATACCTGCA GGGTCCTATTTGGGCCAAAATTCCTCACACGGATGGACACTTTCACCCGTCTCCTCT CATGGGCGGCTTTGGACTTAAGCACCCGCCTCCTCAGATCCTCATCAAAAACACGCC TGTTCCTGCGAATCCTCCGGCAGAGTTTTCGGCTACAAAGTTTGCTTCATTCATCACC CAGTATTCCACAGGACAAGTGAGCGTGGAGATTGAATGGGAGCTGCAGAAAGAAAA CAGCAAACGCTGGAATCCCGAAGTGCAGTATACATCTAACTATGCAAAATCTGCCA ACGTTGATTTCACTGTGGACAACAATGGACTTTATACTGAGCCTCGCCCCATTGGCA CCCGTTACCTCACCCGTCCCCTGTAAGCATGCGGTACCGGGAGATGGGGGAGGCTA ACTGAAACACGGAAGGAGACAATACCGGAAGGAACCCGCGCTATGACGGCAATAA AAAGACAGAATAAAACGCACGGGTGTTGGGTCGTTTGTTCATAAACGCGGGGTTCG GTCCCAGGGCTGGCACTCTGTCGATACCCCACCGAGACCCCATTGGGACCAATACGC CCGCGTTTCTTCCTTTTCCCCACCCCAACCCCCAAGTTCGGGTGAAGGCCCAGGGCT CGCAGCCAACGTCGGGGCGGCAAGCCCTGCCATAGCCACTACGGGTACGTACTATT AATATTCCGGAG SEQ ID NO: 030 Primer 9557 GGAACTGAAGCAGAATCAGTCCGGAAGTGC SEQ ID NO: 031 Primer 9468 CTCCGGAATATTAATAGTAC - 111 - 12961406v1Attorney Docket No.2014202-0027 SEQ ID NO: 032 V705 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCGCCACCATGACGGCTCCTGGAAAGAAACGTCCGGTA GAGCAGTCGCCACAAGAGCCAGACTCCTCCTCGGGCATTGGCAAGACAGGCCAGCA GCCCGCTAAAAAGAGACTCAATTTTGGTCAGACTGGCGACTCAGAGTCAGTCCCCG ACCCACAACCTCTCGGAGAACCTCCAGCAACCCCCGCTGCTGTGGGACCTACTACAA TGGCTTCAGGCGGTGGCGCACCAATGGCAGACAATAACGAAGGCGCCGACGGAGTG GGTAATGCCTCAGGAAATTGGCATTGCGATTCCACATGGCTGGGCGACAGAGTCATC ACCACCAGCACCCGAACATGGGCCTTGCCCACCTATAACAACCACCTCTACAAGCA AATCTCCAGTGCTTCAACGGGGGCCAGCAACGACAACCACTACTTCGGCTACAGCA CCCCCTGGGGGTATTTTGATTTCAACAGATTCCACTGCCATTTCTCACCACGTGACTG GCAGCGACTCATCAACAACAATTGGGGATTCCGGCCCAAGAGACTCAACTTCAAGC TCTTCAACATCCAAGTCAAGGAGGTCACGACGAATGATGGCGTCACGACCATCGCT AATAACCTTACCAGCACGGTTCAAGTCTTCTCGGACTCGGAGTACCAGTTGCCGTAC GTCCTCGGCTCTGCGCACCAGGGCTGCCTCCCTCCGTTCCCGGCGGACGTGTTCATG ATTCCGCAGTACGGCTACCTAACGCTCAACAATGGCAGCCAGGCAGTGGGACGGTC ATCCTTTTACTGCCTGGAATATTTCCCATCGCAGATGCTGAGAACGGGCAATAACTT TACCTTCAGCTACACCTTCGAGGACGTGCCTTTCCACAGCAGCTACGCGCACAGCCA GAGCCTGGACCGGCTGATGAATCCTCTCATCGACCAGTACCTGTATTACCTGAACAG AACTGGCGGTGGAGGAAGCGGTGGAGGTGGCAGCGGTGGAGGTGGCAGCCAGGTC CAGCTGCAGCAGTCTGGTGCTGAATTGGCTCGTCCTGGTGCTTCCGTGAAGATGTCC TGCAAGACCTCCGGTTACACCTTCACTCGTTACACCATGCACTGGGTCAAGCAGCGT CCTGGACAAGGCCTGGAATGGATCGGTTACATCAACCCCTCCAGGGGTTACACCAA CTACAACCAGAAGTTCAAGGACAAGGCTACCCTGACTACCGACAAGTCCTCCTCCA CCGCTTACATGCAGCTGTCCTCACTGACCTCCGAGGACTCCGCTGTGTACTACTGCG CTCGTTACTACGACGACCACTACTCCCTGGACTACTGGGGCCAGGGTACTACTCTGA CCGTGTCCTCCGGTTCTACCTCTGGTTCCGGAAAGCCTGGTTCTGGCGAGGGTTCCA CCAAGGGACAGATCGTGCTGACCCAGTCTCCTGCTATCATGTCCGCTTCTCCCGGCG AGAAAGTGACTATGACCTGCCGTGCTTCCTCCTCCGTGTCCTACATGAACTGGTATC AGCAGAAGTCCGGCACCTCTCCTAAGCGTTGGATCTACGACACCTCCAAGGTGGCCT CTGGTGTCCCCTACCGTTTCTCCGGATCAGGTTCCGGTACTTCCTACTCTCTGACCAT CTCCTCTATGGAAGCTGAGGACGCCGCTACCTACTACTGCCAGCAGTGGTCTAGCAA CCCTCTGACCTTCGGTGCTGGCACCAAGCTGGAACTGAAGCAGAATCAGTCCGGAA GTGCCCAAAACAAGGACTTGCTGTTTAGCCGGGGGTCTCCAGCTGGCATGTCTGTTC - 112 - 12961406v1Attorney Docket No.2014202-0027 AGCCCAAAAACTGGCTACCTGGACCCTGTTACCGGCAGCAGCGCGTTTCTAAAACA AAAACAGACAACAACAACAGCAACTTTACCTGGACTGGTGCTTCAAAATATAACCT TAATGGGCGTGAATCTATAATCAACCCTGGCACTGCTATGGCCTCACACAAAGACGA CAAAGACAAGTTCTTTCCCATGAGCGGTGTCATGATTTTTGGAAAGGAGAGCGCCGG AGCTTCAAACACTGCATTGGACAATGTCATGATCACAGACGAAGAGGAAATCAAAG CCACTAACCCCGTGGCCACCGAAAGATTTGGGACTGTGGCAGTCAATCTCCAGAGC AGCAGCACAGACCCTGCGACCGGAGATGTGCATGTTATGGGAGCCTTACCTGGAAT GGTGTGGCAAGACAGAGACGTATACCTGCAGGGTCCTATTTGGGCCAAAATTCCTCA CACGGATGGACACTTTCACCCGTCTCCTCTCATGGGCGGCTTTGGACTTAAGCACCC GCCTCCTCAGATCCTCATCAAAAACACGCCTGTTCCTGCGAATCCTCCGGCAGAGTT TTCGGCTACAAAGTTTGCTTCATTCATCACCCAGTATTCCACAGGACAAGTGAGCGT GGAGATTGAATGGGAGCTGCAGAAAGAAAACAGCAAACGCTGGAATCCCGAAGTG CAGTATACATCTAACTATGCAAAATCTGCCAACGTTGATTTCACTGTGGACAACAAT GGACTTTATACTGAGCCTCGCCCCATTGGCACCCGTTACCTCACCCGTCCCCTGTAA GCATGCGGTACCGGGAGATGGGGGAGGCTAACTGAAACACGGAAGGAGACAATAC CGGAAGGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAACGCACGGGTG TTGGGTCGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCACTCTGTCGATA CCCCACCGAGACCCCATTGGGACCAATACGCCCGCGTTTCTTCCTTTTCCCCACCCC AACCCCCAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGGGCGGCAAGC CCTGCCATAGCCACTACGGGTACGTACTATTAATATTCCGGAGTATACGTCACGTGT AATCAATGTGTATTTTACAATAAAAACATTTTTATTTAAATAATTATTTTATTTTAGT AAATGACATCATCCCTTGATTGTGTTTTACACGTAGAATTCTACTCGTAAAGCGAGT TTAGTTTTGAAAAACAAATGACATCATTTTTAAAATGACATCATCCCTTGATTGTGTT TTACAAGTAGAATTCTACTCGTAAAGCGAGTTCAGTTTTGAAAACAAATGAGTCATT ATTTGATTGTGTTTATAATCGTGTGCAAAAGATGACATCAGCTTTGCACAAATAAAT GACATCATCCCTTGATCGTGCGTTACAAGTAGAATTCTACTCGTAAAGCGAGTTTAG TTTTGAAAAACAAATGAGTCATACATAAACACGTTTATAATCATGTGTAAAGGATGA CATCATCCCTCGATTGTGTTTTACAAGTAGAATTCTACCCGTAAAGCGAGTTTAGTTT TGAAAAACAAATGACATCATTTGTATAATGACATCATCCCTTGATTGTGTTTTACAA GTAGAATTCTACTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACTCATCCAATG ACATCATCCCTTGATCATGCGTTACAAGTAGAATTCTACTCGTAAAGCCAGTTCAGT TTTGAAAAACAAATGACATCATTTCTTGAGTTCAGTTTTGAAAAACAAATGACATCA TCTCTTGATTATGTTTTACACGTAGAATTCTACTCGTAAAGCGAGTTAGAACGATGA CATCATTTACATATGGCAATTATTCCGCCCAGTTGACATAAGCCTGTTCGGTTCGTAA ACTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTGACCGAAC GCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGTTTTTTTGT ACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGTCGATGTTT GATGTTATGGAGCAGCAACGATGTTACGCAGCAGCAACGATGTTACGCAGCAGGGC AGTCGCCCTAAAACAAAGTTAGGTGGCTCAAGTATGGGCATCATTCGCACATGTAG GCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGA - 113 - 12961406v1Attorney Docket No.2014202-0027 GTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAA CTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGT TGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTGAGATCTA TATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCACCGCGCT CATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCA AGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGG AAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAACAATTCGTTCAAG CCGAGATCGGCTTCCCGGCCGCGGAGTTGTTCGGTAAATTGTCACAACGCCGCGAAT ATAGTCTTTACCATGCCCTTGGCCACGCCCCTCTTTAATACGACGGGCAATTTGCACT TCAGAAAATGAAGAGTTTGCTTTAGCCATAACAAAAGTCCAGTATGCTTTTTCACAG CATAACTGGACTGATTTCAGTTTACAACTATTCTGTCTAGTTTAAGACTTTATTGTCA TAGTTTAGATCTATTTTGTTCAGTTTAAGACTTTATTGTCCGCCCACACCCGCTTACG CAGGGCATCCATTTATTACTCAACCGTAACCGATTTTGCCAGGTTACGCGGCTGGTC TGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGCTCT TCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTA TCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGG AAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCG TTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGC TCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCC TGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCC GCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGGTATCTCA GTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGC CCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACG ACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTA GGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGAC AGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAG CTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCA GCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGG GTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATC AAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTA AAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACC TATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAG ATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCG AGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGG CCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTT GCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCA TTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGG TTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAG CTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATG GTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTG - 114 - 12961406v1Attorney Docket No.2014202-0027 TGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTT GCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAA GTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTG TTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTA CTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAG GGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTAT TGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAG AAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGAAAT TGTAAACGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCAGCTCATTT TTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAATAGACCGA GATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAGAACGTGGA CTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACTACGTGAAC CATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAATCGGAACC CTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGA AAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGG TCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACAGGGCGCGT CCCATTCGCCATTCAGGCTGCAAATAAGCGTTGATATTCAGTCAATTACAAACATTA ATAACGAAGAGATGACAGAAAAATTTTCATTCTGTGACAGAGAA SEQ ID NO: 033 V288 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCCGCCACCATGGCTGCCGATGGTTATCTTCCAGGTAAG TACTCCCTATCAGTGATAGAGATCTATCATGGAGATAATTAAAATGATAACCATCTC GCAAATAAATAAGTATTTTACTGTTTTCGTAACAGTTTTGTAATAAAAAAACCTATA AATATTCCGGATTATTCATACCGTCCCACCATCGGGCGCGAAGGGGGAGACCTGTAG TCAGAGCCCCCGGGCAGCACACACTGACATCCACTCCCTTCCTATTGTTTCAGATTG GCTCGAGGACAACCTCTCTGAGGGCATTCGCGAGTGGTGGGCGCTGAAACCTGGAG CCCCGAAGCCCAAAGCCAACCAGCAAAAGCAGGACGACGGCCGGGGTCTGGTGCTT CCTGGCTACAAGTACCTCGGACCCTTCAACGGACTCGACAAGGGGGAGCCCGTCAA CGCGGCGGACGCAGCGGCCCTCGAGCACGACAAGGCCTACGACCAGCAGCTGCAGG CGGGTGACAATCCGTACCTGCGGTATAACCACGCCGACGCCGAGTTTCAGGAGCGT CTGCAAGAAGATACGTCTTTTGGGGGCAACCTCGGGCGAGCAGTCTTCCAGGCCAA GAAGCGGGTTCTCGAACCTCTCGGTCTGGTTGAGGAAGGCGCTAAGACGGCTCCTG GAAAGAAGAGACCGGTAGAGCCATCACCCCAGCGTTCTCCAGACTCCTCTACGGGC - 115 - 12961406v1Attorney Docket No.2014202-0027 ATCGGCAAGAAAGGCCAACAGCCCGCCAGAAAAAGACTCAATTTTGGTCAGACTGG CGACTCAGAGTCAGTTCCAGACCCTCAACCTCTCGGAGAACCTCCAGCAGCGCCCTC TGGTGTGGGACCTAATACAATGGCTGCAGGCGGTGGCGCACCAATGGCAGACAATA ACGAAGGCGCCGACGGAGTGGGTAGTTCCTCGGGAAATTGGCATTGCGATTCCACA TGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTA CAACAACCACCTCTACAAGCAAATCTCCAACGGGACATCGGGAGGAGCCACCAACG ACAACACCTACTTCGGCTACAGCACCCCCTGGGGGTATTTTGACTTTAACAGATTCC ACTGCCACTTTTCACCACGTGACTGGCAGCGACTCATCAACAACAACTGGGGATTCC GGCCCAAGAGACTCAGCTTCAAGCTCTTCAACATCCAGGTCAAGGAGGTCACGCAG AATGAAGGCACCAAGACCATCGCCAATAACCTCACCAGCACCATCCAGGTGTTTAC GGACTCGGAGTACCAGCTGCCGTACGTTCTCGGCTCTGCCCACCAGGGCTGCCTGCC TCCGTTCCCGGCGGACGTGTTCATGATTCCCCAGTACGGCTACCTAACACTCAACAA CGGTAGTCAGGCCGTGGGACGCTCCTCCTTCTACTGCCTGGAATACTTTCCTTCGCA GATGCTGAGAACCGGCAACAACTTCCAGTTTACTTACACCTTCGAGGACGTGCCTTT CCACAGCAGCTACGCCCACAGCCAGAGCTTGGACCGGCTGATGAATCCTCTGATTG ACCAGTACCTGTACTACTTGTCTCGGACTCAAACAACAGGAGGCACGGCAAATACG CAGACTCTGGGCTTCAGCCAAGGTGGGCCTAATACAATGGCCAATCAGGCAAAGAA CTGGCTGCCAGGACCCTGTTACCGCCAACAACGCGTCTCAACGACAACCGGGCAAA ACAACAATAGCAACTTTGCCTGGACTGCTGGGACCAAATACCATCTGAATGGAAGA AATTCATTGGCTAATCCTGGCATCGCTATGGCAACACACAAAGACGACGAGGAGCG TTTTTTTCCCAGTAACGGGATCCTGATTTTTGGCAAACAAAATGCTGCCAGAGACAA TGCGGATTACAGCGATGTCATGCTCACCAGCGAGGAAGAAATCAAAACCACTAACC CTGTGGCTACAGAGGAATACGGTATCGTGGCAGATAACTTGCAGCAGCAAAACACG GCTCCTCAAATTGGAACTGTCAACAGCCAGGGGGCCTTACCCGGTATGGTCTGGCAG AACCGGGACGTGTACCTGCAGGGTCCCATCTGGGCCAAGATTCCTCACACGGACGG CAACTTCCACCCGTCTCCGCTGATGGGCGGCTTTGGCCTGAAACATCCTCCGCCTCA GATCCTGATCAAGAACACGCCTGTACCTGCGGATCCTCCGACCACCTTCAACCAGTC AAAGCTGAACTCTTTCATCACGCAATACAGCACCGGACAGGTCAGCGTGGAAATTG AATGGGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCCGAGATCCAGTACAC CTCCAACTACTACAAATCTACAAGTGTGGACTTTGCTGTTAATACAGAAGGCGTGTA CTCTGAACCCCGCCCCATTGGCACCCGTTACCTCACCCGTAATCTGTAAGCGGTCGA CTCTAGAGCCTGCAGTCTCGACAAGCTTGTCGAGAAGTACTAGAGGATCATAATCAG CCATACCACATTTGTAGAGGTTTTACTTGCTTTAAAAAACCTCCCACACCTCCCCCTG AACCTGAAACATAAAATGAATGCAATTGTTGTTGTTAACTTGTTTATTGCAGCTTAT AATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCA CTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGATCT GATCACTGCTTGAGCCTAGATTCGAAAGCTACGGACCTTTAATTCAACCCAACACAA TATATTATAGTTAAATAAGAATTATTATCAAATCATTTGTATATTAATTAAAATACTA TACTGTAAATTACATTTTATTTACAATCACTCGACGAAGACTTGATCACCCTACCCGC CATGCCGGGGTTTTACGAGATTGTGATTAAGGTCCCCAGCGACCTTGACGAGCATCT - 116 - 12961406v1Attorney Docket No.2014202-0027 GCCCGGCATTTCTGACAGCTTTGTGAACTGGGTGGCCGAGAAGGAATGGGAGTTGC CGCCAGATTCTGACATGGATCTGAATCTGATTGAGCAGGCACCCCTGACCGTGGCCG AGAAGCTGCAGCGCGACTTTCTGACGGAATGGCGCCGTGTGAGTAAGGCCCCGGAG GCCCTTTTCTTTGTGCAATTTGAGAAGGGAGAGAGCTACTTCCACATGCACGTGCTC GTGGAAACCACCGGGGTGAAATCCATGGTTTTGGGACGTTTCCTGAGTCAGATTCGC GAAAAACTGATTCAGAGAATTTACCGCGGGATCGAGCCGACTTTGCCAAACTGGTTC GCGGTCACAAAGACCAGAAATGGCGCCGGAGGCGGGAACAAGGTGGTGGATGAGT GCTACATCCCCAATTACTTGCTCCCCAAAACCCAGCCTGAGCTCCAGTGGGCGTGGA CTAATATGGAACAGTATTTAAGGTAAGTACTCCCTATCAGTGATAGAGATCTATCAT GGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGTTTTCGT AACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTCCCACC ATCGGGCGCGAAGGGGGAGACCTGTAGTCAGAGCCCCCGGGCAGCACACACTGACA TCCACTCCCTTCCTATTGTTTCAGCGCCTGTTTGAATCTCACGGAGCGTAAACGGTTG GTGGCGCAGCATCTGACGCACGTGTCGCAGACGCAGGAGCAGAACAAAGAGAATC AGAATCCCAATTCTGATGCGCCGGTGATCAGATCAAAAACTTCAGCCAGGTACATG GAGCTGGTCGGGTGGCTCGTGGACAAGGGGATTACCTCGGAGAAGCAGTGGATCCA GGAGGACCAGGCCTCATACATCTCCTTCAATGCGGCCTCCAACTCGCGGTCCCAAAT CAAGGCTGCCTTGGACAATGCGGGAAAGATTATGAGCCTGACTAAAACCGCCCCCG ACTACCTGGTGGGCCAGCAGCCCGTGGAGGACATTTCCAGCAATCGGATTTATAAA ATTTTGGAACTAAACGGGTACGATCCCCAATATGCGGCTTCCGTCTTTCTGGGATGG GCCACGAAAAAGTTCGGCAAGAGGAACACCATCTGGCTGTTTGGGCCTGCAACTAC CGGGAAGACCAACATCGCGGAGGCCATAGCCCACACTGTGCCCTTCTACGGGTGCG TAAACTGGACCAATGAGAACTTTCCCTTCAACGACTGTGTCGACAAGATGGTGATCT GGTGGGAGGAGGGGAAGATGACCGCCAAGGTCGTGGAGTCGGCCAAAGCCATTCTC GGAGGAAGCAAGGTGCGCGTGGACCAGAAATGCAAGTCCTCGGCCCAGATAGACCC GACTCCCGTGATCGTCACCTCCAACACCAACATGTGCGCCGTGATTGACGGGAACTC AACGACCTTCGAACACCAGCAGCCGTTGCAAGACCGGATGTTCAAATTTGAACTCA CCCGCCGTCTGGATCATGACTTTGGGAAGGTCACCAAGCAGGAAGTCAAAGACTTTT TCCGGTGGGCAAAGGATCACGTGGTTGAGGTGGAGCATGAATTCTACGTCAAAAAG GGTGGAGCCAAGAAAAGACCCGCCCCCAGTGACGCAGATATAAGTGAGCCCAAAC GGGTGCGCGAGTCAGTTGCGCAGCCATCGACGTCAGACGCGGAAGCTTCGATCAAC TACGCAGACAGGTACCAAAACAAATGTTCTCGTCACGTGGGCATGAATCTGATGCTG TTTCCCTGCAGACAATGCGAGAGAATGAATCAGAATTCAAATATCTGCTTCACTCAC GGACAGAAAGACTGTTTAGAGTGCTTTCCCGTGTCAGAATCTCAACCCGTTTCTGTC GTCAAAAAGGCGTATCAGAAACTGTGCTACATTCATCATATCATGGGAAAGGTGCC AGACGCTTGCACTGCCTGCGATCTGGTCAATGTGGATTTGGATGACTGCATCTTTGA ACAATAAATGATTTAAATCAGGTATGGCTGCCGATGGTATTGTTTCAGATTGGCTCG AGGACACTCTCTCTGAAGGAGCATGCGGTACCGGGAGATGGGGGAGGCTAACTGAA ACACGGAAGGAGACAATACCGGAAGGAACCCGCGCTATGACGGCAATAAAAAGAC AGAATAAAACGCACGGGTGTTGGGTCGTTTGTTCATAAACGCGGGGTTCGGTCCCAG - 117 - 12961406v1Attorney Docket No.2014202-0027 GGCTGGCACTCTGTCGATACCCCACCGAGACCCCATTGGGACCAATACGCCCGCGTT TCTTCCTTTTCCCCACCCCAACCCCCAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCC AACGTCGGGGCGGCAAGCCCTGCCATAGCCACTACGGGTACGTACTATTAATATTCC GGAGTATACGTCACGTGTAATCAATGTGTATTTTACAATAAAAACATTTTTATTTAA ATAATTATTTTATTTTAGTAAATGACATCATCCCTTGATTGTGTTTTACACGTAGAAT TCTACTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACATCATTTTTAAAATGAC ATCATCCCTTGATTGTGTTTTACAAGTAGAATTCTACTCGTAAAGCGAGTTCAGTTTT GAAAACAAATGAGTCATTATTTGATTGTGTTTATAATCGTGTGCAAAAGATGACATC AGCTTTGCACAAATAAATGACATCATCCCTTGATCGTGCGTTACAAGTAGAATTCTA CTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGAGTCATACATAAACACGTTTATA ATCATGTGTAAAGGATGACATCATCCCTCGATTGTGTTTTACAAGTAGAATTCTACC CGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACATCATTTGTATAATGACATCATC CCTTGATTGTGTTTTACAAGTAGAATTCTACTCGTAAAGCGAGTTTAGTTTTGAAAA ACAAATGACTCATCCAATGACATCATCCCTTGATCATGCGTTACAAGTAGAATTCTA CTCGTAAAGCCAGTTCAGTTTTGAAAAACAAATGACATCATTTCTTGAGTTCAGTTTT GAAAAACAAATGACATCATCTCTTGATTATGTTTTACACGTAGAATTCTACTCGTAA AGCGAGTTAGAACGATGACATCATTTACATATGGCAATTATTCCGCCCAGTTGACAT AAGCCTGTTCGGTTCGTAAACTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGT CCAGAACCTTGACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTT GTTATGACTGTTTTTTTGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTT ACGCCGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGCAA CGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGTGGCTCAAGTATGG GCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTC TTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGG ACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTT CGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCA GCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCA GGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGC TTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATAC AAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCA CCTAACAATTCGTTCAAGCCGAGATCGGCTTCCCGGCCGCGGAGTTGTTCGGTAAAT TGTCACAACGCCGCGAATATAGTCTTTACCATGCCCTTGGCCACGCCCCTCTTTAAT ACGACGGGCAATTTGCACTTCAGAAAATGAAGAGTTTGCTTTAGCCATAACAAAAG TCCAGTATGCTTTTTCACAGCATAACTGGACTGATTTCAGTTTACAACTATTCTGTCT AGTTTAAGACTTTATTGTCATAGTTTAGATCTATTTTGTTCAGTTTAAGACTTTATTGT CCGCCCACACCCGCTTACGCAGGGCATCCATTTATTACTCAACCGTAACCGATTTTG CCAGGTTACGCGGCTGGTCTGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAA ATACCGCATCAGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTT CGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGA ATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGG - 118 - 12961406v1Attorney Docket No.2014202-0027 AACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAG CATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAG ATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCG CTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCT CACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGC ACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGT CCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATT AGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTA CGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTT CGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTG GTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATC CTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGA TTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAAT GAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAAT GCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGC CTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAG TGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAA CCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCA TCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTT TGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTAT GGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTT GTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGC CGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCA TCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAG TGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCA CATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTC TCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAAC TGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGG CAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACT CTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATAC ATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGA AAAGTGCCACCTGAAATTGTAAACGTTAATATTTTGTTAAAATTCGCGTTAAATTTTT GTTAAATCAGCTCATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAAT CAAAAGAATAGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCA CTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGA TGGCCCACTACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAA AGCACTAAATCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGC CGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGC GCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATG CGCCGCTACAGGGCGCGTCCCATTCGCCATTCAGGCTGCAAATAAGCGTTGATATTC - 119 - 12961406v1Attorney Docket No.2014202-0027 AGTCAATTACAAACATTAATAACGAAGAGATGACAGAAAAATTTTCATTCTGTGAC AGAGAA SEQ ID NO: 034 AAV8 VP3 DNA sequence ATCGGGCGCGGATCCGCCACCATGGCTGCAGGCGGTGGCGCACCAATGGCAGACAA TAACGAAGGCGCCGACGGAGTGGGTAGTTCCTCGGGAAATTGGCATTGCGATTCCA CATGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACC TACAACAACCACCTCTACAAGCAAATCTCCAACGGGACATCGGGAGGAGCCACCAA CGACAACACCTACTTCGGCTACAGCACCCCCTGGGGGTATTTTGACTTTAACAGATT CCACTGCCACTTTTCACCACGTGACTGGCAGCGACTCATCAACAACAACTGGGGATT CCGGCCCAAGAGACTCAGCTTCAAGCTCTTCAACATCCAGGTCAAGGAGGTCACGC AGAATGAAGGCACCAAGACCATCGCCAATAACCTCACCAGCACCATCCAGGTGTTT ACGGACTCGGAGTACCAGCTGCCGTACGTTCTCGGCTCTGCCCACCAGGGCTGCCTG CCTCCGTTCCCGGCGGACGTGTTCATGATTCCCCAGTACGGCTACCTAACACTCAAC AACGGTAGTCAGGCCGTGGGACGCTCCTCCTTCTACTGCCTGGAATACTTTCCTTCG CAGATGCTGAGAACCGGCAACAACTTCCAGTTTACTTACACCTTCGAGGACGTGCCT TTCCACAGCAGCTACGCCCACAGCCAGAGCTTGGACCGGCTGATGAATCCTCTGATT GACCAGTACCTGTACTACTTGTCTCGGACTCAAACAACAGGAGGCACGGCAAATAC GCAGACTCTGGGCTTCAGCCAAGGTGGGCCTAATACAATGGCCAATCAGGCAAAGA ACTGGCTGCCAGGACCCTGTTACCGCCAACAACGCGTCTCAACGACAACCGGGCAA AACAACAATAGCAACTTTGCCTGGACTGCTGGGACCAAATACCATCTGAATGGAAG AAATTCATTGGCTAATCCTGGCATCGCTATGGCAACACACAAAGACGACGAGGAGC GTTTTTTTCCCAGTAACGGGATCCTGATTTTTGGCAAACAAAATGCTGCCAGAGACA ATGCGGATTACAGCGATGTCATGCTCACCAGCGAGGAAGAAATCAAAACCACTAAC CCTGTGGCTACAGAGGAATACGGTATCGTGGCAGATAACTTGCAGCAGCAAAACAC GGCTCCTCAAATTGGAACTGTCAACAGCCAGGGGGCCTTACCCGGTATGGTCTGGCA GAACCGGGACGTGTACCTGCAGGGTCCCATCTGGGCCAAGATTCCTCACACGGACG GCAACTTCCACCCGTCTCCGCTGATGGGCGGCTTTGGCCTGAAACATCCTCCGCCTC AGATCCTGATCAAGAACACGCCTGTACCTGCGGATCCTCCGACCACCTTCAACCAGT CAAAGCTGAACTCTTTCATCACGCAATACAGCACCGGACAGGTCAGCGTGGAAATT GAATGGGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCCGAGATCCAGTACA CCTCCAACTACTACAAATCTACAAGTGTGGACTTTGCTGTTAATACAGAAGGCGTGT ACTCTGAACCCCGCCCCATTGGCACCCGTTACCTCACCCGTAATCTGTAAGCGGTCG ACTCTAGAGCATGCGGTACCGGGAGATGGG SEQ ID NO: 035 Primer 9624 ATCGGGCGCGGATCCGCCACCATGGCTGCAGGCGGTGGCG SEQ ID NO: 036 Primer 9625 CCCATCTCCCGGTACCGCATGCTCTAGAGTCGACCGCTTACAGAT - 120 - 12961406v1Attorney Docket No.2014202-0027 SEQ ID NO: 037 V734 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCGCCACCATGGCTGCAGGCGGTGGCGCACCAATGGCA GACAATAACGAAGGCGCCGACGGAGTGGGTAGTTCCTCGGGAAATTGGCATTGCGA TTCCACATGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGC CCACCTACAACAACCACCTCTACAAGCAAATCTCCAACGGGACATCGGGAGGAGCC ACCAACGACAACACCTACTTCGGCTACAGCACCCCCTGGGGGTATTTTGACTTTAAC AGATTCCACTGCCACTTTTCACCACGTGACTGGCAGCGACTCATCAACAACAACTGG GGATTCCGGCCCAAGAGACTCAGCTTCAAGCTCTTCAACATCCAGGTCAAGGAGGT CACGCAGAATGAAGGCACCAAGACCATCGCCAATAACCTCACCAGCACCATCCAGG TGTTTACGGACTCGGAGTACCAGCTGCCGTACGTTCTCGGCTCTGCCCACCAGGGCT GCCTGCCTCCGTTCCCGGCGGACGTGTTCATGATTCCCCAGTACGGCTACCTAACAC TCAACAACGGTAGTCAGGCCGTGGGACGCTCCTCCTTCTACTGCCTGGAATACTTTC CTTCGCAGATGCTGAGAACCGGCAACAACTTCCAGTTTACTTACACCTTCGAGGACG TGCCTTTCCACAGCAGCTACGCCCACAGCCAGAGCTTGGACCGGCTGATGAATCCTC TGATTGACCAGTACCTGTACTACTTGTCTCGGACTCAAACAACAGGAGGCACGGCAA ATACGCAGACTCTGGGCTTCAGCCAAGGTGGGCCTAATACAATGGCCAATCAGGCA AAGAACTGGCTGCCAGGACCCTGTTACCGCCAACAACGCGTCTCAACGACAACCGG GCAAAACAACAATAGCAACTTTGCCTGGACTGCTGGGACCAAATACCATCTGAATG GAAGAAATTCATTGGCTAATCCTGGCATCGCTATGGCAACACACAAAGACGACGAG GAGCGTTTTTTTCCCAGTAACGGGATCCTGATTTTTGGCAAACAAAATGCTGCCAGA GACAATGCGGATTACAGCGATGTCATGCTCACCAGCGAGGAAGAAATCAAAACCAC TAACCCTGTGGCTACAGAGGAATACGGTATCGTGGCAGATAACTTGCAGCAGCAAA ACACGGCTCCTCAAATTGGAACTGTCAACAGCCAGGGGGCCTTACCCGGTATGGTCT GGCAGAACCGGGACGTGTACCTGCAGGGTCCCATCTGGGCCAAGATTCCTCACACG GACGGCAACTTCCACCCGTCTCCGCTGATGGGCGGCTTTGGCCTGAAACATCCTCCG CCTCAGATCCTGATCAAGAACACGCCTGTACCTGCGGATCCTCCGACCACCTTCAAC CAGTCAAAGCTGAACTCTTTCATCACGCAATACAGCACCGGACAGGTCAGCGTGGA AATTGAATGGGAGCTGCAGAAGGAAAACAGCAAGCGCTGGAACCCCGAGATCCAGT ACACCTCCAACTACTACAAATCTACAAGTGTGGACTTTGCTGTTAATACAGAAGGCG TGTACTCTGAACCCCGCCCCATTGGCACCCGTTACCTCACCCGTAATCTGTAAGCGG TCGACTCTAGAGCATGCGGTACCGGGAGATGGGGGAGGCTAACTGAAACACGGAAG GAGACAATACCGGAAGGAACCCGCGCTATGACGGCAATAAAAAGACAGAATAAAA CGCACGGGTGTTGGGTCGTTTGTTCATAAACGCGGGGTTCGGTCCCAGGGCTGGCAC - 121 - 12961406v1Attorney Docket No.2014202-0027 TCTGTCGATACCCCACCGAGACCCCATTGGGACCAATACGCCCGCGTTTCTTCCTTTT CCCCACCCCAACCCCCAAGTTCGGGTGAAGGCCCAGGGCTCGCAGCCAACGTCGGG GCGGCAAGCCCTGCCATAGCCACTACGGGTACGTACTATTAATATTCCGGAGTATAC GTCACGTGTAATCAATGTGTATTTTACAATAAAAACATTTTTATTTAAATAATTATTT TATTTTAGTAAATGACATCATCCCTTGATTGTGTTTTACACGTAGAATTCTACTCGTA AAGCGAGTTTAGTTTTGAAAAACAAATGACATCATTTTTAAAATGACATCATCCCTT GATTGTGTTTTACAAGTAGAATTCTACTCGTAAAGCGAGTTCAGTTTTGAAAACAAA TGAGTCATTATTTGATTGTGTTTATAATCGTGTGCAAAAGATGACATCAGCTTTGCAC AAATAAATGACATCATCCCTTGATCGTGCGTTACAAGTAGAATTCTACTCGTAAAGC GAGTTTAGTTTTGAAAAACAAATGAGTCATACATAAACACGTTTATAATCATGTGTA AAGGATGACATCATCCCTCGATTGTGTTTTACAAGTAGAATTCTACCCGTAAAGCGA GTTTAGTTTTGAAAAACAAATGACATCATTTGTATAATGACATCATCCCTTGATTGTG TTTTACAAGTAGAATTCTACTCGTAAAGCGAGTTTAGTTTTGAAAAACAAATGACTC ATCCAATGACATCATCCCTTGATCATGCGTTACAAGTAGAATTCTACTCGTAAAGCC AGTTCAGTTTTGAAAAACAAATGACATCATTTCTTGAGTTCAGTTTTGAAAAACAAA TGACATCATCTCTTGATTATGTTTTACACGTAGAATTCTACTCGTAAAGCGAGTTAGA ACGATGACATCATTTACATATGGCAATTATTCCGCCCAGTTGACATAAGCCTGTTCG GTTCGTAAACTGTAATGCAAGTAGCGTATGCGCTCACGCAACTGGTCCAGAACCTTG ACCGAACGCAGCGGTGGTAACGGCGCAGTGGCGGTTTTCATGGCTTGTTATGACTGT TTTTTTGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGCCGTGGGT CGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGCAACGATGTTACGCA GCAGGGCAGTCGCCCTAAAACAAAGTTAGGTGGCTCAAGTATGGGCATCATTCGCA CATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCG GTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACC TCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAG CGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAGGTTTGAGCAGCCGCGTAGTG AGATCTATATCTATGATCTCGCAGTCTCCGGCGAGCACCGGAGGCAGGGCATTGCCA CCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCT ACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGC ATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAACAATTC GTTCAAGCCGAGATCGGCTTCCCGGCCGCGGAGTTGTTCGGTAAATTGTCACAACGC CGCGAATATAGTCTTTACCATGCCCTTGGCCACGCCCCTCTTTAATACGACGGGCAA TTTGCACTTCAGAAAATGAAGAGTTTGCTTTAGCCATAACAAAAGTCCAGTATGCTT TTTCACAGCATAACTGGACTGATTTCAGTTTACAACTATTCTGTCTAGTTTAAGACTT TATTGTCATAGTTTAGATCTATTTTGTTCAGTTTAAGACTTTATTGTCCGCCCACACC CGCTTACGCAGGGCATCCATTTATTACTCAACCGTAACCGATTTTGCCAGGTTACGC GGCTGGTCTGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCA GGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCG AGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATA ACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAA - 122 - 12961406v1Attorney Docket No.2014202-0027 GGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAA TCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGT TTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATA CCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCAATGCTCACGCTGTAGG TATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCC GTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTA AGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAG GTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAG AAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGT TGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTG CAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTT CTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGA GATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAAT CAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTG AGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGT CGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGAT ACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCG GAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTA ATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTG TTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAG CTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGC GGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATC ACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATG CTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCG ACCGAGTTGCTCTTGCCCGGCGTCAATACGGGATAATACCGCGCCACATAGCAGAA CTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCT TACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAG CATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCG CAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTC AATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAAT GTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCA CCTGAAATTGTAAACGTTAATATTTTGTTAAAATTCGCGTTAAATTTTTGTTAAATCA GCTCATTTTTTAACCAATAGGCCGAAATCGGCAAAATCCCTTATAAATCAAAAGAAT AGACCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTCCACTATTAAAG AACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATGGCCCACT ACGTGAACCATCACCCTAATCAAGTTTTTTGGGGTCGAGGTGCCGTAAAGCACTAAA TCGGAACCCTAAAGGGAGCCCCCGATTTAGAGCTTGACGGGGAAAGCCGGCGAACG TGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAG TGTAGCGGTCACGCTGCGCGTAACCACCACACCCGCCGCGCTTAATGCGCCGCTACA - 123 - 12961406v1Attorney Docket No.2014202-0027 GGGCGCGTCCCATTCGCCATTCAGGCTGCAAATAAGCGTTGATATTCAGTCAATTAC AAACATTAATAACGAAGAGATGACAGAAAAATTTTCATTCTGTGACAGAGAA SEQ ID NO: 038 V733 DNA sequence AAAGTAGCCGAAGATGACGGTTTGTCACATGGAGTTGGCAGGATGTTTGATTAAAA ACATAACAGGAAGAAAAATGCCCCGCTGTGGGCGGACAAAATAGTTGGGAACTGGG AGGGGTGGAAATGGAGTTTTTAAGGATTATTTAGGGAAGAGTGACAAAATAGATGG GAACTGGGTGTAGCGTCGTAAGCTAATACGAAAATTAAAAATGACAAAATAGTTTG GAACTAGATTTCACTTATCTGGTTCGGATCTCCTAGGTATACTCCGGAATATTAATA GATCATGGAGATAATTAAAATGATAACCATCTCGCAAATAAATAAGTATTTTACTGT TTTCGTAACAGTTTTGTAATAAAAAAACCTATAAATATTCCGGATTATTCATACCGTC CCACCATCGGGCGCGGATCCCGCCACCATGGCTGCCGATGGTTATCTTCCAGGTAAG TACTCCCTATCAGTGATAGAGATCTATCATGGAGATAATTAAAATGATAACCATCTC GCAAATAAATAAGTATTTTACTGTTTTCGTAACAGTTTTGTAATAAAAAAACCTATA AATATTCCGGATTATTCATACCGTCCCACCATCGGGCGCGAAGGGGGAGACCTGTAG TCAGAGCCCCCGGGCAGCACACACTGACATCCACTCCCTTCCTATTGTTTCAGATTG GCTCGAGGACAACCTCTCTGAGGGCATTCGCGAGTGGTGGGCGCTGAAACCTGGAG CCCCGAAGCCCAAAGCCAACCAGCAAAAGCAGGACGACGGCCGGGGTCTGGTGCTT CCTGGCTACAAGTACCTCGGACCCTTCAACGGACTCGACAAGGGGGAGCCCGTCAA CGCGGCGGACGCAGCGGCCCTCGAGCACGACAAGGCCTACGACCAGCAGCTGCAGG CGGGTGACAATCCGTACCTGCGGTATAACCACGCCGACGCCGAGTTTCAGGAGCGT CTGCAAGAAGATACGTCTTTTGGGGGCAACCTCGGGCGAGCAGTCTTCCAGGCCAA GAAGCGGGTTCTCGAACCTCTCGGTCTGGTTGAGGAAGGCGCTAAGACGGCTCCTG GAAAGAAGAGACCGGTAGAGCCATCACCCCAGCGTTCTCCAGACTCCTCTACGGGC ATCGGCAAGAAAGGCCAACAGCCCGCCAGAAAAAGACTCAATTTTGGTCAGACTGG CGACTCAGAGTCAGTTCCAGACCCTCAACCTCTCGGAGAACCTCCAGCAGCGCCCTC TGGTGTGGGACCTAATACAATCGCTGCAGGCGGTGGCGCACCAATCGCAGACAATA ACGAAGGCGCCGACGGAGTGGGTAGTTCCTCGGGAAATTGGCATTGCGATTCCACA TGGCTGGGCGACAGAGTCATCACCACCAGCACCCGAACCTGGGCCCTGCCCACCTA CAACAACCACCTCTACAAGCAAATCTCCAACGGGACATCGGGAGGAGCCACCAACG ACAACACCTACTTCGGCTACAGCACCCCCTGGGGGTATTTTGACTTTAACAGATTCC ACT...

Claims

Attorney Docket No.2014202-0027 CLAIMS 1. A non-naturally-occurring modified adeno-associated virus (AAV) capsid protein comprising (i) a VP1 capsid protein, a VP2 capsid protein, or a VP3 capsid protein; and (ii) an antibody or antibody fragment.

2. The modified AAV capsid protein of claim 1, wherein the antibody or antibody fragment comprises a single-chain fragment variable (scFv) or a Fab fragment.

3. The modified AAV capsid protein of claim 1 or 2, comprising (i) a fusion protein comprising the VP1 capsid protein and the antibody or antibody fragment; (ii) a fusion protein comprising the VP2 capsid protein and the antibody or antibody fragment; or (iii) a fusion protein comprising the VP3 capsid protein and the antibody or antibody fragment.

4. The modified AAV capsid protein of claim 3, comprising (i) a fusion protein comprising the antibody or antibody fragment at the N-terminus, at the C-terminus, or at a surface- exposed loop of the VP1 capsid protein; (ii) a fusion protein comprising the antibody or antibody fragment at the N-terminus, at the C-terminus, or at a surface-exposed loop of the VP2 capsid protein; or (iii) a fusion protein comprising the antibody or antibody fragment at the N-terminus, at the C-terminus, or at a surface-exposed loop of the VP3 capsid protein.

5. The modified AAV capsid protein of any one of claims 1-4, wherein one or more of the VP1, VP2, or VP3 capsid proteins are AAV6 capsid proteins.

6. The modified AAV capsid protein of any one of claims 1-4, wherein one or more of the VP1, VP2, or VP3 capsid proteins are AAV1, AAV5, AAV8, AAV2, AAVrh10, or AAVDJ capsid proteins.

7. The modified AAV capsid protein of any one of claims 1-6, wherein the antibody or antibody fragment is specific for CD3, CD19, or GD2.

8. A polynucleotide comprising a nucleic acid sequence encoding the modified AAV capsid protein of any one of claims 1–7.

9. An expression vector comprising the polynucleotide of claim 8, wherein the nucleic acid sequence encoding the modified AAV capsid protein is operably linked to a promoter sequence.

10. A cell comprising the expression vector of claim 9.

11. The cell of claim 10, further comprising a polynucleotide encoding a therapeutic protein. - 239 - 12961406v1Attorney Docket No.2014202-0027 12. The cell of claim 10 or claim 11, further comprising a polynucleotide encoding a rep protein.

13. A recombinant virus or viral vector comprising the modified AAV capsid protein of any one of claims 1–7.

14. A non-naturally-occurring modified adeno-associated virus (AAV) capsid comprising a VP1 capsid protein, a VP2 capsid protein, and a VP3 capsid protein; and comprising an antibody or antibody fragment.

15. The modified AAV capsid of claim 14, wherein the antibody or antibody fragment comprises a single-chain fragment variable (scFv) or a Fab fragment.

16. The modified AAV capsid of claim 14 or 15, comprising (i) a fusion protein comprising the VP1 capsid protein and the antibody or antibody fragment; (ii) a fusion protein comprising the VP2 capsid protein and the antibody or antibody fragment; and / or (iii) a fusion protein comprising the VP3 capsid protein and the antibody or antibody fragment.

17. The modified AAV capsid of claim 16, comprising (i) a fusion protein comprising the antibody or antibody fragment at the N-terminus, at the C-terminus, or at a surface- exposed loop of the VP1 capsid protein; (ii) a fusion protein comprising the antibody or antibody fragment at the N-terminus, at the C-terminus, or at a surface-exposed loop of the VP2 capsid protein; and / or (iii) a fusion protein comprising the antibody or antibody fragment at the N-terminus, at the C-terminus, or at a surface-exposed loop of the VP3 capsid protein.

18. The modified AAV capsid of any one of claims 14-17, wherein one or more of the VP1, VP2, or VP3 capsid proteins are AAV6 capsid proteins.

19. The modified AAV capsid of any one of claims 14-18, wherein one or more of the VP1, VP2, or VP3 capsid proteins are AAV1, AAV5, AAV8, AAV2, AAVrh10, or AAVDJ capsid proteins.

20. The modified AAV capsid of any one of claims 14-19, wherein the VP2 capsid protein is an AAV6 VP2 capsid protein.

21. The modified AAV capsid of claim 20, comprising a fusion protein comprising the antibody or antibody fragment at the N-terminus, at the C-terminus, or at a surface- exposed loop of the AAV6 VP2 capsid protein.

22. The modified AAV capsid of claim 20 or 21, wherein the VP1 and VP3 capsid proteins are AAV1, AAV5, AAV8, AAV2, AAVrh10, or AAVDJ capsid proteins. - 240 - 12961406v1Attorney Docket No.2014202-0027 23. The modified AAV capsid of any one of claims 20-22, wherein the VP1 and VP3 capsid proteins are AAV2 capsid proteins.

24. The modified AAV capsid of any one of claims 14-23, wherein the antibody or antibody fragment is specific for CD3, CD19, or GD2.

25. A polynucleotide comprising a nucleic acid sequence encoding the modified AAV capsid of any one of claims 14–24.

26. An expression vector comprising the polynucleotide of claim 25, wherein the nucleic acid sequence encoding the modified AAV capsid is operably linked to a promoter sequence.

27. A cell comprising the expression vector of claim 26.

28. The cell of claim 27, further comprising a polynucleotide encoding a therapeutic protein.

29. The cell of claim 27 or claim 28, further comprising a polynucleotide encoding a rep protein.

30. A recombinant virus or viral vector comprising the modified AAV capsid of any one of claims 14–24.

31. A mosaic recombinant adeno-associated virus (AAV) particle comprising capsid proteins from two or more different AAV serotypes, wherein the AAV particle comprises at least one fusion protein comprising a capsid protein and a single-chain fragment variable (scFv) or Fab fragment at a surface-exposed loop, at the N-terminus, or at the C-terminus of the capsid protein, thereby conferring altered cellular tropism compared to parental AAV.

32. The mosaic recombinant AAV particle of claim 31, comprising VP1, VP2, and VP3 capsid proteins.

33. The mosaic recombinant AAV particle of claim 32, wherein the VP1 and VP3 capsid proteins are AAV2 capsid proteins, and the VP2 capsid protein is an AAV6 capsid protein.

34. The mosaic recombinant AAV particle of claim 33, comprising a fusion protein comprising a VP2 capsid protein comprising the amino acid sequence of SEQ ID NO: 097 and a CD3-specific scFv inserted at amino acid position 449 of SEQ ID NO: 097, thereby forming a functional virion.

35. The recombinant virus or viral vector of any one of claims 13 and 30-34, wherein the recombinant virus or viral vector is an AAV selected from AAV3, AAV4, AAV5, - 241 - 12961406v1Attorney Docket No.2014202-0027 AAV7, AAV8, AAV9, AAVrh10, AAVDJ, AAVphp.eB, avian AAV, bovine AAV, canine AAV, equine AAV, primate AAV, non-primate AAV, or ovine AAV.

36. The recombinant virus or viral vector of any one of claims 13 and 30-35, comprising a polynucleotide encoding a therapeutic gene product.

37. The recombinant virus or viral vector of claim 36, wherein the therapeutic gene product is a CD19-specific chimeric antigen receptor (CAR), a GD2-specific CAR, a transgenic T cell receptor (TCR) specific for a tumor-associated antigen (PRAME, NY-ESO-1, MAGE-A4), alpha-1 antitrypsin, factor IX, factor VIII, C1-esterase inhibitor, β-globin, γ- globin, a toxin, a bi-specific T-cell engager, or a tri-specific T-cell engager.

38. The recombinant virus or viral vector of any one of claims 13 and 30-37, wherein the recombinant virus or viral vector exhibits altered cellular tropism compared to a parental or wildtype AAV, preferentially transducing T cells, B cells, or myeloid cells.

39. The recombinant virus or viral vector of claim 38, wherein the altered cellular tropism enhances infectivity of hematopoietic cells compared to a parental or wildtype AAV.

40. The recombinant virus or viral vector of claim 38, wherein the altered cellular tropism facilitates selective delivery to bone marrow, spleen, lymph nodes, or solid tumors.

41. A pharmaceutical composition comprising a pharmaceutically acceptable excipient and the recombinant virus or viral vector of any one of claims 13 and 30-40.

42. A method of treating or preventing a cancerous disease or a genetic disorder in a subject in need thereof, comprising administering to the subject the pharmaceutical composition of claim 41.

43. The method of claim 42, wherein the recombinant virus or viral vector comprises an AAV6, AAV5, AAV8, AAV1, or AAV2 capsid protein comprising an scFv or a Fab fragment.

44. The method of claim 42 or claim 43, wherein the scFv or Fab fragment is specific for CD3, CD19, or GD2.

45. A method of treating or preventing a disease or disorder in a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a recombinant AAV viral vector, wherein the vector comprises (i) an AAV capsid protein comprising an scFv or a Fab fragment and (ii) a polynucleotide encoding a therapeutic gene product.

46. The method of claim 45, wherein the AAV serotype is AAV5, AAV6, or AAV8. - 242 - 12961406v1Attorney Docket No.2014202-0027 47. The method of claim 45 or claim 46, wherein the disease or disorder is a hematological malignancy or a solid tumor.

48. The method of any one of claims 45-46, wherein the scFv or Fab fragment retains antigen-binding activity after incorporation into the AAV capsid and packaging.

49. The method of any one of claims 45–48, wherein the recombinant AAV vector delivers a gene encoding a therapeutic T cell receptor (TCR) or a chimeric antigen receptor (CAR).

50. The recombinant virus or viral vector of any one of claims 13 and 30-40, wherein the capsid protein comprises two or more distinct scFv or Fab fragment insertions.

51. The modified AAV capsid protein of any one of claims 3-7, wherein the scFv or Fab is fused to the VP1, VP2, or VP3 capsid protein via a cleavable peptide linker and / or further comprises a detectable tag.

52. The modified AAV capsid protein of any one of claims 1-7, further comprising one or more amino acid mutations, e.g., that reduce binding to one or more neutralizing antibodies.

53. The recombinant virus or viral vector of any one of claims 36-40 and 50, further comprising a polynucleotide encoding a CRISPR-associated nuclease or a guide RNA specific to an endogenous locus.

54. The method of any one of claims 42-49, wherein the pharmaceutical composition is administered intravenously, intratumorally, or intrathecally at a dose of 1 × 108to 2 × 1014viral genomes per kg of body weight.

55. The method of claim 49, wherein the pharmaceutical composition is administered intraperitoneally to generate CAR-T cells in vivo, and the antibody fragment is a CD3- specific Fab. - 243 - 12961406v1

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