Recombinant AAV capsid proteins

Abstract

Provided herein are recombinant adeno-associated virus (AAV) capsid proteins, compositions (e.g., rAAV) comprising the capsid proteins, nucleic acids encoding the capsid proteins, and methods of making and using the capsid proteins.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Application No. 63 / 362,419, filed on April 4, 2022, and U.S. Provisional Application No. 63 / 366,428, filed on June 15, 2022, each of which is incorporated herein by reference in its entirety. Background Art

[0003] Adeno-associated virus (AAV) has unique characteristics that make it an attractive vector for delivering foreign DNA into cells for gene therapy purposes. For example, AAV infection of cells in culture is non-cytopathic, and natural infection in humans and other animals is silent. In addition, because the signals directing AAV replication, genome encapsidation, and integration are contained within the inverted terminal repeats (ITRs) of the AAV genome, essentially all of the internal 4.7 kb of the AAV genome (encoding replication and structural capsid proteins, rep-cap) can be replaced with heterologous nucleic acid sequences (e.g., transgenes for achieving gene therapy).

[0004] Recombinant AAV (rAAV) gene therapy vectors are typically composed of a recombinant AAV genome containing a gene therapy transgene, contained within an AAV capsid. The AAV capsid, formed by the assembly of 60 AAV capsid proteins, interacts with the cell surface of target cells and determines the tropism and transduction efficiency of rAAV. For tissue-specific genetic diseases, it would be very advantageous if rAAV could be specifically and efficiently targeted to tissues in need of gene therapy.

[0005] Therefore, there is a need in the art for tissue-specific AAV capsids for gene therapy applications. Summary of the invention

[0006] Provided herein are recombinant adeno-associated virus (AAV) capsid proteins, compositions (e.g., rAAV) comprising the capsid proteins, nucleic acids encoding the capsid proteins, and methods for preparing and using the capsid proteins. Provided herein are recombinant AAV capsid proteins engineered to include peptides within parent AAV capsid proteins. In one aspect, provided herein are recombinant AAV capsid proteins engineered to include peptides within loop IV and / or loop VIII of parent AAV capsid proteins. Compared to parent AAV capsid proteins, these recombinant AAV capsid proteins exhibit significantly increased myocyte transduction.

[0007] In one aspect, the present invention provides a recombinant adeno-associated virus (AAV) capsid protein comprising a peptide and a parent capsid protein, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one aspect, the present invention provides a recombinant adeno-associated virus (AAV) capsid protein comprising a peptide and a parent capsid protein, wherein the peptide is contained within loop IV and / or loop VIII of the parent capsid protein, and wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide is contained within loop VIII of the parent capsid protein.

[0008] In one embodiment, the parent capsid protein is a clade A, clade B, clade C, clade D, clade E, clade F, clade G, clade H, clade I, AAVgo.1, AAV3, AAV4, AAV10, AAV11, AAV12, rh.32, rh32.33, rh.33, rh.34, BAAV, or AAV5 capsid protein, or an engineered variant thereof.

[0009] In one embodiment, the parent capsid protein comprises an amino acid sequence that is at least 95% identical to amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence that is at least 99% identical to amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence of amino acids 193-725 of SEQ ID NO: 11.

[0010] In one embodiment, the parent capsid protein comprises an amino acid sequence that is at least 95% identical to amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence that is at least 99% identical to amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence of amino acids 138-725 of SEQ ID NO: 11.

[0011] In one embodiment, the parent capsid protein comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises the amino acid sequence of SEQ ID NO: 10 or 11.

[0012] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 440, 441, 442, 443, 444, 445, 446, 447, 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583 or 584 of SEQ ID NO: 11.

[0013] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 440 of SEQ ID NO:11.

[0014] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 441 of SEQ ID NO:11.

[0015] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 442 of SEQ ID NO:11.

[0016] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 443 of SEQ ID NO:11.

[0017] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 444 of SEQ ID NO:11.

[0018] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 445 of SEQ ID NO:11.

[0019] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 446 of SEQ ID NO:11.

[0020] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 447 of SEQ ID NO:11.

[0021] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 572 of SEQ ID NO:11.

[0022] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 573 of SEQ ID NO:11.

[0023] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 574 of SEQ ID NO:11.

[0024] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 575 of SEQ ID NO:11.

[0025] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 576 of SEQ ID NO:11.

[0026] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 577 of SEQ ID NO:11.

[0027] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 578 of SEQ ID NO:11.

[0028] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 579 of SEQ ID NO:11.

[0029] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 580 of SEQ ID NO:11.

[0030] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 581 of SEQ ID NO:11.

[0031] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 582 of SEQ ID NO:11.

[0032] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 583 of SEQ ID NO:11.

[0033] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid protein corresponding to amino acid 584 of SEQ ID NO:11.

[0034] In one embodiment, the recombinant AAV capsid protein comprises: an amino acid sequence having at least 95% sequence identity to amino acids 194-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 95% sequence identity to amino acids 194-763 of SEQ ID NO: 13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 95% sequence identity to amino acids 194-734 of SEQ ID NO: 14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); an amino acid sequence having at least 95% sequence identity to amino acids 194-734 of SEQ ID NO: 15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 3). an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:21); ID NO:5); an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 95% sequence identity to amino acids 194-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 95% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 95% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 95% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:8); an amino acid sequence having at least 95% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:9); NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 95% sequence identity to amino acids 194-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 95% sequence identity to amino acids 194-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0035] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 99% sequence identity to amino acids 194-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 194-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 194-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 99% sequence identity to amino acids 194-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:3). an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:21); ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:9); NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 194-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 99% sequence identity to amino acids 194-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0036] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having 100% sequence identity to amino acids 194-763 of SEQ ID NO:12; an amino acid sequence having 100% sequence identity to amino acids 194-763 of SEQ ID NO:13; an amino acid sequence having 100% sequence identity to amino acids 194-734 of SEQ ID NO:14; an amino acid sequence having 100% sequence identity to amino acids 194-734 of SEQ ID NO:15; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:16; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:17; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:18; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:19; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:11 an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:20; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:21; an amino acid sequence having 100% sequence identity to amino acids 194-742 of SEQ ID NO:22; an amino acid sequence having 100% sequence identity to amino acids 194-742 of SEQ ID NO:23; an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:24; an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:25; an amino acid sequence having 100% sequence identity to amino acids 194-735 of SEQ ID NO:26; an amino acid sequence having 100% sequence identity to amino acids 194-735 of SEQ ID NO:27; an amino acid sequence having 100% sequence identity to amino acids 194-737 of SEQ ID NO:28; or an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:29. Amino acids 194-737 of NO:29 have an amino acid sequence with 100% sequence identity.

[0037] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 95% sequence identity to amino acids 138-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 95% sequence identity to amino acids 138-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 95% sequence identity to amino acids 138-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 95% sequence identity to amino acids 138-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:3). an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:21); ID NO:5); an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 95% sequence identity to amino acids 138-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 95% sequence identity to amino acids 138-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 95% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 95% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:9); NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 95% sequence identity to amino acids 138-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 95% sequence identity to amino acids 138-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0038] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 99% sequence identity to amino acids 138-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 138-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 138-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 99% sequence identity to amino acids 138-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:3). an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:21); ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 99% sequence identity to amino acids 138-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 138-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:9); NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 138-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 99% sequence identity to amino acids 138-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0039] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:12; an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:13; an amino acid sequence having 100% sequence identity to amino acids 138-734 of SEQ ID NO:14; an amino acid sequence having 100% sequence identity to amino acids 138-734 of SEQ ID NO:15; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:16; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:17; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:18; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:19; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:11 an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:20; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:21; an amino acid sequence having 100% sequence identity to amino acids 138-742 of SEQ ID NO:22; an amino acid sequence having 100% sequence identity to amino acids 138-742 of SEQ ID NO:23; an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:24; an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:25; an amino acid sequence having 100% sequence identity to amino acids 138-735 of SEQ ID NO:26; an amino acid sequence having 100% sequence identity to amino acids 138-735 of SEQ ID NO:27; an amino acid sequence having 100% sequence identity to amino acids 138-737 of SEQ ID NO:28; or an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:29. Amino acids 138-737 of NO:29 have an amino acid sequence with 100% sequence identity.

[0040] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 95% sequence identity to amino acids 1-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 95% sequence identity to amino acids 1-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 95% sequence identity to amino acids 1-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); an amino acid sequence having at least 95% sequence identity to amino acids 1-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:21, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:6); an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 95% sequence identity to amino acids 1-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 95% sequence identity to amino acids 1-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 95% sequence identity to amino acids 1-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 95% sequence identity to amino acids 1-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 95% sequence identity to amino acids 1-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 95% sequence identity to amino acids 1-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0041] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 99% sequence identity to amino acids 1-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 1-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 99% sequence identity to amino acids 1-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 1-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 1-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 1-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:9); NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 1-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 99% sequence identity to amino acids 1-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0042] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12-29. In one embodiment, the amino acid sequence of the AAV capsid protein consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 12-29.

[0043] On the one hand, the present invention provides a recombinant adeno-associated virus (AAV) capsid protein, which comprises a first peptide, a second peptide and a parent capsid protein, wherein the first peptide is contained in loop IV of the parent capsid protein, the second peptide is contained in loop VIII of the parent capsid protein, and wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35 and 36, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35 and 36.

[0044] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:1.

[0045] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:2.

[0046] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:3.

[0047] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:4.

[0048] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:5.

[0049] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:6.

[0050] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:7.

[0051] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:8.

[0052] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:9.

[0053] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:35.

[0054] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:36.

[0055] In one embodiment, the parent capsid protein is a clade A, clade B, clade C, clade D, clade E, clade F, clade G, clade H, clade I, AAVgo.1, AAV3, AAV4, AAV10, AAV11, AAV12, rh.32, rh32.33, rh.33, rh.34, BAAV, or AAV5 capsid protein, or an engineered variant thereof.

[0056] In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 95% identity with amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence of amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 95% identity with amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence of amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 95% identity with SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises the amino acids of SEQ ID NO: 10 or 11.

[0057] In one embodiment, (a) the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440, 441, 442, 443, 444, 445, 446 or 447 of SEQ ID NO: 11; and (b) the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583 or 584 of SEQ ID NO: 11.

[0058] In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 441 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 442 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 443 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 444 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 445 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 446 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 447 of SEQ ID NO:11.

[0059] In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 573 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 574 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 575 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 576 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 577 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 578 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 579 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 580 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 581 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 582 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 583 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 584 of SEQ ID NO: 11.

[0060] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 95% sequence identity to amino acids 194-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 194-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0061] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 99% sequence identity to amino acids 194-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 194-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0062] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 100% sequence identity to amino acids 194-745 of SEQ ID NO:30; (b) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:31; (c) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:32; (d) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:33; or (e) an amino acid sequence having at least 100% sequence identity to amino acids 194-744 of SEQ ID NO:34.

[0063] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 95% sequence identity to amino acids 138-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 138-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0064] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 99% sequence identity to amino acids 138-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 138-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0065] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:30;

[0066] (b) an amino acid sequence that has 100% sequence identity with amino acids 138-763 of SEQ ID NO:31; (c) an amino acid sequence that has 100% sequence identity with amino acids 138-734 of SEQ ID NO:32; (d) an amino acid sequence that has 100% sequence identity with amino acids 138-734 of SEQ ID NO:33; or (e) an amino acid sequence that has 100% sequence identity with amino acids 138-732 of SEQ ID NO:34.

[0067] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 95% sequence identity to amino acids 1-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 1-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0068] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 99% sequence identity to amino acids 1-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 1-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0069] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 30-34. In one embodiment, the amino acid sequence of the recombinant AAV capsid protein consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 30-34.

[0070] In one aspect, provided herein are isolated polynucleotides encoding the recombinant AAV capsid proteins disclosed herein.

[0071] In one aspect, provided herein are vectors comprising a polynucleotide disclosed herein.

[0072] In one embodiment, the vector is a plasmid or a viral vector. In one embodiment, the viral vector is a retroviral vector, a herpes virus vector, a baculovirus vector or an adenovirus vector. In one embodiment, the vector is an expression vector.

[0073] In one aspect, provided herein is a recombinant cell comprising a polynucleotide or a vector disclosed herein.

[0074] In one aspect, provided herein is a method for producing an AAV capsid protein, the method comprising culturing a recombinant cell disclosed herein under conditions for expressing a polynucleotide, and producing the capsid protein.

[0075] In one aspect, provided herein is a recombinant adeno-associated virus (rAAV) comprising a capsid comprising one or more recombinant capsid proteins disclosed herein; and a rAAV genome.

[0076] In one embodiment, the rAAV genome comprises a transgene. In one embodiment, the transgene encodes a polypeptide or a non-coding RNA. In one embodiment, the transgene encodes an antibody or a fragment thereof. In one embodiment, the transgene encodes a scFv, a nanobody, or a VHH.

[0077] In one aspect, the present invention provides a method of delivering a transgene to a cell, the method comprising contacting the cell with a rAAV disclosed herein under conditions that transduce the cell and express the transgene. In one embodiment, the cell is a myocyte, a microglia, an astrocyte, a neuron or a cardiomyocyte.

[0078] In one aspect, provided herein is a method of delivering a transgene to muscle tissue in a subject in need thereof, the method comprising administering a rAAV disclosed herein. In one embodiment, the subject suffers from a disease associated with muscle tissue.

[0079] In one aspect, provided herein is a method of treating a neuromuscular or neurodegenerative disease or disorder in a subject in need thereof, comprising administering to the subject an effective amount of a rAAV disclosed herein.

[0080] In one embodiment, the neuromuscular disease or disorder is selected from the group consisting of: myopathy, hereditary cardiomyopathy, metabolic myopathy, distal myopathy, muscular dystrophy, congenital myopathy, spinal muscular atrophy (SMA), motor neuron disease, congenital myopathy, congenital muscular dystrophy, motor neuron disease, Duchenne muscular dystrophy, Becker muscular dystrophy, limb-girdle muscular dystrophy, myotonic dystrophy, myotubular myopathy, centronuclear myopathy, nemaline myopathy, selenoprotein N-related myopathy, Pompe disease, glycogen storage disease III, spinal muscular atrophy, amyotrophic lateral sclerosis, Charcot-Marie-Tooth disease, multiple sclerosis, myositis, polymyositis and dermatomyositis.

[0081] In one embodiment, the neurodegenerative disease or disorder is selected from the group consisting of motor neuron disease (MND), parkinsonism syndrome, Alzheimer's dementia, progressive supranuclear palsy (PSP), Huntington's disease, multiple system atrophy (MSA), spinocerebellar ataxia (SCA) and Friedreich's ataxia.

[0082] In one embodiment, the rAAV is administered to the subject intravenously, intraperitoneally, subcutaneously, intramuscularly, intrathecally, or intradermally. BRIEF DESCRIPTION OF THE DRAWINGS

[0083] Figure 1A is a series of images showing the amount of GFP protein expression in human myotubes transduced with recombinant AAV capsid protein. Figure 1B is a graph showing quantification of GFP fluorescence in human myotubes after transduction with recombinant AAV capsid proteins comprising VyPep7, VyPep8, VyPep9, VyPep11, VyPep12, or VyPep13 compared to the parental capsid protein without a peptide insert.

[0084] Figure 2 is a graph showing the average intensity of GFP expression in myotubes after transduction with recombinant AAV capsid proteins comprising VYPep14 or VYPep15 in loop IV or loop VIII compared to the parental capsid protein without peptide insertion.

[0085] Figure 3A is a graph showing the average intensity of GFP expression in myotubes after transduction with recombinant AAV capsid proteins containing VYPep14 in loop IV or loop VIII compared to the parental capsid protein without peptide insertion. Figure 3B is a graph showing GFP protein expression in human myotubes after transduction with recombinant AAV capsid proteins containing VYPep7 in loop IV or loop VIII compared to the parental capsid protein without peptide insertion.

[0086] Figure 4A It is displayed in 1×10 4 Figure 3 Plot of the percentage of GFP-positive human astrocytes after transduction with recombinant AAV capsid proteins containing peptide insertions in loop IV or loop VIII compared to the parental capsid protein without peptide insertion at different MOIs. Figure 4B It is displayed in 1×10 6 Figure 3 Plot of the percentage of GFP-positive human astrocytes after transduction with recombinant AAV capsid proteins containing peptide insertions in loop IV or loop VIII compared to the parental capsid protein without peptide insertion at different MOIs.

[0087] Figure 5 is a graph showing the percentage of positive GFP iPSC-derived motor neurons after transduction with recombinant AAV capsid proteins comprising VYPep7 inserted into loop IV or loop VIII compared to the parental capsid protein without peptide insertion.

[0088] Fig. 6A is a graph showing the percentage of GFP-positive astrocytes transduced with recombinant AAV9-GFP capsids in a time course experiment. Figure 6B is a graph showing the percentage of GFP-positive astrocytes transduced with recombinant AAV5.2-GFP capsid in a time course experiment. Figure 6C is a graph showing the percentage of GFP-positive astrocytes transduced with recombinant AAV5.2-VYPep7-loop IV-GFP capsid in a time course experiment.

[0089] Figure 7 is a graph showing the percentage of GFP-positive NHP-derived astrocytes transduced with recombinant AAV capsids.

[0090] Fig. 8A -B shows the 6th day after transduction ( Fig. 8A ) and Day 11 ( Figure 8B ) Graph of the percentage of GFP-positive cells on the blood side (endothelial compartment) or brain side (astrocyte compartment) of the blood-brain barrier in a microfluidic model of the blood-brain barrier. Cells were transduced with recombinant AAV capsid proteins containing VYPep14 or VYPep7 in the blood compartment, compared to the parental capsid protein without peptide insertion, and the transduction level was assessed in the blood (endothelial) and brain (astrocyte) compartments. Figure 8C is a graph showing the average intensity of GFP expression in cells on the blood side or brain side 6 or 13 days after transduction of astrocytes with recombinant AAV capsid proteins containing VYPep14 or VYPep7 compared to the parental capsid protein without peptide insertion in a microfluidic model of the blood-brain barrier.

[0091] Fig. 9A is a graph showing the percentage of GFP-positive astrocytes transduced with recombinant AAV capsids. Fig. 9B is a graph showing intact virus titer (viral genomes / mL) of recombinant AAV capsids. Fig. 9C is a graph of the percentage of whole capsids (dark grey bars) and percentage of aggregates (light grey bars) of recombinant AAV capsids. DETAILED DESCRIPTION

[0092] Provided herein are recombinant adeno-associated virus (AAV) capsid proteins, compositions (e.g., rAAV) comprising the capsid proteins, nucleic acids encoding the capsid proteins, and methods for preparing and using the capsid proteins. Provided herein are recombinant AAV capsid proteins engineered to include peptides within parent AAV capsid proteins. In one aspect, provided herein are recombinant AAV capsid proteins engineered to include peptides within loop IV and / or loop VIII of parent AAV capsid proteins. Compared to parent AAV capsid proteins, these recombinant AAV capsid proteins exhibit significantly increased myocyte transduction.

[0093] definition

[0094] As used herein, the term "AAV" is the standard abbreviation for adeno-associated virus.

[0095] As used herein, the term "parent capsid protein" refers to an AAV capsid protein into which a peptide is inserted. The parent capsid protein can be a naturally occurring AAV capsid protein (e.g., a branch or clone described in Bello et al., Sci. Rep. 2014 4 (6644), 1-11. doi: 10.1038 / srep06644 or Gao et al., J Virol. 2004 June; 78 (12): 6381-6388. doi: 10.1128 / JVI. 78.12.6381-6388.2004) or a non-naturally occurring AAV capsid protein. Suitable non-naturally occurring AAV capsid proteins include, but are not limited to, chimeric capsid proteins and engineered capsid proteins containing one or more amino acid insertions, deletions or substitutions relative to naturally occurring AAV capsid proteins.

[0096] As used herein, the term "recombinant adeno-associated virus" or "rAAV" refers to an AAV comprising a genome lacking functional rep and cap genes.

[0097] As used herein, the term "cap gene" refers to a nucleic acid sequence encoding a capsid protein. For AAV, the capsid protein may be VP1, VP2 or VP3. VP1, VP2 and / or VP3 capsid proteins are assembled into a capsid surrounding the rAAV genome.

[0098] As used herein, the term "rep gene" refers to a nucleic acid sequence encoding non-structural proteins required for AAV replication and production (eg, rep78, rep68, rep52, and rep40).

[0099] As used herein, the term "loop IV" refers to the amino acids in the AAV capsid protein corresponding to amino acids 434-466 of SEQ ID NO: 10 or amino acids 435-467 of SEQ ID NO: 11. Or the corresponding amino acids of a different parent capsid protein. In one embodiment, amino acid 444 of SEQ ID NO: 11 corresponds to amino acid 451 of the AAV2 capsid, amino acid 443 of the AAV5 capsid, or amino acid 444 of the AAV9 capsid.

[0100] As used herein, the term "loop VIII" refers to the amino acids in the AAV capsid protein corresponding to amino acids 563-592 of SEQ ID NO: 10 or amino acids 564-593 of SEQ ID NO: 11, or the corresponding amino acids of a different parent capsid protein.

[0101] As used herein, the term "rAAV genome" refers to a nucleic acid molecule (e.g., DNA and / or RNA) comprising the genomic sequence of rAAV. One skilled in the art will appreciate that where the rAAV genome comprises a transgene (e.g., an antibody heavy chain or light chain encoding sequence operably linked to a transcriptional regulatory element), the rAAV genome may be in a sense or antisense orientation relative to the transcription direction of the transgene.

[0102] As used herein, an "isolated polynucleotide" refers to a polynucleotide that has been separated from one or more nucleic acid molecules present in the natural source of the polynucleotide.

[0103] As used herein, "vector" refers to a nucleic acid molecule that serves as a vehicle for introducing a nucleic acid molecule (eg, a polynucleotide disclosed herein) into a cell.

[0104] As used herein, "expression vector" refers to a vector comprising transcriptional regulatory elements operably linked to a gene of interest (eg, a polynucleotide disclosed herein), which promotes expression of the gene of interest in a cell and / or a cell-free expression system.

[0105] As used herein, the term "transgene" refers to a non-AAV nucleic acid sequence encoding a polypeptide (e.g., an antibody or scFv) or a non-coding RNA (e.g., a miRNA, shRNA, siRNA, antisense RNA, gRNA, antagomir, miRNA sponge, RNA aptamer, or RNA aptamer).

[0106] As used herein, the term "transcription regulatory element" or "TRE" refers to a cis-acting nucleotide sequence, such as a DNA sequence, that regulates (e.g., controls, increases or decreases) the transcription of an operably linked nucleotide sequence to form an RNA molecule by RNA polymerase. The TRE may comprise one or more promoter elements and / or enhancer elements. It will be appreciated by the skilled artisan that the promoter and enhancer elements in a gene may be close in position, and the term "promoter" may refer to a sequence comprising a promoter element and an enhancer element. Therefore, the term "promoter" does not exclude enhancer elements in a sequence. Promoter and enhancer elements do not need to be derived from the same gene or species, and the sequence of each promoter or enhancer element may be identical or substantially identical to the corresponding endogenous sequence in the genome.

[0107] As used herein, the term "operably linked" is used to describe the connection between a TRE and a polynucleotide sequence to be transcribed (e.g., a transgene disclosed herein). Typically, gene expression is placed under the control of a TRE comprising one or more promoter and / or enhancer elements. A transgene is "operably linked" to a TRE if transcription of the transgene is controlled or influenced by the TRE. The promoter and enhancer elements of the TRE can be in any orientation and / or distance from the transgene as long as the desired transcriptional activity is obtained. In one embodiment, the TRE is upstream of the transgene.

[0108] As used herein, the "percent identity" between two nucleotide sequences or between two amino acid sequences is calculated by multiplying the number of matches between the aligned sequence pairs by 100 and dividing by the length of the aligned region (including internal gaps). The identity score only calculates perfect matches and does not take into account the degree of similarity between the amino acids. When a sequence is described herein as having a certain percentage identity to a reference sequence, the percentage identity to the reference sequence is determined over the entire length of the reference sequence.

[0109] As used herein, in the context of administering an AAV to a subject, the term "effective amount" refers to the amount of AAV that achieves the desired prophylactic or therapeutic effect.

[0110] AAV capsid protein

[0111] In one aspect, the present invention provides a recombinant adeno-associated virus (AAV) capsid protein comprising a peptide and a parent AAV capsid protein. The amino acid sequences of exemplary peptides used in the recombinant AAV capsid proteins disclosed herein are provided in Table 1 below.

[0112] Table 1. Amino acid sequences of exemplary peptides

[0113] describe SEQ ID NO Amino acid sequence VYPep1 1 ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA VYPep7 2 APLTNTVKA VYPep8 3 DDTRHWG VYPep9 4 EYHHYNK VYPep10 5 QLFPLFR VYPep12 6 FNPVTGEVPPRYPLDAR VYPep13 7 SV TEQGAELSNEER VYPep14 8 RGDLGL VYPep15 9 ASSLNIA

[0114] In one aspect, the present invention provides a recombinant adeno-associated virus (AAV) capsid protein, comprising a peptide and a parent capsid protein, wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9. In one aspect, the present invention provides a recombinant adeno-associated virus (AAV) capsid protein, comprising a peptide and a parent capsid protein, wherein the peptide is contained within loop IV and / or loop VIII of the parent capsid protein, and wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9.

[0115] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 6, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 7, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 8, and the peptide is contained within loop IV of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 9, and the peptide is contained within loop IV of the parent capsid protein.

[0116] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 6, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 7, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 8, and the peptide is contained within the loop VIII of the parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 9, and the peptide is contained within the loop VIII of the parent capsid protein.

[0117] In one embodiment, the recombinant AAV capsid protein comprises a first peptide within loop IV of a parent capsid protein and a second peptide within loop VIII of a parent capsid protein. In one embodiment, the first peptide and the second peptide are the same peptide. In one embodiment, the first peptide and the second peptide are different peptides. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 1, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 2, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 3, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 4, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 5, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 6, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 7, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 8, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9. In one embodiment, the first peptide comprises the amino acid sequence of SEQ ID NO: 9, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9.

[0118] In one embodiment, the parent capsid protein is VP1 capsid protein. In one embodiment, the parent capsid protein is VP2 capsid protein. In one embodiment, the parent capsid protein is VP3 capsid protein.

[0119] In one embodiment, the parent capsid protein is a clade A, clade B, clade C, clade D, clade E, clade F, clade G, clade H, clade I, AAVgo.1, AAV3, AAV4, AAV10, AAV11, AAV12, rh.32, rh32.33, rh.33, rh.34, BAAV, or AAV5 capsid protein, or an engineered variant thereof.

[0120] In one embodiment, the parent capsid protein is an engineered variant capsid protein comprising amino acid sequences from at least two different AAV capsid proteins. In one embodiment, the parent capsid protein comprises amino acid sequences from AAV2 capsid protein and AAV5 capsid protein. In one embodiment, the parent capsid protein comprises the VP1 amino acid sequence from AAV2 and the VP2 and VP3 amino acid sequences from AAV5.

[0121] In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% identity with amino acids 193-724 of SEQ ID NO: 10 or amino acids 194-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with amino acids 193-724 of SEQ ID NO: 10 or amino acids 194-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having 100% identity with amino acids 193-724 of SEQ ID NO: 10 or amino acids 194-725 of SEQ ID NO: 11.

[0122] In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% identity with amino acids 137-724 of SEQ ID NO: 10 or amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with amino acids 137-724 of SEQ ID NO: 10 or amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having 100% identity with amino acids 137-724 of SEQ ID NO: 10 or amino acids 138-725 of SEQ ID NO: 11.

[0123] In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% identity to the amino acid sequence of SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity to the amino acid sequence of SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having 100% identity to the amino acid sequence of SEQ ID NO: 10 or 11.

[0124] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440, 441, 442, 443, 444, 445, 446, 447, 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583 or 584 of SEQ ID NO: 11.

[0125] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440 of SEQ ID NO:11.

[0126] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 440 of SEQ ID NO: 11.

[0127] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 441 of SEQ ID NO:11.

[0128] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 441 of SEQ ID NO:11.

[0129] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 442 of SEQ ID NO:11.

[0130] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 442 of SEQ ID NO:11.

[0131] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 443 of SEQ ID NO:11.

[0132] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 443 of SEQ ID NO:11.

[0133] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 444 of SEQ ID NO:11.

[0134] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO:11 or the corresponding amino acid of a different parent capsid protein. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 444 of SEQ ID NO:11.

[0135] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 445 of SEQ ID NO:11.

[0136] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 445 of SEQ ID NO: 11.

[0137] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 446 of SEQ ID NO:11.

[0138] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 446 of SEQ ID NO: 11.

[0139] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 447 of SEQ ID NO:11.

[0140] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 447 of SEQ ID NO: 11.

[0141] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572 of SEQ ID NO:11.

[0142] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 572 of SEQ ID NO: 11.

[0143] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 573 of SEQ ID NO:11.

[0144] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 573 of SEQ ID NO:11.

[0145] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 574 of SEQ ID NO:11.

[0146] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 574 of SEQ ID NO: 11.

[0147] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 575 of SEQ ID NO:11.

[0148] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 575 of SEQ ID NO: 11.

[0149] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 576 of SEQ ID NO:11.

[0150] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 576 of SEQ ID NO: 11.

[0151] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 577 of SEQ ID NO:11.

[0152] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 577 of SEQ ID NO: 11.

[0153] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 578 of SEQ ID NO:11.

[0154] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 578 of SEQ ID NO: 11.

[0155] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 579 of SEQ ID NO:11.

[0156] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 579 of SEQ ID NO: 11.

[0157] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 580 of SEQ ID NO:11.

[0158] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 580 of SEQ ID NO: 11.

[0159] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 581 of SEQ ID NO:11.

[0160] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 581 of SEQ ID NO: 11.

[0161] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 582 of SEQ ID NO:11.

[0162] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 582 of SEQ ID NO:11.

[0163] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 583 of SEQ ID NO:11.

[0164] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 583 of SEQ ID NO:11.

[0165] In one embodiment, the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 584 of SEQ ID NO:11.

[0166] In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 1, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 2, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 3, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 4, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO: 5, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO: 11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:6, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:7, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:8, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO:11. In one embodiment, the peptide comprises the amino acid sequence of SEQ ID NO:9, and the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to the amino acid 584 of SEQ ID NO:11.

[0167] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); NO:13 has an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFER QLGA (SEQ ID NO:1); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3);an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 5 an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6 ... NO:23 amino acids 194-742 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); NO:7); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:9); NO:8); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 194-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0168] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 99% sequence identity to amino acids 194-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 194-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 194-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 99% sequence identity to amino acids 194-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:3). an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:21); ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:9); NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 194-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 99% sequence identity to amino acids 194-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0169] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having 100% sequence identity to amino acids 194-763 of SEQ ID NO:12; an amino acid sequence having 100% sequence identity to amino acids 194-763 of SEQ ID NO:13; an amino acid sequence having 100% sequence identity to amino acids 194-734 of SEQ ID NO:14; an amino acid sequence having 100% sequence identity to amino acids 194-734 of SEQ ID NO:15; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:16; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:17; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:18; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:19; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:11 an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:20; an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:21; an amino acid sequence having 100% sequence identity to amino acids 194-742 of SEQ ID NO:22; an amino acid sequence having 100% sequence identity to amino acids 194-742 of SEQ ID NO:23; an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:24; an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:25; an amino acid sequence having 100% sequence identity to amino acids 194-735 of SEQ ID NO:26; an amino acid sequence having 100% sequence identity to amino acids 194-735 of SEQ ID NO:27; an amino acid sequence having 100% sequence identity to amino acids 194-737 of SEQ ID NO:28; or an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:29. Amino acids 194-737 of NO:29 have an amino acid sequence with 100% sequence identity.

[0170] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); NO:13 has an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFER QLGA (SEQ ID NO:1); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3);an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 5 an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6 ... NO:23 amino acids 138-742 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); NO:7); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 138-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0171] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 99% sequence identity to amino acids 138-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 138-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 138-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 99% sequence identity to amino acids 138-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:3). an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:21); ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 99% sequence identity to amino acids 138-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 138-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:9); NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 138-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 99% sequence identity to amino acids 138-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0172] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:12; an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:13; an amino acid sequence having 100% sequence identity to amino acids 138-734 of SEQ ID NO:14; an amino acid sequence having 100% sequence identity to amino acids 138-734 of SEQ ID NO:15; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:16; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:17; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:18; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:19; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:11 an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:20; an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:21; an amino acid sequence having 100% sequence identity to amino acids 138-742 of SEQ ID NO:22; an amino acid sequence having 100% sequence identity to amino acids 138-742 of SEQ ID NO:23; an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:24; an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:25; an amino acid sequence having 100% sequence identity to amino acids 138-735 of SEQ ID NO:26; an amino acid sequence having 100% sequence identity to amino acids 138-735 of SEQ ID NO:27; an amino acid sequence having 100% sequence identity to amino acids 138-737 of SEQ ID NO:28; or an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:29. Amino acids 138-737 of NO:29 have an amino acid sequence with 100% sequence identity.

[0173] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); NO:13 has an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-763, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO:1); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO:2); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3);an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 4); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); NO:22 having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); NO:6);an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); NO:7); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% sequence identity to amino acids 1-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9).;

[0174] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence having at least 99% sequence identity to amino acids 1-763 of SEQ ID NO:12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 1-763 of SEQ ID NO:13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO:14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO:15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO:3); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO:4); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6);an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); an amino acid sequence having at least 99% sequence identity to amino acids 1-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 1-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); an amino acid sequence having at least 99% sequence identity to amino acids 1-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 1-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:9); NO:8); an amino acid sequence having at least 99% sequence identity to amino acids 1-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or an amino acid sequence having at least 99% sequence identity to amino acids 1-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9). ;

[0175] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 12-29. In one embodiment, the amino acid sequence of the recombinant AAV capsid protein consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 12-29.

[0176] In one embodiment, the peptide is directly connected to the amino acids of the parent AAV capsid protein (i.e., connected by a peptide bond). In one embodiment, the peptide is connected to the amino acids of the parent AAV capsid protein by an amino acid linker. In one embodiment, the peptide comprises an N-terminal linker. In one embodiment, the peptide comprises a C-terminal linker. In one embodiment, the peptide comprises an N-terminal linker and a C-terminal linker. Suitable linkers include, but are not limited to, a G linker (AG), a GS linker (GS), and a GG linker (GG), TG, GLS, or GQSG.

[0177] In one embodiment, the recombinant AAV capsid protein disclosed herein binds to a protein expressed on the cell membrane of a muscle cell. In one embodiment, the recombinant AAV capsid protein disclosed herein binds to a protein selected from the group consisting of integrin α7 (ITGA7), insulin receptor (INSR / CD220), ADP-ribosyltransferase 1 (ART1), and cadherin 15 (CDH15).

[0178] The amino acid sequences of exemplary recombinant AAV capsid proteins provided herein are listed in Table 2 below.

[0179] Table 2. Amino acid sequences of exemplary recombinant AAV capsid proteins

[0180]

[0181]

[0182]

[0183]

[0184]

[0185]

[0186]

[0187]

[0188] On the one hand, the present invention provides a recombinant adeno-associated virus (AAV) capsid protein, which comprises a first peptide, a second peptide and a parent capsid protein, wherein the first peptide is contained in loop IV of the parent capsid protein, the second peptide is contained in loop VIII of the parent capsid protein, and wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35 and 36, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35 and 36.

[0189] The amino acid sequences of exemplary recombinant AAV capsid proteins ("bicyclic capsid proteins") comprising the first and second peptides provided herein are listed in Table 3 below.

[0190] Table 3. Amino acid sequences of exemplary recombinant AAV bicyclic capsid proteins

[0191]

[0192]

[0193]

[0194] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:1.

[0195] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:2.

[0196] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:3.

[0197] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:4.

[0198] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:5.

[0199] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:6.

[0200] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:7.

[0201] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:8.

[0202] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:9.

[0203] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:35.

[0204] In one embodiment, the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:36.

[0205] In one embodiment, the parent capsid protein is a clade A, clade B, clade C, clade D, clade E, clade F, clade G, clade H, clade I, AAVgo.1, AAV3, AAV4, AAV10, AAV11, AAV12, rh.32, rh32.33, rh.33, rh.34, BAAV, or AAV5 capsid protein, or an engineered variant thereof.

[0206] In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 95% identity with amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence of amino acids 193-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 95% identity with amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence of amino acids 138-725 of SEQ ID NO: 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 95% identity with SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises an amino acid sequence having at least 99% identity with SEQ ID NO: 10 or 11. In one embodiment, the parent capsid protein comprises the amino acids of SEQ ID NO: 10 or 11.

[0207] In one embodiment, (a) the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440, 441, 442, 443, 444, 445, 446 or 447 of SEQ ID NO: 11; and (b) the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583 or 584 of SEQ ID NO: 11.

[0208] In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 441 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 442 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 443 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 444 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 445 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 446 of SEQ ID NO: 11. In one embodiment, the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 447 of SEQ ID NO:11.

[0209] In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 573 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 574 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 575 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 576 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 577 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 578 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 579 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 580 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 581 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 582 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 583 of SEQ ID NO: 11. In one embodiment, the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 584 of SEQ ID NO: 11.

[0210] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 95% sequence identity to amino acids 194-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 194-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0211] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 99% sequence identity to amino acids 194-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 194-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0212] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 100% sequence identity to amino acids 194-745 of SEQ ID NO:30; (b) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:31; (c) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:32; (d) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:33; or (e) an amino acid sequence having at least 100% sequence identity to amino acids 194-744 of SEQ ID NO:34.

[0213] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 95% sequence identity to amino acids 138-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 138-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0214] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 99% sequence identity to amino acids 138-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 138-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0215] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:30;

[0216] (b) an amino acid sequence that has 100% sequence identity with amino acids 138-763 of SEQ ID NO:31; (c) an amino acid sequence that has 100% sequence identity with amino acids 138-734 of SEQ ID NO:32; (d) an amino acid sequence that has 100% sequence identity with amino acids 138-734 of SEQ ID NO:33; or (e) an amino acid sequence that has 100% sequence identity with amino acids 138-732 of SEQ ID NO:34.

[0217] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 95% sequence identity to amino acids 1-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 1-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0218] In one embodiment, the recombinant AAV capsid protein comprises: (a) an amino acid sequence having at least 99% sequence identity to amino acids 1-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8) and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36) and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 1-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2) and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

[0219] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 30-34. In one embodiment, the amino acid sequence of the recombinant AAV capsid protein consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 30-34.

[0220] Additional examples of recombinant AAV capsid proteins disclosed herein are listed in Table 4 below.

[0221] Table 4. Amino acid sequences of additional exemplary recombinant AAV capsid proteins

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231] In one embodiment, the recombinant AAV capsid protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 37-59. In one embodiment, the amino acid sequence of the recombinant AAV capsid protein consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 37-59.

[0232] Polynucleotides, vectors and methods for producing AAV capsid proteins

[0233] In one aspect, provided herein are isolated polynucleotides encoding the recombinant AAV capsid proteins disclosed herein.

[0234] In one embodiment, polynucleotides are optimized, for example, by codon / RNA optimization, replacement with heterologous signal sequences and / or elimination of mRNA instability elements. The method of producing optimized polynucleotides for recombinant expression by introducing codon changes and / or eliminating inhibitory regions in mRNA can be achieved by adjusting, for example, the optimization methods described in U.S. Patent Nos. 5,965,726; 6,174,666; 6,291,664; 6,414,132; and 6,794,498, and accordingly, all of these patents are incorporated herein by reference as a whole. For example, potential splicing sites and instability elements (e.g., elements rich in A / T or A / U) in RNA can be mutated without changing the amino acids encoded by the nucleic acid sequence to increase the stability of RNA for recombinant expression. The change utilizes the degeneracy of the genetic code, such as using alternative codons of the same amino acid. In one embodiment, one or more codons may need to be changed to encode conservative mutations, such as similar amino acids with chemical structures and properties and / or functions similar to the original amino acids. Such methods can increase expression of the encoded capsid protein relative to expression of a capsid encoded by a polynucleotide that has not been optimized.

[0235] In one aspect, provided herein are vectors comprising polynucleotides disclosed herein. Suitable vectors include, but are not limited to, plasmids, viruses, cosmids, artificial chromosomes, linear DNA, and mRNA. In one embodiment, the vector is a plasmid or a viral vector. In one embodiment, the vector is a retroviral vector, a herpes virus vector, a baculovirus vector, or an adenovirus vector. In one embodiment, the vector is an expression vector.

[0236] The vector (e.g., expression vector) can be introduced into a cell (using any technique known in the art) for propagation of the vector and / or for expression of the AAV capsid protein encoded by the vector. Therefore, in another aspect, the present disclosure provides a recombinant cell comprising a polynucleotide or vector (e.g., expression vector) disclosed herein. In addition, in another aspect, the present disclosure provides a method for producing an AAV capsid protein, the method comprising culturing a recombinant cell under conditions where the polynucleotide is expressed and a capsid is produced.

[0237] A variety of host cells and expression vector systems can be used to express the capsid proteins described herein. In these expression systems, the coding sequence of interest can be produced and subsequently purified. These expression systems also represent cells that can in situ express the capsid proteins described herein when transformed or transfected with appropriate nucleotide coding sequences. These include, but are not limited to, microorganisms such as bacteria (e.g., E. coli and B. subtilis) transformed with, for example, a recombinant phage DNA, plasmid DNA, or cosmid DNA expression vector containing a capsid protein coding sequence; yeast (e.g., Saccharomyces Pichia) transformed with, for example, a recombinant yeast expression vector containing a capsid protein coding sequence; insect cell systems infected with, for example, a recombinant virus expression vector (e.g., baculovirus) containing a capsid protein coding sequence; plant cell systems (e.g., green algae, such as Chlamydomonas reinhardtii) infected with, for example, a recombinant virus expression vector (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with, for example, a recombinant plasmid expression vector (e.g., Ti plasmid) containing a capsid protein coding sequence; or mammalian cell systems (e.g., COS (e.g., COS1 or COS), CHO, BHK, MDCK, HEK 293, NS0, PER.C6, VERO, CRL7030, HsS78Bst, HeLa and NIH 3T3, HEK-293T, HepG2, SP210, R1.1, BW, LM, BSC1, BSC40, YB / 20 and BMT10 cells), which have, for example, a recombinant expression construct containing a promoter derived from a mammalian cell genome (e.g., a metallothionein promoter) or derived from a mammalian virus (e.g., adenovirus late promoter; vaccinia virus 7.5K promoter). In one embodiment, the cell used to express the capsid protein described herein is a human cell, such as a human cell line. In one embodiment, the mammalian expression vector is pOptiVEC TMOr pcDNA3.3. In one embodiment, bacterial cells such as Escherichia coli or eukaryotic cells (e.g., mammalian cells) are used to express capsid proteins. For example, mammalian cells such as CHO or HEK293 cells combined with vectors (e.g., major intermediate early gene promoter elements from human cytomegalovirus) are effective expression systems for capsid proteins disclosed herein. In one embodiment, insect cells (e.g., Sf9 cells) are used to express capsid proteins.

[0238] In insect systems, for example, Autographa californica nuclear polyhedrosis virus (AcNPV) can be used as a vector for expressing foreign genes. The virus is grown in Spodoptera frugiperda cells. The capsid protein coding sequence can be cloned separately into a non-essential region of the virus (e.g., a polyhedrin gene) and placed under the control of an AcNPV promoter (e.g., a polyhedrin promoter).

[0239] In mammalian host cells, many virus-based expression systems can be used. In the case where adenovirus is used as an expression vector, the capsid protein coding sequence of interest can be connected to an adenovirus transcription / translation control complex, such as a late promoter and a tripartite leader sequence. The chimeric gene can then be inserted into the adenovirus genome by in vitro or in vivo recombination. Insertion of a non-essential region of the viral genome (e.g., region E1 or E3) will illustrate a recombinant virus that has viability in the infected host and can express capsid protein molecules (e.g., see Logan J & Shenk T (1984) PNAS 81 (12): 3655-9, which is incorporated herein by reference in its entirety). The effective translation of the inserted capsid protein coding sequence may also require specific initiation signals. These signals include an ATG initiation codon and adjacent sequences. In addition, the initiation codon must be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert. These exogenous translation control signals and initiation codons can be of various origins, natural and synthetic. Expression efficiency can be enhanced by including appropriate transcription enhancer elements, transcription terminators, etc. (see, e.g., Bitter G et al., (1987) Methods Enzymol. 153:516-544, which is incorporated herein by reference in its entirety). Such mammalian host cells include, but are not limited to, CHO, VERO, BHK, Hela, MDCK, HEK 293, NIH 3T3, W138, BT483, Hs578T, HTB2, BT20 and T47D, NSO, CRL7030, COS (e.g., COS1 or COS), PER.C6, VERO, HsS78Bst, HEK-293T, HepG2, SP210, R1.1, BW, LM, BSC1, BSC40, YB / 20, BMT10 and HsS78Bst cells.

[0240] For long-term, high-yield production of recombinant proteins, stable expressing cells can be generated. For example, cell lines that stably express the capsid proteins described herein can be engineered.

[0241] In one embodiment, instead of using an expression vector containing a viral origin of replication, a polynucleotide (e.g., DNA or RNA) and a selection marker controlled by an appropriate transcriptional regulatory element (e.g., a promoter, an enhancer, a sequence, a transcription terminator, a polyadenylation site, etc.) can be used to transform a host cell. After the polynucleotide is introduced, the engineered cells can be grown in an enriched medium for 1-2 days and then converted to a selective medium. The selection marker in the recombinant plasmid confers resistance to selection and allows the cell to stably integrate the plasmid into its chromosome and grow to form a transformation focus, which can be cloned and amplified into a cell line. The method can be advantageously used for engineered cell lines expressing capsid proteins or fragments thereof as described herein.

[0242] A number of selection systems may be used, including but not limited to the herpes simplex virus thymidine kinase (Wigler M et al., (1977) Cell 11(1):223-32), hypoxanthine guanine phosphoribosyltransferase (Szybalska EH & Szybalski W (1962) PNAS 48(12):2026-2034), and adenine phosphoribosyltransferase (Lowy I et al., (1980) Cell 22(3):817-23) genes in tk-, hgprt-, or aprt- cells, respectively, all of which are incorporated herein by reference in their entirety. In addition, antimetabolite resistance can be used as the basis for selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler M et al., (1980) PNAS 77(6):3567-70; O'Hare K et al., (1981) PNAS 78:1527-31); gpt, which confers resistance to mycophenolic acid (Mulligan RC & Berg P (1981) PNAS 78(4):2072-6); neo, which confers resistance to the aminoglycoside G-418 (Wu GY and Wu CH (1991) Biotherapy 3:87-95; Tolstoshev P (1993) Ann Rev Pharmacol Toxicol 32:573-596; Mulligan RC (1993) Science 260:926-932; and Morgan RA and Anderson WF (1993) Ann Rev Biochem 62: 191-217; Nabel GJ and Felgner PL (1993) Trends Biotechnol 11(5):211-5); and hygro, which confers resistance to hygromycin (Santerre RF et al., (1984) Gene 30(1-3): 147-56), all of which are incorporated herein by reference in their entirety.Methods generally known in the field of recombinant DNA technology can be routinely applied to select the desired recombinant clones, and such methods are described in, for example, Ausubel FM et al. (eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993); Kriegler M, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990); and in Chapters 12 and 13, Dracopoli NC et al. (eds.), Current Protocols in Human Genetics, John Wiley & Sons, NY (1994); Colbère-Garapin F et al., (1981) J Mol Biol 150: 1-14, all of which are incorporated herein by reference in their entirety.

[0243] AAV composition

[0244] In one aspect, provided herein are rAAV compositions comprising a capsid comprising a recombinant AAV capsid protein disclosed herein.

[0245] In one aspect, provided herein is a rAAV comprising a capsid comprising one or more recombinant capsid proteins disclosed herein; and a rAAV genome.

[0246] In one embodiment, the rAAV genome comprises a transgene. In one embodiment, the transgene encodes a polypeptide. In one embodiment, the transgene encodes an antibody or fragment thereof. In one embodiment, the transgene encodes a scFv.

[0247] The rAAV disclosed herein generally comprises a recombinant genome (e.g., a rAAV genome) packaged in a capsid. The rAAV genome can be any type that can be packaged in an AAV capsid disclosed herein. For example, in one embodiment, the rAAV genome is a single-stranded DNA genome. In one embodiment, the rAAV genome is a self-complementary genome, such as described in US7790154, which is incorporated herein by reference in its entirety.

[0248] In one embodiment, the rAAV genome comprises a transgene. In one embodiment, the transgene encodes a therapeutic protein. In one embodiment, the transgene encodes an antibody or a fragment thereof (e.g., a Fab, scFv, or a full-length antibody). In one embodiment, the transgene encodes a scFv, a nanobody, or a VHH. In one embodiment, the transgene encodes a non-coding RNA.

[0249] In one embodiment, the transgenic coding reporter sequence produces a detectable signal when expressed. Such reporter sequences include, but are not limited to, DNA sequences encoding: β-lactamase, β-galactosidase (LacZ), alkaline phosphatase, thymidine kinase, green fluorescent protein (GFP), red fluorescent protein (RFP), chloramphenicol acetyltransferase (CAT), luciferase, membrane-bound proteins (including, for example, CD2, CD4, CD8, influenza hemagglutinin protein and other proteins well known in the art, for which there are or can be produced by conventional methods High affinity antibodies), and fusion proteins comprising membrane-bound proteins appropriately fused to antigen tag domains from hemagglutinin or Myc.

[0250] In one embodiment, the rAAV genome comprises a TRE operably connected to a transgene to control the expression of RNA or polypeptides encoded by the transgene. In one embodiment, TRE comprises a constitutive promoter. In one embodiment, TRE is active in any mammalian cell (e.g., any human cell). In one embodiment, TRE is active in a wide range of human cells. Such TRE may comprise a constitutive promoter and / or enhancer element, including any of those described herein and any of those known to those skilled in the art. In one embodiment, TRE comprises an inducible promoter. In one embodiment, TRE is a tissue-specific TRE, i.e., it is active in specific tissues and / or organs. Tissue-specific TRE comprises one or more tissue-specific promoters and / or enhancer elements, and optionally one or more constitutive promoters and / or enhancer elements. Tissue-specific promoters and / or enhancer elements can be separated from genes specifically expressed in tissues by methods well known in the art.

[0251] Suitable promoters include, for example, the cytomegalovirus promoter (CMV) (Stinski et al. (1985) Journal of Virology 55(2):431-441), CMV early enhancer / chicken β-actin (CBA) promoter / rabbit β-globin intron (CAG) (Miyazaki et al. (1989) Gene 79(2):269-277), a hybrid form of the CBA promoter (CBh) (Gray et al. (2011) Hum Gene Ther. 22(9):1143-53), CB SB(Jacobson et al. (2006) Molecular Therapy 13(6):1074-1084), human elongation factor 1α promoter (EF1α) (Kim et al. (1990) Gene 91(2):217-223), human phosphoglycerate kinase promoter (PGK) (Singer-Sam et al. (1984) Gene 32(3):409-417), mitochondrial heavy chain promoter (Lodeiro et al. (2012) PNAS 109(17):6513-6518), ubiquitin promoter (Wulff et al. (1990) FEBS Letters 261:101-105).

[0252] In one aspect, provided herein is a pharmaceutical composition comprising a rAAV disclosed herein and a pharmaceutically acceptable excipient, adjuvant, diluent, vehicle or carrier or a combination thereof. A "pharmaceutically acceptable carrier" includes any material that, when combined with the active ingredient of the composition, allows the ingredient to retain biological activity and does not cause a disruptive physiological response (e.g., an unintended immune response). Pharmaceutically acceptable carriers include water, phosphate-buffered saline, emulsions (e.g., oil / water emulsions), and wetting agents. Compositions containing such carriers are formulated by well-known conventional methods, such as those described in Remington's Pharmaceutical Sciences, current Ed., Mack Publishing Co., Easton Pa. 18042, USA; A. Gennaro (2000) "Remington: The Science and Practice of Pharmacy", 20th edition, Lippincott, Williams, & Wilkins; Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) HC Ansel et al., 7th edition, Lippincott, Williams, & Wilkins; and Handbook of Pharmaceutical Excipients (2000) AK Kibbe et al., 3rd edition. Amer. Pharmaceutical Assoc.

[0253] How to use

[0254] In one aspect, provided herein is a method of transducing a cell, the method comprising contacting the cell with a rAAV disclosed herein under conditions that transduce the cell and express a transgene. In one embodiment, the cell is a muscle cell.

[0255] In one embodiment, a method for expressing a transgene in a cell is provided herein, the method generally comprising contacting the cell with such rAAV under conditions of transducing the cell, and transgene expression. As described herein, the transgene can encode a polypeptide, such as an antibody or a fragment thereof (e.g., scFv). Therefore, in one embodiment, a method for producing a polypeptide, such as an antibody or a fragment thereof (e.g., scFv) in a cell is provided herein, the method generally comprising contacting the cell with such rAAV under conditions of transducing the cell and producing a polypeptide, such as an antibody or a fragment thereof (e.g., scFv).

[0256] In one aspect, a method for delivering a transgene to a cell is provided herein, the method comprising contacting the cell with a rAAV disclosed herein under conditions of transducing the cell and expressing the transgene. The rAAV disclosed herein can be used for transducing cells in vitro, in vivo, and ex vivo. Cells suitable for transduction by the rAAV disclosed herein include, but are not limited to, blood, liver, heart, joint tissue, muscle, brain, kidney, or lung cells. In one embodiment, the cell is a myocyte, microglia, astrocyte, neuron, or cardiomyocyte. In one embodiment, the rAAV provided herein is used to transduce cells in vivo in a subject suffering from a disease associated with muscle tissue.

[0257] In one aspect, provided herein is a method of delivering a transgene to muscle tissue in a subject in need thereof, the method comprising administering to the subject a disclosed rAAV.

[0258] In one aspect, provided herein is a method of treating a neuromuscular or neurodegenerative disease or condition in a subject in need thereof, comprising administering to the subject an effective amount of a rAAV disclosed herein. In one embodiment, the neuromuscular disease is selected from the group consisting of myopathy, hereditary cardiomyopathy, metabolic myopathy, distal myopathy, muscular dystrophy, congenital myopathy, spinal muscular atrophy (SMA), motor neuron disease, congenital myopathy, congenital muscular dystrophy, motor neuron disease, Duchenne muscular dystrophy, Becker muscular dystrophy, limb-girdle muscular dystrophy, myotonic dystrophy, myotubular myopathy, centronuclear myopathy, nemaline myopathy, selenoprotein N-related myopathy, Pompe disease, glycogen storage disease III, spinal muscular atrophy, amyotrophic lateral sclerosis, Charcot-Marie-Tooth disease, multiple sclerosis, myositis, polymyositis, and dermatomyositis.

[0259] In one embodiment, the neurodegenerative disease or disorder is selected from the group consisting of motor neuron disease (MND), Parkinson's syndrome, Alzheimer's dementia, progressive supranuclear palsy (PSP), Huntington's disease, multiple system atrophy (MSA), spinocerebellar ataxia (SCA) and Friedreich's ataxia.

[0260] In one embodiment, the rAAV disclosed herein can be administered to a subject (e.g., a human subject) by all routes applicable to rAAV, including, but not limited to, intravenous, intraperitoneal, subcutaneous, intramuscular, intrathecal, or intradermal.

[0261] In one aspect, provided herein is a rAAV as disclosed herein for use in medicine. In one aspect, provided herein is a rAAV as disclosed herein for use as therapy. In one aspect, provided herein is a rAAV as disclosed herein for use as medicine.

[0262] Adeno-associated virus packaging system

[0263] In one aspect, the present invention provides a packaging system for recombinantly preparing a recombinant adeno-associated virus (rAAV) disclosed herein. Such packaging systems generally include: a first nucleotide sequence encoding one or more AAV Rep proteins; a second nucleotide sequence encoding a capsid protein of any recombinant AAV capsid disclosed herein; and a third nucleotide sequence comprising any AAV genome sequence disclosed herein, wherein the packaging system is operable in a cell to encapsulate the AAV genome in a capsid to form rAAV.

[0264] In one embodiment, the packaging system comprises a first vector and a second vector, wherein the first vector comprises a first nucleotide sequence encoding one or more AAV Rep proteins and a second nucleotide sequence encoding an AAV capsid protein, and the second vector comprises a third nucleotide sequence containing an rAAV genome. As used in the context of a packaging system as described herein, "vector" refers to a nucleic acid molecule (e.g., a plasmid, a virus, a cosmid, an artificial chromosome) that is a medium for introducing nucleic acid into a cell.

[0265] Any AAV Rep protein can be used in the packaging system disclosed herein. In one embodiment of the packaging system, the Rep nucleotide sequence encodes an AAV2 Rep protein. Suitable AAV2 Rep proteins may include, but are not limited to, Rep 78 / 68 or Rep68 / 52.

[0266] In one embodiment, the packaging system further comprises a fourth nucleotide sequence comprising one or more helper virus genes. In one embodiment, the fourth nucleotide sequence comprises adenovirus E2, E4 and VA genes. In one embodiment, the packaging system further comprises a third vector (e.g., a helper virus vector) comprising a fourth nucleotide sequence. The third vector can be an independent third vector, integrated with the first vector, or integrated with the second vector.

[0267] In one embodiment of the packaging system, the helper virus is selected from the group consisting of adenovirus, herpes virus (including herpes simplex virus (HSV)), poxvirus (e.g., vaccinia virus), cytomegalovirus (CMV), and baculovirus. In embodiments where the helper virus is an adenovirus, the adenovirus genome comprises one or more adenovirus RNA genes selected from the group consisting of E1, E2, E4, and VA. In one embodiment, the adenovirus genome comprises one or more adenovirus RNA genes selected from the group consisting of E2, E4, and VA. In embodiments where the helper virus is HSV, the HSV genome comprises one or more HSV genes selected from the group consisting of UL5 / 8 / 52, ICPO, ICP4, ICP22, and UL30 / UL42.

[0268] In one embodiment of the packaging system, the first, second and / or third vectors are contained within one or more plasmids. In one embodiment, the first vector and the third vector are contained within a first plasmid. In one embodiment, the second vector and the third vector are contained within a second plasmid.

[0269] In one embodiment of the packaging system, the first, second and / or third vectors are contained within one or more recombinant helper viruses. In one embodiment, the first vector and the third vector are contained within a recombinant helper virus. In one embodiment, the second vector and the third vector are contained within a recombinant helper virus.

[0270] In one aspect, the present invention provides a method for recombinantly preparing rAAV as described herein, wherein the method comprises transfecting or transducing cells with a packaging system as described herein under conditions operable to encapsulate the rAAV genome in a capsid to form an rAAV as described herein. Exemplary methods for recombinantly preparing rAAV include transient transfection (e.g., with one or more transfection plasmids containing a first vector and a second vector as described herein and an optional third vector), viral infection (e.g., with one or more recombinant helper viruses containing a first vector and a second vector as described herein and an optional third vector, such as adenovirus, poxvirus (e.g., vaccinia virus), herpes virus (including HSV, cytomegalovirus or baculovirus)) and stable production cell line transfection or infection (e.g., with a stable production cell, such as a mammalian or insect cell, which contains a Rep nucleotide sequence encoding one or more AAV Rep proteins as described herein and / or a Cap nucleotide sequence encoding one or more capsid proteins as described herein, and has a transfer genome as described herein delivered in the form of a plasmid or a recombinant helper virus).

[0271] Thus, the present disclosure provides a packaging system for preparing rAAV, wherein the packaging system comprises: a first nucleotide sequence encoding one or more AAV Rep proteins; a second nucleotide sequence encoding an AAV capsid protein disclosed herein; a third nucleotide sequence comprising the rAAV genome sequence described herein; and optionally a fourth nucleotide sequence comprising one or more helper virus genes (e.g., adenovirus E2, E4 and VA genes).

[0272] Examples

[0273] Example 1: Generation of modified capsids

[0274] Recombinant AAV capsids that specifically target myocytes were developed by inserting peptides within the amino acid sequence of the parental AAV capsid protein.

[0275] a. AAV2 / 5 capsid

[0276] Chimeric AAV capsid (AAV5.2, SEQ ID NO: 11) was used as a parent capsid protein to produce recombinant AAV capsid protein. The AAV capsid is encoded by three viral proteins (VPs) VP1, VP2, and VP3, which are incorporated into the 60-subunit capsid at a ratio of 1:1:10. The AAV5.2 capsid is an engineered variant of the AAV2 and AAV5 capsids, which includes the VP1 sequence from AAV2 and the VP2 / 3 sequence from AAV5.

[0277] b. Capsid Design

[0278] The VP1 and VP2 unique sequences are located inside the capsid and contain a phospholipase (PLA2) domain (in VP1) and a nuclear localization signal (VP1 and VP2). The VP3 sequence (which is contained in all three VPs) is the main protein sequence that constitutes the surface topology of the AAV capsid, which in turn determines tropism and antigenicity. The VP3 topology contains a core eight-strand (βB to βI) barrel motif and a large loop insertion between the β-strands that form the capsid surface. Conformational changes in these nine surface loops (VR-I to VR-IX) define the unique surface features of different AAV serotypes. Therefore, the peptide insertions described herein are inserted into these outward-facing loops. Specifically, loops IV and VIII are selected for peptide insertion sites.

[0279] Position 443, located at the apex of loop IV on the AAV5 capsid (SEQ ID NO: 10) or position 444 on the AAV5.2 capsid (SEQ ID NO: 11), was selected as the insertion site in loop IV, and the peptide was inserted directly C-terminal to these amino acid positions. Position 576, located at the apex of loop VIII on the AAV5 capsid (SEQ ID NO: 10) or position 577 on the AAV5.2 capsid (SEQ ID NO 11), was selected as the insertion site in loop VIII, and the peptide was inserted directly C-terminal to these amino acid positions. Insertion of the peptide immediately C-terminally to these positions should have minimal impact on capsid integrity, as the remaining VP residues interact to remain intact at the base of the capsid.

[0280] Nine different peptides (SEQ ID NO: 1-9, see Table 1) were used to engineer recombinant AAV capsid proteins. Specifically, polynucleotides encoding capsid proteins comprising parent AAV5.2 capsid proteins with peptide insertions from Table 1 were synthesized and inserted into plasmid vectors. According to AAV5.2 (SEQ ID NO: 11), each AAV capsid protein was engineered to contain only one peptide insertion, adjacent to the C-terminus of position 444 in loop IV or position 577 in loop VIII, i.e., position 443 in loop IV and position 576 in loop VIII of AAV5 capsid (SEQ ID NO: 10). The amino acid sequences of the resulting recombinant AAV capsid proteins (SEQ ID NO: 12-29) are disclosed in Table 2.

[0281] Example 2: Transduction of human muscle bundles

[0282] The ability of recombinant AAV capsids to target myocytes was analyzed by transducing myocytes in human muscle bundles.

[0283] Myocyte transduction was analyzed in a 3D muscle microtissue model generated using human skeletal muscle myoblasts (CC-2580, Lonza). Briefly, cells were seeded within the hydrogel mixture, and an appropriate volume of the seeded hydrogel mixture containing cells was added to each muscle chamber of the OMEGAMP device, ensuring that the cells were evenly spread within the seeded hydrogel and throughout the chamber. After seeding, myoblasts were maintained in myoblast seeding medium for 2 days. The medium was then replaced with 2 mL of freshly prepared myoblast differentiation medium. On day 11 of differentiation, the myoblasts were seeded with 1×10 6AAV transduction of muscle microtissues was performed by adding myoblast differentiation medium containing different AAV capsids VYAAV-5.2-GFP, AAV-VYPep15-loopIV-GFP, AAV-VYPep14-loopIV-GFP or AAV9-GFP (HEK) at an MOI of 1:1. Two days after transduction, the medium containing AAV capsids was replaced with fresh myoblast differentiation medium. Myoblast differentiation medium was partially replaced every 2 days until the day of harvest on day 18.

[0284] At day 18 after differentiation, myoblast differentiation medium was replaced with PBS containing Hoechst 3342 (1:10,000), and muscle microtissues were imaged using ImageXpico at 4× magnification. GFP expression from muscle microtissues was quantified as mean fluorescence intensity using ImageJ, and the results are expressed as the fold increase of GFP expression from different capsids relative to GFP expression from AAV9.

[0285] The results summarized in Table 5 below demonstrate that recombinant AAV capsids with a peptide inserted at position 443 in loop IV exhibited improved myocyte transduction compared to the parental AAV capsid.

[0286] Table 5. Quantification of GFP expression in muscle bundles

[0287]

[0288] Example 3: Transduction of human myotubes

[0289] The transduction efficiency of the recombinant AAV capsids was further analyzed by transducing human myotubes with the capsids.

[0290] Human skeletal muscle cells (CC-2580, Lonza) were seeded in 24-well plates at a cell density of 50,000 cells / well. When the cells reached 85-90% confluence, myoblast differentiation was induced by replacing the myoblast proliferation medium with myoblast (low serum) differentiation medium. Myotube differentiation was performed for 7 days, with partial replacement of differentiation medium every 2 days. AAV transduction of myotubes was performed at 1×10 per cell. 6 The MOI of 100 μL was carried out, mixed with 50 μL PBS at the beginning of differentiation, and the negative control was treated with 50 μL PBS only. One day after transduction, the culture medium was renewed. In order to analyze the transduction efficacy, GFP+ cells were monitored over time using the EVOS FLoid imaging system (Cat. No. 447116). One week after transduction, the transduced myotubes were imaged at 20 times magnification using the EVOS FLoid imaging system (Cat. No. 447116), then harvested and frozen for further quantitative analysis.

[0291] Figure 1A and Figure 1B The GFP imaging results shown in confirm that several recombinant AAV capsids successfully transduced human myotubes, and some capsids showed increased myotube transduction when the peptide was inserted into loop IV compared to loop VIII. Figure 2 The results shown in demonstrated that recombinant AAV capsids containing VYPep14 or VYPep15 at position 443 in loop IV (AAV5.2-VYPep14-loop IV and AAV5.2-VYPep15-loop IV) or position 576 in loop VIII (AAV5.2-VYPep14-loop VIII and AAV5.2-VYPep15-loop VIII) successfully transduced myotubes; Figure 3A and Figure 3B The results shown in indicate that recombinant AAV capsids containing VYPep14 or VYPep7 at position 443 in loop IV (AAV5.2-VYPep14-loop IV and AAV5.2-VYPep7-loop IV, respectively) showed improved myotube transduction compared to the parental AAV5.2 capsid.

[0292] Example 4: Transduction of human astrocytes and motor neurons

[0293] The transduction efficiency of the recombinant AAV capsids was further analyzed by transducing human astrocytes and motor neurons with the capsids.

[0294] Human astrocytes and iPSC-derived motor neurons were seeded at a cell density of 50,000 cells / well in 24-well plates. Parental AAV9, AAV9 with VYPep14 inserted into loop VIII (AAV9-VYPep14-loop VIII), parental AAV5.2, AAV5.2-VYPep14-loop IV, AAV5.2-VYPep14-loop VIII, AAV5.2-VYPep15-loop VIII, AAV5.2-VYPep7-loop IV, AAV5.2-VYPep7-loop VIII were used at a density of 1×10 per cell. 4 Astrocytes were transduced with AAV9, parental AAV5.2, AAV5.2-VYPep7-loop IV, AAV5.2-VYPep7-loop VIII at an MOI of 1×10 per cell. 6Transduce motor neurons. One day after transduction, update culture medium. In order to analyze the transduction efficacy of astrocytes, GFP+ cells were monitored over time using EVOS FLoid imaging system (Cat. No. 447116). One week after transduction, EVOS FLoid imaging system (Cat. No. 447116) was used to image transduced astrocytes at 20 times magnification, then harvested and frozen for further quantitative analysis. Motor neurons were stained with Hoechst one week after transduction, and Hoechst and GFP fluorescence were analyzed. Positive motor neurons% were determined using the cell scoring analysis in PICOImageXpress software, and the software scored the nuclei that were positive for Hoechst and GFP.

[0295] GFP expression of astrocytes was quantified and expressed as the percentage of GFP-positive cells to the total number of cells. Figure 4A and Figure 4B The results presented in show that several recombinant AAV capsids successfully transduced human astrocytes when the peptide was inserted into loop IV compared to loop VIII, and some capsids showed increased transduction of astrocytes. Figure 5 The results shown in demonstrate that recombinant AAV capsids containing VYPep7 at position 443 in loop IV successfully transduced human motor neurons.

[0296] The transduction efficiency of AAV5.2-VYPep7-loop IV in astrocytes was further analyzed at different MOIs over the course of eight days. iPSC-derived astrocytes were seeded on day 0 and transduced with AAV5.2-GFP, AAV9-GFP, or AAV5.2-VYPep7-loop IV at 3 different MOIs: 1 × 10 4 , 1×10 5 or 1×10 6 Fluorescence images were acquired at 2, 4, and 8 days post-transduction. The % positive cells were determined using cell scoring analysis within PICO ImageXpress software, which scored nuclei positive for both Hoechst and GFP.

[0297] Figures 6A-6C The results in Figure 2 showed that the transduction efficiency of each AAV capsid increased with increasing MOI and was similar to that of the parental AAV9 ( Fig. 6A ) or AAV5.2( Figure 6B ) compared to AAV5.2-VYPep7-loop IV ( Figure 6C ) more efficiently transduced astrocytes.

[0298] Example 5: Transduction of non-human primate astrocytes

[0299] The transduction efficiency of recombinant AAV capsids was further analyzed by transducing non-human primate (NHP) astrocytes.

[0300] Primary NHP astrocytes were seeded on day 0 and grown to 70-90% confluence. Four days after seeding, cells were incubated with 1 × 10 5 and 1×10 6 Cells were transduced at an MOI of 1.5 ng / mL. Images were acquired 3 days after transduction. The % positive cells were determined using cell scoring analysis within PICO ImageXpress software, which scored nuclei as positive for both Hoechst and GFP.

[0301] Figure 7 The results in show that the AAV5.2-VYPep7-loop IV capsid is significantly more efficient in transducing NHP astrocytes compared to the parental AAV5.2 or AAV9 capsids.

[0302] Example 6: Transduction across the blood-brain barrier (BBB)

[0303] The ability of recombinant AAV capsids to penetrate the blood-brain barrier (BBB) ​​was analyzed using a microfluidic model of the BBB.

[0304] The BBB model used to test capsids contains human cell lines of brain endothelial cells (blood side) and astrocytes (brain side) in a three-channel microfluidic platform with 40 chips in a 384-well plate format ( 3-lane 40). In each chip, perfused blood vessels of brain endothelial cells were grown against an extracellular matrix gel that was patterned by surface tension technology. Astrocytes were added on the other side of the gel to complete the BBB-on-chip model. The barrier function of the model was studied using a fluorescent barrier integrity assay. Different AAV capsids, VVAAV-5.2-GFP, VYAAV-5.2-GFP, AAV5.2-VYPep7-loop IV-GFP, AAV5.2-VYPep14-loop IV-GFP, AAV5.2-VYPep14-loop VIII-GFP, and AAV9-VYPep14-loop VIII-GFP, were inoculated at 1×10 per cell on day 10 after endothelial cell inoculation. 6The MOI of 1:1 was added to the chip on the blood side. The culture medium on the blood side and the brain side was replaced with fresh human brain microvascular endothelial cell culture medium and astrocyte culture medium, respectively. On the 6th day after transduction with AAV, the culture medium was replaced with PBS containing Hoescht 33342, 1:10,000 (Catalog No. 62249, Thermofisher Scientific), and the Organoplate was imaged at 4 times magnification using ImageXpico.

[0305] GFP expression from cells transduced with AAV5.2-GFP, AAV5.2VYPep7-loop IV, and AAV5.2VYPep14-loop IV was quantified on the blood side (endothelium) and brain side (astrocytes) and expressed as Fig. 8A ) and Day 11 ( Figure 8B ) The percentage of GFP-positive cells in the total number of cells. GFP expression of cells transduced with AAV5.2, AAV9, AAV5.2VYPep14-loop IV, and AAV5.2VYPep14-loop VIII was quantified from the blood and brain sides on day 13 after transduction and compared with the GFP expression of cells on day 6 after transduction. The results are expressed as the average GFP intensity per cell ( Figure 8C ). ImageJ software was used to determine the percentage of positive cells.

[0306] Fig. 8A , Figure 8B and Figure 8C The results in show that recombinant AAV capsids successfully penetrated the BBB and transduced astrocytes on the brain side of the BBB.

[0307] Example 7: Transduction efficiency of bicyclic capsids

[0308] Recombinant AAV5.2 capsids with two peptide insertions (one in loop IV and one in loop VIII) ("bicyclic capsids") were analyzed for their ability to transduce iPSC-derived cortical astrocytes.

[0309] Design the following recombinant AAV5.2 bicyclic capsid:

[0310] Table 6. Bicyclic capsid design

[0311]

[0312] *According to AAV5.2 (SEQ ID NO: 11), the peptide was inserted immediately C-terminally at position 444 in loop IV, i.e. position 443 in loop IV of the AAV5 capsid (SEQ ID NO: 10).

[0313] **According to AAV5.2 (SEQ ID NO: 11), the peptide was inserted directly C-terminally at position 577 in loop VIII, ie position 576 in loop VIII of AAV5 capsid (SEQ ID NO: 10).

[0314] To test these bicyclic capsids, iPSC-derived cortical astrocytes were seeded on day 0 and transduced 4 days after seeding with AAV5.2-GFP, AAV9-GFP, AAV5.2-VYPep14-IV, AAV5.2-VYPep4-IV, AAV5.2-VYPep7-IV, AAV5.2-VYPep14-IV-VYPep14-VIII-GFP, AAV5.2-VYPep14-IV-VYPep4-VIII-GFP, AAV5.2-VYPep4-IV-VYPep14-VIII-GFP, AAV5.2-VYPep5-IV-VYPep14-VIII-GFP, AAV5.2-VYPep7-IV-VYPep14-VIII-GFP at an MOI of 10^5. Images were acquired 3 days after transduction. The % positive cells were determined using cell scoring analysis within PICO ImageXpress software, which scored nuclei as positive for both Hoechst and GFP. Fig. 9A The results in show that most of the bicyclic capsids tested transduced astrocytes more efficiently than the parental AAV5.2 or AAV9 capsids.

[0315] The complete virus titers of AAV5.2-GFP, AAV9-GFP, AAV5.2-VYPep14-IV-VYPep14-VIII-GFP, AAV5.2-VYPep14-IV-VYPep4-VIII-GFP, AAV5.2-VYPep4-IV-VYPep14-VIII-GFP, AAV5.2-VYPep5-IV-VYPep14-VIII-GFP, and AAV5.2-VYPep7-IV-VYPep14-VIII-GFP were measured using the Stunner system (Unchained Labs) and are expressed as viral genomes / mL. Virus titer results are shown in Fig. 9B middle.

[0316] The complete capsid and aggregates of the double capsid preparations were also tested. The Stunner system (Unchained Labs) was used to determine the complete capsid percentage (%) and aggregation percentage values ​​of AAV5.2-GFP, AAV9-GFP, AAV5.2-VYPep14-IV-VYPep14-VIII-GFP, AAV5.2-VYPep14-IV-VYPep4-VIII-GFP, AAV5.2-VYPep4-IV-VYPep14-VIII-GFP, AAV5.2-VYPep5-IV-VYPep14-VIII-GFP, AAV5.2-VYPep7-IV-VYPep14-VIII-GFP. Fig. 9C The results in show that the capsids have a high percentage of whole capsids and a low percentage of aggregates.

[0317] ***

[0318] The scope of the present invention is not limited by the specific embodiments described herein. In fact, various modifications of the present invention, in addition to those described, will become apparent to those skilled in the art from the foregoing description and the accompanying drawings. These modifications are intended to fall within the scope of the appended claims.

[0319] All references (e.g., publications or patents or patent applications) cited herein are incorporated by reference in their entirety and for all purposes to the same extent as if each individual reference (e.g., publication or patent or patent application) was specifically and individually indicated to be incorporated by reference in its entirety for all purposes. Other embodiments are within the following claims.

Claims

1. A recombinant adeno-associated virus (AAV) capsid protein, comprising a peptide and a parent capsid protein, wherein the peptide is contained within loop IV and / or loop VIII of the parent capsid protein, and wherein the peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9.

2. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein is a clade A, clade B, clade C, clade D, clade E, clade F, clade G, clade H, clade I, AAVgo.1, AAV3, AAV4, AAV10, AAV11, AAV12, rh.32, rh32.33, rh.33, rh.34, BAAV or AAV5 capsid protein or an engineered variant thereof.

3. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence that is at least 95% identical to amino acids 193-725 of SEQ ID NO:

11.

4. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence that is at least 99% identical to amino acids 193-725 of SEQ ID NO:

11.

5. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence of amino acids 193-725 of SEQ ID NO:

11.

6. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence that is at least 95% identical to amino acids 138-725 of SEQ ID NO:

11.

7. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence that is at least 99% identical to amino acids 138-725 of SEQ ID NO:

11.

8. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence of amino acids 138-725 of SEQ ID NO:

11.

9. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 10 or 11.

10. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 10 or 11.

11. The recombinant AAV capsid protein of claim 1, wherein the parent capsid protein comprises the amino acids of SEQ ID NO: 10 or 11.

12. The recombinant AAV capsid protein of any one of claims 1 to 11, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440, 441, 442, 443, 444, 445, 446, 447, 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583 or 584 of SEQ ID NO:

11.

13. A recombinant AAV capsid protein according to claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440 of SEQ ID NO:

11.

14. A recombinant AAV capsid protein according to claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 441 of SEQ ID NO:

11.

15. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 442 of SEQ ID NO:

11.

16. A recombinant AAV capsid protein according to claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 443 of SEQ ID NO:

11.

17. A recombinant AAV capsid protein according to claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 444 of SEQ ID NO:

11.

18. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 445 of SEQ ID NO:

11.

19. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 446 of SEQ ID NO:

11.

20. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 447 of SEQ ID NO:

11.

21. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572 of SEQ ID NO:

11.

22. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 573 of SEQ ID NO:

11.

23. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 574 of SEQ ID NO:

11.

24. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 575 of SEQ ID NO:

11.

25. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 576 of SEQ ID NO:

11.

26. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 577 of SEQ ID NO:

11.

27. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 578 of SEQ ID NO:

11.

28. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 579 of SEQ ID NO:

11.

29. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 580 of SEQ ID NO:

11.

30. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 581 of SEQ ID NO:

11.

31. A recombinant AAV capsid protein according to claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 582 of SEQ ID NO:

11.

32. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 583 of SEQ ID NO:

11.

33. The recombinant AAV capsid protein of claim 12, wherein the peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 584 of SEQ ID NO:

11.

34. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having at least 95% sequence identity to amino acids 194-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (b) an amino acid sequence having at least 95% sequence identity to amino acids 194-763 of SEQ ID NO: 13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (c) an amino acid sequence having at least 95% sequence identity to amino acids 194-734 of SEQ ID NO: 14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); (d) an amino acid sequence having at least 95% sequence identity to amino acids 194-734 of SEQ ID NO: 15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); (e) an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO: 16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO: 3); (f) an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO: 17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO: 3); (g) an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); (h) an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 4); (i) an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); (j) an amino acid sequence having at least 95% sequence identity to amino acids 194-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); (k) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (l) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (m) an amino acid sequence having at least 95% sequence identity to amino acids 194-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); (n) an amino acid sequence having at least 95% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); (o) an amino acid sequence having at least 95% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); (p) an amino acid sequence having at least 95% sequence identity to amino acids 194-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); (q) an amino acid sequence having at least 95% sequence identity to amino acids 194-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or (r) an amino acid sequence having at least 95% sequence identity to amino acids 194-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9).

35. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having at least 99% sequence identity to amino acids 194-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (b) an amino acid sequence having at least 99% sequence identity to amino acids 194-763 of SEQ ID NO: 13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (c) an amino acid sequence having at least 99% sequence identity to amino acids 194-734 of SEQ ID NO: 14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); (d) an amino acid sequence having at least 99% sequence identity to amino acids 194-734 of SEQ ID NO: 15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); (e) an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO: 16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO: 3); (f) an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO: 17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO: 3); (g) an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); (h) an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 4); (i) an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); (j) an amino acid sequence having at least 99% sequence identity to amino acids 194-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); (k) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (l) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (m) an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); (n) an amino acid sequence having at least 99% sequence identity to amino acids 194-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); (o) an amino acid sequence having at least 99% sequence identity to amino acids 194-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); (p) an amino acid sequence having at least 99% sequence identity to amino acids 194-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); (q) an amino acid sequence that has at least 99% sequence identity to amino acids 194-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or (r) an amino acid sequence having at least 99% sequence identity to amino acids 194-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9).

36. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having 100% sequence identity to amino acids 194-763 of SEQ ID NO: 12; (b) an amino acid sequence having 100% sequence identity to amino acids 194-763 of SEQ ID NO: 13; (c) an amino acid sequence having 100% sequence identity to amino acids 194-734 of SEQ ID NO: 14; (d) an amino acid sequence having 100% sequence identity to amino acids 194-734 of SEQ ID NO: 15; (e) an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO: 16; (f) an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO: 17; (g) an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO: 18; (h) an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO: 19; (i) an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:20; (j) an amino acid sequence having 100% sequence identity to amino acids 194-732 of SEQ ID NO:21; (k) an amino acid sequence having 100% sequence identity to amino acids 194-742 of SEQ ID NO:22; (l) an amino acid sequence having 100% sequence identity to amino acids 194-742 of SEQ ID NO:23; (m) an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:24; (n) an amino acid sequence having 100% sequence identity to amino acids 194-739 of SEQ ID NO:25; (o) an amino acid sequence having 100% sequence identity to amino acids 194-735 of SEQ ID NO:26; (p) an amino acid sequence having 100% sequence identity to amino acids 194-735 of SEQ ID NO:27; (q) an amino acid sequence that has 100% sequence identity to amino acids 194-737 of SEQ ID NO:28; or (r) an amino acid sequence having 100% sequence identity to amino acids 194-737 of SEQ ID NO:

29.

37. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having at least 95% sequence identity to amino acids 138-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (b) an amino acid sequence having at least 95% sequence identity to amino acids 138-763 of SEQ ID NO: 13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (c) an amino acid sequence having at least 95% sequence identity to amino acids 138-734 of SEQ ID NO: 14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); (d) an amino acid sequence having at least 95% sequence identity to amino acids 138-734 of SEQ ID NO: 15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); (e) an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO: 16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO: 3); (f) an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO: 17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO: 3); (g) an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); (h) an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 4); (i) an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); (j) an amino acid sequence having at least 95% sequence identity to amino acids 138-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); (k) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (l) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (m) an amino acid sequence having at least 95% sequence identity to amino acids 138-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); (n) an amino acid sequence having at least 95% sequence identity to amino acids 138-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); (o) an amino acid sequence having at least 95% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); (p) an amino acid sequence having at least 95% sequence identity to amino acids 138-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); (q) an amino acid sequence that has at least 95% sequence identity to amino acids 138-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or (r) an amino acid sequence having at least 95% sequence identity to amino acids 138-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9).

38. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having at least 99% sequence identity to amino acids 138-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (b) an amino acid sequence having at least 99% sequence identity to amino acids 138-763 of SEQ ID NO: 13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (c) an amino acid sequence having at least 99% sequence identity to amino acids 138-734 of SEQ ID NO: 14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); (d) an amino acid sequence having at least 99% sequence identity to amino acids 138-734 of SEQ ID NO: 15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); (e) an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO: 16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO: 3); (f) an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO: 17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO: 3); (g) an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); (h) an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 4); (i) an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); (j) an amino acid sequence having at least 99% sequence identity to amino acids 138-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); (k) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (l) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (m) an amino acid sequence having at least 99% sequence identity to amino acids 138-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); (n) an amino acid sequence having at least 99% sequence identity to amino acids 138-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); (o) an amino acid sequence having at least 99% sequence identity to amino acids 138-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); (p) an amino acid sequence having at least 99% sequence identity to amino acids 138-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); (q) an amino acid sequence that has at least 99% sequence identity to amino acids 138-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or (r) an amino acid sequence having at least 99% sequence identity to amino acids 138-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9).

39. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO: 12; (b) an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO: 13; (c) an amino acid sequence having 100% sequence identity to amino acids 138-734 of SEQ ID NO: 14; (d) an amino acid sequence having 100% sequence identity to amino acids 138-734 of SEQ ID NO: 15; (e) an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO: 16; (f) an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO: 17; (g) an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO: 18; (h) an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO: 19; (i) an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:20; (j) an amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:21; (k) an amino acid sequence having 100% sequence identity to amino acids 138-742 of SEQ ID NO:22; (l) an amino acid sequence having 100% sequence identity to amino acids 138-742 of SEQ ID NO:23; (m) an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:24; (n) an amino acid sequence having 100% sequence identity to amino acids 138-739 of SEQ ID NO:25; (o) an amino acid sequence having 100% sequence identity to amino acids 138-735 of SEQ ID NO:26; (p) an amino acid sequence having 100% sequence identity to amino acids 138-735 of SEQ ID NO:27; (q) an amino acid sequence that has 100% sequence identity to amino acids 138-737 of SEQ ID NO:28; or (r) an amino acid sequence having 100% sequence identity to amino acids 138-737 of SEQ ID NO:

29.

40. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having at least 95% sequence identity to amino acids 1-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (b) an amino acid sequence having at least 95% sequence identity to amino acids 1-763 of SEQ ID NO: 13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (c) an amino acid sequence having at least 95% sequence identity to amino acids 1-734 of SEQ ID NO: 14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); (d) an amino acid sequence having at least 95% sequence identity to amino acids 1-734 of SEQ ID NO: 15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); (e) an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO: 16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO: 3); (f) an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO: 17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO: 3); (g) an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); (h) an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 4); (i) an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); (j) an amino acid sequence having at least 95% sequence identity to amino acids 1-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); (k) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (l) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (m) an amino acid sequence having at least 95% sequence identity to amino acids 1-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); (n) an amino acid sequence having at least 95% sequence identity to amino acids 1-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); (o) an amino acid sequence having at least 95% sequence identity to amino acids 1-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); (p) an amino acid sequence having at least 95% sequence identity to amino acids 1-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); (q) an amino acid sequence that has at least 95% sequence identity to amino acids 1-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or (r) an amino acid sequence having at least 95% sequence identity to amino acids 1-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9).

41. The recombinant AAV capsid protein of claim 1, comprising: (a) an amino acid sequence having at least 99% sequence identity to amino acids 1-763 of SEQ ID NO: 12, wherein amino acids 445-482 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (b) an amino acid sequence having at least 99% sequence identity to amino acids 1-763 of SEQ ID NO: 13, wherein amino acids 578-615 are ASLEEEWAQVECEVYGRGCPSGSLDESFYDWFERQLGA (SEQ ID NO: 1); (c) an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO: 14, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO: 2); (d) an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO: 15, wherein amino acids 578-586 are APLTNTVKA (SEQ ID NO: 2); (e) an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO: 16, wherein amino acids 445-451 are DDTRHWG (SEQ ID NO: 3); (f) an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO: 17, wherein amino acids 578-584 are DDTRHWG (SEQ ID NO: 3); (g) an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO: 18, wherein amino acids 445-451 are EYHHYNK (SEQ ID NO: 4); (h) an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO: 19, wherein amino acids 578-584 are EYHHYNK (SEQ ID NO: 4); (i) an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:20, wherein amino acids 445-451 are QLFPLFR (SEQ ID NO:5); (j) an amino acid sequence having at least 99% sequence identity to amino acids 1-732 of SEQ ID NO:21, wherein amino acids 578-584 are QLFPLFR (SEQ ID NO:5); (k) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:22, wherein amino acids 445-461 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (l) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:23, wherein amino acids 578-594 are FNPVTGEVPPRYPLDAR (SEQ ID NO:6); (m) an amino acid sequence having at least 99% sequence identity to amino acids 1-739 of SEQ ID NO:24, wherein amino acids 445-458 are SVTEQGAELSNEER (SEQ ID NO:7); (n) an amino acid sequence having at least 99% sequence identity to amino acids 1-739 of SEQ ID NO:25, wherein amino acids 578-591 are SVTEQGAELSNEER (SEQ ID NO:7); (o) an amino acid sequence having at least 99% sequence identity to amino acids 1-735 of SEQ ID NO:26, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8); (p) an amino acid sequence having at least 99% sequence identity to amino acids 1-735 of SEQ ID NO:27, wherein amino acids 578-584 are RGDLGL (SEQ ID NO:8); (q) an amino acid sequence that has at least 99% sequence identity to amino acids 1-737 of SEQ ID NO:28, wherein amino acids 447-453 are ASSLNIA (SEQ ID NO:9); or (r) an amino acid sequence having at least 99% sequence identity to amino acids 1-737 of SEQ ID NO:29, wherein amino acids 580-586 are ASSLNIA (SEQ ID NO:9).

42. The recombinant AAV capsid protein of claim 1, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 12-29.

43. The recombinant AAV capsid protein of claim 1, wherein the amino acid sequence of the recombinant AAV capsid protein consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 12-29.

44. A recombinant adeno-associated virus (AAV) capsid protein, comprising a first peptide, a second peptide and a parent capsid protein, wherein the first peptide is contained within loop IV of the parent capsid protein and the second peptide is contained within loop VIII of the parent capsid protein, and wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35 and 36, and the second peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35 and 36.

45. The recombinant AAV capsid protein of claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:

1.

46. ​​A recombinant AAV capsid protein according to claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9, 35 and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:

2.

47. A recombinant AAV capsid protein according to claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9, 35 and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:

3.

48. A recombinant AAV capsid protein according to claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35 and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:

4.

49. A recombinant AAV capsid protein according to claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9, 35 and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:

5.

50. The recombinant AAV capsid protein of claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises an amino acid sequence of SEQ ID NO:

6.

51. A recombinant AAV capsid protein according to claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9, 35 and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:

7.

52. The recombinant AAV capsid protein of claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises an amino acid sequence of SEQ ID NO:

8.

53. The recombinant AAV capsid protein of claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises an amino acid sequence of SEQ ID NO:

9.

54. A recombinant AAV capsid protein according to claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 1-9, 35 and 36, and the second peptide comprises the amino acid sequence of SEQ ID NO:

35.

55. The recombinant AAV capsid protein of claim 44, wherein the first peptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 1-9, 35, and 36, and the second peptide comprises an amino acid sequence of SEQ ID NO:

36.

56. A recombinant AAV capsid protein according to any one of claims 45 to 55, wherein the parent capsid protein is a clade A, clade B, clade C, clade D, clade E, clade F, clade G, clade H, clade I, AAVgo.1, AAV3, AAV4, AAV10, AAV11, AAV12, rh.32, rh32.33, rh.33, rh.34, BAAV or AAV5 capsid protein or an engineered variant thereof.

57. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises an amino acid sequence that is at least 95% identical to amino acids 193-725 of SEQ ID NO:

11.

58. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises an amino acid sequence that is at least 99% identical to amino acids 193-725 of SEQ ID NO:

11.

59. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises the amino acid sequence of amino acids 193-725 of SEQ ID NO:

11.

60. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises an amino acid sequence that is at least 95% identical to amino acids 138-725 of SEQ ID NO:

11.

61. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises an amino acid sequence that is at least 99% identical to amino acids 138-725 of SEQ ID NO:

11.

62. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises an amino acid sequence of amino acids 138-725 of SEQ ID NO:

11.

63. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises an amino acid sequence that is at least 95% identical to SEQ ID NO: 10 or 11.

64. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises an amino acid sequence that is at least 99% identical to SEQ ID NO: 10 or 11.

65. The recombinant AAV capsid protein of any one of claims 45 to 55, wherein the parent capsid protein comprises the amino acids of SEQ ID NO: 10 or 11.

66. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein: (a) the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440, 441, 442, 443, 444, 445, 446 or 447 of SEQ ID NO: 11; and (b) the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572, 573, 574, 575, 576, 577, 578, 579, 580, 581, 582, 583 or 584 of SEQ ID NO:

11.

67. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 440 of SEQ ID NO:

11.

68. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 441 of SEQ ID NO:

11.

69. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 442 of SEQ ID NO:

11.

70. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 443 of SEQ ID NO:

11.

71. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 444 of SEQ ID NO:

11.

72. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 445 of SEQ ID NO:

11.

73. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 446 of SEQ ID NO:

11.

74. The recombinant AAV capsid protein of any one of claims 45 to 65, wherein the first peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 447 of SEQ ID NO:

11.

75. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 572 of SEQ ID NO:

11.

76. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 573 of SEQ ID NO:

11.

77. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 574 of SEQ ID NO:

11.

78. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 575 of SEQ ID NO:

11.

79. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 576 of SEQ ID NO:

11.

80. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 577 of SEQ ID NO:

11.

81. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 578 of SEQ ID NO:

11.

82. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 579 of SEQ ID NO:

11.

83. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 580 of SEQ ID NO:

11.

84. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 581 of SEQ ID NO:

11.

85. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 582 of SEQ ID NO:

11.

86. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 583 of SEQ ID NO:

11.

87. The recombinant AAV capsid protein of any one of claims 45 to 74, wherein the second peptide is positioned immediately C-terminal to the amino acid in the parent capsid corresponding to amino acid 584 of SEQ ID NO:

11.

88. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having at least 95% sequence identity to amino acids 194-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 194-742 of SEQ ID NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 194-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2), and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

89. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having at least 99% sequence identity to amino acids 194-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 194-742 of SEQ ID NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 194-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2), and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

90. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having at least 100% sequence identity to amino acids 194-745 of SEQ ID NO:30; (b) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:31; (c) an amino acid sequence having at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:32; (d) an amino acid sequence that has at least 100% sequence identity to amino acids 194-742 of SEQ ID NO:33; or (e) an amino acid sequence having at least 100% sequence identity to amino acids 194-744 of SEQ ID NO:

34.

91. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having at least 95% sequence identity to amino acids 138-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 138-742 of SEQ ID NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 138-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2), and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

92. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having at least 99% sequence identity to amino acids 138-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 138-742 of SEQ ID NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 138-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2), and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

93. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:30; (b) an amino acid sequence having 100% sequence identity to amino acids 138-763 of SEQ ID NO:31; (c) an amino acid sequence having 100% sequence identity to amino acids 138-734 of SEQ ID NO:32; (d) an amino acid sequence that has 100% sequence identity to amino acids 138-734 of SEQ ID NO:33; or (e) An amino acid sequence having 100% sequence identity to amino acids 138-732 of SEQ ID NO:

34.

94. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having at least 95% sequence identity to amino acids 1-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 95% sequence identity to amino acids 1-742 of SEQ ID NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 95% sequence identity to amino acids 1-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2), and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

95. The recombinant AAV capsid protein of claim 45, comprising: (a) an amino acid sequence having at least 99% sequence identity to amino acids 1-745 of SEQ ID NO:30, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 592-597 are RGDLGL (SEQ ID NO:8); (b) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:31, wherein amino acids 449-454 are RGDLGL (SEQ ID NO:8), and amino acids 588-594 are RTIGPSV (SEQ ID NO:35); (c) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:32, wherein amino acids 445-451 are RTIGPSV (SEQ ID NO:35), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); (d) an amino acid sequence having at least 99% sequence identity to amino acids 1-742 of SEQ ID NO:33, wherein amino acids 445-451 are LSSRLDA (SEQ ID NO:36), and amino acids 589-594 are RGDLGL (SEQ ID NO:8); or (e) an amino acid sequence having at least 99% sequence identity to amino acids 1-744 of SEQ ID NO:34, wherein amino acids 445-453 are APLTNTVKA (SEQ ID NO:2), and amino acids 591-596 are RGDLGL (SEQ ID NO:8).

96. The recombinant AAV capsid protein of claim 45, comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 30-34.

97. The recombinant AAV capsid protein of claim 45, wherein the amino acid sequence of the recombinant AAV capsid protein consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 30-34.

98. An isolated polynucleotide encoding the recombinant AAV capsid protein of any one of claims 1 to 97.

99. A vector comprising the polynucleotide according to claim 98.

100. The vector of claim 99, which is a plasmid or a viral vector.

101. The vector of claim 100, wherein the viral vector is a retroviral vector, a herpes virus vector, a baculovirus vector or an adenovirus vector.

102. The vector according to any one of claims 99 to 101, which is an expression vector.

103. A recombinant cell comprising the polynucleotide of claim 98 or the vector of any one of claims 99 to 102.

104. A method for producing a recombinant AAV capsid protein, the method comprising culturing the recombinant cell according to claim 103 under conditions where a polynucleotide is expressed and the capsid protein is produced.

105. A recombinant adeno-associated virus (rAAV), comprising: (a) a capsid comprising one or more of the recombinant AAV capsid proteins of any one of claims 1 to 97; and (b) rAAV genome.

106. The rAAV of claim 105, wherein the rAAV genome comprises a transgene.

107. The rAAV of claim 106, wherein the transgene encodes a polypeptide or a non-coding RNA.

108. The rAAV of claim 106 or 107, wherein the transgene encodes an antibody or fragment thereof.

109. The rAAV of claim 108, wherein the transgene encodes a scFv, a nanobody, or a VHH.

110. A method of delivering a transgene to a cell, the method comprising contacting the cell with the rAAV of any one of claims 105 to 109 under conditions that transduce the cell and express the transgene.

111. The method of claim 110, wherein the cell is a myocyte, a microglia, an astrocyte, a neuron, or a cardiomyocyte.

112. A method of delivering a transgene to muscle tissue of a subject in need thereof, the method comprising administering to the subject the rAAV of any one of claims 105 to 109.

113. The method of claim 112, wherein the subject suffers from a disease associated with muscle tissue.

114. A method of treating a neuromuscular or neurodegenerative disease or condition in a subject in need thereof, the method comprising administering to the subject an effective amount of the rAAV of any one of claims 105 to 109.

115. The method of claim 114, wherein the neuromuscular disease or disorder is selected from the group consisting of: a myopathy, a hereditary cardiomyopathy, a metabolic myopathy, a distal myopathy, a muscular dystrophy, a congenital myopathy, a spinal muscular atrophy (SMA), a motor neuron disease, a congenital myopathy, a congenital muscular dystrophy, a motor neuron disease, a Duchenne muscular dystrophy, a Becker muscular dystrophy, a limb-girdle muscular dystrophy, a myotonic dystrophy, a myotubular myopathy, a centronuclear myopathy, a nemaline myopathy, a selenoprotein N-related myopathy, a Pompe disease, a glycogen storage disease III, a spinal muscular atrophy, an amyotrophic lateral sclerosis, a Charcot-Marie-Tooth disease, multiple sclerosis, a myositis, a polymyositis, and a dermatomyositis.

116. The method of claim 114, wherein the neurodegenerative disease or disorder is selected from the group consisting of motor neuron disease (MND), parkinsonism syndrome, Alzheimer's dementia, progressive supranuclear palsy (PSP), Huntington's disease, multiple system atrophy (MSA), spinocerebellar ataxia (SCA), and Friedreich's ataxia.

117. The method of any one of claims 112 to 116, wherein the rAAV is administered to the subject intravenously, intraperitoneally, subcutaneously, intramuscularly, intrathecally, or intradermally.

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