Capsid polypeptides and methods of use thereof

Abstract

The disclosure is directed in part to dependoparvovirus capsid polypeptides that can be used to deliver payloads.

Description

CAPSID POLYPEPTIDES AND METHODS OF USE THEREOF1. CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the priority benefit of U.S. provisional application no. 63 / 696,218, filed September 18, 2024, the contents of which are incorporated herein in their entireties by reference thereto.2. SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML Sequence Listing, created on September 16, 2025, is named DYN-012WO_SL.xml and is 189,885 bytes in size.3. BACKGROUND

[0003] Dependoparvoviruses, e.g., adeno-associated dependoparvoviruses, e.g., adeno-associated viruses (AAVs), are of interest as vectors for delivering various payloads to cells, including in human subjects.4. SUMMARY

[0004] The present disclosure relates, in part, to improved dependoparvovirus capsid polypeptides, such as VP1 , VP2 and / or VP3 capsid polypeptides, methods of producing a dependoparvovirus comprising capsid polypeptides, compositions for use in the same, as well as viral particles produced by the same In certain aspects, the present disclosure relates to viral particles comprising the improved dependoparvovirus capsid polypeptides, with increased muscle biodistribution and / or transduction as compared to viral particles, e.g., without the mutations in the improved dependoparvovirus capsid polypeptides.

[0005] In certain aspects, the disclosure provides capsid polypeptides comprising the sequence motif “RGD” and one or both of (i) an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7 and (ii) a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7. Optionally, the capsid polypeptides comprise an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NOV. The “RGD” sequence motif is typically inserted into loop VIII (VR-VIII variable region) of the capsid polypeptide, e.g., immediately subsequent (C-terminal) to a position corresponding to Q585, S586, A587, Q588, A589, Q590, A591 , Q592, or T593 of the VP1 capsid polypeptide of SEQ ID NO:7

[0006] In some aspects, the disclosure provides capsid polypeptides comprising the amino acid sequence TRGDYAS (SEQ ID NO:16). The amino acid sequence TRGDYAS (SEQ ID NO:16) is typically inserted into loop VIII (VR-VIII variable region) of the capsid polypeptide, e.g., immediately subsequent (C-terminal) to a position corresponding to Q585, S586, A587, Q588, A589, Q590, A591 , Q592, or T593 of the VP1 capsid polypeptide of SEQ ID NOV. In some embodiments, the amino acid sequence TRGDYAS (SEQ ID NO:16) is immediately subsequent to a position corresponding to A587of the VP1 capsid polypeptide of SEQ ID NO:7. In some embodiments, the capsid polypeptide further comprises an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7, a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7, an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NO:7, or any combination of two or all three of the foregoing.

[0007] Also disclosed, in some embodiments, are capsid polypeptides comprising the amino acid sequence RGDYSMT (SEQ ID NO:17) The amino acid sequence RGDYSMT (SEQ ID NO:17) is typically inserted into loop VIII (VR-VIII variable region) of the capsid polypeptide, e.g., immediately subsequent (C-terminal) to a position corresponding to Q585, S586, A587, Q588, A589, Q590, A591 , Q592, or T593 of the VP1 capsid polypeptide of SEQ ID NOV. In some embodiments, the amino acid sequence RGDYSMT (SEQ ID NO:17) is immediately subsequent to a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NOV. In some embodiments, the capsid polypeptide further comprises a histidine at a position corresponding to T582 of the VP1 capsid polypeptide of the VP1 capsid polypeptide of SEQ ID NOV

[0008] Accordingly, the present disclosure provides a capsid polypeptide described herein. In some embodiments, the capsid polypeptide comprises an amino acid sequence having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity to a VP1 polypeptide of SEQ ID NO: 12 or 14, or to a VP2 or VP3 portion thereof. In some embodiments, a capsid polypeptide of the disclosure comprises one or more mutation differences or a mutation set present in any one of V1 or V2.

[0009] In some embodiments, the percentage sequence identity is calculated excluding any targeting peptide sequence insertion(s) in the capsid polypeptide sequence. In other embodiments, the percentage sequence identity is calculated including any targeting peptide sequence insertion(s) in the capsid polypeptide sequence

[0010] Exemplary VP1 amino acid sequences of the disclosure are set forth in SEQ ID NOS:12 or 14. Additional exemplary capsid polypeptides are disclosed in Section 6.2 and numbered embodiments 1 to 240

[0011] The present disclosure further provides a nucleic acid comprising a nucleotide sequence encoding a capsid polypeptide as provided for herein, e.g., a capsid polypeptide disclosed in Section 6.2 or any one of numbered embodiments 1 to 240. In some embodiments, the nucleic acid molecule comprises a nucleotide sequence of SEQ ID NO:13 or 15, a fragment thereof (e.g., a fragment thereof encoding a VP2 or VP3 polypeptide), or a variant of any of the foregoing having at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity thereto. In some embodiments, the percentage sequence identity is calculated excluding any nucleotide sequence(s) encoding targeting peptide sequence insertion(s) In some embodiments, the percentage sequence identity is calculated including any nucleotide sequence(s) encoding targeting peptide sequence insertion(s). In some embodiments, the nucleic acid is a vector, e.g., a plasmid. Exemplary nucleic acids are disclosed in Section 6.2 and numbered embodiments 241 to 276.- Z -

[0012] The present disclosure further provides a dependoparvovirus particle comprising a capsid polypeptide, a capsid polypeptide disclosed in Section 6.2 or any one of numbered embodiments 1 to 240 and / or a nucleic acid described herein, e.g., a nucleic acid disclosed in Section 6.2 or any one of numbered embodiments 241 to 276 or a nucleic acid comprising a transgene as disclosed in Section 6.7.1. In some embodiments, the dependoparvovirus is an adeno-associated dependoparvovirus (AAV). In some embodiments, the AAV is AAV9, e.g., a variant AAV9. Exemplary virus particles are disclosed in Section 6.4 and numbered embodiments 277 to 303 In some embodiments, the virus particles have one or more characteristics disclosed in Section 6.5 and numbered embodiments 290 to 295.

[0013] In some embodiments, the disclosure is directed, in part, to a cell, cell-free system, or other translation system comprising a nucleic acid or vector described herein, e.g., comprising a sequence encoding a capsid polypeptide having one or more mutations described herein, for example a capsid polypeptide disclosed in Section 6.2 or any one of numbered embodiments 1 to 240. In some embodiments, the cell, cell-free system, or other translation system comprises a dependoparvovirus particle described herein, e.g., wherein the particle comprises a nucleic acid comprising a sequence encoding a capsid polypeptide, e.g., a capsid polypeptide disclosed in Section 6.2 or any one of numbered embodiments 1 to 240 and / or a nucleic acid described herein, e.g., a nucleic acid disclosed in Section 6.2 or any one of numbered embodiments 241 to 276 or a nucleic acid comprising a transgene as disclosed in Section 6.7.1 Exemplary cells, cell-free and other translation systems and their use to produce dependoparvovirus particles are disclosed in Section 6.6 and in numbered embodiments 556 to 567

[0014] The present disclosure further provides methods of using a dependoparvovirus disclosed herein, e.g., for delivering a payload to a cell or treating a disease or condition in a subject. The methods typically comprise contacting the cell or administering to the subject a dependoparvovirus particle described herein in an amount effective to treat the disease or condition. Exemplary methods are disclosed in Section 6.7 and numbered embodiments 304 to 555. The dependoparvovirus particles may be in the form of a composition, e.g., a pharmaceutical composition comprising the dependoparvovirus particles and a pharmaceutically acceptable carrier or excipient, for example as described in Section 6.7.2 and numbered embodiment 568.

[0015] Additional features, advantages and applications of the capsid polypeptides, nucleic acids, dependoparvovirus particles of the disclosure and methods of their production and use are more particularly described below.5. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] FIGS. 1 A-1C. Illustration of exemplary AAV serotype alignments. Amino acids that are present only in VP1 polypeptides are in normal text; amino acids that are present only in VP1 and VP2 polypeptides are in bold; amino acids that are present in VP1 , VP2 and VP3 polypeptides are underlined. Figure discloses SEQ ID NOS 3, 1 , 7, 5, and 9, respectively, in order of appearance.6. DETAILED DESCRIPTION6.1. Definitions

[0017] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure. In case of conflict, the present specification, including definitions, will control. Generally, nomenclature used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics, analytical chemistry, synthetic organic chemistry, medicinal and pharmaceutical chemistry, and protein and nucleic acid chemistry and hybridization described herein are those well-known and commonly used in the art. Enzymatic reactions and purification techniques are performed according to manufacturer’s specifications, as commonly accomplished in the art or as described herein. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. Throughout this specification and embodiments, the words “have” and “comprise,” or variations such as “has,” “having,” “comprises,” or “comprising,” will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers. All publications and other references mentioned herein are incorporated by reference in their entirety. Although a number of documents are cited herein, this citation does not constitute an admission that any of these documents forms part of the common general knowledge in the art

[0018] A, An, The: As used herein, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.

[0019] About, Approximately: As used herein, the terms “about” and “approximately” shall generally mean an acceptable degree of error for the quantity measured given the nature or precision of the measurements. Exemplary degrees of error are within 15 percent (%), typically, within 10%, and more typically, within 5% of a given value or range of values. Any disclosure herein of a value preceded by the term “about” or “approximately” is also a disclosure of the value per se. For example, disclosure of “about 10 pg / ml” is a disclosure of the value “10 pg / ml.”

[0020] Corresponds to: As used herein, the term “corresponds to” as used in reference to a position in a sequence, such as an amino acid or nucleic acid sequence, can be used in reference to an entire capsid polypeptide or polynucleotide sequence, such as the full-length sequence of the capsid polypeptide that comprises a VP1 , VP2, and VP3 polypeptide, or a nucleic acid molecule encoding the same. In some embodiments, the term “corresponds to” can be used in reference to a region or domain of the capsid polypeptide. For example, a position that corresponds to a position in the VP1 section of the reference capsid polypeptide can correspond to the VP1 portion of the polypeptide of the variant capsid polypeptide. Thus, when aligning the two sequences to determine whether a position corresponds to another position the full-length polypeptide can be used or domains (regions) can be used to determine whether a position corresponds to a specific position. In some embodiments, the region is the VP1 polypeptide. In some embodiments, the region is the VP2polypeptide. In some embodiments, the region is the VP3 polypeptide. In some embodiments, when the reference polypeptide is the wild-type sequence (e.g , full-length or region) of a certain serotype of AAV, the variant polypeptide can be of the same serotype with a mutation made at such corresponding position as compared to the reference sequence (e.g., full-length or region). In some embodiments, the variant capsid polypeptide is a different serotype as compared to the reference sequence

[0021] Dependoparvovirus capsid: As used herein, the term “dependoparvovirus capsid” refers to an assembled viral capsid comprising dependoparvovirus polypeptides In some embodiments, a dependoparvovirus capsid is a functional dependoparvovirus capsid, e.g., is fully folded and / or assembled, is competent to infect a target cell, or remains stable (e.g., folded / assembled and / or competent to infect a target cell) for at least a threshold time.

[0022] Dependoparvovirus particle: As used herein, the term “dependoparvovirus particle” refers to an assembled viral capsid comprising dependoparvovirus polypeptides and a packaged nucleic acid, e.g., comprising a payload, one or more components of a dependoparvovirus genome (e.g., a whole dependoparvovirus genome), or both In some embodiments, a dependoparvovirus particle is a functional dependoparvovirus particle, e.g., comprises a desired payload, is fully folded and / or assembled, is competent to infect a target cell, or remains stable (e.g., folded / assembled and / or competent to infect a target cell) for at least a threshold time.

[0023] Dependoparvovirus X particle / capsid: As used herein, the term “dependoparvovirus X particle / capsid” refers to a dependoparvovirus particle / capsid comprising at least one polypeptide or polypeptide encoding nucleic acid sequence derived from a naturally occurring dependoparvovirus X species or serotype. For example, a dependoparvovirus B particle refers to a dependoparvovirus particle comprising at least one polypeptide or polypeptide encoding nucleic acid sequence derived from a naturally occurring dependoparvovirus B sequence. Derived from, as used in this context, means having at least 70, 75, 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identity to the sequence in question. Correspondingly, an AAVX particle / capsid, as used herein, refers to an AAV particle / capsid comprising at least one polypeptide or polypeptide encoding nucleic acid sequence derived from a naturally occurring AAV X serotype For example, an AAV9 particle refers to an AAV particle comprising at least one polypeptide or polypeptide encoding nucleic acid sequence derived from a naturally occurring AAV9 sequence Sometimes, a dependoparvovirus X capsid is referred to as “Wild Type” or“wt” when such capsid comprises capsid polypeptides from a specified sequence identifier associated with such dependoparvovirus X capsid. Thus, for example, the terms wild-type AAV9 capsid or wtAAV9 capsid (or simply wtAAV9) are used interchangeably and refer to a capsid that comprises capsid polypeptides of SEQ ID NOV (e.g., a VP1 capsid of SEQ ID NOV and VP2 and VP3 portions thereof).

[0024] Edit Distance: Sequences disclosed herein may be described in terms of “edit distance.” The minimum number of sequence edits, i.e. , additions, substitutions, or deletions of a single amino acid (for amino acid sequence) or a single nucleotide (for nucleotide sequences), which change onesequence into another sequence is the edit distance between the two sequences. The term “edit distance” is often used interchangeably with the term “Levenshtein distance.”

[0025] Exogenous: As used herein, the term “exogenous” refers to a feature, sequence, or component present in a circumstance (e.g., in a nucleic acid, polypeptide, or cell) that does not naturally occur in said circumstance. For example, a nucleic acid sequence encoding a polypeptide can comprise an exogenous codon (e.g., codon encoding for an amino acid that does not naturally occur in that position, for example in a reference sequence), such as provided for herein Use of the term exogenous in this fashion means that the codon in question at this position does not occur naturally, e.g., is not present in AAV9, e g., is not present in SEQ ID NO:7. In some embodiments, the codon replaces an endogenous codon. In some embodiments, the exogenous codon is inserted into the nucleic acid sequence, for example, relative to a reference sequence. A person of skill will readily understand that a sequence (e.g., a codon) can be exogenous when provided in a particular sequence (e.g., that does not naturally comprise the codon at the site in question) but may not be exogenous in a second sequence (e g , that does naturally comprise that particular codon at the site in question).

[0026] Functional: As used herein in reference to a polypeptide component of a dependoparvovirus capsid (e.g., Cap (e.g., VP1 , VP2, and / or VP3) or Rep), the term “functional” refers to a polypeptide which provides at least 50, 60, 70, 80, 90, or 100% of the activity of a naturally occurring version of that polypeptide component (e.g., when present in a host cell). For example, a functional VP1 polypeptide can stably fold and assemble into a dependoparvovirus capsid (e g., that is competent for packaging and / or secretion). As used herein in reference to a dependoparvovirus capsid or particle, “functional” refers to a capsid or particle comprising one or more of the following production characteristics: comprises a desired payload, is fully folded and / or assembled, is competent to infect a target cell, or remains stable (e.g , folded / assembled and / or competent to infect a target cell) for at least a threshold time.

[0027] Mutation Difference: As used herein with respect to a polypeptide sequence, means a single amino acid mutation (e.g., substitution, insertion or deletion) present in a subject polypeptide sequence, relative to a reference polypeptide sequence. In various embodiments, the reference polypeptide sequence is a polypeptide of any one of SEQ ID NOU , SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, or a VP2 or VP3 portion thereof. In some embodiments, the reference polypeptide is any one of SEQ ID NO:7, 12, 14, or a VP2 or VP3 portion thereof. In a preferred embodiment, the reference polypeptide is a polypeptide of SEQ ID NO:7. In various embodiments, the subject polypeptide is SEQ ID NO:12, 14, or a VP2 or VP3 portion thereof.

[0028] Mutation Set: As used herein, the term “mutation set” refers to the complete set of single amino acid mutations (substitutions, deletions and / or insertions) in a variant capsid polypeptide sequence (e.g., a polypeptide sequence of SEQ ID NO:12, 14, or a VP2 or VP3 portion thereof) relative to a reference sequence (e.g., a wild-type reference sequence). In some embodiments, the reference sequence is wild-type AAV9 VP1 capsid polypeptide (SEQ ID NO:7) or a VP2 or VP3portion thereof. In some cases, part of the mutation set (i.e. , more than one single amino acid mutation) is notated collectively, however, it will be understood that even when referred to in this way, the mutation set is a collection of single amino acid mutations. For example, an insertion of amino acid 1, 2, and 3 between amino acid N at position nn and amino acid W at position ww of a reference sequence may be notated as "Nnn_3aa_Www_123," and it will be understood that each of amino acids 1 , 2 and 3 represent separate single amino acid mutations within the mutation set. The mutation sets for certain variant capsid polypeptides described herein are found, for example, in Table 1. In some embodiments, a variant capsid polypeptide of the disclosure comprises a mutation set not consisting solely of a mutation set present in a capsid polypeptide of SEQ ID NO:12 or 14.

[0029] Nucleic Acid: As used herein, in its broadest sense, the term “nucleic acid” refers to any compound and / or substance that is or can be incorporated into an oligonucleotide chain In some embodiments, a nucleic acid is a compound and / or substance that is or can be incorporated into an oligonucleotide chain via a phosphodiester linkage. As will be clear from context, in some embodiments, “nucleic acid" refers to an individual nucleic acid monomer (e g , a nucleotide and / or nucleoside); in some embodiments, “nucleic acid’ refers to an oligonucleotide chain comprising individual nucleic acid monomers or a longer polynucleotide chain comprising many individual nucleic acid monomers. In some embodiments, a “nucleic acid” is or comprises RNA; in some embodiments, a “nucleic acid" is or comprises DNA. In some embodiments, a nucleic acid is, comprises, or consists of one or more natural nucleic acid residues. In some embodiments, a nucleic acid is, comprises, or consists of one or more nucleic acid analogs. In some embodiments, a nucleic acid is, comprises, or consists of one or more modified, synthetic, or non-naturally occurring nucleotides In some embodiments, a nucleic acid analog differs from a nucleic acid in that it does not utilize a phosphodiester backbone. For example, in some embodiments, a nucleic acid is, comprises, or consists of one or more “peptide nucleic acids”, which are known in the art and have peptide bonds instead of phosphodiester bonds in the backbone, are considered within the scope of the present invention. Alternatively or additionally, in some embodiments, a nucleic acid has one or more phosphorothioate and / or 5'-N-phosphoramidite linkages rather than phosphodiester bonds. In some embodiments, a nucleic acid has a nucleotide sequence that encodes a functional gene product such as an RNA or protein. In some embodiments, a nucleic acid is partly or wholly single stranded; in some embodiments, a nucleic acid is partly or wholly double stranded.

[0030] Or: Unless indicated otherwise, an “or” conjunction is intended to be used in its correct sense as a Boolean logical operator, encompassing both the selection of features in the alternative (A or B, where the selection of A is mutually exclusive from B) and the selection of features in conjunction (A or B, where both A and B are selected). In some places in the text, the term “and / or” is used for the same purpose, which shall not be construed to imply that “or” is used with reference to mutually exclusive alternatives.

[0031] Percent Identity: Sequences disclosed herein may be described in terms of “percent identity” (% identity). For calculating percent identity between two amino acid sequences or two nucleic acid sequences, the two sequences to be compared are aligned using the EMBOSS Needle PairwiseSequence Alignment software tool based on the Needleman and Wunsch algorithm (Needleman & Wunsch, 1970, J. Mol. Biol 48(3):443-53) (available at www.ebi.ac uk / Tools / psa / emboss_needle / ) using the following parameters: Matrix: BLOSUM62 (for amino acid sequences) or DNAfull (for DNA sequences); Gap Open: 10; Gap Extend: 0.5; End Gap Penalty: false; End Gap Open: 10; and End Gap Extend: 0.5. Percent identity is determined by dividing the number of amino acid or nucleotide matches in the alignment by the length of the alignment and multiplying by 100. For example, if an alignment of two amino acid sequences has 95 matching amino acids and an alignment length of 100 amino acids, the two sequences have 95% identity.

[0032] When calculating percent identity of two capsid polypeptides, one or both of which contain(s) one or more targeting peptide insertions, percent identity can be determined without removing the targeting peptide insertion sequence(s) from the capsid polypeptide sequence(s) or, alternatively, percent identity can be determined after removing the targeting peptide insertion sequence(s) from the capsid polypeptide sequence(s). For example, if a first capsid polypeptide has an identical sequence to a second capsid polypeptide, except that the first capsid polypeptide has a 7-mer targeting peptide insertion, the two capsid polypeptides have less than 100% sequence identity when percent identity is determined without removal of the targeting peptide insertion sequence from the first capsid polypeptide sequence, whereas the two capsid polypeptides have 100% sequence identity when the targeting peptide insertion sequence is removed from the first capsid polypeptide sequence prior to calculating percent identity. References herein to percent identity of capsid polypeptides without mention of a targeting peptide refer to percent identity of the capsid polypeptides determined following removal of targeting polypeptide insertion sequence(s), if any, present in both capsid polypeptides, unless required otherwise by context. References herein to percent identity calculated “taking targeting peptide insertions into account" means that the percent identity is calculated without removal of targeting polypeptide insertion sequence(s), if any, present in both capsid polypeptides. References herein to percent identity calculated “without taking targeting peptide insertions into account” means that the percent identity is calculated following removal of targeting polypeptide insertion sequence(s), if any, present in both capsid polypeptides.

[0033] Polypeptide, peptide, and protein: The terms “polypeptide,” “peptide” and “protein” are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear or branched, it may comprise modified amino acids, and it may be interrupted by non-amino acids.

[0034] Targeting Peptide: As used herein, a “targeting peptide” refers to a peptide inserted into, or attached to, a capsid polypeptide to alter the tropism of the capsid polypeptide. A targeting peptide can be inserted into an AAV capsid sequence for enhanced targeting to a desired cell-type, tissue, or organ, for example for enhanced targeting to muscle tissue. A targeting peptide is typically 3 to 20 amino acids in length, for example, 3 to 12 amino acids, 5 to 12 amino acids, 5 to 10 amino acids, or 7 to 10 amino acids in length

[0035] Treating: As used herein, the term “treating a disease or condition” refers to treating a manifest disease or condition, for example, where the subject is already suffering from one or moresymptoms of the disease or condition, or refers to treating a pre-manifest disease or condition, for example, where the subject is identified as having a disease or condition but is not yet exhibiting one or more symptoms of the disease or condition. Pre-manifest conditions may be identified by, for example, genetic testing.

[0036] Variant: As used herein, a “variant capsid polypeptide” refers to a polypeptide that differs from a reference sequence (e.g, SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, or SEQ ID NO:11 , preferably SEQ ID NO:7), or sequence subunit thereof such as a VP2 or VP3 portion thereof). The variant capsid polypeptide can, for example, comprise a mutation (e.g., substitution, deletion, or insertion). In some embodiments, the variant is about, or at least, 70%, 75%, 80%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the reference sequence. It will be clear to the skilled artisan from this disclosure that any capsid polypeptide, for example any capsid polypeptide disclosed herein, for example, a capsid polypeptide of SEQ ID NO: 12 or 1 , is a variant capsid polypeptide with respect to another capsid polypeptide having a different amino acid sequence, e g, another capsid polypeptide with a reference sequence as set forth above. Thus, the term “variant capsid polypeptide” herein means, and is used interchangeably with, “capsid polypeptide,” and does not require any comparison to a specific reference sequence. In some embodiments, the reference sequence is a polypeptide comprising SEQ ID NO:7. In some embodiments, the reference sequence comprises or consists of a VP1 , VP2 or VP3 polypeptide, e g, of SEQ ID NO:7. In some contexts used herein, the term “variant” refers to a virus particle that includes a variant capsid polypeptide, e g, described herein.6.2. Capsid Polypeptides

[0037] The disclosure is directed, in part, to a variant capsid polypeptide, wherein the variant capsid polypeptide comprises a mutation (insertion, deletion, or substitution) as compared to the wild-type sequence In some embodiments, the wild-type sequence is SEQ ID NO:7. The disclosure is directed, in part, to a variant capsid polypeptide comprising SEQ ID NO:7 with one or more mutations as compared to SEQ ID NO:7. The mutation can be, for example, an insertion, deletion, or substitution as compared to the wild-type sequence In some embodiments, the wild-type sequence is SEQ ID NO:7.

[0038] In some embodiments, disclosed are variant capsid polypeptides that increase muscle (cardiac and / or skeletal muscle) biodistribution and / or transduction when incorporated into a viral particle as compared to reference capsid polypeptides, e.g, wild-type capsid polypeptides. The variant capsid polypeptides of the disclosure are engineered to include an amino acid sequence comprising the integrin-targeting sequence motif “RGD”, e.g, via a peptide insertion.

[0039] Certain aspects of the disclosure relate to variant capsid polypeptides comprising the sequence motif “RGD” and one or both of (i) an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7 and (ii) a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7. Optionally, the capsid polypeptides comprise an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NO:7. At least a portion of the RGD-containing sequence motif is typically in a VR-VIII variable region and / or in a surface loopof the capsid polypeptide. For example, the RGD-containing sequence motif can be positioned (a) C- terminal to a position corresponding to Q585, S586, or A587 of the VP1 capsid polypeptide of SEQ ID NO:7, e g., immediately C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NO:7, and / or (b) N-terminal to a position corresponding to Q588, A589, or Q590 of the VP1 capsid polypeptide of SEQ ID NO:7. In some embodiments, the RGD sequence motif comprises a subsequence Y or F amino acid to produce an RGDY (SEQ ID NO: 513) or RGDF (SEQ ID NO: 514) motif. In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDX1X2X3X4, with Xi to X4 each being any amino acid. In various aspects of the motif RGDX1X2X3X4, Xi, X2, and X3 are each independently selected from L, G, V, and A and / or X4 is S, V, A, G, or L. In some embodiments, at least one of X2 and X3 is G. In some embodiments, the RGD sequence motif comprises the amino acid sequence TRGDYAS (SEQ ID NO:16). In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDYSMT (SEQ ID NO:17). In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDLGLS (SEQ ID NO:504). In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDLSTP (SEQ ID NO:505). In some embodiments, the RGD sequence motif comprises the amino acid sequence SNSRGDYNSL (SEQ ID NO:506). In some embodiments, the RGD sequence motif comprises the amino acid sequence ENRRGDFNNT (SEQ ID NQ:507). In some embodiments, the RGD sequence motif comprises the amino acid sequence SRGDYNSL (SEQ ID NO:508). In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDYNSL (SEQ ID NO:509). In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDLST (SEQ ID NO:510). In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDYVGL (SEQ ID NO:511 ). In some embodiments, the RGD sequence motif comprises the amino acid sequence RGDAVGV (SEQ ID NO:512). Additional RGD- containing sequence motifs are disclosed in WO 2020 / 206189A1, WO 2022 / 226374 A1 , WO 2022 / 053630 A1, and WO 2019 / 207132 A1 , which are incorporated by reference herein.

[0040] In some embodiments, the disclosure provides a variant capsid polypeptide comprising the (a) the amino acid sequence X1X2RGDX3X4X5X6, where Xi is any amino acid or absent, each of X2-X5 is any amino acid, and XB is any amino acid or absent (b) one or both of (i) an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7 and (ii) a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7. Optionally, the capsid polypeptide comprise an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NO:7. At least a portion of the amino acid sequence X1X2RGDX3X4X5X6 is typically in a VR- VIII variable region and / or in a surface loop of the capsid polypeptide For example, the amino acid sequence X1X2RGDX3X4X5X6 can be positioned (a) C-terminal to a position corresponding to Q585, S586, or A587 of the VP1 capsid polypeptide of SEQ ID NOV, e.g, immediately C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV, and / or (b) N-terminal to a position corresponding to Q588, A589, or Q590 of the VP1 capsid polypeptide of SEQ ID NOV. In various embodiments of the amino acid sequence X1X2RGDX3X4X5X6, (a) Xi is any amino acid or absent; (b) X2 is threonine, (c) X3 is tyrosine, (d) X4 is alanine, (e) X5 is serine, (f) Xs is any amino acid(optionally isoleucine) or absent, or (g) any combination of two or more (or optionally all) of (a) through (f).

[0041] Other aspects of the disclosure relate to variant capsid polypeptides comprising the amino acid sequence TRGDYAS (SEQ ID NO:16). At least a portion of SEQ ID NO:16 is typically in a VR- VIII variable region and / or in a surface loop of the capsid polypeptide For example, the amino acid sequence of SEQ ID NO:16 can be positioned (a) C-terminal to a position corresponding to Q585, S586, or A587 of the VP1 capsid polypeptide of SEQ ID NOV, e.g., immediately C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV, and / or (b) N-terminal to a position corresponding to Q588, A589, or Q590 of the VP1 capsid polypeptide of SEQ ID NOV, e.g., immediately N-terminal to a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NOV. In some embodiments, the variant capsid polypeptide comprises a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NOV and / or an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NOV.

[0042] In certain embodiments of variant capsid polypeptides comprising the amino acid sequence TRGDYAS (SEQ ID NO:16), the disclosure provides a variant capsid polypeptide comprising the amino acid sequence TRGDYASI (SEQ ID NO:18). At least a portion of SEQ ID NO:18 is typically in a VR-VIII variable region and / or in a surface loop of the capsid polypeptide. For example, the amino acid sequence of SEQ ID NO: 18 can be positioned (a) C-terminal to a position corresponding to Q585, S586, or A587 of the VP1 capsid polypeptide of SEQ ID NOV, e.g., immediately C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV, and / or (b) N- terminal to a position corresponding to A589, or Q590 of the VP1 capsid polypeptide of SEQ ID NOV, e.g., immediately N-terminal to a position corresponding to A589 of the VP1 capsid polypeptide of SEQ ID NOV. In some embodiments, the variant capsid polypeptide lacks an amino acid at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NOV and / or comprises a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NOV and / or comprises an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NOV.

[0043] In certain embodiments of variant capsid polypeptides comprising the amino acid sequence TRGDYAS (SEQ ID NO:16), the disclosure provides a variant capsid polypeptide comprising the amino acid sequence TRGDYASIAQAQTQ (SEQ ID NO:19), or an amino acid sequence having one, two, or three amino acid differences as compared to the amino acid sequence of SEQ ID NO:19 The amino acid of SEQ ID NO: 19 or the amino acid sequence having one, two or three amino acid differences as compared to the amount acid sequence of SEQ ID NO:19 can be positioned N-terminal to a position corresponding to Q597, V596 or W595 of the VP1 capsid polypeptide of SEQ ID NOV, for example immediately N-terminal to a position corresponding to W595 of the VP1 capsid polypeptide of SEQ ID NOV.

[0044] In certain embodiments of variant capsid polypeptides comprising the amino acid sequence TRGDYAS (SEQ ID NO:16), the disclosure provides a variant capsid polypeptide comprising the amino acid sequence TRGDYASIAQAQTQWVQNQGA (SEQ ID NO:20), or an amino acid sequencehaving one, two, or three amino acid differences as compared to the amino acid sequence of SEQ ID NO:20.

[0045] In certain embodiments of variant capsid polypeptides comprising the amino acid sequence TRGDYAS (SEQ ID NO:16), the disclosure provides a variant capsid polypeptide comprising the amino acid sequence SATRGDYASIAQAQTQWVQNQGAL (SEQ ID NO:21), or an amino acid sequence having one, two, or three amino acid differences as compared to the amino acid sequence of SEQ ID NO:21.

[0046] Other aspects of the disclosure relate to variant capsid polypeptides comprising the amino acid sequence RGDYSMT (SEQ ID NO:17) At least a portion of SEQ ID NO:17 is typically in a VR- VIII variable region and / or in a surface loop of the capsid polypeptide For example, the amino acid sequence of SEQ ID NO: 17 can be positioned (a) C-terminal to a position corresponding to Q585, S586, or A587 of the VP1 capsid polypeptide of SEQ ID NOV, e.g., immediately C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV, and / or (b) N-terminal to a position corresponding to Q588, A589, or Q590 of the VP1 capsid polypeptide of SEQ ID NOV, , e.g., immediately N-terminal to a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NOV. In some embodiments, the variant capsid polypeptide comprises a histidine at a position corresponding to T582 of the VP1 capsid polypeptide of SEQ ID NOV.

[0047] In certain embodiments of variant capsid polypeptides comprising the amino acid sequence RGDYSMT (SEQ ID NO: 17), the disclosure provides a variant capsid polypeptide comprising the amino acid sequence HNHQSAQRGDYSMT (SEQ ID NO:22), or an amino acid sequence having one, two, or three amino acid differences as compared to the amino acid sequence of SEQ ID NO:21.

[0048] Variant capsid polypeptides of the disclosure comprising the sequence motif “RGD” (e.g., the amino acid sequence of SEQ ID NO:16 or SEQ ID NO:17) typically comprise an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a reference AAV serotype, e.g., as described herein, e.g., to SEQ ID NOV. In some embodiments, the percent sequence identity is calculated not taking into account any peptide insertions (e.g., not taking into account the amino acid sequence of SEQ ID NO:16 or SEQ ID NO:17).

[0049] Exemplary variant capsid polypeptides comprising the amino acid sequence TRGDYAS (SEQ ID NO: 16) include the VP1 capsid polypeptide referred to herein as V1, corresponding to a capsid polypeptide of SEQ ID NO: 12, as well as VP2 and VP3 polypeptides thereof.

[0050] Exemplary variant capsid polypeptides comprising the amino acid sequence RGDYSMT (SEQ ID NO:17) include the VP1 capsid polypeptide referred to herein as V2, corresponding to a capsid polypeptide of SEQ ID NO: 14, as well as VP2 and VP3 polypeptides thereof.

[0051] Mutation sets, including peptide insertions, associated with V1 (corresponding to a capsid polypeptide of SEQ ID NO:12) and V2 (corresponding to a capsid polypeptide of SEQ ID NO:14) in relation to a VP1 polypeptide of SEQ ID NOV are shown in Table 1.

[0052] In some embodiments, a variant capsid polypeptide of the disclosure comprises the entire mutation set of V1 or the entire mutation set of V2, e.g., as compared to a VP1 capsid polypeptide of SEQ ID NOG

[0053] In various embodiments, the variant capsid polypeptide is, but for the amino acid sequence TRGDYAS (SEQ ID NO:16) or the amino acid sequence RGDYSMT (SEQ ID NO:17), at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a reference AAV serotype described herein. In various embodiments, the variant capsid polypeptide is, but for the mutation differences or mutation set associated with V1 or V2, at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:1 (e.g., a VP1, VP2 or VP3 sequence of SEQ ID NO:1 ).

[0054] In various embodiments, the variant capsid polypeptide is, but for the amino acid sequence TRGDYAS (SEQ ID NO:16) or the amino acid sequence RGDYSMT (SEQ ID NO:17), at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NOG (e g , a VP1 , VP2 or VP3 sequence of SEQ ID NOG).

[0055] In various embodiments, the variant capsid polypeptide is, but for the amino acid sequence TRGDYAS (SEQ ID NO:16) or the amino acid sequence RGDYSMT (SEQ ID NO:17), at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NOG (e.g., a VP1 , VP2 or VP3 sequence of SEQ ID NOG).

[0056] In various embodiments, the variant capsid polypeptide is, but for the amino acid sequence TRGDYAS (SEQ ID NO:16) or the amino acid sequence RGDYSMT (SEQ ID NO:17), at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NOG (e.g., a VP1 , VP2 or VP3 sequence of SEQ ID NOG)

[0057] In various embodiments, the variant capsid polypeptide is, but for the amino acid sequence TRGDYAS (SEQ ID NO:16) or the amino acid sequence RGDYSMT (SEQ ID NO:17), at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NOG (e.g., a VP1 , VP2 or VP3 sequence of SEQ ID NOG).

[0058] In various embodiments, the variant capsid polypeptide is, but for the amino acid sequence TRGDYAS (SEQ ID NO:16) or the amino acid sequence RGDYSMT (SEQ ID NO:17), at least 90%, atleast 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:11 (e.g., a VP1 , VP2 or VP3 sequence of SEQ ID NO:11).

[0059] In some embodiments, a variant capsid polypeptide is provided that comprises a variant capsid polypeptide that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a variant capsid polypeptide as provided herein, e g., a VP1 capsid polypeptide of SEQ ID NO:12 (and optionally comprising the amino acid sequence TRGDYAS (SEQ ID NO:16)) or SEQ ID NO:14 (and optionally comprising the amino acid sequence RGDYSMT (SEQ ID NO:17)), or a VP2 or VP3 portion thereof.

[0060] In some embodiments, the variant capsid polypeptide comprises a VP1 , VP2, VP3, or any combination thereof, that each has about 1 to about 20 mutations as compared to a VP1 polypeptide of SEQ ID NO:12 or SEQ ID NO:14, and comprises one or more of the mutation differences or the entire mutation set of V1 or V2.

[0061] In some embodiments, the variant capsid polypeptide comprises a VP1 , VP2, VP3, or any combination thereof, that each has about 1 to about 10 mutations as compared to a VP1 polypeptide of SEQ ID NO:12 or SEQ ID NO:14, and comprises one or more of the mutation differences or the entire mutation set of V1 or V2

[0062] In some embodiments, the variant capsid polypeptide comprises a VP1 , VP2, VP3, or any combination thereof, that each has about 1 to about 5 mutations as compared to a VP1 polypeptide of SEQ ID NO:12 or SEQ IDNO:14, and comprises one or more of the mutation differences or the entire mutation set of V1 or V2.

[0063] The mutations to capsid polypeptide sequences described herein are described in relation to a position and / or amino acid at a position within a reference sequence, e.g., SEQ ID NO:7. Thus, in some embodiments, the capsid polypeptides described herein are variant capsid polypeptides of the reference sequence, e.g., SEQ ID NO:7, e.g., include capsid polypeptides comprising at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the reference capsid polypeptide sequence (e.g., reference capsid polypeptide VP1, VP2 and / or VP3 sequence), e g., SEQ ID NOY (or VP2 or VP3 sequence comprised therein) and include the amino acid sequence TRGDYAS (SEQ ID NO:16) or the amino acid sequence RGDYSMT (SEQ ID NO:17), optionally together with one or more mutations described herein.

[0064] It will be understood by the skilled artisan, and without being bound by theory, that each amino acid position within a reference sequence corresponds to a position within the sequence of other reference capsid polypeptides such as capsid polypeptides derived from dependoparvoviruses with different serotypes. Such corresponding positions are identified using sequence alignment tools known in the art. A particularly preferred sequence alignment tool is EMBOSS Needle Pairwise Sequence Alignment software tool based on the Needleman and Wunsch algorithm (Needleman & Wunsch, 1970, J. Mol. Biol 48(3):443-53) (available on the World Wide Web at ebi.ac.uk / Tools / psa / emboss_needle / ) An alignment of exemplary reference capsid polypeptides isshown in FIGS. 1A-1C Thus, in some embodiments, the variant capsid polypeptides of the invention include variants of reference capsid polypeptides that include one or more mutations described herein in such reference capsid polypeptides at positions corresponding to the position of the mutation described herein in relation to a different reference capsid polypeptide. Thus, for example, a mutation described as XnnnY relative to SEQ ID NO:7 (where X is the amino acid present at position nnn in SEQ ID NOV and Y is the amino acid mutation at that position, e.g., described herein), the disclosure provides variant capsid polypeptides comprising at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a reference capsid polypeptide sequence (e.g., reference capsid polypeptide VP1 , VP2 and / or VP3 sequence) other than SEQ ID NO:7 (or VP2 or VP3 sequence comprised therein) and further comprising the disclosed mutation at a position corresponding to position nnn of SEQ ID NO:7 (e.g., comprising Y at the position in the new variant capsid polypeptide sequence that corresponds to position nnn of SEQ ID NOV). As described above, such corresponding position is determined using a sequence alignment tool, such as, for example, the clustal omega tool described above. Examples of corresponding amino acid positions of exemplary known AAV serotypes is provided in FIGS. 1 A-1 C. In some embodiments, the variant is a variant of the AAV9 capsid polypeptide, which can be referred to as a “AAV9 variant capsid polypeptide” or “variant AAV9 capsid polypeptide ”

[0065] Thus, in some embodiments, the disclosure provides variant capsid polypeptide sequences that are variants of a reference sequence other than SEQ ID NOV, e g., a reference sequence other than SEQ ID NOV as described herein, which include one or more mutation corresponding to the mutations described herein. In some embodiments, such variants include mutations corresponding to all of the mutations associated with SEQ ID NO:12 or SEQ ID NO:14.

[0066] The variant capsid polypeptides described herein are optionally variants of reference capsids serotypes known in the art. Non-limiting examples of such reference AAV serotypes include AAV1 , AAVrhI O, AAV-DJ, AAV-DJ8, AAV5, AAVPHP.B (PHP.B), AAVPHP.A (PHP.A), AAVG2B-26, AAVG2B- 13, AAVTH1.1-32, AAVTH1.1- 35, AAVPHP.B2 (PHP B2), AAVPHP.B3 (PHP.B3), AAVPHP.N / PHP.B- DGT, AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T, AAVPHP.B-DGT-T, AAVPHRB-GGT-T, AAVPHP.B-SGS, AAVPHP B-AQP, AAVPHRB-QQP, AAVPHRB-SNP(3), AAVPHP.B-SNP, AAVPHRB-QGT, AAVPHRB-NQT, AAVPHP.B- EGS, AAVPHP. B-SGN, AAVPHP.B- EGT, AAVPHP.B-DST, AAVPHP.B-DST, AAVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B-SQP, AAVPHP.B-QLP, AAVPHP.B-TMP, AAVPHP.B-TTP, AAVPHP.eB, AAVPHP S / G2A12, AAVG2A15 / G2A3 (G2A3), AAVG2B4 (G2B4), AAVG2B5 (G2B5), PHP.S, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV4, AAV4-4, AAV6, AAV6.1 , AAV6.2, AAV6.1.2, AAV7, AAV7 2, AAV8, AAV9.11 , AAV9.13, AAV9, AAV9 K449R (or K449R AAV9), AAV9.16, AAV9 24, AAV9.45, AAbiodisV9.47, AAV9 61 , AAV9.68, AAV9 84, AAV9.9, AAV10, AAV11 , AAV12, AAV16.3, AAV24.1 , AAV27.3, AAV42.12, AAV42- 1 b, AAV42-2, AAV42-3a, AAV42-3b, AAV42-4, AAV42-5a, AAV42-5b, AAV42-6b, AAV42-8, AAV42-10, AAV42-11 , AAV42-12, AAV42-13, AAV42-15, AAV42-aa, AAV43-1 , AAV43-12, AAV43-20, AAV43-21 , AAV43-23, AAV43-25, AAV43-5, AAV44.1 , AAV44.2, AAV44.5, AAV223.1 , AAV223.2, AAV223.4, AAV223.5, AAV223.6, AAV223.7, AAV1-7 / rh.48, AAV1-8 / rh 49, AAV2-15 / rh.62, AAV2-3 / rh.61 , AAV2-4 / rh.5O, AAV2-5 / rh.51 , AAV3.1 / hu.6, AAV3.1 / hu.9, AAV3-9 / rh 52, AAV3-11 / rh.53, AAV4- 8 / r11.64, AAV4-9 / rh.54, AAV4-19 / rh.55, AAV5-3 / rh.57, AAV5-22 / rh.58, AAV7.3 / hu.7, AAV16 8 / hu.1O, AAV16.12 / hu,11 , AAV29.3 / bb.1, AAV29.5 / bb.2, AAV106.1 / hu 37, AAV114.3 / hu.4O, AAV127.2 / hu.41 , AAV127.5Zhu.42, AAV128.3 / hu.44, AAV130.4 / hu.48, AAV145. 1 / hu.53, AAV145.5 / hu.54, AAV145.6 / hu.55, AAV161.10 / hu.60, AAV161.6 / hu.61, AAV33.12 / hu.17, AAV33.4 / hu.15, AAV33.8 / hu 16, AAV52 / hu.19, AAV52.1 / hu.2O, AAV58.2 / hu.25, AAVA3.3, AAVA3.4, AAVA3 5, AAVA3.7, AAVC1 , AAVC2, AAVC5, AAVF3, AAVF5, AAVH2, AAVrh.72, AAVhu 8, AAVrh.68, AAVrh.70, AAVpi.1 , AAVpi.3, AAVpi.2, AAVrh.60, AAVrh.44, AAVrh.65, AAVrh.55, AAVrh.47, AAVrh.69, AAVrh.45, AAVrh.59, AAVhu.12, AAVH6, AAVH-1 / hu.1, AAVH-5 / hu.3, AAVLG- 10 / rh.40, AAVLG-4 / rh.38, AAVLG- 9 / hu.39, AAVN721-8 / rh.43, AAVCh.5, AAVCh.5R1 , AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVCy.5R1 , AAVCy.5R2, AAVCy.5R3, AAVCy.5R4, AAVcy.6, AAVhu.1 , AAVhu.2, AAVhu.3, AAVhu.4, AAVhu.5, AAVhu.6, AAVhu.7, AAVhu 9, AAVhu.10, AAVhu 11 , AAVhu.13, AAVhu.15, AAVhu.16, AAVhu.17, AAVhu.18, AAVhu.2O, AAVhu.21 , AAVhu.22, AAVhu.23.2, AAVhu 24, AAVhu.25, AAVhu.27, AAVhu.28, AAVhu.29, AAVhu.29R, AAVhu 31 , AAVhu.32, AAVhu.34, AAVhu.35, AAVhu.37, AAVhu.39, AAVhu.40, AAVhu.41 , AAVhu.42, AAVhu.43, AAVhu.44, AAVhu.44R1 , AAVhu.44R2, AAVhu.44R3, AAVhu.45, AAVhu.46, AAVhu.47, AAVhu.48, AAVhu.48R1 , AAVhu.48R2, AAVhu.48R3, AAVhu.49, AAVhu.51 , AAVhu.52, AAVhu.54, AAVhu.55, AAVhu.56, AAVhu.57, AAVhu.58, AAVhu.60, AAVhu.61 , AAVhu 63, AAVhu.64, AAVhu.66, AAVhu.67, AAVhu.14 / 9, AAVhu.t 19, AAVrh.2, AAVrh.2R, AAVrh 8, AAVrh.8R, AAVrh.10, AAVrh.12, AAVrh.13, AAVrh.13R, AAVrh.14, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.20, AAVrh.21 , AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.31 , AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, AAVrh.37R2, AAVrh.38, AAVrh.39, AAVrh.4O, AAVrh.46, AAVrh.48, AAVrh.48.1 , AAVrh.48.1 .2, AAVrh.48.2, AAVrh 49, AAVrh 51 , AAVrh 52, AAVrh 53, AAVrh.54, AAVrh.56, AAVrh.57, AAVrh.58, AAVrh.61 , AAVrh.64, AAVrh.64R1, AAVrh.64R2, AAVrh.67, AAVrh.73, AAVrh.74 (also referred to as AAVrh74), AAVrh8R, AAVrh8R A586R mutant, AAVrh8R R533A mutant, AAAV, BAAV, caprine AAV, bovine AAV, AAVhE1.1 , AAVhEr1.5, AAVhER1.14, AAVhEr1.8, AAVhEr1.16, AAVhEr1.18, AAVhEr1.35, AAVhEr1.7, AAVhEr1.36, AAVhEr2.29, AAVhEr2 4, AAVhEr2.16, AAVhEr2.30, AAVhEr2.31 , AAVhEr2.36, AAVhER1.23, AAVhEr3.1 , AAV2.5T , AAV-PAEC, AAV-LK01 , AAV-LK02, AAV- LK03, AAV-LK04, AAV-LK05, AAV-LK06, AAV-LK07, AAV-LK08, AAV-LK09, AAV- LK10, AAV-LK11, AAV-LK12, AAV-LK13, AAV-LK14, AAV-LK15, AAV-LK16, AAV- LK17, AAV-LK18, AAV-LK19, AAV-PAEC2, AAV-PAEC4, AAV-PAEC6, AAV-PAEC7, AAV-PAEC8, AAV-PAEC 11 , AAV- PAEC^, AAV-2-pre-miRNA-101 , AAV-8h, AAV- 8b, AAV-h, AAV-b, AAV SM 10-2 , AAV Shuffle 100-1 , AAV Shuffle 100-3, AAV Shuffle 100-7, AAV Shuffle 10-2, AAV Shuffle 10-6, AAV Shuffle 10-8, AAV Shuffle 100- 2, AAV SM 10-1. AAV SM 10-8 . AAV SM 100-3, AAV SM 100-10, BNP61 AAV, BNP62 AAV, BNP63 AAV, AAVrh.50, AAVrh.43, AAVrh.62, AAVrh.48, AAVhu.19, AAVhu.11 , AAVhu.53, AAV4- 8 / rh.64, AAVLG-9 / hu.39, AAV54.5 / hu.23, AAV54.2 / hu.22, AAV54.7 / hu.24, AAV54. 1 / hu 21 , AAV54.4R / hu.27, AAV46.2 / hu.28, AAV46.6 / hu.29, AAV128.1 / hu.43, true type AAV (ttAAV), UPENN AAV 10, Japanese AAV 10 serotypes, AAV CBr-7 1 , AAV CBr-7 10, AAV CBr-7 2, AAV CBr-7 3, AAV CBr-7.4, AAV CBr-7.5, AAV CBr-7.7, AAV CBr-7 8, AAV CBr-B7.3, AAV CBr-B7.4, AAV CBr-E1 , AAV CBr- E2, AAV CBr-E3, AAV CBr-E4, AAV CBr-E5, AAV CBr-e5, AAV CBr-E6, AAV CBr-E7, AAV CBr- E8, AAV CHt-1 , AAV CHt-2, AAV CHt-3, AAV CHt-6.1 , AAV CHt-6.10, AAV CHt-6.5, AAV CHt-6 6, AAVCHt-6.7, AAV CHt-6.8, AAV CHt-P1, AAV CHt-P2, AAV CHt-P5, AAV CHt-P6, AAV CHt-P8, AAV CHt- P9, AAV CKd-1 , AAV CKd-10, AAV CKd-2, AAV CKd-3, AAV CKd-4, AAV CKd-6, AAV CKd-7, AAV CKd-8, AAV CKd-B1, AAV CKd-B2, AAV CKd-B3, AAV CKd-B4, AAV CKd-B5, AAV CKd-B6, AAV CKd- B7, AAV CKd-B8, AAV CKd-H1, AAV CKd-H2, AAV CKd-H3, AAV CKd-H4, AAV CKd- H5, AAV CKd- H6, AAV CKd-N3, AAV CKd-N4, AAV CKd-N9, AAV CLg-F1 , AAV CLg-F2, AAV CLg-F3, AAV CLg-F4, AAV CLg-F5, AAV CLg-F6, AAV CLg-F7, AAV CLg-F8, AAV CLv-1 , AAV CLv1-1 , AAV Clv1-10, AAV CL.vl-2, AAV CLv-12, AAV CLv1-3, AAV CLv-13, AAV CLv1-4, AAV Clv1 -7, AAV Clv1 -8, AAV Clv1 -9, AAV CLv-2, AAV CLv-3, AAV CLv-4, AAV CLv-6, AAV CLv-8, AAV CL.V-D1, AAV CLv-D2, AAV CLv-D3, AAV CLv-D4, AAV CLv-D5, AAV CLv-D6, AAV CLv-D7, AAV CLv-D8, AAV CL.V-E1, AAV CLv-K1 , AAV CLV-K3, AAV CLV-K6, AAV CLv-L4, AAV CLv-L5, AAV CLv-L6, AAV CL.V-M1, AAV CL.V-M11 , AAV CLv- M2, AAV CLV-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-R1 , AAV CLv-R2, AAV CLV-R3, AAV CLV-R4, AAV CLV-R5, AAV CLV-R6, AAV CLV-R7, AAV CLV-R8, AAV CLV-R9, AAV CSp-1 , AAV CSp-10, AAV CSp-11 , AAV CSp-2, AAV CSp-3, AAV CSp-4, AAV CSp-6, AAV CSp-7, AAV CSp-8, AAV CSp-8.10, AAV CSp-8.2, AAV CSp-8.4, AAV CSp-8.5, AAV CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1 , AAVF11 / HSC11 , AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, and / or AAVF9 / HSC9, 7m8, Sparkl OO, AAVMYO and variants thereof.

[0067] In some embodiments, the reference AAV capsid sequence comprises an AAV2 sequence In some embodiments, the reference AAV capsid sequence comprises an AAV5 sequence In some embodiments, the reference AAV capsid sequence comprises an AAV8 sequence In some embodiments, the reference AAV capsid sequence comprises an AAV9 sequence In some embodiments, the reference AAV capsid sequence comprises an AAVrh74 sequence. While not wishing to be bound by theory, it is understood that a reference AAV capsid sequence comprises a VP1 region. In certain embodiments, a reference AAV capsid sequence comprises a VP1 , VP2 and / or VP3 region, or any combination thereof A reference VP1 sequence may be considered synonymous with a reference AAV capsid sequence

[0068] An exemplary reference sequence of wild-type AAV2, SEQ ID NO: 1 (wild-type AAV2) is as follows:MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNGLDKGEPV NEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRVLEPLGL VEEPVKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVPDPQPLGQPPAAP SGLGTNTMATGSGAPMADNNEGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQI SSQSGASNDNHYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQ NDGTTTIANNLTSTVQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMVPQYGYLTLNNGSQAVGRSSF YCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTNTPSGTTTQSRL QFSQAGASDIRDQSRNWLPGPCYRQQRVSKTSADNNNSEYSWTGATKYHLNGRDSLVNPGPAMA SHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQRGNRQAAT ADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQILIKNTPVPANPST TFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYNKSVNVDFTVDTNGVYSEPRPIG TRYLTRNL (SEQ ID NO:1 )

[0069] In the sequence above, the sequence found in VP1 , VP2 and VP3 is underlined (e.g., a VP3 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 203-735 of SEQ ID NO:1), the sequence found in both VP1 and VP2 is in bold (e.g., a VP2 capsid polypeptide includes, e.g., consists of, the sequence corresponding to amino acids 138-735 of SEQ ID NO:1) and the sequence that is not underlined or bold is found only in VP1 (e.g., a VP1 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 1-735 of SEQ ID NO:1).

[0070] An example nucleic acid sequence encoding SEQ ID NO:1 is SEQ ID NO:2

[0071] An exemplary reference sequence of wild-type AAV5, SEQ ID NO:3 (wild-type AAV5), is as follows:MSFVDHPPDWLEEVGEGLREFLGLEAGPPKPKPNQQHQDQARGLVLPGYNYLGPGNGLDRGEPVN RADEVAREHDISYNEQLEAGDNPYLKYNHADAEFQEKLADDTSFGGNLGKAVFQAKKRVLEPFGLVE EGAKTAPTGKRIDDHFPKRKKARTEEDSKPSTSSDAEAGPSGSQQLQIPAQPASSLGADTMSAGG GGPLGDNNQGADGVGNASGDWHCDSTWMGDRVVTKSTRTWVLPSYNNHQYREIKSGSVDGSNA NAYFGYSTPWGYFDFNRFHSHWSPRDWQRLINNYWGFRPRSLRVKIFNIQVKEVTVQDSTTTIANNL TSTVQVFTDDDYQLPYVVGNGTEGCLPAFPPQVFTLPQYGYATLNRDNTENPTERSSFFCLEYFPSK MLRTGNNFEFTYNFEEVPFHSSFAPSQNLFKLANPLVDQYLYRFVSTNNTGGVQFNKNLAGRYANTY KNWFPGPMGRTQGWNLGSGVNRASVSAFATTNRMELEGASYQVPPQPNGMTNNLQGSNTYALEN TMIFNSQPANPGTTATYLEGNMLITSESETQPVNRVAYNVGGQMATNNQSSTTAPATGTYNLQEIVP GSVWMERDVYLQGPIWAKIPETGAHFHPSPAMGGFGLKHPPPMMLIKNTPVPGNITSFSDVPVSSFI TQYSTGQVTVEMEWELKKENSKRWNPEIQYTNNYNDPQFVDFAPDSTGEYRTTRPIGTRYLTRPL (SEQ ID NO:3)

[0072] Unless otherwise noted, SEQ ID NO:3 is the reference sequence. In the sequence above, the sequence found in VP1, VP2 and VP3 is underlined (e.g., a VP3 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 193-724 of SEQ ID NO:3), the sequence found in both VP1 and VP2 is in bold (e.g , a VP2 capsid polypeptide includes, e.g., consists of, the sequence corresponding to amino acids 137-724 of SEQ ID NO:3) and the sequence that is not underlined or bold is found only in VP1 (e g., a VP1 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 1-724 of SEQ ID NO:3).

[0073] The wild-type reference sequence of SEQ ID NO:3 can be encoded by a reference nucleic acid molecule sequence of SEQ ID NO:4.

[0074] An exemplary reference sequence of wild-type AAV8, SEQ ID NO:5 (wild-type AAV8), is as follows:MAADGYLPDWLEDNLSEGIREWWALKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFNGLDKGEPV NAADAAALEHDKAYDQQLQAGDNPYLRYNHADAEFQERLQEDTSFGGNLGRAVFQAKKRVLEPLGL VEEGAKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPARKRLNFGQTGDSESVPDPQPLGEPPAAP SGVGPNTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQI SNGTSGGATNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLSFKLFNIQVKEVT QNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSF YCLEYFPSQMLRTGNNFQFTYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQTTGGTANTQT LGFSQGGPNTMANQAKNWLPGPCYRQQRVSTTTGQNNNSNFAWTAGTKYHLNGRNSLANPGIAM ATHKDDEERFFPSNGILIFGKQNAARDNADYSDVMLTSEEEIKTTNPVATEEYGIVADNLQQQNTAPQI GTVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPT TFNQSKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTSVDFAVNTEGVYSEPRPIG TRYLTRNL (SEQ ID NO:5)

[0075] In the sequence above, the sequence found in VP1 , VP2 and VP3 is underlined (e.g., a VP3 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 204-738 of SEQ ID NO:5), the sequence found in both VP1 and VP2 is in bold (e.g., a VP2 capsid polypeptide includes, e.g., consists of, the sequence corresponding to amino acids 138-738 of SEQ ID NO:5) and the sequence that is not underlined or bold is found only in VP1 (e.g., a VP1 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 1-738 of SEQ ID NO:5).

[0076] An example nucleic acid sequence encoding SEQ ID NO:5 is SEQ ID NO:6

[0077] An exemplary reference sequence of wild-type AAV9, SEQ ID NO:7 (wild-type AAV9) is as follows:MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPGNGLDKGEP VNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRLLEPLG LVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPDPQPIGEPPAAPS GVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQIS NSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTD NNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSF YCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGSGQNQQTL KFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSLMNPGPAMA SHKEGEDRFFPLSGSLIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADP PTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPR PIGTRYLTRNL (SEQ ID NO:7)

[0078] In the sequence above, the sequence found in VP1 , VP2 and VP3 is underlined (e.g., a VP3 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 203-736 ofSEQ ID NO:7), the sequence found in both VP1 and VP2 is in bold (e.g., a VP2 capsid polypeptide includes, e.g., consists of, the sequence corresponding to amino acids 138-736 of SEQ ID NO:7) and the sequence that is not underlined or bold is found only in VP1 (e.g., a VP1 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 1-736 of SEQ ID NO:7).

[0079] An example nucleic acid sequence encoding SEQ ID NO:7 is SEQ ID NO:8

[0080] An exemplary reference sequence of wild-type AAVrh74, SEQ ID NO:9 (wild-type AAVrh74), is as follows:MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDNGRGLVLPGYKYLGPFNGLDKGEPV NAADAAALEHDKAYDQQLQAGDNPYLRYNHADAEFQERLQEDTSFGGNLGRAVFQAKKRVLEPLGL VESPVKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPAKKRLNFGQTGDSESVPDPQPIGEPPAGP SGLGSGTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQI SNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVT QNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSF YCLEYFPSQMLRTGNNFEFSYNFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQQ LLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAM ATHKDDEERFFPSSGVLMFGKQGAGKDNVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAA PIVGAVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEP RPIGTRYLTRNL (SEQ ID NO:9)

[0081] An alternative exemplary reference sequence of SEQ ID NO:11 (alternate wild-type AAVrh74) is as follows:MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDNGRGLVLPGYKYLGPFNGLDKGEPV NAADAAALEHDKAYDQQLQAGDNPYLRYNHADAEFQERLQEDTSFGGNLGRAVFQAKKRVLEPLGL VESPVKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPAKKRLNFGQTGDSESVPDPQPIGEPPAGP SGLGSGTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQI SNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVT QNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQAVGRSSF YCLEYFPSQMLRTGNNFEFSYNFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQQ LLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAM ATHKDDEERFFPSSGVLMFGKQGAGKDNVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAA PIVGAVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPAD PPTTFTKAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEPR PIGTRYLTRNL (SEQ ID NO:11 )

[0082] In the sequences above (SEQ ID NO:9 or SEQ ID NO:11), the sequence found in VP1, VP2 and VP3 is underlined (e.g., a VP3 capsid polypeptide includes, e.g., consists of, amino acids corresponding to amino acids 204-738 of SEQ ID NO:9), the sequence found in both VP1 and VP2 is in bold (e.g., a VP2 capsid polypeptide includes, e.g , consists of, the sequence corresponding to amino acids 137-738 of SEQ ID NO:9) and the sequence that is not underlined or bold is found only in VP1 (e.g., a VP1 capsid polypeptide includes, e g., consists of, amino acids corresponding to amino acids 1-738 of SEQ ID NO:9).

[0083] An example nucleic acid sequence encoding SEQ ID NO:9 is SEQ ID NO:10.

[0084] The present disclosure refers to structural capsid proteins (including VP1 , VP2 and VP3) which are encoded by capsid (Cap) genes. These capsid proteins form an outer protein structural shell (i.e. capsid) of a viral vector such as AAV. VP capsid proteins synthesized from Cap polynucleotides generally include a methionine as the first amino acid in the peptide sequence (Met1 ), which is associated with the start codon (AUG or ATG) in the corresponding Cap nucleotide sequence However, it is common for a first-methionine (Met1 ) residue or generally any first amino acid (AA1) to be cleaved off after or during polypeptide synthesis by protein processing enzymes such as Met-aminopeptidases. This “Met / AA-clipping" process often correlates with a corresponding acetylation of the second amino acid in the polypeptide sequence (e.g., alanine, valine, serine, threonine, etc.). Met-clipping commonly occurs with VP1 and VP3 capsid proteins but can also occur with VP2 capsid proteins. Where the Met / AA-clipping is incomplete, a mixture of one or more (one, two or three) VP capsid proteins comprising the viral capsid can be produced, some of which include a Met1 / AA1 amino acid (Met+ / AA+) and some of which lack a Met1 / AA1 amino acid as a result of Met / AA-clipping (Met- / AA-). For further discussion regarding Met / AA-clipping in capsid proteins, see Jin, et al. Direct Liquid Chromatography / Mass Spectrometry Analysis for Complete Characterization of Recombinant Adeno-Associated Virus Capsid Proteins. Hum Gene Ther Methods 2017 Oct 28(5):255-267; Hwang, et al. N- Terminal Acetylation of Cellular Proteins Creates Specific Degradation Signals Science. 2010 February 19.327(5968): 973-977; the contents of which are each incorporated herein by reference in their entirety. According to the present disclosure, references to capsid polypeptides is not limited to either clipped (Met- / AA-) or unclipped (Met+ / AA+) and, in context, also refer to independent capsid polypeptides, viral capsids comprised of a mixture of capsid proteins, and / or polynucleotide sequences (or fragments thereof) which encode, describe, produce or result in capsid polypeptides of the present disclosure. A direct reference to a “capsid polypeptide” (such asVP1 , VP2 or VP3) also comprises VP capsid proteins which include a Met1 / AA1 amino acid (Met+ / AA+) as well as corresponding VP capsid polypeptide which lack the Met1 / AA1 amino acid as a result of Met / AA-clipping (Met- / AA-). Further according to the present disclosure, a reference to a specific SEQ ID NO: which comprises one or more capsid polypeptides which include a Met1 / AA1 amino acid (Met+ / AA+) should be understood to teach the VP capsid polypeptides which lack the Met1 / AA1 amino acid as upon review of the sequence, it is readily apparent any sequence which merely lacks the first listed amino acid (whether or not Met1 / AA1 ). As a non-limiting example, reference to a VP1 polypeptide sequence which is 736 amino acids in length and which includes a “Met1” amino acid (Met+) encoded by the AUG / ATG start codon is also understood to teach a VP1 polypeptide sequence which is 735 amino acids in length and which does not include the “Met1” amino acid (Met-) of the 736 amino acid Met+ sequence As a second non-limiting example, reference to a VP1 polypeptide sequence which is 736 amino acids in length and which includes an “AA1” amino acid (AA1+) encoded by any NNN initiator codon can also be understood to teach a VP1 polypeptide sequence which is 735 amino acids in length and which does not include the “AA1” amino acid (AA1-) of the 736 amino acid AA1+ sequence. References to viral capsids formed from VP capsid proteins (such as reference to specific AAV capsid serotypes), can incorporate VP capsid proteins which include a Met1 / AA1 amino acid (Met+ / AA1+), corresponding VP capsid proteins which lack the Met1 / AA1 amino acid as a result of Met / AA1 -clipping (Met- / AA1-), and combinations thereof (Met+ / AA1+ and Met- / AA1-). As a non-limiting example, an AAV capsid serotype can include VP1 (Met+ / AA1+), VP1 (Met- / AA1-), or a combination of VP1 (Met+ / AA1+) and VP1 (Met- / AA1-). An AAV capsid serotype can also include VP3 (Met+ / AA1+), VP3 (Met- / AA1-), or a combination of VP3 (Met+ / AA1+) and VP3 (Met- / AA1-); and can also include similar optional combinations of VP2 (Met+ / AA1) and VP2 (Met- / AA1-).

[0085] In some embodiments, the reference AAV capsid sequence comprises an amino acid sequence with 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of the those described above

[0086] In some embodiments, a variant capsid polypeptide of the disclosure is an isolated or purified polypeptide (e.g , isolated or purified from a cell, other biological component, or contaminant). In some embodiments, the variant capsid polypeptide is present in a dependoparvovirus particle, e g., described herein. In some embodiments, the variant capsid polypeptide is present in a cell, cell-free system, or translation system, e.g., described herein.

[0087] In some embodiments, the variant capsid polypeptide is present in a dependoparvovirus B (e.g., AAV9) particle. In some embodiments, the capsid particle has increased muscle transduction as compared to a reference capsid polypeptide, e.g., a wild-type capsid polypeptide such as a VP1 capsid polypeptide of SEQ ID NO:7 or a VP2 or VP3 portion thereof6.3. Nucleic Acids

[0088] The disclosure is further directed, in part, to a nucleic acid comprising a sequence encoding a variant capsid polypeptide as provided for herein, e.g., as described in Section 6.2. In some embodiments the nucleic acid encodes a VP1 variant capsid polypeptide. In some embodiments, the nucleic acid encodes a VP2 variant capsid polypeptide. In some embodiments, the nucleic acid encodes a VP3 variant capsid polypeptide. In some embodiments, the nucleic acid encodes a VP1, VP2 and VP3 variant capsid polypeptide.

[0089] Accordingly, in some embodiments, the disclosure provides a nucleic acid comprising a nucleotide sequence encoding a variant capsid polypeptide that comprises one or more of the mutation differences or the entire mutation set associated with the capsid polypeptide of SEQ ID NO:12 or 14.

[0090] Typically, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide comprising one or more mutation differences or the entire mutation set associated with the capsid polypeptide of SEQ ID NO:12 or 14, and has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity to a reference AAV serotype, e.g., as described herein, e.g., to SEQ ID NO:7.

[0091] In various embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which, but for the mutation differences or mutation set associated with the capsid polypeptide of SEQ ID NO: 12 or 14, is at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a reference AAV serotype described herein

[0092] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which, but for the mutation differences associated with the capsid polypeptide of SEQ ID NO:12 or 14, is at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:1 (e.g., a VP1 , VP2 or VP3 sequence of SEQ ID NO:1).

[0093] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which, but for the mutation differences associated with the capsid polypeptide of SEQ ID NO:12 or 14, is at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:3 (e g , a VP1 , VP2 or VP3 sequence of SEQ ID NO:3).

[0094] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which, but for the mutation differences associated with the capsid polypeptide of SEQ ID NO:12 or 14, is at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:5 (e.g., a VP1 , VP2 or VP3 sequence of SEQ ID NO:5).

[0095] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which, but for the mutation differences associated with the capsidpolypeptide of SEQ ID NO:12 or 14, is at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:7 (e.g. , a VP1 , VP2 or VP3 sequence of SEQ ID NO:7).

[0096] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which, but for the mutation differences associated with the capsid polypeptide of SEQ ID NO:12 or 14, is at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:9 (e g , a VP1 , VP2 or VP3 sequence of SEQ ID NO:9).

[0097] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which, but for the mutation differences associated with the capsid polypeptide of SEQ ID NO:12 or 14, is at least 90%, at least 95%, 96%, 97%, 98%, 99%, or 100% identical to a capsid polypeptide of SEQ ID NO:11 (e.g., a VP1 , VP2 or VP3 sequence of SEQ ID NO:11 ).

[0098] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide which is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence of a variant capsid polypeptide as provided herein, e.g., a VP1 capsid polypeptide of the capsid polypeptide of SEQ ID NO:12 or 14, or a VP2 or VP3 portion thereof.

[0099] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide that is at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a variant capsid polypeptide as provided herein.

[0100] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide comprising a VP1 , VP2, VP3, or any combination thereof, that each has about 1 to about 20 mutations as compared to a VP1 polypeptide of the capsid polypeptide of SEQ ID NO: 12 or 14, and comprises one or more of the mutation differences or the entire mutation set of the capsid polypeptide of SEQ ID NO:12 or 14.

[0101] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide comprising a VP1 , VP2, VP3, or any combination thereof, that each has about 1 to about 10 mutations as compared to a VP1 polypeptide of the capsid polypeptide of SEQ ID NO: 12 or 14, and comprises one or more of the mutation differences or the entire mutation set of the capsid polypeptide of SEQ ID NO:12 or 14.

[0102] In some embodiments, the nucleic acid comprises a nucleotide sequence that encodes a variant capsid polypeptide comprising a VP1 , VP2, VP3, or any combination thereof, that each has about 1 to about 5 mutations as compared to a VP1 polypeptide of the capsid polypeptide of SEQ ID NO: 12 or 14, and comprises one or more of the mutation differences or the entire mutation set of the capsid polypeptide of SEQ ID NO:12 or 14.

[0103] In some embodiments, the nucleic acid comprises a nucleotide sequence of SEQ ID NO:13 (which encodes a capsid polypeptide of SEQ ID NO:12). In some embodiments, the nucleic acidcomprises a nucleotide sequence of SEQ ID NO:15 (which encodes a capsid polypeptide of SEQ ID NO:14).

[0104] In some embodiments, a nucleic acid of the disclosure (e.g., encoding a variant capsid polypeptide as described in Section 6.2) comprises conventional control elements or sequences which are operably linked to the nucleic acid molecule in a manner which permits transcription, translation and / or expression in a cell transfected with the nucleic acid (e.g., a plasmid vector comprising said nucleic acid) or infected with a virus comprising said nucleic acid As used herein, “operably linked” sequences include both expression control sequences that are contiguous with the gene of interest and expression control sequences that act in trans or at a distance to control the gene of interest.6.4. Dependoparvovirus Particles

[0105] The disclosure is also directed, in part, to a dependoparvovirus particle (e.g , a functional dependoparvovirus particle) comprising a nucleic acid or polypeptide described herein or produced by a method described herein.

[0106] In some embodiments, the viral particle comprising a variant capsid polypeptide, e.g., a variant capsid polypeptide described herein, exhibits increased muscle (cardiac and / or skeletal) transduction as compared to a viral particle with the wild-type capsid polypeptide (SEQ ID NO:7)

[0107] In some embodiments, a dependoparvovirus particle comprises an amino acid sequence that has at least 80, 85, 90, 91 , 92, 93, 94, 95, 96, 97, 98, 99, or 100% identity to the amino acid sequences provided for herein (e.g., a capsid polypeptide described in Section 6.2 such as the VP1 capsid polypeptide of SEQ ID NO:12 or 14 or a VP2 or VP3 portion thereof). In some embodiments, the variant capsid polypeptide comprises an amino acid sequence that differs by no more than 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11 , 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acids from the amino acid sequence of a variant capsid polypeptide provided for herein.

[0108] In some embodiments, the additional alteration improves a production characteristic of a dependoparvovirus particle or method of making the same. In some embodiments, the additional alteration improves or alters another characteristic of a dependoparvovirus particle, e.g., tropism

[0109] Dependoparvovirus is a single-stranded DNA parvovirus that grows only in cells in which certain functions are provided, e.g., by a co-infecting helper virus. Several species of dependoparvovirus are known, including dependoparvovirus A and dependoparvovirus B, which include serotypes known in the art as adeno-associated viruses (AAV). At least thirteen serotypes of AAV that have been characterized. General information and reviews of AAV can be found in, for example, Carter, Handbook of Parvoviruses, Vol. 1 , pp. 169-228 (1989), and Berns, Virology, pp. 1743-1764, Raven Press, (New York, 1990). AAV serotypes, and to a degree, dependoparvovirus species, are significantly interrelated structurally and functionally. (See, for example, Blacklowe, pp. 165-174 of Parvoviruses and Human Disease, J. R. Pattison, ed. (1988); and Rose, Comprehensive Virology 3:1-61 (1974)). For example, all AAV serotypes apparently exhibit very similar replication properties mediated by homologous rep genes; and all bear three related capsid proteins. In addition,heteroduplex analysis reveals extensive cross-hybridization between serotypes along the length of the genome, further suggesting interrelatedness. Dependoparvoviruses genomes also comprise selfannealing segments at the termini that correspond to “inverted terminal repeat sequences” (ITRs).

[0110] The genomic organization of naturally occurring dependoparvoviruses, e.g., AAV serotypes, is very similar. For example, the genome of AAV is a linear, single-stranded DNA molecule that is approximately 5,000 nucleotides (nt) in length or less. Inverted terminal repeats (ITRs) flank the unique coding nucleotide sequences for the non-structural replication (Rep) proteins and the structural capsid (Cap) proteins. Three different viral particle (VP) proteins form the capsid. The terminal approximately 145 nt of the genome are self-complementary and are organized so that an energetically stable intramolecular duplex forming a T-shaped hairpin may be formed. These hairpin structures function as an origin for viral DNA replication, serving as primers for the cellular DNA polymerase complex. The Rep genes encode the Rep proteins: Rep78, Rep68, Rep52, and Rep40. Rep78 and Rep68 are transcribed from the p5 promoter, and Rep 52 and Rep40 are transcribed from the p19 promoter The cap genes encode the VP proteins, VP1, VP2, and VP3 The cap genes are transcribed from the p40 promoter.

[0111] In some embodiments, a dependoparvovirus particle of the disclosure comprises a nucleic acid comprising a variant capsid polypeptide provided for herein. In some embodiments, the particle comprises a polypeptide as provided for herein.

[0112] In some embodiments, the dependoparvovirus particle of the disclosure is an AAV9 particle. In some embodiments, the AAV9 particle comprises a variant capsid polypeptide as provided for herein or a nucleic acid molecule encoding the same.

[0113] In some embodiments the dependoparvovirus particle comprises a variant capsid comprising a variant capsid polypeptide described herein. In some embodiments, the dependoparvovirus particle comprises variant capsid polypeptide described herein and a nucleic acid molecule. In some embodiments, the dependoparvovirus particle comprises variant capsid polypeptide described herein and a nucleic acid molecule comprising one or more inverted terminal repeat sequences (ITRs), for example, ITRs derived from an AAV9 dependoparvovirus or an AAV2 dependoparvovirus, one or more regulatory elements (for example, a promoter), and a payload (e.g., as described herein, e.g., a heterologous transgene) In some embodiments, at least one of the ITRs is modified. In some embodiments, the nucleic acid molecule is single-stranded. In some embodiments, the nucleic acid molecule is double stranded, for example, self-complementary. Various ITRs and their use in self- complementary AAV vectors are recognized in the art and include those described in U.S. Patent Nos. 7,465,583, 8,298,818 and 9,150,882; in U.S. Patent Application Publication Nos. 2004 / 0029106 A1, 2023 / 0139985 A1 , 2022 / 0175887 A1 ; and in Wilmott et al , 2019, Hum Gene Then Methods, 30(6):206-213 and McCarty, 2008, Mol. Then., 16( 10): 1648-1656, each incorporated herein by reference.6.5. Improved Biodistribution and Transduction Characteristics

[0114] The disclosure is directed, in part, to nucleic acids, polypeptides, cells, cell free systems, translation systems, viral particles, and methods associated with using and making the same toproduce viral particles that have increased distribution to tissues and cells of the muscle and / or muscle tissue transduction as compared to a viral particle comprising a reference sequence that does not otherwise comprise the mutations described herein (or mutations corresponding thereto), for example, as compared with a viral particle comprising a capsid polypeptide sequence of SEQ ID NO:7. In some embodiments, a use of a viral particle (e.g., a viral particle as described in Section 6.4) comprising the variant capsid polypeptides as described in Section 6.2 or any one of numbered embodiments 1 to 240 (e.g., V1 or V2) leads to increased muscle biodistribution of the viral particle and / or increased transduction of a transgene virus particle in the cells of the muscle, and, therefore, increased expression of the payload (transgene) in the muscle (e.g., increased cardiac muscle expression and / or increased skeletal muscle expression). In some embodiments, use of a viral particle (e.g., a viral particle as described in Section 6.4) comprising the variant capsid polypeptides as described in Section 6.2 or any one of numbered embodiments 1 to 240 further leads to reduced (or non-increased) biodistribution of the viral particle and / or reduced (or non-increased) transduction of the transgene in one or more peripheral tissues, e.g., (i) liver, (ii) spleen, (Hi) dorsal root ganglia, (iv) brain / central nervous system, or any combination of two, three, or all four of (i), (ii), (Hi), and (iv).

[0115] In some embodiments, biodistribution and transduction (e.g., of the tissue types described in this section) are measured as described herein, for example as described in Section 8.1 (e.g., Example 1) for example by relative quantification (e.g., via qPCR) of transgene mRNA in one or more samples isolated from the relevant tissue type, e.g., muscle. In some embodiments, biodistribution and / or transduction of a virus particle having a variant capsid polypeptide can be measured using virus particles having a transgene operably linked to a promoter that is active in a target cell or tissue type of interest. In some embodiments, the promoter is a ubiquitous promoter In other embodiments, the promoter is selective or specific to a target cell or tissue type (e.g , muscle), and, optionally, is less (or not) active in a cell or tissue type where transgene expression is not desired (e.g., liver). A promoter that is selective for a first cell or tissue type over a second cell or tissue type is active in the first cell or tissue type and less active or silent in the second cell or tissue type. In some embodiments, the promoter is a muscle-specific promoter-specific or muscle-selective promoter. In some embodiments, biodistribution and / or transduction of a virus particle having a variant capsid polypeptide can be measured using virus particles having a transgene operably linked to a ubiquitous promoter or a muscle-specific promoter In various embodiments, the transgene is a transgene encoding a capsid polypeptide or any other suitable heterologous transgene, for example a nucleic acid sequence encoding a synthetic, mammalian or human therapeutic protein or nucleic acid (e g., mRNA or RNAi) or reporter gene such as, for example a nucleic acid encoding a GFP or mCherry reporter

[0116] In some embodiments, the virus particle, e.g., as described herein, e.g., comprising a variant capsid polypeptide described herein, exhibits increased transduction of myocytes (muscle cells) relative to a virus particle comprising a reference capsid polypeptide, e.g., a reference capsid polypeptide of SEQ ID NO:7.

[0117] In some embodiments, a viral particle comprising the variant capsid polypeptide, e.g., the variant capsid polypeptide described herein, exhibits improved properties, e.g., improved biodistribution, transduction and / or production. Unless indicated otherwise, improvement rates are presented as fold-improvement over the rates exhibited by a virus particle comprising capsid polypeptides of SEQ ID NO:7. In some embodiments, improvement means an increase, e.g., in the case of muscle biodistribution or muscle transduction. In other embodiments, improvement means a decrease, e.g., in the case of liver biodistribution or liver transduction A virus particle having increased biodistribution or transduction in target cells or target tissue types, e.g., muscle, and / or decreased biodistribution or transduction in off-target cells or off-target tissue types, e.g., liver, may have improved specificity for a target cell or target tissue type. This improvement may be beneficial in the use of the viral particle to deliver a therapeutic transgene to the target cell or target tissue type in a subject afflicted with a disease affecting the target cell or target tissue type, for example.

[0118] In some embodiments, one or more improved properties (e.g., increased or decreased biodistribution and / or transduction) is exhibited in a mammal, e g , a primate, e g , a human In some embodiments, the increased or decreased biodistribution and / or transduction is exhibited upon administration of the virus particle or pharmaceutical composition comprising the virus particle, e g., as described herein, by systemic administration, e.g , intravenous administration.6.6. Methods of Making Compositions Described Herein

[0119] The disclosure is directed, in part, to a method of making a dependoparvovirus particle, e.g., a dependoparvovirus particle described herein. In some embodiments, a method of making dependoparvovirus particle comprises providing a cell, cell-free system, or other translation system, comprising a nucleic acid described encoding a variant capsid polypeptide provided for herein, or a polypeptide provided for herein (e.g., a variant capsid polypeptide); and cultivating the cell, cell-free system, or other translation system under conditions suitable for the production of the dependoparvovirus particle, thereby making the dependoparvovirus particle.

[0120] In some embodiments, a nucleic acid or polypeptide described herein is produced by a method known to one of skill in the art. In some embodiments, the nucleic acids, polypeptides, and fragments thereof of the disclosure are produced by any suitable means, including recombinant production, chemical synthesis, or other synthetic means. Such production methods are within the knowledge of those of skill in the art and are not a limitation of the present invention6.6.1. Host Cells

[0121] Aspects of the disclosure are directed to a host cell comprising a nucleic acid of the disclosure (e.g., encoding a variant capsid polypeptide as described in Section 6.2). A host cell of the disclosure, e.g., a host cell useful to general AAV virus particles comprising a variant AAV capsid as described herein, generally comprises one or more nucleic acids comprising a coding sequence encoding a variant capsid polypeptide of the disclosure (e.g., as described in Section 6.2), together with a payload (e g , transgene) and one or more coding sequences encoding additional components useful for promoting packaging of the payload into a dependoparvovirus capsid. Additional components include, for example, coding sequences for a rep protein and dependoparvovirusinverted terminal repeats (ITRs), as well as helper sequences which promote dependoparvovirus particle production and / or secretion. Examples of helper sequences include E1a, E1b, E2a, E4, and VA. Such helper sequences may be included endogenously within the host cell (e g., the host cell may be engineered to express such helper sequences, e g., integrated into the host cell genome) or may be provided exogenously (e.g., transduced on the same or a different nucleic acid as the variant capsid polypeptide, payload, rep, and / or ITRs).

[0122] In some embodiments, the helper sequences include AdV5 helper sequences An exemplary AdV5 genome is disclosed in the art as NCBI Reference Sequence AC_000008.1 (disclosed as SEQ ID NO:1 of PCT Patent Application Publication No WO / 2022 / 079429 A1 ). In some embodiments, a host cell disclosed herein comprises a portion of an AdV5 genome encoding for one or more helper protein sequences (e.g., E1a, E1 b, E2a, E4) or RNA sequences (e.g., VA). In some embodiments, a host cell disclosed herein comprises a nucleic acid sequence encoding one or more AdV5 helper protein sequences. Certain exemplary AdV5 helper protein sequences are provided below

[0123] AdV5 E1A:MRHIICHGGVITEEMAASLLDQLIEEVLADNLPPPSHFEPPTLHELYDLDVTAPEDPNEEAVSQIFPDSV MLAVQEGIDLLTFPPAPGSPEPPHLSRQPEQPEQRALGPVSMPNLVPEVIDLTCHEAGFPPSDDEDEE GEEFVLDYVEHPGHGCRSCHYHRRNTGDPDIMCSLCYMRTCGMFVYSPVSEPEPEPEPEPEPARPT RRPKMAPAILRRPTSPVSRECNSSTDSCDSGPSNTPPEIHPVVPLCPIKPVAVRVGGRRQAVECIEDLL NEPGQPLDLSCKRPRP (SEQ ID NO: 515)

[0124] AdV5 E1B 19K:MEAWECLEDFSAVRNLLEQSSNSTSWFWRFLWGSSQAKLVCRIKEDYKWEFEELLKSCGELFDSLN LGHQALFQEKVIKTLDFSTPGRAAAAVAFLSFIKDKWSEETHLSGGYLLDFLAMHLWRAVVRHKNRLL LLSSVRPAIIPTEEQQQQQEEARRRRQEQSPWNPRAGLDPRE (SEQ ID NO: 516)

[0125] AdV5 E1B 55K:MERRNPSERGVPAGFSGHASVESGCETQESPATVVFRPPGDNTDGGAAAAAGGSQAAAAGAEPME PESRPGPSGMNVVQVAELYPELRRILTITEDGQGLKGVKRERGACEATEEARNLAFSLMTRHRPECIT FQQIKDNCANELDLLAQKYSIEQLTTYWLQPGDDFEEAIRVYAKVALRPDCKYKISKLVNIRNCCYISGN GAEVEIDTEDRVAFRCSMINMWPGVLGMDGWIMNVRFTGPNFSGTVFLANTNLILHGVSFYGFNNT CVEAWTDVRVRGCAFYCCWKGWCRPKSRASIKKCLFERCTLGILSEGNSRVRHNVASDCGCFMLV KSVAVIKHNMVCGNCEDRASQMLTCSDGNCHLLKTIHVASHSRKAWPVFEHNILTRCSLHLGNRRGV FLPYQCNLSHTKILLEPESMSKVNLNGVFDMTMKIWKVLRYDETRTRCRPCECGGKHIRNQPVMLDV TEELRPDHLVLACTRAEFGSSDEDTD (SEQ ID NO: 517)

[0126] AdV5 E3 12.5K:MLSGEAEQLRLKHLVHCRRHKCFARDSGEFCYFELPEDHIEGPAHGVRLTAQGELARSLIREFTQRPL LVERDRGPCVLTVICNCPNLGLHQDLCCHLCAEYNKYRN (SEQ ID NO: 518)

[0127] AdV5 E3 CR1-alpha0:MNNSSNSTGYSNSGFSRIGVGVILCLVILFILILTLLCLRLAACCVHICIYCQLFKRWGRHPR (SEQ ID NO: 519)

[0128] AdV5 E3 gp19K:MIRYIILGLLTLASAHGTTQKVDFKEPACNVTFAAEANECTTLIKCTTEHEKLLIRHKNKIGKYAVYAIWQ PGDTTEYNVTVFQGKSHKTFMYTFPFYEMCDITMYMSKQYKLWPPQNCVENTGTFCCTAMLITVLAL VCTLLYIKYKSRRSFIEEKKMP (SEQ ID NO: 520)

[0129] AdV5 E3 CR1-beta0:MTNTTNAAAATGLTSTTNTPQVSAFVNNWDNLGMWWFSIALMFVCLIIMWLICCLKRKRARPPIYSPIIVLHPNNDGIHRLDGLKHMFFSLTV (SEQ ID NO: 521 )

[0130] AdV5 E3 RID-alpha:MIPRVFILLTLVALFCACSTLAAVSHIEVDCIPAFTVYLLYGFVTLTLICSLITVVIAFIQCIDWVCVRFAYLRHHPQYRDRTIAELLRIL (SEQ ID NO: 522)

[0131] AdV5 E3 RID-beta:MKFTVTFLLIICTLSAFCSPTSKPQRHISCRFTRIWNIPSCYNEKSDLSEAWLYAIISVMVFCSTILALAIY PYLDIGWKRIDAMNHPTFPAPAMLPLQQVVAGGFVPANQPRPTSPTPTEISYFNLTGGDD (SEQ ID NO: 523)

[0132] AdV5 E3 14.7K:MTDTLDLEMDGIITEQRLLERRRAAAEQQRMNQELQDMVNLHQCKRGIFCLVKQAKVTYDSNTTGHR LSYKLPTKRQKLVVMVGEKPITITQHSVETEGCIHSPCQGPEDLCTLIKTLCGLKDLIPFN (SEQ ID NO: 524)

[0133] AdV5 E4 ORF6 / 7:MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNAWTSPSPPVK QPQVGQQPVAQQLDSDMNLSELPGEFINITDERLARQETVWNITPKNMSVTHDMMLFKASRGERTV YSVCWEGGGRLNTRVL (SEQ ID NO: 525)

[0134] AdV5 E4 34K:MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNVSYVRGLPCS VGFTLIQEWVVPWDMVLTREELVILRKCMHVCLCCANIDIMTSMMIHGYESWALHCHCSSPGSLQCIA GGQVLASWFRMVVDGAMFNQRFIWYREVVNYNMPKEVMFMSSVFMRGRHLIYLRLWYDGHVGSV VPAMSFGYSALHCGILNNIVVLCCSYCADLSEIRVRCCARRTRRLMLRAVRIIAEETTAMLYSCRTERR RQQFIRALLQHHRPILMHDYDSTPM (SEQ ID NO: 526)

[0135] AdV5 E4 ORF4:MVLPALPAPPVCDSQNECVGWLGVAYSAVVDVIRAAAHEGVYIEPEARGRLDALREWIYYNYYTERSKRRDRRRRSVCHARTWFCFRKYDYVRRSIWHDTTTNTISVVSAHSVQ (SEQ ID NO: 527)

[0136] AdV5 E4 ORF3:MIRCLRLKVEGALEQIFTMAGLNIRDLLRDILRRWRDENYLGMVEGAGMFIEEIHPEGFSLYVHLDVRAVCLLEAIVQHLTNAIICSLAVEFDHATGGERVHLIDLHFEVLDNLLE (SEQ ID NO: 528)

[0137] AdV5 E4 ORFB:MFERKMVSFSVVVPELTCLYLHEHDYDVLSFLREALPDFLSSTLHFISPPMQQAYIGATLVSIAPSMRVIISVGSFVMVPGGEVAALVRADLHDYVQLALRRDLRDRGIFVNVPLLNLIQVCEEPEFLQS (SEQ ID NO: 529)

[0138] AdV5 E4 ORF1 :MAAAVEALYVVLEREGAILPRQEGFSGVYVFFSPINFVIPPMGAVMLSLRLRVCIPPGYFGRFLALTDV NQPDVFTESYIMTPDMTEELSVVLFNHGDQFFYGHAGMAVVRLMLIRVVFPVVRQASNV (SEQ ID NO: 530)

[0139] In some embodiments, the helper sequences include AdV2 helper sequences. An exemplary AdV2 genome is disclosed in the art as NCBI Reference Sequence AC_000007.1 (disclosed as SEQ ID NO:2 of PCT Patent Application Publication No WQ / 2022 / 079429 A1 ). In some embodiments, a host cell disclosed herein comprises a portion of an AdV2 genome encoding for one or more helper protein sequences (e.g., E1a, E1 b, E2a, E4) or RNAsequences (e.g., VA). In some embodiments, a host cell disclosed herein comprises a nucleic acid sequence encoding one or more AdV2 helper protein sequences. Certain exemplary AdV2 helper protein sequences are provided below

[0140] AdV2 E1A:MRHIICHGGVITEEMAASLLDQLIEEVLADNLPPPSHFEPPTLHELYDLDVTAPEDPNEEAVSQIFPESV MLAVQEGIDLFTFPPAPGSPEPPHLSRQPEQPEQRALGPVSMPNLVPEVIDLTCHEAGFPPSDDEDE EGEEFVLDYVEHPGHGCRSCHYHRRNTGDPDIMCSLCYMRTCGMFVYSPVSEPEPEPEPEPEPARP TRRPKLVPAILRRPTSPVSRECNSSTDSCDSGPSNTPPEIHPVVPLCPIKPVAVRVGGRRQAVECIEDL LNESGQPLDLSCKRPRP (SEQ ID NO: 531)

[0141] AdV2 E1B 19K:MEAWECLEDFSAVRNLLEQSSNSTSWFWRFLWGSSQAKLVCRIKEDYKWEFEELLKSCGELFDSLN LGHQALFQEKVIKTLDFSTPGRAAAAVAFLSFIKDKWSEETHLSGGYLLDFLAMHLWRAVVRHKNRLL LLSSVRPAIIPTEEQQQEEARRRRRQEQSPWNPRAGLDPRE (SEQ ID NO: 532)

[0142] AdV2 E1B 55K:MERRNPSERGVPAGFSGHASVESGGETQESPATVVFRPPGNNTDGGATAGGSQAAAAAGAEPMEP ESRPGPSGMNVVQVAELFPELRRILTINEDGQGLKGVKRERGASEATEEARNLTFSLMTRHRPECVT FQQIKDNCANELDLLAQKYSIEQLTTYWLQPGDDFEEAIRVYAKVALRPDCKYKISKLVNIRNCCYISGN GAEVEIDTEDRVAFRCSMINMWPGVLGMDGWIMNVRFTGPNFSGTVFLANTNLILHGVSFYGFNNT CVEAWTDVRVRGCAFYCCWKGVVCRPKSRASIKKCLFERCTLGILSEGNSRVRHNVASDCGCFMLV KSVAVIKHNMVCGNCEDRASQMLTCSDGNCHLLKTIHVASHSRKAWPVFEHNILTRCSLHLGNRRGV FLPYQCNLSHTKILLEPESMSKVNLNGVFDMTMKIWKVLRYDETRTRCRPCECGGKHIRNQPVMLDV TEELRPDHLVLACTRAEFGSSDEDTD (SEQ ID NO: 533)

[0143] AdV2 E3 12.5K:MTSGEAERLRLTHLDHCRRHKCFARGSGEFCYFELPEEHIEGPAHGVRLTTQVELTRSLIREFTKRPL LVERERGPCVLTVVCNCPNPGLHQDLCCHLCAEYNKYRN (SEQ ID NO: 534)

[0144] AdV2 E3 CR1-alphap0:MSNSSNSTSLSNFSGIGVGVILTLVILFILILALLCLRVAACCTHVCTYCQLFKRWGQHPR (SEQ ID NO: 535)

[0145] AdV2 E3 gp19K:MRYMILGLLALAAVCSAAKKVEFKEPACNVTFKSEANECTTLIKCTTEHEKLIIRHKDKIGKYAVYAIWQP GDTNDYNVTVFQGENRKTFMYKFPFYEMCDITMYMSKQYKLWPPQKCLENTGTFCSTALLITALALV CTLLYLKYKSRRSFIDEKKMP (SEQ ID NO: 536)

[0146] AdV2 E3 CR1-beta0:MTGSTIAPTTDYRNTTATGLTSALNLPQVHAFVNDWASLDMWWFSIALMFVCLIIMWLICCLKRRRARPPIYRPIIVLNPHNEKIHRLDGLKPCSLLLQYD (SEQ ID NO: 537)

[0147] AdV2 E3 RID alpha:MIPRVLILLTLVALFCACSTLAAVAHIEVDCIPPFTVYLLYGFVTLILICSLVTVVIAFIQFIDWVCVRIAYLRHHPQYRDRTIADLLRIL (SEQ ID NO: 538)

[0148] AdV2 E3 RID beta:MKRSVIFVLLIFCALPVLCSQTSAPPKRHISCRFTQIWNIPSCYNKQSDLSEAWLYAIISVMVFCSTIFAL AIYPYLDIGWNAIDAMNHPTFPVPAVIPLQQVIAPINQPRPPSPTPTEISYFNLTGGDD (SEQ ID NO:539)

[0149] AdV2 E3 14.7K:MTESLDLELDGINTEQRLLERRKAASERERLKQEVEDMVNLHQCKRGIFCVVKQAKLTYEKTTTGNRL SYKLPTQRQKLVLMVGEKPITVTQHSAETEGCLHFPYQGPEDLCTLIKTMCGIRDLIPFN (SEQ ID NO:540)

[0150] AdV2 E4 ORF6 / 7:MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNAWTSPSPPVE QPQVGQQPVAQQLDSDMNLSELPGEFINITDERLARQETVWNITPKNMSVTHDMMLFKASRGERTV YSVCWEGGGRLNTRVL (SEQ ID NO: 541 )

[0151] AdV2 E4 34K:MTTSGVPFGMTLRPTRSRLSRRTPYSRDRLPPFETETRATILEDHPLLPECNTLTMHNVSYVRGLPCS VGFTLIQEWVVPWDMVLTREELVILRKCMHVCLCCANIDIMTSMMIHGYESWALHCHCSSPGSLQCIA GGQVLASWFRMVVDGAMFNQRFIWYREVVNYNMPKEVMFMSSVFMRGRHLIYLRLWYDGHVGSV VPAMSFGYSALHCGILNNIVVLCCSYCADLSEIRVRCCARRTRRLMLRAVRIIAEETTAMLYSCRTERRRQQFIRALLQHHRPILMHDYDSTPM (SEQ ID NO: 526)

[0152] AdV2 E4 ORF4:MVLPALPAPPVCDSQNECVGWLGVAYSAVVDVIRAAAHEGVYIEPEARGRLDALREWIYYNYYTERAKRRDRRRRSVCHARTWFCFRKYDYVRRSIWHDTTTNTISVVSAHSVQ (SEQ ID NO: 542)

[0153] AdV2 E4 ORF3:MIRCLRLKVEGALEQIFTMAGLNIRDLLRDILIRWRDENYLGMVEGAGMFIEEIHPEGFSLYVHLDVRAVCLLEAIVQHLTNAIICSLAVEFDHATGGERVHLIDLHFEVLDNLLE (SEQ ID NO: 543)

[0154] AdV2 E4 ORF2:MFERKMVSFSVVVPELTCLYLHEHDYDVLAFLREALPDFLSSTLHFISPPMQQAYIGATLVSIAPSMRVIISVGSFVMVPGGEVAALVRADLHDYVQLALRRDLRDRGIFVNVPLLNLIQVCEEPEFLQS (SEQ ID NO: 544)

[0155] AdV2 E4 ORF1 :MAAAVEALYVVLEREGAILPRQEGFSGVYVFFSPINFVIPPMGAVMLSLRLRVCIPPGYFGRFLALTDV NQPDVFTESYIMTPDMTEELSVVLFNHGDQFFYGHAGMAVVRLMLIRVVFPVVRQASNV (SEQ ID NO: 530)

[0156] Additional AAV helper sequences are recognized in the art and include, for example, those described in U.S. Patent Application Publication Nos. 2004 / 0248288 A1 and 2022 / 0259572A1, and in PCT Patent Application Publication Nos. WO / 1997 / 017458 A1 , WO / 2024 / 143429 A1, and WO / 2020 / 208379 A1, each of which is incorporated herein by reference.

[0157] Expression control sequences include efficient RNA processing signals such as splicing and polyadenylation (polyA) signals; appropriate transcription initiation, termination, promoter and enhancer sequences; sequences that stabilize cytoplasmic mRNA; sequences that enhance protein stability; sequences that enhance translation efficiency (e.g., Kozak consensus sequence); and in some embodiments, sequences that enhance secretion of the encoded transgene product.Expression control sequences, including promoters which are native, constitutive, inducible and / or tissue-specific, are known in the art and can be utilized with the compositions and methods disclosed herein.

[0158] In some embodiments, the native promoter for the transgene is used. Without wishing to be bound by theory, the native promoter can mimic native expression of the transgene, or provide temporal, developmental, or tissue-specific expression, or expression in response to specific transcriptional stimuli. In some embodiments, the transgene is operably linked to other native expression control elements, such as enhancer elements, polyadenylation sites or Kozak consensus sequences, e.g., to mimic the native expression

[0159] In some embodiments, the transgene is operably linked to a tissue-specific promoter, e.g., a promoter active specifically in one or more muscle cell types.

[0160] In some embodiments, a vector, e.g., a plasmid, carrying a transgene includes a selectable marker or a reporter gene. Such selectable reporters or marker genes can be used to signal the presence of the vector, e.g , plasmid, in bacterial cells. Other components of the vector, e.g., plasmid, include an origin of replication. Selection of these and other promoters and vector elements are conventional and many such sequences are available (see, e.g., Sambrook et al, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, NY, and references cited therein).

[0161] In some embodiments, an insect cell may be used in production of the compositions described herein or in the methods of making a dependoparvovirus particle described herein For example, an insect cell line used can be from Spodoptera frugiperda, such as Sf9, SF21, SF900+, drosophila cell lines, mosquito cell lines, e.g., Aedes albopictus derived cell lines, domestic silkworm cell lines, e.g., Bombyxmori cell lines, Trichoplusia ni cell lines such as High Five cells or Lepidoptera cell lines such as Ascalapha odorata cell lines. In some embodiments, the insect cells are susceptibleto baculovirus infection, including High Five, Sf9, Se301 , SelZD2109, SeUCRI , SP900+, Sf21 , BTI- TN-5B1-4, MG-1, Tn368, HzAml, BM-N, Ha2302, Hz2E5 and Ao38. Use of insect cells for expression of heterologous proteins is well recognized in the art, as are methods of introducing nucleic acids, such as vectors, e g., insect-cell compatible vectors, into such cells and methods of maintaining such cells in culture. See, for example, O'Reilly et al., 1994, Baculovirus Expression Vectors, A Laboratory Manual. Oxford Univ. Press; Satnulski et al., 1989, J. Vir.63: 3822-8; Kajigaya et al., 1991 PNAS 88:4646-50; Ruffin et al., 1992, J. Vir. 66:6922-30; Kimbauer et al ., 1996, Vir.21.9:37-44; Zhao et al., 2000, Vir.272:382-93; and U.S. Pat No. 6,204,059, the contents of each of which are incorporated herein by reference in their entireties.

[0162] In certain embodiments, insect host cell systems, in combination with baculoviral systems (e.g., as described by Luckow et al., 1988, Bio / Technology 6:47) is used. In certain embodiments, the expression system is a Trichoplusia ni, Tn 5B1-4 insect cells / baculoviral system, which can be used for production of high levels of proteins, as described in U.S. Pat. No. 6,660,521, incorporated herein by reference in its entirety

[0163] Expansion, culture, transfection, infection, and storage of insect cells can be carried out in any cell culture media, cell transfection media or storage media known in the art. Nonlimiting examples of media are Hyclone SFX Insect Cell Culture Media, Expression System ESF AF Insect Cell Culture Medium, Basal IPL-41 Insect Cell Culture Media, ThermoFisher Sf90011 media, ThermoFisher Sf900111 media, and ThermoFisher Grace's Insect Media. Insect cell mixtures and / or media can also comprise appropriate formulation additives or elements, including but not limited to salts, acids, bases, buffers, and surfactants (such as Poloxatner 188 / Pluronic F-68).

[0164] In some embodiments, the methods of the disclosure can be carried out with a mammalian cell type which allows for replication of dependoparvovirus or production of biologic products, and which can be maintained in culture. Host cells include cells derived from mammalian species including but not limited to, human, monkey, mouse, rat, rabbit, and hamster. Host cells can be of any suitable cell type, including but not limited to cell lines, fibroblasts, hepatocytes, tumor cells, and transformed cells. The mammalian cells used can be HEK293, HEK293T, HeLa, CHO, NS0, SP2 / 0, PER C6, Vero, RD, BHK, HT 1080, A549, Cos-7, ARPE-19, MRC-5, WEH1, 3T3, 1.0T1 / 2, MDCK, COS 1 , COS 7, BSC 1 , BSC 40, BMT 10, W138, Saos, C2C12, HepG2, L cells, primary fibroblast, hepatocyte and myoblast cells derived from mammals, COS cells, C127, 3T3, CHO, HeLa cells, KB cells, BHK, and other mammalian cell lines as described in U.S. Pat. Nos. 6,156,303, 5,387,484, 5,741 ,683, 5,691 ,176, 6,428,988 and 5,688,676, 6,541 ,258, the contents of each of which are incorporated herein by reference.

[0165] In some embodiments, the host cell comprises a nucleic acid encoding a variant capsid polypeptide disclosed herein, where the nucleic acid is integrated into the host cell genome. Such host cells include adenovirus rep and cap genes integrated into the genome. T ranscription of the integrated rep and cap genes may be dependent upon introduction of certain helper virus sequences (e.g., adenovirus E4, E2a and / or VA RNA) into the cell by transduction or other suitable means.Example plasmid free host cells are described in U.S. Patent Application Publication No. 2022 / 0025396 A1 , and U.S. Patent No. US 5,658,785, incorporated herein by reference.

[0166] In some embodiments, the host cells are trans-complementing packaging cell lines that provide functions deleted from a replication-defective helper virus, e.g., HEK293 cells or other Ea trans-complementing cells. In some embodiments, the packaging cell line 293-10-3 is used as described in U.S. Pat. No. 6,281 ,010, incorporated herein by reference.

[0167] In some embodiments, mammalian host cells (e g. 293T cells) can be in an adherent state (e.g., adhered / attached to a suitable surface of a cell culture flask, vial, tray, well, tube, etc.). In other embodiments, mammalian host cells can be in a suspended state (e.g., suspended in a medium).6.6.2. Viral Particle Production

[0168] In some embodiments, the nucleic acids of the disclosure are situated as a part of any genetic element (vector) which can be delivered to a host cell, e.g., naked DNA, a plasmid, phage, transposon, cosmid, episome, a protein in a non-viral delivery vehicle (e.g., a lipid-based carrier), virus, etc , which transfer the sequences carried thereon Such a vector can be delivered into a host cell by any suitable method, including transfection, liposome delivery, electroporation, membrane fusion techniques, viral infection, high velocity DNA- coated pellets, and protoplast fusion. A person of skill in the art possesses the knowledge and skill in nucleic acid manipulation to construct any embodiment of this invention and said skills include genetic engineering, recombinant engineering, and synthetic techniques. See, e.g., Sambrook et al, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, NY.

[0169] In some embodiments, a vector of the disclosure comprises sequences encoding a dependoparvovirus variant capsid polypeptide as provided for herein or a fragment thereof. In some embodiments, a vector of the disclosure comprises sequences encoding a dependoparvovirus Rep protein or a fragment thereof. Such a Rep coding region encodes at least for AAV Rep78, Rep68, Rep52, and Rep40, or functional homologs thereof. The Rep coding region is not required to include all wild-type genes but may be altered (e.g., by insertion, deletion, or mutation of one or more nucleotides) so long as the rep genes present provide for sufficient replication functions when expressed in the recombinant cell. The Rep coding region may be derived from any AAV serotype. In some embodiments, the Rep coding region is or comprises a rep gene encoding AAV2 Rep proteins, exemplary sequences of which are provided below.

[0170] AAV2 Rep78:MPGFYEIVIKVPSDLDGHLPGISDSFVNWVAEKEWELPPDSDMDLNLIEQAPLTVAEKLQRDFLTEWR RVSKAPEALFFVQFEKGESYFHMHVLVETTGVKSMVLGRFLSQIREKLIQRIYRGIEPTLPNWFAVTKT RNGAGGGNKVVDECYIPNYLLPKTQPELQWAWTNMEQYLSACLNLTERKRLVAQHLTHVSQTQEQN KENQNPNSDAPVIRSKTSARYMELVGWLVDKGITSEKQWIQEDQASYISFNAASNSRSQIKAALDNAG KIMSLTKTAPDYLVGQQPVEDISSNRIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGKT NIAEAIAHTVPFYGCVNWTNENFPFNDCVDKMVIWWEEGKMTAKVVESAKAILGGSKVRVDQKCKSS AQIDPTPVIVTSNTNMCAVIDGNSTTFEHQQPLQDRMFKFELTRRLDHDFGKVTKQEVKDFFRWAKD HVVEVEHEFYVKKGGAKKRPAPSDADISEPKRVRESVAQPSTSDAEASINYADRYQNKCSRHVGMNLMLFPCRQCERMNQNSNICFTHGQKDCLECFPVSESQPVSVVKKAYQKLCYIHHIMGKVPDACTACDLVNVDLDDCIFEQ (SEQ ID NO: 545)

[0171] AAV2 Rep68:MPGFYEIVIKVPSDLDGHLPGISDSFVNWVAEKEWELPPDSDMDLNLIEQAPLTVAEKLQRDFLTEWR RVSKAPEALFFVQFEKGESYFHMHVLVETTGVKSMVLGRFLSQIREKLIQRIYRGIEPTLPNWFAVTKT RNGAGGGNKVVDECYIPNYLLPKTQPELQWAWTNMEQYLSACLNLTERKRLVAQHLTHVSQTQEQN KENQNPNSDAPVIRSKTSARYMELVGWLVDKGITSEKQWIQEDQASYISFNAASNSRSQIKAALDNAG KIMSLTKTAPDYLVGQQPVEDISSNRIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGKT NIAEAIAHTVPFYGCVNWTNENFPFNDCVDKMVIWWEEGKMTAKVVESAKAILGGSKVRVDQKCKSS AQIDPTPVIVTSNTNMCAVIDGNSTTFEHQQPLQDRMFKFELTRRLDHDFGKVTKQEVKDFFRWAKD HVVEVEHEFYVKKGGAKKRPAPSDADISEPKRVRESVAQPSTSDAEASINYADRLARGHSL (SEQ ID NO: 546)

[0172] AAV2 Rep52:MELVGWLVDKGITSEKQWIQEDQASYISFNAASNSRSQIKAALDNAGKIMSLTKTAPDYLVGQQPVEDI SSNRIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGKTNIAEAIAHTVPFYGCVNWTNE NFPFNDCVDKMVIWWEEGKMTAKVVESAKAILGGSKVRVDQKCKSSAQIDPTPVIVTSNTNMCAVID GNSTTFEHQQPLQDRMFKFELTRRLDHDFGKVTKQEVKDFFRWAKDHVVEVEHEFYVKKGGAKKRP APSDADISEPKRVRESVAQPSTSDAEASINYADRYQNKCSRHVGMNLMLFPCRQCERMNQNSNICFT HGQKDCLECFPVSESQPVSVVKKAYQKLCYIHHIMGKVPDACTACDLVNVDLDDCIFEQ (SEQ ID NO: 547)

[0173] AAV2 Rep40:MELVGWLVDKGITSEKQWIQEDQASYISFNAASNSRSQIKAALDNAGKIMSLTKTAPDYLVGQQPVEDI SSNRIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGKTNIAEAIAHTVPFYGCVNWTNE NFPFNDCVDKMVIWWEEGKMTAKVVESAKAILGGSKVRVDQKCKSSAQIDPTPVIVTSNTNMCAVID GNSTTFEHQQPLQDRMFKFELTRRLDHDFGKVTKQEVKDFFRWAKDHVVEVEHEFYVKKGGAKKRP APSDADISEPKRVRESVAQPSTSDAEASINYADRLARGHSL (SEQ ID NO: 548)

[0174] In some embodiments the Rep coding region encodes for Rep sequences having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to one or more AAV2 Rep proteins. In some embodiments, the Rep coding region comprises a nucleotide sequence encoding for a Rep78 protein having at least 80%, at least about 85%, at least about 90%, at least about 95%, at least 98%, at least 99%, or 100% sequence identity to AAV2 Rep78. In some embodiments, the Rep coding region comprises a nucleotide sequence encoding for a Rep68 protein having at least 80%, at least about 85%, at least about 90%, at least about 95%, at least 98%, at least 99%, or 100% sequence identity to AAV2 Rep68. In some embodiments, the Rep coding region comprises a nucleotide sequence encoding for a Rep52 protein having at least 80%, at least about 85%, at least about 90%, at least about 95%, at least 98%, at least 99%, or 100% sequence identity to AAV2 Rep52. In some embodiments, the Rep coding region comprises a nucleotide sequence encoding for a Rep40 protein having at least 80%, at least about 85%, at least about 90%, at least about 95%, at least 98%, at least 99%, or 100% sequence identity to AAV2 Rep40.

[0175] In some embodiments, the Rep coding sequence encodes for wild type Rep proteins. Alternatively, the Rep coding sequence may encode for one or more mutant Rep proteins having improved properties compared with wild type Rep proteins Examples of such mutant Rep proteins are described in U.S. Patent No. 11,060,070, incorporated herein by reference.

[0176] Additional exemplary Rep gene and protein sequences are disclosed as SEQ ID NOs: 7 to 15 and 20 to 22 of PCT Patent Application Publication No. WO / 2022 / 079429 A1 , incorporated herein by reference

[0177] In some embodiments, such vectors contain both dependoparvovirus cap and rep proteins. In vectors in which both AAV rep and cap are provided, in some embodiments, the dependoparvovirus rep and dependoparvovirus cap sequences are both of the same dependoparvovirus species or serotype origin. Alternatively, the present embodiments also provide vectors in which the rep sequences are from a dependoparvovirus species or serotype which differs from that which is providing the cap sequences (e.g., AAV2 rep sequences and AAV9 cap sequences). In some embodiments, the rep and cap sequences are expressed from separate sources (e.g., separate vectors, or a host cell genome and a vector). In some embodiments, the rep sequences are fused in frame to cap sequences of a different dependoparvovirus species or serotype to form a chimeric dependoparvovirus vector.

[0178] In some embodiments, a vector of the disclosure comprises one or more helper sequences. Examples of helper sequences include E1a, E1b, E2a, E4, and VA. Certain exemplary helper protein sequences are provided in Section 6.6.1. Additional AAV helper sequences are recognized in the art and include, for example, those described in U.S. Patent Application Publication Nos. 2004 / 0248288 A1 and 2022 / 0259572A1, and in PCT Patent Application Publication Nos. WO / 1997 / 017458 A1 , WO / 2024 / 143429 A1, and WO / 2020 / 208379 A1 , each of which is incorporated herein by reference

[0179] In some embodiments, the vectors of the disclosure further contain a payload, e.g , a minigene comprising a selected transgene, e.g., flanked by dependoparvovirus 5' ITR and dependoparvovirus 3' ITR. In some embodiments, the ITR is from the same serotype as the variant capsid polypeptide. In some embodiments, the ITR is of a different serotype than the variant capsid polypeptide. In some embodiments, the viral genome comprises two ITR sequence regions, wherein the ITRs are of the same serotype as one another In some embodiments, the viral genome comprises two ITR sequence regions, wherein the ITRs are of different serotypes. Non-limiting examples include zero, one or both of the ITRs having the same serotype as the capsid. In one embodiment both ITRs of the viral genome of the AAV particle are AAV2 ITRs Independently, each ITR may be about 100 to about 150 nucleotides in length. An ITR may be about 100-105 nucleotides in length, 106-110 nucleotides in length, 111-115 nucleotides in length, 116-120 nucleotides in length, 121-125 nucleotides in length, 126-130 nucleotides in length, 131-135 nucleotides in length, 136-140 nucleotides in length, 141-145 nucleotides in length or 146- 150 nucleotides in length. In one embodiment, the ITRs are 140-142 nucleotides in length. Nonlimiting examples of ITR lengths are 102, 105, 130, 140, 141 , 142, 145 nucleotides in length.

[0180] The vectors described herein, e.g., a plasmid, are useful for a variety of purposes, but are particularly well suited for use in production of recombinant dependoparvovirus particles comprising dependoparvovirus sequences or a fragment thereof, and in some embodiments, a payload.

[0181] In one aspect, the disclosure provides a method of making a dependoparvovirus particle (e.g., a dependoparvovirus B particle, e.g., an AAV9 particle), or a portion thereof. In some embodiments, the method comprises culturing a host cell which contains a nucleic acid sequence encoding a dependoparvovirus variant capsid protein as provided for herein, or fragment thereof; a functional rep gene (e g., encoding Rep proteins as described herein); a payload, e g., a minigene comprising dependoparvovirus inverted terminal repeats (ITRs) and a transgene; and sufficient helper functions to promote packaging of the payload, e.g., minigene, into the dependoparvovirus capsid. In some embodiments, the components necessary to be cultured in the host cell to package a payload, e.g., minigene, in a dependoparvovirus capsid are provided to the host cell in trans. In some embodiments, any one or more of the required components (e.g., payload (e.g., minigene), rep sequences, cap sequences, and / or helper functions) are provided by a host cell which has been engineered to stably comprise one or more of the required components using methods known to those of skill in the art. In some embodiments, a host cell which has been engineered to stably comprise the required component(s) comprises it under the control of an inducible promoter. In some embodiments, the required components are under the control of a constitutive promoter Examples of suitable inducible and constitutive promoters are provided herein, and further examples are known to those of skill in the art. In some embodiments, a selected host cell which has been engineered to stably comprise one or more components comprises a component under the control of a constitutive promoter and another component under the control of one or more inducible promoters For example, a host cell which has been engineered to stably comprise the required components is generated from HEK 293 cells (e.g., which comprise helper functions under the control of a constitutive promoter), which comprises the rep and / or cap proteins under the control of one or more inducible promoters.

[0182] In some embodiments, the payload (e.g., minigene), rep sequences, cap sequences, and helper functions required for producing a dependoparvovirus particle of the disclosure are delivered to the packaging host cell in the form of any genetic element which transfers the sequences carried thereon (e.g., in a vector or combination of vectors). The genetic element may be delivered by any suitable method, including those described herein. Methods used to construct genetic elements, vectors, and other nucleic acids of the disclosure are known to those with skill and include genetic engineering, recombinant engineering, and synthetic techniques. See, e.g., Sambrook et al, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press, Cold Spring Harbor, NY Similarly, methods of generating rAAV virions are well known and the selection of a suitable method is not a limitation on the present invention. See, e g., K. Fisher et al, J. Virol, 70:520-532 (1993) and US Patent 5,478,745. Unless otherwise specified, the dependoparvovirus ITRs, and other selected dependoparvovirus components described herein, are readily selected from among any dependoparvovirus species and serotypes, e.g., AAV1 , AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAVrh74, or AAV9.

[0183] ITRs or other dependoparvovirus components may be readily isolated using techniques available to those of skill in the art from a dependoparvovirus species or serotype. Dependoparvovirus species and serotypes may be isolated or obtained from academic, commercial, or public sources (e.g., the American Type Culture Collection, Manassas, VA). In some embodiments, the dependoparvovirus sequences may be obtained through synthetic or other suitable means by reference to published sequences such as are available in the literature or in databases such as, e.g., GenBank or PubMed.

[0184] Methods of expressing proteins (e.g., recombinant or heterologous proteins, e.g., dependoparvovirus polypeptides) in insect cells are well documented, as are methods of introducing nucleic acids, such as vectors, e.g., insect-cell compatible vectors, into such cells and methods of maintaining such cells in culture. See, for example, METHODS IN MOLECULAR BIOLOGY, ed. Richard, Humana Press, N J (1995); O'Reilly et al , BACULOVIRUS EXPRESSION VECTORS, A LABORATORY MANUAL, Oxford Univ. Press (1994); Samulski et al., J. Vir. 63:3822-8 (1989); Kajigaya et al , Proc Natl Acad Sci USA 88:4646-50 (1991 ); Ruffing et al , J Vir 66:6922-30 (1992); Kirnbauer et al., Vir 219:37-44 (1996); Zhao et al., Vir. 272:382-93 (2000); Samulski et al., and U.S. Pat. No. 6,204,059. In some embodiments, a nucleic acid construct encoding dependoparvovirus polypeptides (e.g., a dependoparvovirus genome) in insect cells is an insect cellcompatible vector. An “insect cell-compatible vector” as used herein refers to a nucleic acid molecule capable of productive transformation or transfection of an insect or insect cell. Exemplary biological vectors include plasmids, linear nucleic acid molecules, and recombinant viruses. Any vector can be employed as long as it is insect cell compatible The vector may integrate into the insect cell's genome or remain present extra-chromosomally. The vector may be present permanently or transiently, e.g , as an episomal vector. Vectors may be introduced by any means known in the art. Such means include but are not limited to chemical treatment of the cells, electroporation, or infection. In some embodiments, the vector is a baculovirus, a viral vector, or a plasmid. Methods of dependoparvovirus capsid (e.g., AAV) production in insect cells include, for example, those described in U.S. Patent Application Publication No. 2024 / 0093231 A1 , U.S. Patent No. 11 ,306,291 , Joshi et al., 2024, Methods Mol Biol, 2829:203-214, and Marwidi et al , 2024, Mol Ther Methods Clin Dev , 32(2)101228, incorporated herein by reference.

[0185] In some embodiments, a nucleic acid sequence encoding a dependoparvovirus polypeptide is operably linked to regulatory expression control sequences for expression in a specific cell type, such as Sf9 or HEK cells. Techniques known to one skilled in the art for expressing foreign genes in insect host cells or mammalian host cells can be used with the compositions and methods of the disclosure. Methods for molecular engineering and expression of polypeptides in insect cells are described, for example, in Summers and Smith. A Manual of Methods for Baculovirus Vectors and Insect Culture Procedures, Texas Agricultural Experimental Station Bull. No. 7555, College Station, Tex. (1986); Luckow. 1991 . In Prokop et al., Cloning and Expression of Heterologous Genes in Insect Cells with Baculovirus Vectors' Recombinant DNA Technology and Applications, 97-152 (1986); King, L. A. and R. D. Possee, The baculovirus expression system, Chapman and Hall, United Kingdom (1992); O'Reilly, D. R., L. K. Miller, V. A Luckow, Baculovirus Expression Vectors: A Laboratory Manual, NewYork (1992); W. H. Freeman and Richardson, C. D., Baculovirus Expression Protocols, Methods in Molecular Biology, volume 39 (1995); U.S. Pat. No. 4,745,051 ; US2003148506; and WO 03 / 074714. Promoters suitable for transcription of a nucleotide sequence encoding a dependoparvovirus polypeptide include the polyhedron, p10, p35 or IE-1 promoters and further promoters described in the above references are also contemplated.

[0186] In some embodiments, providing a cell comprising a nucleic acid described herein comprises acquiring a cell comprising the nucleic acid

[0187] Methods of cultivating host cells, cell-free systems, and other translation systems are known to those of skill in the art. In some embodiments, cultivating a host cell comprises providing the cell with suitable media and incubating the cell and media for a time suitable to achieve viral particle production.

[0188] In some embodiments, a method of making a dependoparvovirus particle further comprises a purification step comprising isolating the dependoparvovirus particle from one or more other components (e.g., from a cell or media component).

[0189] In some embodiments, production of the dependoparvovirus particle comprises one or more (e.g., all) of: expression of dependoparvovirus polypeptides, assembly of a dependoparvovirus capsid, expression (e.g., duplication) of a dependoparvovirus genome, and packaging of the dependoparvovirus genome into the dependoparvovirus capsid to produce a dependoparvovirus particle. In some embodiments, production of the dependoparvovirus particle further comprises secretion of the dependoparvovirus particle into the media. The dependoparvovirus particle can be isolated from the collected media. In other embodiments, dependoparvovirus particles are isolated from host cells. For instance, adherent host cells can subsequently be collected by scraping and / or pelleting and suspended cells can be collected by pelleting and transferred into a receptacleCollection steps can be repeated as necessary for full collection of produced cells. If necessary, host cell lysis can be achieved by consecutive freeze-thaw cycles (-80°C to 37°C), chemical lysis (such as adding detergent, e.g., triton), mechanical lysis, or by allowing the cell culture to degrade after reaching ~0% viability. Cellular debris can be removed by centrifugation and / or depth filtration.

[0190] In some embodiments, and as described elsewhere herein, the nucleic acid molecule encoding the variant capsid polypeptide is disposed in a dependoparvovirus genome. In some embodiments, and as described elsewhere herein, the nucleic acid molecule encoding the variant capsid polypeptide is packaged into a dependoparvovirus particle along with the dependoparvovirus genome as part of a method of making a dependoparvovirus particle described herein In other embodiments, the nucleic acid molecule encoding the variant capsid polypeptide is not packaged into a dependoparvovirus particle made by a method described herein.

[0191] In some embodiments, a method of making a dependoparvovirus particle described herein produces a dependoparvovirus particle comprising a payload (e.g., a payload described herein) and the variant capsid polypeptide. In some embodiments, the payload comprises a second nucleic acid (e.g., in addition to the dependoparvovirus genome), and production of the dependoparvovirus particlecomprises packaging the second nucleic acid into the dependoparvovirus particle. In some embodiments, a cell, cell-free system, or other translation system for use in a method of making a dependoparvovirus particle comprises the second nucleic acid. In some embodiments, the second nucleic acid comprises an exogenous sequence (e.g., exogenous to the dependoparvovirus, the cell, or to a target cell or subject who will be administered the dependoparvovirus particle). In some embodiments, the exogenous sequence encodes an exogenous polypeptide. In some embodiments, the exogenous sequence encodes a therapeutic product.

[0192] In some embodiments, virus particles of the disclosure have a similar production efficiency to viral particles with a reference capsid polypeptide, for example, with the wild-type capsid polypeptide (SEQ ID NOV). In some embodiments, production efficiency of viral particles of the disclosure is (a) at least 0.1-fold, at least 0.2-fold, at least 0.3-fold, at least 0.4-fold, at least 0.5-fold, at least 0.6-fold, at least 0.7-fold, at least 0.8-fold, or at least 0.9-fold and / or (b) up to 1-fold, e.g., as compared to a viral particle with a reference capsid polypeptide, for example, with the wild-type capsid polypeptide (SEQ ID NO:1), or the production efficiency is within any range bounded by a value in (a) and a value in (b) In some embodiments, production efficiency is at least 0 5-fold as compared to a viral particle with the wild-type capsid polypeptide (SEQ ID NOV).

[0193] Production efficiency can be evaluated by producing viral particles having a variant capsid with a genome encoding a unique barcode and a fluorescent reporter gene under the control of a ubiquitous (e.g., CBh) or a muscle-specific promoter via transient triple transfection of adherent HEK293T cells followed by iodixanol gradient purification.

[0194] Various methods and systems for dependoparvovirus production in host cells are recognized in the art and are contemplated herein including, for example, those described in U.S. Patent Application Publication Nos 20220064671 A1, 20220259572 A1, 20220025396 A1 and PCT Patent Application Publication Nos. WO / 1999 / 011764 A2, WO / 2023 / 178220 A1 , WQ / 2020 / 208379 A1, WQ / 2023 / 143063 A1, WO / 2023 / 239627 A2, WQ / 2021 / 156609 A1, and WO / 2021 / 113767 A1, each of which is incorporated herein by reference in its entirety.6.7. Applications

[0195] The disclosure is directed, in part, to compositions comprising a nucleic acid, polypeptide, or particles described herein. The disclosure is further directed, in part, to methods utilizing a composition, nucleic acid, polypeptide, or particles described herein. As will be apparent based on the disclosure, nucleic acids, polypeptides, particles, and methods disclosed herein have a variety of utilities.

[0196] The disclosure is directed, in part, to a vector comprising a nucleic acid described herein, e.g., a nucleic acid encoding a variant capsid polypeptide. Many types of vectors are known to those of skill in the art. In some embodiments, a vector comprises a plasmid. In some embodiments, the vector is an isolated vector, e.g., removed from a cell or other biological components.

[0197] The disclosure is directed, in part to a cell, cell-free system, or other translation system, comprising a nucleic acid or vector described herein, e.g., a nucleic acid or vector comprising anucleic acid molecule encoding a variant capsid polypeptide. In some embodiments, the cell, cell-free system, or other translation system is capable of producing dependoparvovirus particles comprising the variant capsid polypeptides. In some embodiments, the cell, cell-free system, or other translation system comprises a nucleic acid comprising a dependoparvovirus genome or components of a dependoparvovirus genome sufficient to promote production of dependoparvovirus particles comprising the variant capsid polypeptides.

[0198] In some embodiments, the cell, cell-free system, or other translation system further comprises one or more non-dependoparvovirus nucleic acid sequences that promote dependoparvovirus particle production and / or secretion. Said sequences are referred to herein as helper sequences. In some embodiments, a helper sequence comprises one or more genes from another virus, e.g., an adenovirus or herpes virus In some embodiments, the presence of a helper sequence is necessary for production and / or secretion of a dependoparvovirus particle. In some embodiments, a cell, cell- free system, or other translation system comprises a vector, e.g., plasmid, comprising one or more helper sequences

[0199] In some embodiments, a cell, cell-free system, or other translation system comprises a first nucleic acid and a second nucleic acid, wherein the first nucleic acid comprises sequences encoding one or more dependoparvovirus genes (e.g , a Cap gene, a Rep gene, or a complete dependoparvovirus genome) and a helper sequence, and wherein the second nucleic acid comprises a payload In some embodiments, a cell, cell-free system, or other translation system comprises a first nucleic acid and a second nucleic acid, wherein the first nucleic acid comprises sequences encoding one or more dependoparvovirus genes (e.g , a Cap gene, a Rep gene, or a complete dependoparvovirus genome) and a payload, and wherein the second nucleic acid comprises a helper sequence In some embodiments, a cell, cell-free system, or other translation system comprises a first nucleic acid and a second nucleic acid, wherein the first nucleic acid comprises a helper sequence and a payload, and wherein the second nucleic acid comprises sequences encoding one or more dependoparvovirus genes (e g., a Cap gene, a Rep gene, or a complete dependoparvovirus genome). In some embodiments, a cell, cell-free system, or other translation system comprises a first nucleic acid, a second nucleic acid, and a third nucleic acid, wherein the first nucleic acid comprises sequences encoding one or more dependoparvovirus genes (e.g., a Cap gene, a Rep gene, or a complete dependoparvovirus genome), the second nucleic acid comprises a helper sequence, and the third nucleic acid comprises a payload.

[0200] In some embodiments, the first nucleic acid, second nucleic acid, and optionally third nucleic acid are situated in separate molecules, e.g., separate vectors or a vector and genomic DNA. In some embodiments, one, two, or all of the first nucleic acid, second nucleic acid, and optionally third nucleic acid are integrated (e.g., stably integrated) into the genome of a cell.

[0201] In some embodiments, a cell of the disclosure is generated by transfecting a suitable cell with a nucleic acid described herein. In some embodiments, a method of making a dependoparvovirus particle comprising a variant capsid polypeptide as provided for herein or improving a method of making a dependoparvovirus particle comprises providing a cell described herein. In someembodiments, providing a cell comprises transfecting a suitable cell with one or more nucleic acids described herein.

[0202] Many types and kinds of cells suitable for use with the nucleic acids and vectors described herein are known in the art. In some embodiments, the cell is a human cell. In some embodiments, the cell is an immortalized cell or a cell from a cell line known in the art. In some embodiments, the cell is an HEK293 cell. In some embodiments, the cell is an HEK293T cell.6.7.1 . Methods of delivering a payload

[0203] The disclosure is directed, in part, to a method of delivering a payload to a cell, e.g., a cell in a subject or in a sample. In some embodiments, a method of delivering a payload to a cell comprises contacting the cell with a dependoparvovirus particle comprising a variant capsid polypeptide (e.g., described herein) comprising the payload The disclosure also includes a dependoparvovirus particle comprising a variant capsid polypeptide (e.g., described herein) comprising a payload described herein for use in the methods of delivering a payload described herein. In some embodiments, the dependoparvovirus particle is a dependoparvovirus particle described herein and comprises a payload described herein. In some embodiments, the cell is a myocyte (muscle cell). Non-limiting examples of skeletal muscles include the biceps, the triceps, the quadriceps, the tibialis interior, the gastrocnemius muscle, and diaphragm. In some embodiments, the method is conducted ex vivo. In some embodiments, the cell is a cell in an ex vivo sample that has been obtained from a subject.

[0204] In some embodiments, the payload comprises a transgene. In some embodiments, the transgene is a nucleic acid sequence heterologous to the vector sequences flanking the transgene which encodes a polypeptide, RNA (e g , a miRNA or siRNA) or other product of interest In some embodiments, the nucleic acid of the transgene is operatively linked to a regulatory component in a manner sufficient to promote transgene transcription, translation, and / or expression in a host cell.

[0205] In aspects, a transgene is any polypeptide- or RNA-encoding sequence and the transgene selected will depend upon the use envisioned In some embodiments, a transgene comprises a reporter sequence, which upon expression produces a detectable signal. Such reporter sequences include, without limitation, DNA sequences encoding colorimetric reporters (e.g., P-lactamase, p- galactosidase (LacZ), alkaline phosphatase), cell division reporters (e.g., thymidine kinase), fluorescent or luminescence reporters (e.g., green fluorescent protein (GFP) or luciferase), resistance conveying sequences (e.g., chloramphenicol acetyltransferase (CAT)), or membrane bound proteins including to which high affinity antibodies directed thereto exist or can be produced by conventional means, e.g., comprising an antigen tag, e.g., hemagglutinin or Myc.

[0206] In some embodiments, a reporter sequence operably linked with regulatory elements which drive their expression, provide signals detectable by conventional means, including enzymatic, radiographic, colorimetric, fluorescence or other spectrographic assays, fluorescent activating cell sorting assays and immunological assays, including enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA) and immunohistochemistry In some embodiments, the transgene encodes a product which is useful in biology and medicine, such as RNA, proteins, peptides, enzymes, dominant negative mutants. In some embodiments, the RNA comprises a tRNA, ribosomal RNA, dsRNA,catalytic RNAs, small hairpin RNA, siRNA, trans-splicing RNA, and antisense RNAs. In some embodiments, the RNA inhibits or abolishes expression of a targeted nucleic acid sequence in a treated subject (e.g., a human or animal subject).

[0207] In some embodiments, the transgene is used to correct or ameliorate gene deficiencies. In some embodiments, gene deficiencies include deficiencies in which normal genes are expressed at less than normal levels or deficiencies in which the functional gene product is not expressed In some embodiments, the transgene encodes a therapeutic protein or polypeptide which is expressed in a host cell. In some embodiments, a dependoparvovirus particle comprises or delivers multiple transgenes, e.g., to correct or ameliorate a gene defect caused by a multi-subunit protein. In some embodiments, a different transgene (e.g., each situated / delivered in a different dependoparvovirus particle, or in a single dependoparvovirus particle) is used to encode each subunit of a protein, or to encode different peptides or proteins, e.g., when the size of the DNA encoding the protein subunit is large, e.g., for immunoglobulin, platelet-derived growth factor, or dystrophin protein. In some embodiments, different subunits of a protein are encoded by the same transgene, e.g, a single transgene encoding each of the subunits with the DNA for each subunit separated by an internal ribozyme entry site (IRES). In some embodiments, the DNA is separated by sequences encoding a 2A peptide, which self-cleaves in a post-translational event. See, e.g., Donnelly et al, J. Gen. Virol., 78(Pt 1 ): 13-21 (January 1997); Furler, et al, Gene Then, 8(11 ):864-873 (June 2001 ); Klump et al., Gene Then 8( 10): 811 -817 (May 2001 ) (incorporated herein by reference in their entirety).

[0208] In some embodiments, virus particles comprising a genome are provided, wherein the genome includes a nucleic acid expression construct. The nucleic acid expression construct can include a heterologous transgene and one or more regulatory elements.

[0209] In some embodiments, the regulatory elements include a promotor, e g, a promoter that is active in a target cell or tissue type of interest. In some embodiments, the promoter is a ubiquitous promoter. In other embodiments, the promoter is selective or specific to a target cell or tissue type (e.g., muscle (either broad muscle expression or skeletal muscle)), and, optionally, is less (or not) active in a cell or tissue type where transgene expression is not desired (e.g., liver). A promoter that is selective for a first cell or tissue type over a second cell or tissue type is active in the first cell or tissue type and less active or silent in the second cell or tissue type. In some embodiments, the promoter is a muscle-specific promoter-specific or muscle-selective promoter

[0210] In some embodiments, the promoter is a ubiquitous or constitutive promoter active in a mammalian cell, for example a human cell, for example, in a human cell type of interest. In some embodiments, the cell type is a muscle cell such as, for example, a cardiac myocyte, skeletal muscle myocyte, or smooth muscle myocyte Examples of ubiquitous promoters include, but are not limited, to a CAG promoter (hybrid from a cytomegalovirus early enhancer element, a chicken-beta actin promoter, e.g., the first exon and the first intron of the chicken beta actin gene, and the splice acceptor of the rabbit beta globin gene), chicken-beta actin promoter, CBA promoter, OBh promoter, CB6 promoter, CMV promoter, human EF1-alpha promoter, PGK promoter, ubiquitin C (UBC) promoter and fragments thereof. In some embodiments, the promoter is a tissue-specific promoter, for example,a promoter specific in muscle tissue or muscle cells, e.g., a desmin, MCK, TNNT2, or smooth muscle 22 (SM22) promoter. Further exemplary muscle specific promoters are described below.

[0211] In some embodiments, the promoter is a CBh promoter. An exemplary CBh promoter sequence is set forth as SEQ ID NO:500. In some embodiments, the CBh promoter comprises a nucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, at least 99%, or 100% sequence identity to SEQ ID NQ:500.

[0212] In other embodiments, the promoter is a EF1a promoter, for example a human EF1a promoter. An exemplary human EF1a promoter sequence is set forth as SEQ ID NQ:501. In some embodiments, the human EF1a promoter comprises a nucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:501.

[0213] In some embodiments, where preferential expression of a payload (e.g., a transgene) to a specific tissue is desired, the promoter is a tissue-specific promoter. Example tissue-specific promoters include muscle-specific promoters.

[0214] In some embodiments, the promoter is a promoter specific in muscle tissue or muscle cells, e.g., a desmin, MCK, TNNT2, or smooth muscle 22 (SM22) promoter. Further exemplary muscle specific promoters are described below.

[0215] In some embodiments, the nucleic acid expression construct comprises an intron In some embodiments, the intron is disposed between the promoter and the heterologous transgene. In some aspects, the intron is disposed 5’ to the heterologous transgene on the expression construct, for example immediately 5’ to the heterologous transgene or 100 nucleotides or less 5’ to the heterologous transgene. In some aspects, the intron is a chimeric intron derived from human b-globin and Ig heavy chain (also known as b- globin splice donor / immunoglobulin heavy chain splice acceptor intron, or b-globin / IgG chimeric intron; Reed, R., et al. Genes and Development, 1989, incorporated herein by reference in its entirety) In other aspects, the intron is a VH4 intron or a SV40 intron

[0216] As provided herein, in some embodiments, virus particles comprising a payload, wherein the payload includes a nucleic acid that includes a heterologous transgene are provided. In some embodiments, the heterologous transgene encodes an RNA interference agent, for example a siRNA, shRNA or other interfering nucleic acid.

[0217] In some embodiments, the payload includes a heterologous transgene that encodes a therapeutic polypeptide. In some aspects, the heterologous transgene is a human gene or fragment thereof. In some aspects, the therapeutic polypeptide is a human protein. In some embodiments, the heterologous transgene of the virus particle encodes a molecule useful in treating a disease, and the virus particle is administered to a patient in need thereof to treat said disease In some aspects, the payload comprises a molecule that is effective in treating muscle-related disease. Exemplary, nonlimiting muscle-related diseases are described below.

[0218] In some embodiments, the virus particle comprises a heterologous transgene encoding a genome editing system. Examples include a CRISPR genome editing system (e.g., one or morecomponents of a CRISPR genome editing system such as, for example, a guide RNA molecule and / or a RNA-guided nuclease such as a Cas enzyme such as Cas9, Cpf1 and the like), a zinc finger nuclease genome editing system, a TALEN genome editing system or a meganuclease genome editing system. In some embodiments, the genome editing system targets a mammalian, e.g., human, genomic target sequence. In some embodiments, the virus particle includes a heterologous transgene encoding a targetable transcription regulator. Examples include a CRISPR-based trascription regulator (for example, one or more components of a CRISPR-based transcription regulator, for example, a guide RNA molecule and / or a enzymatically-inactive RNA-guided nuclease / transcription factor (“TF”) fusion protein such as a dCas9-TF fusion, dCpf1-TF fusion and the like), a zinc finger transcription factor fusion protein, a TALEN transcription regulator or a meganuclease transcription regulator.

[0219] In some embodiments, components of a therapeutic molecule or system are delivered by more than one unique virus particle (e.g., a population that includes more than one unique virus particles) In other embodiments, the therapeutic molecule or components of a therapeutic molecule or system are delivered by a single unique virus particle (e.g , a population that includes a single unique virus particle).

[0220] In some embodiments, the transgene encodes any biologically active product or other product, e.g , a product desirable for study. Suitable transgenes may be readily selected by persons of skill in the art, such as those, but not limited to, those described herein.

[0221] The disclosure is further directed, in part, to a method of delivering a payload to a subject, e.g., an animal or human subject. In some embodiments, a method of delivering a payload to a subject comprises administering to the subject a dependoparvovirus particle comprising a variant polypeptide (e g , described herein) comprising the payload, e g , in a quantity and for a time sufficient to deliver the payload. In some embodiments, the dependoparvovirus particle is a dependoparvovirus particle described herein and comprises a payload described herein.

[0222] In some embodiments, the particle delivers the payload to muscle tissue (e.g., cardiac muscle or skeletal muscle). In some embodiments, the delivery to muscle is increased as compared to a particle without the variant capsid polypeptide or as compared to a wild-type capsid polypeptide, e.g., a particle with capsid polypeptides of SEQ ID NO:7.

[0223] In some embodiments, virus particles comprising a genome are provided, wherein the genome includes a nucleic acid expression construct including a heterologous transgene and one or more regulatory elements, where the one or more regulatory elements include a muscle specific promoter. Examples of muscle specific promoters that can be used to drive expression of a transgene in muscle include but are not limited to Desmin (DES), CAMK, Mb, myosin (e.g., myo-3), dystrophin, muscle creatine kinase (MCK), MHCK7, CK6, CK7, CK8, CK8e, dMCK, tMCK, MH, SPc-5-12 (also known as 05-12), alpha skeletal actin (ASKA), SP-301, E-syn, myosin light chain (MLC), myosin heavy chain (MHO), four and a half LIM domains protein 1 (FHL1), alpha 2 actinin (ACTN2), filamin-C (FLNC), sarcoplasmic / endoplasmic reticulum calcium ATPase 1 (ATP2A1), troponin I type 1 (TNNI1 ), myosin-1 (MYH1 ), phosphorylatable, fast skeletal muscle myosin light chain (MYLPF), alpha-3 chaintropomyosin (TPM3), Pitx3, and ankyrin repeat domain-containing protein 2 (ANKRD2) promoters. Promoters capable of driving expression in muscle are further described in Skopenkova et al., 2021 , Acta Naturae 13(1 ):47-58, Piekarowicz et al., 2019, Mol Ther Methods Clin Dev. 15:157-169, Wang, 2008 Gene Ther. 15:1489-1499, Coulon ef al., 2007, JBC 282(45):33192-33200, WO 2021 / 127655, and WO 2023 / 006890, the contents of each of which are incorporated herein by reference in their entireties.

[0224] In some aspects, the payload comprises a molecule that is effective in treating a muscle disease, such as, for example, a protein or an antisense oligonucleotide such as an RNA interference nucleotide (e.g., shRNA, siRNA or miRNA).

[0225] In certain aspects, the disclosure provides a virus particle comprising (a) a capsid polypeptide described herein, for example, a capsid polypeptide described in Section 6.2 and (b) an engineered viral genome comprising (i) a nucleotide sequence encoding a molecule for the treatment of Duchenne muscular dystrophy (DMD), for example human Dystrophin (DMD) or a fragment or variant thereof (e.g., a variant thereof having at least 90% or at least 95% sequence identity to human DMD), and (ii) a promoter operably linked to said nucleotide sequence. An exemplary accession number for human DMD is set forth in Table M1. The viral genome can further comprise one or more (e.g., two, three, four, or all five) of (a) a pair of dependoparvovirus ITRs, (b) an intron, (c) an enhancer or repressor sequence, (d) a stuffer sequence, and (e) a polyA sequence. Preferably, the viral genome comprises ITRs flanking the nucleotide sequence and a polyA sequence operably linked to the nucleotide sequence. Typically, the viral genome lacks rep and cap sequences, which are in trans by the host cell in which the virus particle is produced. In some embodiments, the viral genome is self- complementary.

[0226] In certain aspects, the disclosure provides a virus particle comprising (a) a capsid polypeptide described herein, for example, a capsid polypeptide described in Section 6.2 and (b) an engineered viral genome comprising (i) a nucleotide sequence encoding a molecule for the treatment of X-linked myotubular myopathy, for example human myotubularin (MTM1) or a fragment or variant thereof (e.g., a variant thereof having at least 90% or at least 95% sequence identity to human MTM1), and (ii) a promoter operably linked to said nucleotide sequence. An exemplary accession number for human MTM1 is set forth in Table M1. The viral genome can further comprise one or more (e.g., two, three, four, or all five) of (a) a pair of dependoparvovirus ITRs, (b) an intron, (c) an enhancer or repressor sequence, (d) a stuffer sequence, and (e) a polyA sequence. Preferably, the viral genome comprises ITRs flanking the nucleotide sequence and a polyA sequence operably linked to the nucleotide sequence Typically, the viral genome lacks rep and cap sequences, which are in trans by the host cell in which the virus particle is produced. In some embodiments, the viral genome is self- complementary.

[0227] In certain aspects, the disclosure provides a virus particle comprising (a) a capsid polypeptide described herein, for example, a capsid polypeptide described in Section 6.2 and (b) an engineered viral genome comprising (i) a nucleotide sequence encoding a molecule for the treatment of Myotonic Dystrophy Type 1 (DM1 ), for example human myotonin-protein kinase (DMPK) or a fragment orvariant thereof (e.g., a variant thereof having at least 90% or at least 95% sequence identity to human DMPK, and (ii) a promoter operably linked to said nucleotide sequence An exemplary accession number for human DMPK is set forth in Table M1. The viral genome can further comprise one or more (e.g., two, three, four, or all five) of (a) a pair of dependoparvovirus ITRs, (b) an intron, (c) an enhancer or repressor sequence, (d) a stuffer sequence, and (e) a polyA sequence. Preferably, the viral genome comprises ITRs flanking the nucleotide sequence and a polyA sequence operably linked to the nucleotide sequence. Typically, the viral genome lacks rep and cap sequences, which are in trans by the host cell in which the virus particle is produced. In some embodiments, the viral genome is self-complementary.

[0228] In certain aspects, the disclosure provides a virus particle comprising (a) a capsid polypeptide described herein, for example, a capsid polypeptide described in Section 6.2 and (b) an engineered viral genome comprising (i) a nucleotide sequence encoding a molecule for the treatment of Limbgirdle muscular dystrophy (LGMD), for example human Gamma-sarcoglycan, Alpha-sarcoglycan, Beta-sarcoglycan, or Delta-sarcoglycan or a fragment or variant thereof (e g , a variant thereof having at least 90% or at least 95% sequence identity to human Gamma-sarcoglycan, Alpha-sarcoglycan, Beta-sarcoglycan, or Delta-sarcoglycan), and (II) a promoter operably linked to said nucleotide sequence Exemplary accession numbers for human Gamma-sarcoglycan, Alpha-sarcoglycan, Beta- sarcoglycan, and Delta-sarcoglycan are set forth in Table M1. The viral genome can further comprise one or more (e.g., two, three, four, or all five) of (a) a pair of dependoparvovirus ITRs, (b) an intron, (c) an enhancer or repressor sequence, (d) a stuffer sequence, and (e) a polyA sequence. Preferably, the viral genome comprises ITRs flanking the nucleotide sequence and a polyA sequence operably linked to the nucleotide sequence. Typically, the viral genome lacks rep and cap sequences, which are in trans by the host cell in which the virus particle is produced. In some embodiments, the viral genome is self-complementary.

[0229] In certain aspects, the disclosure provides a virus particle comprising (a) a capsid polypeptide described herein, for example, a capsid polypeptide described in Section 6.2 and (b) an engineered viral genome comprising (i) a nucleotide sequence encoding a molecule for the treatment of Pompe disease, for example human acid alpha-glucosidase (GAA) or a fragment (e.g., exon 2) or variant thereof (e.g., a variant thereof having at least 90% or at least 95% sequence identity to human GAA or a portion thereof), and (ii) a promoter operably linked to said nucleotide sequence. An exemplary accession number for human GAA is set forth in Table M1 . The viral genome can further comprise one or more (e.g., two, three, four, or all five) of (a) a pair of dependoparvovirus ITRs, (b) an intron, (c) an enhancer or repressor sequence, (d) a stuffer sequence, and (e) a polyA sequence. Preferably, the viral genome comprises ITRs flanking the nucleotide sequence and a polyA sequence operably linked to the nucleotide sequence. Typically, the viral genome lacks rep and cap sequences, which are in trans by the host cell in which the virus particle is produced. In some embodiments, the viral genome is self-complementary.

[0230] In certain aspects, the disclosure provides a virus particle comprising (a) a capsid polypeptide described herein, for example, a capsid polypeptide described in Section 6.2 and (b) an engineeredviral genome comprising (i) a nucleotide sequence encoding a molecule for the treatment of Facioscapulohumeral muscular dystrophy (FSHD), for example an antisense oligonucleotide targeting human DUX4, and (ii) a promoter operably linked to said nucleotide sequence. The viral genome can further comprise one or more (e.g., two, three, four, or all five) of (a) a pair of dependoparvovirus ITRs, (b) an intron, (c) an enhancer or repressor sequence, (d) a stuffer sequence, and (e) a polyA sequence Preferably, the viral genome comprises ITRs flanking the nucleotide sequence and a polyA sequence operably linked to the nucleotide sequence. Typically, the viral genome lacks rep and cap sequences, which are in trans by the host cell in which the virus particle is produced. In some embodiments, the viral genome is self-complementary

[0231] In certain aspects, the disclosure provides a virus particle comprising (a) a capsid polypeptide described herein, for example, a capsid polypeptide described in Section 6.2 and (b) an engineered viral genome comprising (i) a nucleotide sequence encoding a molecule for the treatment of Facioscapulohumeral muscular dystrophy (FSHD), for example human SMCHD1 or a fragment or variant thereof (e g , a variant thereof having at least 90% or at least 95% sequence identity to human SMCHD1 or a portion thereof), and (ii) a promoter operably linked to said nucleotide sequence. An exemplary accession number for human SMCHD1 is set forth in Table M1. The viral genome can further comprise one or more (e.g., two, three, four, or all five) of (a) a pair of dependoparvovirus ITRs, (b) an intron, (c) an enhancer or repressor sequence, (d) a stuffer sequence, and (e) a polyA sequence Preferably, the viral genome comprises ITRs flanking the nucleotide sequence and a polyA sequence operably linked to the nucleotide sequence. Typically, the viral genome lacks rep and cap sequences, which are in trans by the host cell in which the virus particle is produced In some embodiments, the viral genome is self-complementary

[0232] Exemplary muscle tissue related diseases that can be treated include but are not limited to Acid Maltase Deficiency (AMD), Amyotrophic Lateral Sclerosis (ALS), Andersen-Tawil Syndrome, Barth syndrome (TAZ), Becker Muscular Dystrophy (BMD), Becker Myotonia Congenita, Bethlem Myopathy, Bulbospinal Muscular Atrophy (Spinal- Bulbar Muscular Atrophy), Carnitine Deficiency, Carnitine Palmityl Transferase Deficiency (CPT Deficiency), Central Core Disease (CCD), Centronuclear Myopathy, Charcot-Marie-Tooth Disease (CMT), Congenital Muscular Dystrophy (CMD), Congenital Myasthenic Syndromes (CMS), Congenital Myotonic Dystrophy, Cori Disease (Debrancher Enzyme Deficiency), Danon disease, Debrancher Enzyme Deficiency, Dejerine-Sottas Disease (DSD), Dermatomyositis (DM), Distal Muscular Dystrophy (DD), Distal myopathy with anterior tibial onset, Duchenne Muscular Dystrophy (DMD), Dystrophia Myotonica (Myotonic Muscular Dystrophy), Emery-Dreifuss Muscular Dystrophy (EDMD), Endocrine Myopathies, Eulenberg Disease (Paramyotonia Congenita), Facioscapulohumeral Muscular Dystrophy (FSH or FSHD), Finnish (Tibial) Distal Myopathy, Forbes Disease (Debrancher Enzyme Deficiency), Friedreich's Ataxia (FA), Fukuyama Congenital Muscular Dystrophy, Glycogenosis Type 10, Glycogenosis Type 11 , Glycogenosis Type 2, Glycogenosis Type 3, Glycogenosis Type 5, Glycogenosis Type 7, Glycogenosis Type 9, Gowers-Laing Distal Myopathy, Hauptmann-Thanheuser MD (Emery- Dreifuss Muscular Dystrophy), Hereditary Inclusion-Body Myositis, Hereditary Motor and Sensory Neuropathy (Charcot-Marie-Tooth Disease), Hyperthyroid Myopathy, Hypothyroid Myopathy, Inclusion-BodyMyositis (IBM), Inherited Myopathies, Integrin-Deficient Congenital Muscular Dystrophy, Kennedy Disease (Spinal-Bulbar Muscular Atrophy), Kugelberg-Welander Disease (Spinal Muscular Atrophy), Lactate Dehydrogenase Deficiency, Lambert-Eaton Myasthenic Syndrome (LEMS), Limb-Girdle Muscular Dystrophy (LGMD), Lou Gehrig's Disease (Amyotrophic Lateral Sclerosis), McArdle Disease (Phosphorylase Deficiency), Merosin-Deficient Congenital Muscular Dystrophy, Metabolic Diseases of Muscle, Mitochondrial Myopathy, Miyoshi myopathy, Miyoshi Distal Myopathy, Motor Neurone Disease, Muscle-Eye-Brain Disease, Myasthenia Gravis (MG), Myoadenylate Deaminase Deficiency, Myofibrillar Myopathy, Myophosphorylase Deficiency, Myotonia Congenita (MC), Myotonic Muscular Dystrophy (MMD), Myotubular Myopathy (MTM or MM), Nemaline Myopathy, Nonaka Distal Myopathy, Oculopharyngeal Muscular Dystrophy (OPMD), Paramyotonia Congenita, Pearson Syndrome, Periodic Paralysis, Peroneal Muscular Atrophy (Charcot-Marie-Tooth Disease), Phosphofructokinase Deficiency, Phosphoglycerate Kinase Deficiency, Phosphoglycerate Mutase Deficiency, Phosphorylase Deficiency, Phosphorylase Deficiency Polymyositis (PM), Pompe Disease (Acid Maltase Deficiency), Primary merosin deficiency (LAMA2), Progressive External Ophthalmoplegia (PEO), Rod Body Disease (Nemaline Myopathy), Spinal Muscular Atrophy (SMA), Spinal-Bulbar Muscular Atrophy (SBMA), Steinert Disease (Myotonic Muscular Dystrophy), Tarui Disease (Phosphofructokinase Deficiency), Thomsen Disease (Myotonia Congenita), Ullrich Congenital Muscular Dystrophy, Walker-Warburg Syndrome (Congenital Muscular Dystrophy), Welander Distal Myopathy, Werdnig-Hoffmann Disease (Spinal Muscular Atrophy), and ZASP- Related Myopathy.

[0233] Payloads suitable for treating muscle-related disease are known in the art and include the following disease (suitable payload) combinations: Barth syndrome (TAZ), Primary merosin deficiency (LAMA2), Duchenne muscular dystrophy (DMD), Becker muscular dystrophy (BMD), Danon disease (LAMP2), Limb girdle muscular dystrophy (Subtypes and affected genes: LGMD1A (TTID), LGMD1B (LMNA), LGMD1C (CAV3), LGMD1D (DNAJB6), LGMD1 E (DES), LGMD1 F (TNP03), LGMD1G (HNRPDL), LGMD1H, LGMD2A (CAPN3), LGMD2B (DYSF), LGMD2C (SGCG), LGMD2D (SGCA), LGMD2E (SGCB), LGMD2F (SGCD), LGMD2G (TCAP), LGMD2H (TRIM32), LGMD2I (FKRP), LGMD2J (TTN), LGMD2K (POMT1 ), LGMD2L (AN05), LGMD2M (FKTN), LGMD2N (POMT2), LGMD20 (POMGNT1 ), LGMD2Q (PLEC1 )), Miyoshi myopathy (DYSF), Distal myopathy with anterior tibial onset (DYSF), Welander distal myopathy (TIA1 ), Gowers-Laing distal myopathy (MYH7), Facioscapulohumeral muscular dystrophy (Subtypes and affected genes: Type 1 (DUX4), Type 2 (SMCHD1)), Oculopharyngeal muscular dystrophy (PABPN1 ), myotonic dystrophy (Subtypes and affected genes: DM1 (DMPK) and DM2 (ZNF9)), congenital myotonia (CLCN1 ), paramyotonia congenital (SCN4A), myotubular myopathy (MTM1 ), glycogen storage disease type II (Pompe disease) (GAA)

[0234] In some embodiments, the payload is selected from: TAZ, LAMA2, DMD, LAMP2, CAPN3, DYSF, SGCA, SGCB, FKRP, PABPN1 , MTM1, and GAA.

[0235] Certain exemplary muscle tissue related diseases that can be treated and exemplary heterologous transgenes that can be delivered via the viral particles of the disclosure are provided in Table M1.

[0236] In some embodiments, the payload is human dystrophin.

[0237] In some embodiments, the payload is an antisense oligonucleotide effective in treating a muscle disease, for example by modulating expression of a target gene. Exemplary target genes of such antisense oligonucleotides for treatment of muscle disease include, for example, MSTN, INHBA, ACVR1 B, MLCK1. ACVR1 , FBXO32, TRIM63, MEF2D, KLF15, MED1 , MED13, DUX4, LMNA, SMN, GAA, GYS2, DMD, DMPK, MSTN, and PPP1 R3A

[0238] Non-limiting, example antisense oligonucleotides that can be delivered via the viral particles of the disclosure (along with associated target genes and muscle tissue related diseases that can be treated) and are provided in Table M2. Where such antisense oligonucleotides are indicated by reference to a patent or patent application publication, such patent and patent applications are incorporated herein by reference in their entirety for their disclosed antisense oligonucleotide structures and sequences.6.7.2. Methods of treatment

[0239] The disclosure is directed, in part, to a method of treating a disease or condition in a subject, e.g., an animal or human subject. In some embodiments, a method of treating a disease or condition in a subject comprises administering to the subject a dependoparvovirus particle comprising a variant polypeptide described herein, e.g., comprising a payload described herein. In some embodiments, the dependoparvovirus particle, which comprises a variant polypeptide, comprising a payload described herein is administered in an amount and / or time effective to treat the disease or condition. In some embodiments, the payload is a therapeutic product In some embodiments, the payload is a nucleic acid, e.g., encoding an exogenous polypeptide. The disclosure is also directed to a dependoparvovirus particle comprising a variant polypeptide described herein, e.g., comprising a payload described herein, for use in the methods of treatment described herein. The disclosure is also directed to the use of a dependoparvovirus particle comprising a variant polypeptide described herein, e.g., comprising a payload described herein for the manufacture of a medicament for the treatment of a disease or condition as described herein.

[0240] The dependoparvovirus particles comprising a variant polypeptide described herein or produced by the methods described herein can be used to express one or more therapeutic proteins to treat various diseases or disorders In some embodiments, the disease or disorder is a cancer, e.g., a cancer such as carcinoma, sarcoma, leukemia, lymphoma; or an autoimmune disease, e.g., multiple sclerosis. Non-limiting examples of carcinomas include esophageal carcinoma; bronchogenic carcinoma; colon carcinoma; colorectal carcinoma; gastric carcinoma; hepatocellular carcinoma; basal cell carcinoma, squamous cell carcinoma (various tissues); bladder carcinoma, includingtransitional cell carcinoma; lung carcinoma, including small cell carcinoma and non-small cell carcinoma of the lung; adrenocortical carcinoma; sweat gland carcinoma; sebaceous gland carcinoma; thyroid carcinoma; pancreatic carcinoma; breast carcinoma; ovarian carcinoma; prostate carcinoma; adenocarcinoma; papillary carcinoma; papillary adenocarcinoma; cystadenocarcinoma; medullary carcinoma; renal cell carcinoma; uterine carcinoma; testicular carcinoma; osteogenic carcinoma; ductal carcinoma in situ or bile duct carcinoma; choriocarcinoma; seminoma; embryonal carcinoma; Wilm's tumor; cervical carcinoma; epithelial carcinoma; and nasopharyngeal carcinoma. Non-limiting examples of sarcomas include fibrosarcoma, myxosarcoma, liposarcoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, chondrosarcoma, chordoma, osteogenic sarcoma, osteosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's sarcoma, leiomyosarcoma, rhabdomyosarcoma, and other soft tissue sarcomas. Non-limiting examples of solid tumors include ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, menangioma, melanoma, neuroblastoma, and retinoblastoma. Non-limiting examples of leukemias include chronic myeloproliferative syndromes; T-cell CLL prolymphocytic leukemia, acute myelogenous leukemias; chronic lymphocytic leukemias, including B-cell CLL, hairy cell leukemia; and acute lymphoblastic leukemias Examples of lymphomas include, but are not limited to, B-cell lymphomas, such as Burkitt's lymphoma; and Hodgkin's lymphoma. In some embodiments, the disease or disorder is a genetic disorder. In some embodiments, the genetic disorder is sickle cell anemia, Glycogen storage diseases (GSD, e.g., GSD types I, II, III, IV, V, VI, VII, VIII, IX, X, XI, XII, XIII, and XIV), cystic fibrosis, lysosomal acid lipase (LAL) deficiency 1, Tay-Sachs disease, Phenylketonuria, Mucopolysaccharidoses, Galactosemia, muscular dystrophy (e.g., Duchenne muscular dystrophy), hemophilia such as hemophilia A (classic hemophilia) or hemophilia B (Christmas Disease), Wilson's disease, Fabry Disease, Gaucher Disease hereditary angioedema (HAE), and alpha 1 antitrypsin deficiency. Examples of other diseases or disorders are provided above in Section 6.7.1.

[0241] In some aspects, the disease or condition is a disease of muscle. Exemplary diseases of the muscle include Acid Maltase Deficiency (AMD), Amyotrophic Lateral Sclerosis (ALS), Andersen-Tawil Syndrome, Becker Muscular Dystrophy (BMD), Becker Myotonia Congenita, Bethlem Myopathy, Bulbospinal Muscular Atrophy (Spinal- Bulbar Muscular Atrophy), Carnitine Deficiency, Carnitine Palmityl Transferase Deficiency (CPT Deficiency), Central Core Disease (CCD), Centronuclear Myopathy, Charcot-Marie- Tooth Disease (CMT), Congenital Muscular Dystrophy (CMD), Congenital Myasthenic Syndromes (CMS), Congenital Myotonic Dystrophy, Cori Disease (Debrancher Enzyme Deficiency), Debrancher Enzyme Deficiency, Dejerine-Sottas Disease (DSD), Dermatomyositis (DM), Distal Muscular Dystrophy (DD), Duchenne Muscular Dystrophy (DMD), Dystrophia Myotonica (Myotonic Muscular Dystrophy), Emery-Dreifuss Muscular Dystrophy (EDMD), Endocrine Myopathies, Eulenberg Disease (Paramyotonia Congenita), Facioscapulohumeral Muscular Dystrophy (FSH or FSHD), Finnish (Tibial) Distal Myopathy, Forbes Disease (Debrancher Enzyme Deficiency), Friedreich's Ataxia (FA), Fukuyama Congenital Muscular Dystrophy, Glycogenosis Type 10, Glycogenosis Type 11 , Glycogenosis Type 2, Glycogenosis Type 3, Glycogenosis Type 5, Glycogenosis Type 7, Glycogenosis Type 9, Gowers-Laing Distal Myopathy, Hauptmann-ThanheuserMD (Emery- Dreifuss Muscular Dystrophy), Hereditary Inclusion-Body Myositis, Hereditary Motor and Sensory Neuropathy (Charcot-Marie-Tooth Disease), Hyperthyroid Myopathy, Hypothyroid Myopathy, Inclusion-Body Myositis (IBM), Inherited Myopathies, Integrin-Deficient Congenital Muscular Dystrophy, Kennedy Disease (Spinal-Bulbar Muscular Atrophy), Kugelberg-Welander Disease (Spinal Muscular Atrophy), Lactate Dehydrogenase Deficiency, Lambert-Eaton Myasthenic Syndrome (LEMS), Limb-Girdle Muscular Dystrophy (LGMD), Lou Gehrig's Disease (Amyotrophic Lateral Sclerosis), McArdle Disease (Phosphorylase Deficiency), Merosin-Deficient Congenital Muscular Dystrophy, Metabolic Diseases of Muscle, Mitochondrial Myopathy, Miyoshi Distal Myopathy, Motor Neurone Disease, Muscle-Eye-Brain Disease, Myasthenia Gravis (MG), Myoadenylate Deaminase Deficiency, Myofibrillar Myopathy, Myophosphorylase Deficiency, Myotonia Congenita (MC), Myotonic Muscular Dystrophy (MMD), Myotubular Myopathy (MTM or MM), Nemaline Myopathy, Nonaka Distal Myopathy, Oculopharyngeal Muscular Dystrophy (OPMD), Paramyotonia Congenita, Pearson Syndrome, Periodic Paralysis, Peroneal Muscular Atrophy (Charcot-Marie-Tooth Disease), Phosphofructokinase Deficiency, Phosphoglycerate Kinase Deficiency, Phosphoglycerate Mutase Deficiency, Phosphorylase Deficiency, Phosphorylase Deficiency, Polymyositis (PM), Pompe Disease (Acid Maltase Deficiency), Progressive External Ophthalmoplegia (PEO), Rod Body Disease (Nemaline Myopathy), Spinal Muscular Atrophy (SMA), Spinal-Bulbar Muscular Atrophy (SBMA), Steinert Disease (Myotonic Muscular Dystrophy), Tarui Disease (Phosphofructokinase Deficiency), Thomsen Disease (Myotonia Congenita), Ullrich Congenital Muscular Dystrophy, Walker-Warburg Syndrome (Congenital Muscular Dystrophy), Welander Distal Myopathy, Werdnig-Hoffmann Disease (Spinal Muscular Atrophy), and ZASP-Related Myopathy. Additional examples of muscle diseases and disorders are provided above in Section 6.7.1

[0242] In some embodiments, administration of a dependoparvovirus particle comprising a variant polypeptide and comprising a payload (e.g. , a transgene as described in Section 6.7.1) to a subject induces expression of the payload (e.g., transgene) in a subject. In some embodiments, the expression is induced in muscle tissue. In some embodiments, the production is similar in the muscle tissue as compared to a similar particle with the wild-type capsid protein. In some embodiments, the production is increased in the muscle tissue as compared to a similar particle with the wild-type capsid protein. The amount of a payload, e.g., transgene, e.g., heterologous protein, e.g., therapeutic polypeptide, expressed in a subject (e.g., the serum of the subject) can vary. For example, in some embodiments the payload, e.g., protein or RNA product of a transgene, can be expressed in the serum of the subject in the amount of at least about 9 g / ml, at least about 10 pg / ml, at least about 50 g / ml, at least about 100 pg / ml, at least about 200 pg / ml, at least about 300 pg / ml, at least about 400 g / ml, at least about 500 pg / ml, at least about 600 pg / ml, at least about 700 pg / ml, at least about 800 g / ml, at least about 900 pg / ml, or at least about 1000 pg / ml In some embodiments, the payload, e.g., protein or RNA product of a transgene, is expressed in the serum of the subject in the amount of about 9 pg / ml, about 10 g / ml, about 50 g / ml, about 100 g / ml, about 200 g / ml, about 300 g / ml, about 400 pg / ml, about 500 g / ml, about 600 pg / ml, about 700 pg / ml, about 800 g / ml, about 900 g / ml, about 1000 pg / ml, about 1500 pg / ml, about 2000 pg / ml, about 2500 pg / ml, or a range between any two of these values.

[0243] In some embodiments, for therapeutic applications, a viral particle comprising a capsid polypeptide as described herein is prepared as a pharmaceutical composition. As used herein the term “pharmaceutical composition” refers to a composition comprising at least one active ingredient (e.g., the viral particle) and optionally, one or more pharmaceutically acceptable carriers or excipients.

[0244] Relative amounts of the active ingredient, pharmaceutically acceptable carrier or excipient, and / or any additional ingredients in a pharmaceutical composition in accordance with the present disclosure may vary Differences in the constitution of a pharmaceutical composition may depend upon the identity, size, and / or condition of the subject being treated, the route by which the composition is to be administered, and / or any other factor. The composition may comprise between 0.0001 % and 99% (w / w) of the active ingredient. By way of example, the composition may comprise between 0.0001% and 100%, e.g., between .5 and 50%, between 1-30%, between 5-80%, or at least 80% (w / w) active ingredient. Non limiting examples of carriers and / or excipients include solvents, dispersion media, diluents, or other liquid vehicles, dispersion or suspension aids, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, or combination thereof7. NUMBERED EMBODIMENTS

[0245] While various specific embodiments have been illustrated and described, it will be appreciated that various changes can be made without departing from the spirit and scope of the disclosure(s). The present disclosure is exemplified by the numbered embodiments set forth below. Unless otherwise specified, features of any of the concepts, aspects and / or embodiments described in the detailed description above are applicable mutatis mutandis to any of the following numbered embodiments.1 . A capsid polypeptide comprising the amino acid sequence TRGDYAS (SEQ ID NO:16).2. The capsid polypeptide of embodiment 1 , wherein at least a portion of the amino acid sequence of SEQ ID NO:16 is in a VR-VIII variable region of the capsid polypeptide3. The capsid polypeptide of embodiment 1 , wherein at least a portion of the amino acid sequence of SEQ ID NO:16 is in a surface loop of the capsid polypeptide4. The capsid polypeptide of embodiment 3, wherein the surface loop comprises a VR- VIII variable region.5. The capsid polypeptide of any one of embodiments 1 to 4, wherein the amino acid sequence of SEQ ID NO: 16 is C-terminal to a position corresponding to Q585 of the VP1 capsid polypeptide of SEQ ID NO:76. The capsid polypeptide of any one of embodiments 1 to 5, wherein the amino acid sequence of SEQ ID NO: 16 is C-terminal to a position corresponding to S586 of the VP1 capsid polypeptide of SEQ ID NOV.7. The capsid polypeptide of any one of embodiments 1 to 6, wherein the amino acid sequence of SEQ ID NO: 16 is C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NO:7.8. The capsid polypeptide of any one of embodiments 1 to 7, wherein the amino acid sequence of SEQ ID NO:16 is immediately C-terminal to the position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NO:7.9. The capsid polypeptide of any one of embodiments 1 to 8, wherein the amino acid sequence of SEQ ID NO: 16 is N-terminal to a position corresponding to Q590 of the VP1 capsid polypeptide of SEQ ID NO:7.10. The capsid polypeptide of any one of embodiments 1 to 9, wherein the amino acid sequence of SEQ ID NO: 16 is N-terminal to a position corresponding to A589 of the VP1 capsid polypeptide of SEQ ID NO:7.11 . The capsid polypeptide of any one of embodiments 1 to 10, wherein the amino acid sequence of SEQ ID NO: 16 is N-terminal to a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.12. The capsid polypeptide of any one of embodiments 1 to 11 , wherein the amino acid sequence of SEQ ID NO:16 is immediately N-terminal to the position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.13. The capsid polypeptide of any one of embodiments 1 to 12, which comprises an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.14. The capsid polypeptide of any one of embodiments 1 to 13, which comprises a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7.15. The capsid polypeptide of any one of embodiments 1 to 14, which comprises an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NO:7.16. The capsid polypeptide of any one of embodiments 1 to 15, which comprises the amino acid sequence TRGDYASI (SEQ ID NO:18).17. The capsid polypeptide of any one of embodiments 1 to 16, which comprises an amino acid sequence having one, two, or three amino acid differences as compared to the amino acid sequence of TRGDYASIAQAQTQ (SEQ ID NO:19).18. The capsid polypeptide of any one of embodiments 1 to 16, which comprises the amino acid sequence of SEQ ID NO:1919. The capsid polypeptide of any one of embodiments 1 to 18, which comprises an amino acid sequence having one, two, or three amino acid differences as compared to the amino acid sequence of TRGDYASIAQAQTQWVQNQGA (SEQ ID NO:20).20. The capsid polypeptide of embodiment 17, which comprises the amino acid sequence of SEQ ID NO:20.21. The capsid polypeptide of any one of embodiments 1 to 19, which comprises an amino acid sequence having one, two, or three amino acid differences as compared to the amino acid sequence of SATRGDYASIAQAQTQWVQNQGAL (SEQ ID NO:21 ).22. The capsid polypeptide of embodiment 21 , which comprises the amino acid sequence of SEQ ID NO:21.23. A capsid polypeptide comprising the amino acid sequence TRGDYASI (SEQ ID NO:18).24. The capsid polypeptide of embodiment 23, wherein at least a portion of the amino acid sequence of SEQ ID NO:16 is in a VR-VIII variable region of the capsid polypeptide.25. The capsid polypeptide of embodiment 23, wherein at least a portion of the amino acid sequence of SEQ ID NO: 18 is in a surface loop of the capsid polypeptide.26. The capsid polypeptide of embodiment 25, wherein the surface loop comprises a VR- VIII variable region.27 The capsid polypeptide of any one of embodiments 23 to 26, wherein the amino acid sequence of SEQ ID NO: 18 is C-terminal to a position corresponding to Q585 of the VP1 capsid polypeptide of SEQ ID NOV.28. The capsid polypeptide of any one of embodiments 23 to 27, wherein the amino acid sequence of SEQ ID NO:18 is C-terminal to a position corresponding to S586 of the VP1 capsid polypeptide of SEQ ID NOV.29. The capsid polypeptide of any one of embodiments 23 to 28, wherein the amino acid sequence of SEQ ID NO:18 is C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV.30. The capsid polypeptide of any one of embodiments 23 to 29, wherein the amino acid sequence of SEQ ID NO: 18 is immediately C-terminal to the position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV.31 . The capsid polypeptide of any one of embodiments 23 to 30, wherein the amino acid sequence of SEQ ID NO: 18 is N-terminal to a position corresponding to Q590 of the VP1 capsid polypeptide of SEQ ID NO:7.32. The capsid polypeptide of any one of embodiments 23 to 31 , wherein the amino acid sequence of SEQ ID NO:18 is N-terminal to a position corresponding to A589 of the VP1 capsid polypeptide of SEQ ID NO:7.33. The capsid polypeptide of any one of embodiments 23 to 32, which lacks an amino acid at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.34. The capsid polypeptide of any one of embodiments 23 to 33, which comprises a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7.35. The capsid polypeptide of any one of embodiments 23 to 34, which comprises an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NO:7.36. A capsid polypeptide comprising an amino acid sequence which:(a) comprises the amino acid sequence of SEQ ID NO:16; and(b) has no more than three, two, or one amino acid differences as compared to the amino acid sequence TRGDYASIAQAQTQ (SEQ ID NO:19).37. The capsid polypeptide of embodiment 36, wherein at least a portion of the amino acid sequence of SEQ ID NO:16 is in a VR-VIII variable region of the capsid polypeptide.38 The capsid polypeptide of embodiment 36 or 37, wherein the amino acid sequence of SEQ ID NO:16 is C-terminal to a position corresponding to Q585 of the VP1 capsid polypeptide of SEQ ID NO:7.39. The capsid polypeptide of any one of embodiments 36 to 38, wherein the amino acid sequence of SEQ ID NO: 16 is C-terminal to a position corresponding to S586 of the VP1 capsid polypeptide of SEQ ID NO:7.40. The capsid polypeptide of any one of embodiments 36 to 39, wherein the amino acid sequence of SEQ ID NO: 16 is C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NO:7.41 . The capsid polypeptide of any one of embodiments 36 to 40, wherein the amino acid sequence of SEQ ID NO:16 is immediately C-terminal to the position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NO:7.42. The capsid polypeptide of any one of embodiments 36 to 41 , wherein the amino acid sequence has no more than two amino acid differences as compared to the amino acid sequence of SEQ ID NO:19.43. The capsid polypeptide of any one of embodiments 36 to 42, wherein the amino acid sequence has no more than one amino acid differences as compared to the amino acid sequence of SEQ ID NO:19.44. The capsid polypeptide of any one of embodiments 36 to 43, wherein the amino acid sequence comprises the amino acid sequence of SEQ ID NO: 1945. The capsid polypeptide of embodiment 44, wherein at least a portion of the amino acid sequence of SEQ ID NO: 19 is in a VR-VIII variable region of the capsid polypeptide.46. The capsid polypeptide of embodiment 44, wherein at least a portion of the amino acid sequence of SEQ ID NO: 19 is in a surface loop of the capsid polypeptide.47. The capsid polypeptide of embodiment 46, wherein the surface loop comprises a VR- VIII variable region.48. The capsid polypeptide of any one of embodiments 36 to 47, wherein the amino acid sequence is N-terminal to a position corresponding to Q597 of the VP1 capsid polypeptide of SEQ ID NOV.49. The capsid polypeptide of any one of embodiments 36 to 48, wherein the amino acid sequence of SEQ ID NO: 19 is N-terminal to a position corresponding to V596 of the VP1 capsid polypeptide of SEQ ID NOV.50. The capsid polypeptide of any one of embodiments 36 to 49, wherein the amino acid sequence of SEQ ID NO: 19 is N-terminal to a position corresponding to W595 of the VP1 capsid polypeptide of SEQ ID NOV.51 . The capsid polypeptide of any one of embodiments 36 to 50, wherein the amino acid sequence of SEQ ID NO:19 is immediately N-terminal to a position corresponding to W595 of the VP1 capsid polypeptide of SEQ ID NOV.52. The capsid polypeptide of any one of embodiments 36 to 51 , which comprises an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NOV.53. A capsid polypeptide comprising:(a) the amino acid sequence X1X2RGDX3X4X5X6, where Xi is any amino acid or absent, each of X2-X5 is any amino acid, and X@ is any amino acid or absent; and(b) one or both of:(i) an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7; and(ii) a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7.54. The capsid polypeptide of embodiment 53, which comprises an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.55. The capsid polypeptide of embodiment 53 or 54, which comprises a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7.56. The capsid polypeptide of any one of embodiments 53 to 55, which comprises an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NO:7.57. The capsid polypeptide of any one of embodiments 53 to 56, wherein at least a portion of the amino acid sequence X1X2RGDX3X4X5X6 is in a VR-VIII variable region of the capsid polypeptide.58. The capsid polypeptide of any one of embodiments 53 to 56, wherein at least a portion of the amino acid sequence X1X2RGDX3X4X5X6 is in a surface loop of the capsid polypeptide.59. The capsid polypeptide of embodiment 58, wherein the surface loop comprises a VR- VIII variable region.60 The capsid polypeptide of any one of embodiments 53 to 59, wherein the amino acid sequence X1X2RGDX3X4X5X6 is C-terminal to a position corresponding to Q585 of the VP1 capsid polypeptide of SEQ ID NOV.61 . The capsid polypeptide of any one of embodiments 53 to 60, wherein the amino acid sequence X1X2RGDX3X4X5X6 is C-terminal to a position corresponding to S586 of the VP1 capsid polypeptide of SEQ ID NOV.62. The capsid polypeptide of any one of embodiments 53 to 61 , wherein the amino acid sequence X1X2RGDX3X4X5X6 is C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV.63. The capsid polypeptide of any one of embodiments 53 to 62, wherein the amino acid sequence X1X2RGDX3X4X5X6 is immediately C-terminal to the position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV.64. The capsid polypeptide of any one of embodiments 53 to 63, wherein the amino acid sequence X1X2RGDX3X4X5X6 is N-terminal to a position corresponding to Q590 of the VP1 capsid polypeptide of SEQ ID NO:7.65. The capsid polypeptide of any one of embodiments 53 to 64, wherein the amino acid sequence X1X2RGDX3X4X5X6 is N-terminal to a position corresponding to A589 of the VP1 capsid polypeptide of SEQ ID NO:7.66. The capsid polypeptide of any one of embodiments 53 to 65, wherein the amino acid sequence X1X2RGDX3X4X5X6 is N-terminal to a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.67. The capsid polypeptide of any one of embodiments 53 to 66, wherein the amino acid sequence X1X2RGDX3X4X5X6 is immediately N-terminal to the position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.68. The capsid polypeptide of any one of embodiments 53 to 67, wherein Xi is any amino acid.69. The capsid polypeptide of any one of embodiments 53 to 67, wherein Xi is absent.70. The capsid polypeptide of any one of embodiments 53 to 69, wherein X2 is threonine.71. The capsid polypeptide of any one of embodiments 53 to 70, wherein X3 is tyrosine.72. The capsid polypeptide of any one of embodiments 53 to 71 , wherein X4 is alanine.73. The capsid polypeptide of any one of embodiments 53 to 72, wherein X5 is serine.74. The capsid polypeptide of any one of embodiments 53 to 73, wherein Xe is any amino acid.75. The capsid polypeptide of any one of embodiments 53 to 73, wherein Xe is isoleucine.76. The capsid polypeptide of any one of embodiments 53 to 73, wherein Xe is absent.77. A capsid polypeptide comprising the amino acid sequence RGDYSMT (SEQ ID NO:17).78. The capsid polypeptide of embodiment 77, wherein at least a portion of the amino acid sequence of SEQ ID NO:17 is in a VR-VIII variable region of the capsid polypeptide.79. The capsid polypeptide of embodiment 77, wherein at least a portion of the amino acid sequence of SEQ ID NO: 17 is in a surface loop of the capsid polypeptide.80. The capsid polypeptide of embodiment 79, wherein the surface loop comprises a VR- VIII variable region.81 . The capsid polypeptide of any one of embodiments 77 to 80, wherein the amino acid sequence of SEQ ID NO:17 is C-terminal to a position corresponding to S586 of the VP1 capsid polypeptide of SEQ ID NOV.82. The capsid polypeptide of any one of embodiments 77 to 81 , wherein the amino acid sequence of SEQ ID NO:17 is C-terminal to a position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NOV.83. The capsid polypeptide of any one of embodiments 77 to 82, wherein the amino acid sequence of SEQ ID NO: 17 is C-terminal to a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NOV.84. The capsid polypeptide of any one of embodiments 77 to 83, wherein the amino acid sequence of SEQ ID NO: 17 is immediately C-terminal to the position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NOV.85. The capsid polypeptide of any one of embodiments 77 to 84, wherein the amino acid sequence of SEQ ID NO:17 is N-terminal to a position corresponding to A591 of the VP1 capsid polypeptide of SEQ ID NOV.86. The capsid polypeptide of any one of embodiments 77 to 85, wherein the amino acid sequence of SEQ ID NO: 17 is N-terminal to a position corresponding to Q590 of the VP1 capsid polypeptide of SEQ ID NOV.87. The capsid polypeptide of any one of embodiments 77 to 86, wherein the amino acid sequence of SEQ ID NO:17 is N-terminal to a position corresponding to A589 of the VP1 capsid polypeptide of SEQ ID NOV.88. The capsid polypeptide of any one of embodiments 77 to 87, wherein the amino acid sequence of SEQ ID NO: 17 is immediately N-terminal to the position corresponding to A589 of the VP1 capsid polypeptide of SEQ ID NOV.89. The capsid polypeptide of any one of embodiments 77 to 88, which comprises a histidine at a position corresponding to T582 of the VP1 capsid polypeptide of SEQ ID NOV.90. The capsid polypeptide of any one of embodiments 77 to 89, which comprises the amino acid sequence HNHQSAQRGDYSMT (SEQ ID NO:22).91 . The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof92. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof93. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof94. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof95. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof96. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof97. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof98. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof99. The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof100 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof101 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof102 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof103 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof104 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 1 or the VP2 or VP3 portion thereof105 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof106 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof107 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof108 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof109 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof110 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof111 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof112 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof113 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof114 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof115 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof116 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof117 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof118 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:3 or the VP2 or VP3 portion thereof119 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof120 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof121 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof122 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof123 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof124 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof125 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof126 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof127 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof128 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof129 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof130 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof131 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof132 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:5 or the VP2 or VP3 portion thereof133 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof134 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof135 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof136 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof137 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof138 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof139 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof140 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof141 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof142 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof143 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof144 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof145 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof146 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof147 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof148 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof149 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof150 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof151 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof152 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof153 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof154 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof155 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof156 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof157 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof158 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof159 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof160 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:9 or the VP2 or VP3 portion thereof161 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.162 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.163 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.164 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.165 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof166 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.167 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.168 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.169 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.170 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.171 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.172 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.173 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.174 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:11 or the VP2 or VP3 portion thereof.175 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.176 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof177 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.178 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.179 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.180 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.181 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.182 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.183 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.184 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.185 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.186 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.187 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof188 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.189 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 70% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.190 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 75% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.191 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 80% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.192 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 85% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.193 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.194 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 91% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.195 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 92% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.196 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 93% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.197 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 94% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.198 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof199 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 96% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.200 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 97% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.201 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 98% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.202 The capsid polypeptide of any one of embodiments 1 to 90, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.203 A capsid polypeptide, which is optionally a capsid polypeptide according to any one of embodiments 1 to 202, which comprises the amino acid sequence of a VP1 capsid polypeptide of SEQ ID NO:12 or the VP2 or VP3 portion thereof.204 A capsid polypeptide, which is optionally a capsid polypeptide according to any one of embodiments 1 to 202, which comprises the amino acid sequence of a VP1 capsid polypeptide of SEQ ID NO:14 or the VP2 or VP3 portion thereof.205 A capsid polypeptide which:(a) comprises the mutations in the mutation set present in the amino acid sequence of SEQ ID NO:12 as compared to the amino acid sequence of SEQ ID NO:7; and(b) comprises an amino acid sequence with an edit distance of 15 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).206 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 14 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).207 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 13 or less to the amino acid sequence of SEQ ID NO: 12 (or to the VP2 or VP3 portion thereof).208 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 12 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).209 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 11 or less to the amino acid sequence of SEQ ID NO: 12 (or to the VP2 or VP3 portion thereof).210 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 10 or less to the amino acid sequence of SEQ ID NO: 12 (or to the VP2 or VP3 portion thereof).211 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 9 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).212 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 8 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).213 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 7 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).214 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 6 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).215 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 5 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).216 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 4 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).217 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 3 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).218 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 2 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof)219 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 1 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).220 A capsid polypeptide which:(a) comprises the mutations in the mutation set present in the amino acid sequence of SEQ ID NO:14 as compared to the amino acid sequence of SEQ ID NO:7; and(b) comprises an amino acid sequence with an edit distance of 15 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).221 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 14 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).222 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 13 or less to the amino acid sequence of SEQ ID NO: 14 (or to the VP2 or VP3 portion thereof).223 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 12 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).224 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 11 or less to the amino acid sequence of SEQ ID NO: 14 (or to the VP2 or VP3 portion thereof).225 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 10 or less to the amino acid sequence of SEQ ID NO: 14 (or to the VP2 or VP3 portion thereof).226 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 9 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).227 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 8 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof)228 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 7 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).229 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 6 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).230 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 5 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).231 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 4 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).232 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 3 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).233 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 2 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).234 The capsid polypeptide of embodiment 205, comprising an amino acid sequence with an edit distance of 1 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).235 The capsid polypeptide of any one of embodiments 1 to 234, whose sequence has an edit distance of (a) 12 or lower, (b) 11 or lower, or (c) 10 or lower to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof.236 The capsid polypeptide of any one of embodiments 1 to 235, whose sequence has an edit distance of (a) 12 or lower, (b) 11 or lower, or (c) 10 or lower to:(a) a VP1 capsid polypeptide of SEQ ID NO: 12;(b) a VP2 capsid polypeptide corresponding to the VP2 portion of SEQ ID NO:12; or(c) a VP3 capsid polypeptide corresponding to the VP3 portion of SEQ ID NO:12.237 The capsid polypeptide of any one of embodiments 1 to 235, whose sequence has an edit distance of (a) 12 or lower, (b) 11 or lower, or (c) 10 or lower to:(a) a VP1 capsid polypeptide of SEQ ID NO: 14;(b) a VP2 capsid polypeptide corresponding to the VP2 portion of SEQ ID NO:14; or(c) a VP3 capsid polypeptide corresponding to the VP3 portion of SEQ ID NO:14.238 The capsid polypeptide of any one of embodiments 1 to 237, which is a VP1 capsid polypeptide.239 The capsid polypeptide of any one of embodiments 1 to 237, which is a VP2 capsid polypeptide.240 The capsid polypeptide of any one of embodiments 1 to 237, which is a VP3 capsid polypeptide.241 A nucleic acid molecule encoding a capsid polypeptide of any one of embodiments 1 to 240.242 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 70% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).243 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).244 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 85% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).245 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).246 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 91% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).247 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 92% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).248 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 93% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).249 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 94% sequence identity to the nucleotide sequenceof SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).250 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).251 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 96% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).252 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 97% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).253 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 98% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).254 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 99% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).255 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e.g., a VP1 -encoding, a VP2-encoding or a VP3-encoding fragment thereof).256 The nucleic acid molecule of any one of embodiments 241 to 255, wherein the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:13.257 The nucleic acid molecule of any one of embodiments 241 to 255, wherein the nucleic acid molecule comprises a fragment of the nucleotide sequence of SEQ ID N0:13 that encodes a VP2 capsid polypeptide.258 The nucleic acid molecule of any one of embodiments 241 to 255, wherein the nucleic acid molecule comprises a fragment of the nucleotide sequence of SEQ ID N0:13 that encodes a VP3 capsid polypeptide.259 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 70% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).260 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).261 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 85% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).262 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).263 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 91% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).264 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 92% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).265 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 93% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).266 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 94% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).267 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 95% sequence identity to the nucleotide sequenceof SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).268 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 96% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).269 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 97% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).270 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 98% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).271 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 99% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof).272 The nucleic acid molecule of embodiment 241, wherein the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e.g., a VP1 -encoding, a VP2-encoding or a VP3-encoding fragment thereof).273 The nucleic acid molecule of any one of embodiments 259 to 272, wherein the nucleic acid molecule comprises the nucleotide sequence of SEQ ID NO:15.274 The nucleic acid molecule of any one of embodiments 259 to 272, wherein the nucleic acid molecule comprises a fragment of the nucleotide sequence of SEQ ID N0:15 that encodes a VP2 capsid polypeptide.275 The nucleic acid molecule of any one of embodiments 259 to 272, wherein the nucleic acid molecule comprises a fragment of the nucleotide sequence of SEQ ID N0:15 that encodes a VP3 capsid polypeptide.276 The nucleic acid molecule of any one of embodiments 241 to 275, wherein the nucleic acid molecule is double-stranded or single-stranded, and wherein the nucleic acid molecule is linear or circular, e.g., wherein the nucleic acid molecule is a plasmid277 A virus particle (e.g., adeno-associated virus (“AAV”) particle) comprising a capsid polypeptide of any one of embodiments 1 to 240 or comprising a capsid polypeptide encoded by the nucleic acid molecule of any one of embodiments 241 to 276.278 In a virus particle (e.g., adeno-associated virus (“AAV”) particle) comprising a capsid polypeptide, the improvement comprising including in the capsid polypeptide the amino acid sequence TRGDYAS (SEQ ID NO:16), optionally wherein the capsid polypeptide is the capsid polypeptide of any one of embodiments 1 to 76.279 In a virus particle (e.g., adeno-associated virus (“AAV”) particle) comprising a capsid polypeptide and an encapsulated nucleic acid, the improvement comprising including in the capsid polypeptide the amino acid sequence TRGDYAS (SEQ ID NO:16), optionally wherein the capsid polypeptide is the capsid polypeptide of any one of embodiments 1 to 76.280 In a virus particle (e.g., adeno-associated virus (“AAV”) particle) comprising a capsid polypeptide, the improvement comprising including in the capsid polypeptide the amino acid sequence RGDYSMT (SEQ ID NO:17), optionally wherein the capsid polypeptide is the capsid polypeptide of any one of embodiments 77 to 90.281 In a virus particle (e.g., adeno-associated virus (“AAV”) particle) comprising a capsid polypeptide and an encapsulated nucleic acid, the improvement comprising including in the capsid polypeptide the amino acid sequence RGDYSMT (SEQ ID NO:17), optionally wherein the capsid polypeptide is the capsid polypeptide of any one of embodiments 77 to 90.282 The virus particle of embodiment 277, 278, or 280, comprising a nucleic acid comprising a payload (e.g., a heterologous transgene) and one or more regulatory elements283 The virus particle of embodiment 279 or 281 , wherein the encapsulated nucleic acid comprises a payload (e.g., a heterologous transgene) and one or more regulatory elements.284 The virus particle of any one of embodiments 277 to 283, wherein the payload comprises a heterologous nucleic acid sequence as defined in any one of embodiments 409 to 554.285 The virus particle of any one of embodiments 277 to 284, wherein the one or more regulatory elements comprise a promoter.286 The virus particle of embodiment 285, wherein the promoter is a constitutive promoter.287 The virus particle of embodiment 285 or embodiment 286, wherein the promoter is aCBh promoter288 The virus particle of embodiment 287, wherein the CBh promoter comprises a nucleotide sequence having at least 90%, at least 95%, at least 96%, at least 97%, or at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO:500.289 The virus particle of embodiment 285, wherein the promoter is a muscle-specific promoter.290 The virus particle of any one of embodiments 277 to 289, wherein said virus particle exhibits increased cardiac muscle biodistribution, e.g., as measured in a mammal, e.g., in NHP, e.g., as described herein, relative to wild-type AAV9 (e.g., a virus particle comprising capsid polypeptides of SEQ ID NO:7 or encoded by SEQ ID NO:8), optionally wherein the increased cardiac muscle biodistribution is measured using a method described in Section 8.1.291 The virus particle of any one of embodiments 277 to 290, wherein said virus particle exhibits increased skeletal muscle biodistribution, e.g., as measured in a mammal, e.g., in NHP, e.g., as described herein, relative to wild-type AAV9 (e.g., a virus particle comprising capsid polypeptides of SEQ ID NO:7 or encoded by SEQ ID NO:8), optionally wherein the increased skeletal muscle biodistribution is measured using a method described in Section 8.1.292 The virus particle of any one of embodiments 277 to 291 , wherein said virus particle exhibits increased cardiac muscle transduction, e.g., as measured in a mammal, e.g., in NHP, e.g., as described herein, relative to wild-type AAV9 (e.g., a virus particle comprising capsid polypeptides of SEQ ID NO:7 or encoded by SEQ ID NO:8), optionally wherein the increased cardiac muscle transduction is measured using a method described in Section 8.1.293 The virus particle of any one of embodiments 277 to 292, wherein said virus particle exhibits increased skeletal muscle transduction, e.g., as measured in a mammal, e.g., in NHP, e g., as described herein, relative to wild-type AAV9 (e.g., a virus particle comprising capsid polypeptides of SEQ ID NO:7 or encoded by SEQ ID NO:8), optionally wherein the increased skeletal muscle transduction is measured using a method described in Section 8.1.294 The virus particle of any one of embodiments 277 to 293, wherein said virus particle exhibits reduced liver biodistribution, e.g., as measured in a mammal, e.g., in NHP, e.g., as described herein, relative to wild-type AAV9 (e.g., a virus particle comprising capsid polypeptides of SEQ ID NO:7 or encoded by SEQ ID NO:8), optionally wherein the decreased liver biodistribution is measured using a method described in Section 8.1.295 The virus particle of any one of embodiments 277 to 293, wherein said virus particle exhibits reduced liver transduction, e.g., as measured in a mammal, e.g., in NHP, e.g., as described herein, relative to wild-type AAV9 (e.g., a virus particle comprising capsid polypeptides of SEQ ID NO:7 or encoded by SEQ ID NO:8), optionally wherein the decreased liver transduction is measured using a method described in Section 8.1.296 An adeno-associated virus particle comprising (a) a capsid polypeptide of any one of embodiments 1 to 240 or comprising a capsid polypeptide encoded by the nucleic acid molecule of any one of embodiments 241 to 276 and (b) a nucleic acid comprising a payload (e.g., a heterologous transgene) and one or more regulatory elements.297 The virus particle of embodiment 296, wherein the virus particle comprises a capsid polypeptide comprising the amino acid sequence of SEQ ID NO:12 or the VP2 or VP3 portion thereof.298 The virus particle of embodiment 296, wherein the virus particle comprises a capsid polypeptide comprising the amino acid sequence of SEQ ID NO:14 or the VP2 or VP3 portion thereof.299 The virus particle of any one of embodiments 296 to 298, wherein the payload comprises a heterologous nucleic acid sequence as defined in any one of embodiments 241 to 276.300 The virus particle of any one of embodiments 296 to 299, wherein the one or more regulatory elements comprise a promoter.301 The virus particle of embodiment 300, wherein the promoter is as defined in any one of embodiments 286 to 289.302 The virus particle of any one of embodiments 277 to 301 , wherein the nucleic acid comprises AAV inverted terminal repeat sequences, optionally wherein the AAV inverted repeat sequences are AAV2 inverted terminal repeat sequences.303 The virus particle of any one of embodiments 277 to 302 (suitable) for use in gene therapy.304 In a method of delivering a nucleic acid to a subject via an adeno-associated virus (“AAV”) particle comprising a capsid polypeptide and the nucleic acid, the improvement comprising a capsid polypeptide comprising the amino acid sequence TRGDYAS (SEQ ID NO:16).305 In a method of delivering a nucleic acid to a subject via an adeno-associated virus (“AAV”) particle comprising a capsid polypeptide and the nucleic acid, the improvement comprising a capsid polypeptide comprising the amino acid sequence RGDYSMT (SEQ ID NO:17).306 A method of treating a disease or condition in a subject, comprising administering to the subject in an amount effective to treat the disease or condition:(a) a virus particle:(i) comprising the capsid polypeptide of any one of embodiments 1 to 240 and a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition;(ii) comprising a capsid polypeptide encoded by the nucleic acid molecule of any one of embodiments 241 to 276 and a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition, or(iii) of any one of embodiments 277 to 303 comprising a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition; or(b) a composition, e.g., a pharmaceutical composition, comprising the virus particle of (a) and, optionally, a pharmaceutically acceptable carrier.307 In a method of treating a disease or condition in a subject with an adeno-associated virus (“AAV”) particle comprising a capsid polypeptide and a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition, the improvement comprising a capsid polypeptide comprising the amino acid sequence TRGDYAS (SEQ ID NO:16)308 In a method of treating a disease or condition in a subject with an adeno-associated virus (“AAV”) particle comprising a capsid polypeptide and a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition, the improvement comprising a capsid polypeptide comprising the amino acid sequence RGDYSMT (SEQ ID NO:17).309 The method of embodiment 306, wherein the disease or condition is a muscle disease or condition.310 The method of embodiment 309, wherein the disease or condition is a skeletal muscle disease or condition.311 The method of embodiment 309, wherein the disease or condition is a cardiac muscle disease or condition.312 The method of embodiment 309, wherein the disease or condition is Acid Maltase Deficiency (AMD), Amyotrophic Lateral Sclerosis (ALS), Andersen-Tawil Syndrome, Barth syndrome (TAZ), Becker Muscular Dystrophy (BMD), Becker Myotonia Congenita, Bethlem Myopathy, Bulbospinal Muscular Atrophy (Spinal- Bulbar Muscular Atrophy), Carnitine Deficiency, Carnitine Palmityl Transferase Deficiency (CPT Deficiency), Central Core Disease (CCD), Centronuclear Myopathy, Charcot-Marie-Tooth Disease (CMT), Congenital Muscular Dystrophy (CMD), Congenital Myasthenic Syndromes (CMS), Congenital Myotonic Dystrophy, Cori Disease (Debrancher Enzyme Deficiency), Danon disease, Debrancher Enzyme Deficiency, Dejerine-Sottas Disease (DSD), Dermatomyositis (DM), Distal Muscular Dystrophy (DD), Distal myopathy with anterior tibial onset, Duchenne Muscular Dystrophy (DMD), Dystrophia Myotonica (Myotonic Muscular Dystrophy), Emery- Dreifuss Muscular Dystrophy (EDMD), Endocrine Myopathies, Eulenberg Disease (Paramyotonia Congenita), Facioscapulohumeral Muscular Dystrophy (FSH or FSHD), Finnish (Tibial) DistalMyopathy, Forbes Disease (Debrancher Enzyme Deficiency), Friedreich's Ataxia (FA), Fukuyama Congenital Muscular Dystrophy, Glycogenosis Type 10, Glycogenosis Type 11 , Glycogenosis Type 2, Glycogenosis Type 3, Glycogenosis Type 5, Glycogenosis Type 7, Glycogenosis Type 9, Gowers- Laing Distal Myopathy, Hauptmann-Thanheuser MD (Emery- Dreifuss Muscular Dystrophy), Hereditary Inclusion-Body Myositis, Hereditary Motor and Sensory Neuropathy (Charcot-Marie-Tooth Disease), Hyperthyroid Myopathy, Hypothyroid Myopathy, Inclusion-Body Myositis (IBM), Inherited Myopathies, Integrin-Deficient Congenital Muscular Dystrophy, Kennedy Disease (Spinal-Bulbar Muscular Atrophy), Kugelberg-Welander Disease (Spinal Muscular Atrophy), Lactate Dehydrogenase Deficiency, Lambert-Eaton Myasthenic Syndrome (LEMS), Limb-Girdle Muscular Dystrophy (LGMD), Lou Gehrig's Disease (Amyotrophic Lateral Sclerosis), McArdle Disease (Phosphorylase Deficiency), Merosin-Deficient Congenital Muscular Dystrophy, Metabolic Diseases of Muscle, Mitochondrial Myopathy, Miyoshi myopathy, Miyoshi Distal Myopathy, Motor Neurone Disease, Muscle-Eye-Brain Disease, Myasthenia Gravis (MG), Myoadenylate Deaminase Deficiency, Myofibrillar Myopathy, Myophosphorylase Deficiency, Myotonia Congenita (MC), Myotonic Muscular Dystrophy (MMD), Myotubular Myopathy (MTM or MM), Nemaline Myopathy, Nonaka Distal Myopathy, Oculopharyngeal Muscular Dystrophy (OPMD), Paramyotonia Congenita, Pearson Syndrome, Periodic Paralysis, Peroneal Muscular Atrophy (Charcot-Marie-Tooth Disease), Phosphofructokinase Deficiency, Phosphoglycerate Kinase Deficiency, Phosphoglycerate Mutase Deficiency, Phosphorylase Deficiency, Phosphorylase Deficiency, Polymyositis (PM), Pompe Disease (Acid Maltase Deficiency), Primary merosin deficiency (LAMA2), Progressive External Ophthalmoplegia (PEO), Rod Body Disease (Nemaline Myopathy), Spinal Muscular Atrophy (SMA), Spinal-Bulbar Muscular Atrophy (SBMA), Steinert Disease (Myotonic Muscular Dystrophy), Tarui Disease (Phosphofructokinase Deficiency), Thomsen Disease (Myotonia Congenita), Ullrich Congenital Muscular Dystrophy, Walker- Warburg Syndrome (Congenital Muscular Dystrophy), Welander Distal Myopathy, Werdnig-Hoffmann Disease (Spinal Muscular Atrophy), or ZASP-Related Myopathy.313 The method of any one of embodiments 309 to 312, wherein the disease or condition is Acid Maltase Deficiency (AMD).314 The method of any one of embodiments 309 to 312, wherein the disease or condition is Amyotrophic Lateral Sclerosis (ALS).315 The method of any one of embodiments 309 to 312, wherein the disease or condition is Andersen-Tawil Syndrome.316 The method of any one of embodiments 309 to 312, wherein the disease or condition is Barth syndrome (TAZ).317 The method of any one of embodiments 309 to 312, wherein the disease or condition is Becker Muscular Dystrophy (BMD).The method of any one of embodiments 309 to 312, wherein the disease or conditionnia Congenita. The method of any one of embodiments 309 to 312, wherein the disease or conditionathy. The method of any one of embodiments 309 to 312, wherein the disease or conditionuscular Atrophy (Spinal- Bulbar Muscular Atrophy). The method of any one of embodiments 309 to 312, wherein the disease or condition iency The method of any one of embodiments 309 to 312, wherein the disease or condition ityl Transferase Deficiency (CPT Deficiency). The method of any one of embodiments 309 to 312, wherein the disease or condition isease (CCD) The method of any one of embodiments 309 to 312, wherein the disease or condition Myopathy. The method of any one of embodiments 309 to 312, wherein the disease or condition-Tooth Disease (CMT). The method of any one of embodiments 309 to 312, wherein the disease or conditionscular Dystrophy (CMD). The method of any one of embodiments 309 to 312, wherein the disease or conditionasthenic Syndromes (CMS). The method of any one of embodiments 309 to 312, wherein the disease or conditionotonic Dystrophy. The method of any one of embodiments 309 to 312, wherein the disease or conditionDebrancher Enzyme Deficiency). The method of any one of embodiments 309 to 312, wherein the disease or conditione. The method of any one of embodiments 309 to 312, wherein the disease or conditionnzyme Deficiency. The method of any one of embodiments 309 to 312, wherein the disease or conditions Disease (DSD).The method of any one of embodiments 309 to 312, wherein the disease or conditionitis (DM). The method of any one of embodiments 309 to 312, wherein the disease or condition r Dystrophy (DD). The method of any one of embodiments 309 to 312, wherein the disease or conditionhy with anterito tibial onset. The method of any one of embodiments 309 to 312, wherein the disease or conditionscular Dystrophy (DMD). The method of any one of embodiments 309 to 312, wherein the disease or conditionotonica (Myotonic Muscular Dystrophy). The method of any one of embodiments 309 to 312, wherein the disease or conditions Muscular Dystrophy (EDMD). The method of any one of embodiments 309 to 312, wherein the disease or condition myopathy. The method of any one of embodiments 309 to 312, wherein the disease or conditionease (Paramyotonia Congenita). The method of any one of embodiments 309 to 312, wherein the disease or conditionumeral Muscular Dystrophy (FSH to FSHD) The method of any one of embodiments 309 to 312, wherein the disease or condition) Distal Myopathy. The method of any one of embodiments 309 to 312, wherein the disease or conditione (Debrancher Enzyme Deficiency). The method of any one of embodiments 309 to 312, wherein the disease or conditionaxia (FA). The method of any one of embodiments 309 to 312, wherein the disease or conditionngenital Muscular Dystrophy. The method of any one of embodiments 309 to 312, wherein the disease or condition Type 10. The method of any one of embodiments 309 to 312, wherein the disease or condition Type 11.The method of any one of embodiments 309 to 312, wherein the disease or condition Type 2. The method of any one of embodiments 309 to 312, wherein the disease or condition Type 3. The method of any one of embodiments 309 to 312, wherein the disease or condition Type 5. The method of any one of embodiments 309 to 312, wherein the disease or condition Type 7. The method of any one of embodiments 309 to 312, wherein the disease or condition Type 9. The method of any one of embodiments 309 to 312, wherein the disease or condition Distal Myopathy. The method of any one of embodiments 309 to 312, wherein the disease or conditionhanheuser MD (Emery- Dreifuss Muscular Dystrophy). The method of any one of embodiments 309 to 312, wherein the disease or conditionlusion-Body Myositis. The method of any one of embodiments 309 to 312, wherein the disease or conditiontto and Senstoy Neuropathy (Charcot-Marie-Tooth Disease). The method of any one of embodiments 309 to 312, wherein the disease or condition yopathy. The method of any one of embodiments 309 to 312, wherein the disease or conditionyopathy. The method of any one of embodiments 309 to 312, wherein the disease or condition Myositis (IBM). The method of any one of embodiments 309 to 312, wherein the disease or conditionyopathy The method of any one of embodiments 309 to 312, wherein the disease or conditionent Congenital Muscular Dystrophy. The method of any one of embodiments 309 to 312, wherein the disease or conditionase (Spinal-Bulbar Muscular Atrophy)The method of any one of embodiments 309 to 312, wherein the disease or conditionlander Disease (Spinal Muscular Atrophy). The method of any one of embodiments 309 to 312, wherein the disease or condition rogenase Deficiency. The method of any one of embodiments 309 to 312, wherein the disease or conditionn Myasthenic Syndrome (LEMS). The method of any one of embodiments 309 to 312, wherein the disease or conditionuscular Dystrophy (LGMD). The method of any one of embodiments 309 to 312, wherein the disease or condition isease (Amyotrophic Lateral Sclerosis). The method of any one of embodiments 309 to 312, wherein the disease or condition se (Phosphtoylase Deficiency). The method of any one of embodiments 309 to 312, wherein the disease or conditionient Congenital Muscular Dystrophy The method of any one of embodiments 309 to 312, wherein the disease or condition ases of Muscle. The method of any one of embodiments 309 to 312, wherein the disease or condition Myopathy. The method of any one of embodiments 309 to 312, wherein the disease or conditionathy The method of any one of embodiments 309 to 312, wherein the disease or condition Myopathy The method of any one of embodiments 309 to 312, wherein the disease or conditione Disease. The method of any one of embodiments 309 to 312, wherein the disease or conditionrain Disease. The method of any one of embodiments 309 to 312, wherein the disease or conditionravis (MG). The method of any one of embodiments 309 to 312, wherein the disease or condition Deaminase Deficiency.The method of any one of embodiments 309 to 312, wherein the disease or conditionopathy. The method of any one of embodiments 309 to 312, wherein the disease or conditionlase Deficiency. The method of any one of embodiments 309 to 312, wherein the disease or conditiongenita (MC). The method of any one of embodiments 309 to 312, wherein the disease or conditioncular Dystrophy (MMD). The method of any one of embodiments 309 to 312, wherein the disease or condition opathy (MTM to MM). The method of any one of embodiments 309 to 312, wherein the disease or conditionpathy. The method of any one of embodiments 309 to 312, wherein the disease or condition Myopathy. The method of any one of embodiments 309 to 312, wherein the disease or conditioneal Muscular Dystrophy (OPMD). The method of any one of embodiments 309 to 312, wherein the disease or condition Congenita. The method of any one of embodiments 309 to 312, wherein the disease or conditionrome. The method of any one of embodiments 309 to 312, wherein the disease or conditionysis. The method of any one of embodiments 309 to 312, wherein the disease or conditioncular Atrophy (Charcot-Marie-Tooth Disease). The method of any one of embodiments 309 to 312, wherein the disease or conditionkinase Deficiency. The method of any one of embodiments 309 to 312, wherein the disease or conditionrate Kinase Deficiency. The method of any one of embodiments 309 to 312, wherein the disease or conditionrate Mutase Deficiency.The method of any one of embodiments 309 to 312, wherein the disease or condition Deficiency. The method of any one of embodiments 309 to 312, wherein the disease or conditionPM) The method of any one of embodiments 309 to 312, wherein the disease or conditione (Acid Maltase Deficiency). The method of any one of embodiments 309 to 312, wherein the disease or condition in deficiency (LAMA2). The method of any one of embodiments 309 to 312, wherein the disease or conditionxternal Ophthalmoplegia (PEO). The method of any one of embodiments 309 to 312, wherein the disease or conditionease (Nemaline Myopathy). The method of any one of embodiments 309 to 312, wherein the disease or conditionphy (SMA). The method of any one of embodiments 309 to 312, wherein the disease or condition Muscular Atrophy (SBMA). The method of any one of embodiments 309 to 312, wherein the disease or conditionse (Myotonic Muscular Dystrophy). The method of any one of embodiments 309 to 312, wherein the disease or condition (Phosphofructokinase Deficiency). The method of any one of embodiments 309 to 312, wherein the disease or conditionase (Myotonia Congenita). The method of any one of embodiments 309 to 312, wherein the disease or condition ital Muscular Dystrophy. The method of any one of embodiments 309 to 312, wherein the disease or conditionrg Syndrome (Congenital Muscular Dystrophy). The method of any one of embodiments 309 to 312, wherein the disease or conditional Myopathy The method of any one of embodiments 309 to 312, wherein the disease or condition ann Disease (Spinal Muscular Atrophy).408 The method of any one of embodiments 309 to 312, wherein the disease or condition is ZASP-Related Myopathy.409 The method of any one of embodiments 309 to 312, wherein the disease or condition is Advanced heart failure and / to wherein the heterologous nucleic acid sequence encodes SERCA2a, ATP2A2 (e.g., a polypeptide represented by UniProt Accession number P16615 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).410 The method of any one of embodiments 309 to 312, wherein the disease or condition is Amyotrophic lateral sclerosis (ALS) (Lou Gehrig’s disease) and / to wherein the heterologous nucleic acid sequence encodes Superoxide dismutase-1 (SOD1 ) (e.g., a polypeptide represented by UniProt Accession number P00441 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).411 The method of any one of embodiments 309 to 312, wherein the disease or condition is Andersen-Tawil Syndrome and / to wherein the heterologous nucleic acid sequence encodes KCNJ2 (e.g., a polypeptide represented by UniProt Accession number P63252 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).412 The method of any one of embodiments 309 to 312, wherein the disease or condition is Barth syndrome and / to wherein the heterologous nucleic acid sequence encodes TAFAZZIN (e.g., a polypeptide represented by UniProt Accession number Q16635 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).413 The method of any one of embodiments 309 to 312, wherein the disease or condition is Becker Muscular Dystrophy (BMD) and / to wherein the heterologous nucleic acid sequence encodes DMD (e g., a polypeptide represented by UniProt Accession number P11532 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).414 The method of any one of embodiments 309 to 312, wherein the disease or condition is Becker Myotonia Congenita and / to wherein the heterologous nucleic acid sequence encodes CLCN1 (e.g , a polypeptide represented by UniProt Accession number P35523 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).415 The method of any one of embodiments 309 to 312, wherein the disease or condition is Bethlem Myopathy and / to wherein the heterologous nucleic acid sequence encodes COL6A3 (e.g., a polypeptide represented by UniProt Accession number P12111 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).416 The method of any one of embodiments 309 to 312, wherein the disease or condition is Bethlem Myopathy and / to wherein the heterologous nucleic acid sequence encodes COL6A2 (e.g., a polypeptide represented by UniProt Accession number P12110 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).417 The method of any one of embodiments 309 to 312, wherein the disease or condition is Bethlem Myopathy and / to wherein the heterologous nucleic acid sequence encodes COL6A1 (e g., a polypeptide represented by UniProt Accession number P12109 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).418 The method of any one of embodiments 309 to 312, wherein the disease or condition is Bulbospinal Muscular Atrophy and / to wherein the heterologous nucleic acid sequence encodes AR (e.g., a polypeptide represented by UniProt Accession number P10275 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).419 The method of any one of embodiments 309 to 312, wherein the disease or condition is Carnitine Deficiency, systemic primary and / to wherein the heterologous nucleic acid sequence encodes SLC22A5 (e.g., a polypeptide represented by UniProt Accession number 076082 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).420 The method of any one of embodiments 309 to 312, wherein the disease or condition is Carnitine Palmityl Transferase Deficiency, type 1 and / to wherein the heterologous nucleic acid sequence encodes CPT1A (e.g., a polypeptide represented by UniProt Accession number P50416 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).421 The method of any one of embodiments 309 to 312, wherein the disease or condition is Carnitine Palmityl Transferase Deficiency, type 2 and / to wherein the heterologous nucleic acid sequence encodes CPT2 (e.g., a polypeptide represented by UniProt Accession number P23786 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).422 The method of any one of embodiments 309 to 312, wherein the disease or condition is Catecholaminergic polymtophic ventricular tachycardia 2 (CPVT2) and / to wherein the heterologous nucleic acid sequence encodes Calsequestrin-2 (CASQ2) (e g., a polypeptide represented by UniProt Accession number 014958 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).423 The method of any one of embodiments 309 to 312, wherein the disease or condition is Central Ctoe Disease (congenital myopathy, type 1A) and / to wherein the heterologous nucleic acid sequence encodes RYR1 (e.g., a polypeptide represented by UniProt Accession number P21817 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).424 The method of any one of embodiments 309 to 312, wherein the disease or condition is Centronuclear Myopathy type 1 and / to wherein the heterologous nucleic acid sequence encodes MTMR14 (e g., a polypeptide represented by UniProt Accession number Q8NCE2 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).425 The method of any one of embodiments 309 to 312, wherein the disease or condition is Centronuclear Myopathy type 2 and / to wherein the heterologous nucleic acid sequence encodesDNM2 (e g., a polypeptide represented by UniProt Accession number 014717 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).426 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes PMP22 (e.g., a polypeptide represented by UniProt Accession number Q01453 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).427 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes MPZ (e.g., a polypeptide represented by UniProt Accession number P25189 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).428 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes DNM2 (e.g., a polypeptide represented by UniProt Accession number P50570 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).429 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes MFN2 (e.g., a polypeptide represented by UniProt Accession number 095140 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).430 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes KIF1 B (e.g., a polypeptide represented by UniProt Accession number 060333 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).431 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes SBF2 (e.g., a polypeptide represented by UniProt Accession number Q86WG5 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).432 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes PNKP (e.g., a polypeptide represented by UniProt Accession number Q96T60 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).433 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes GDAP1 (e.g., a polypeptide represented by UniProt Accession number Q8TB36 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).434 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes LMNA (e.g., a polypeptide represented by UniProt Accession number P02545 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).435 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes FGD4 (e.g., a polypeptide represented by UniProt Accession number Q96M96 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).436 The method of any one of embodiments 309 to 312, wherein the disease or condition is Charcot-Marie-Tooth disease and / to wherein the heterologous nucleic acid sequence encodes MTMR2 (e.g., a polypeptide represented by UniProt Accession number Q13614 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).437 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Muscular Dystrophy (Ullrich disease) and / to wherein the heterologous nucleic acid sequence encodes COL6A3 (e g., a polypeptide represented by UniProt Accession number P12111 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).438 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Muscular Dystrophy (Ullrich disease) and / to wherein the heterologous nucleic acid sequence encodes COL6A2 (e g., a polypeptide represented by UniProt Accession number P12110 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).439 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Muscular Dystrophy (Ullrich disease) and / to wherein the heterologous nucleic acid sequence encodes COL6A1 (e g., a polypeptide represented by UniProt Accession number P12109 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).440 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes COLQ (e.g., a polypeptide represented by UniProt Accession number Q9Y215 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).441 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes AGRN (e.g., a polypeptide represented by UniProt Accession number 000468 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).442 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodesRAPSN (e.g., a polypeptide represented by UniProt Accession number Q13702 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).443 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes GFPT1 (e.g., a polypeptide represented by UniProt Accession number Q06210 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).444 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes SCN4A (e.g., a polypeptide represented by UniProt Accession number P35499 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).445 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes ALG2 (e.g., a polypeptide represented by UniProt Accession number Q9H553 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).446 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes ALG14 (e.g., a polypeptide represented by UniProt Accession number Q96F25 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).447 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes DPAGT1 (e.g., a polypeptide represented by UniProt Accession number Q9H3H5 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).448 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes CHRNE (e.g., a polypeptide represented by UniProt Accession number Q04844 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).449 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes CHRNA1 (e g., a polypeptide represented by UniProt Accession number P02708 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).450 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes DOK7 (e.g., a polypeptide represented by UniProt Accession number Q18PE1 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).451 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes CHAT (e.g., a polypeptide represented by UniProt Accession number P28329 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).452 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myopathy and / to wherein the heterologous nucleic acid sequence encodes ACTA1 (e.g., a polypeptide represented by UniProt Accession number P68133 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).453 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myopathy and / to wherein the heterologous nucleic acid sequence encodes STAC3 (e.g., a polypeptide represented by UniProt Accession number Q96MF2 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).454 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myopathy and / to wherein the heterologous nucleic acid sequence encodes TPM3 (e.g., a polypeptide represented by UniProt Accession number P06753 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).455 The method of any one of embodiments 309 to 312, wherein the disease or condition is Congenital Myotonic Dystrophy and / to wherein the heterologous nucleic acid sequence encodes DMPK (e.g., a polypeptide represented by UniProt Accession number Q09013 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).456 The method of any one of embodiments 309 to 312, wherein the disease or condition is Ctoi Disease (Debrancher Enzyme Deficiency to Ftobes Disease) and / to wherein the heterologous nucleic acid sequence encodes AGL (e g., a polypeptide represented by UniProt Accession number P35573 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto)457 The method of any one of embodiments 309 to 312, wherein the disease or condition is Danon disease and / to wherein the heterologous nucleic acid sequence encodes LAMP2 (e.g., a polypeptide represented by UniProt Accession number P13473 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).458 The method of any one of embodiments 309 to 312, wherein the disease or condition is Dejerine-Sottas Disease and / to wherein the heterologous nucleic acid sequence encodes MPZ (e.g., a polypeptide represented by UniProt Accession number P25189 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).459 The method of any one of embodiments 309 to 312, wherein the disease or condition is Dejerine-Sottas Disease and / to wherein the heterologous nucleic acid sequence encodes EGR2(e.g., a polypeptide represented by UniProt Accession number P11161 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).460 The method of any one of embodiments 309 to 312, wherein the disease or condition is Dejerine-Sottas Disease and / to wherein the heterologous nucleic acid sequence encodes PMP22 (e.g., a polypeptide represented by UniProt Accession number Q01453 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).461 The method of any one of embodiments 309 to 312, wherein the disease or condition is Dejerine-Sottas Disease and / to wherein the heterologous nucleic acid sequence encodes PRX (e.g., a polypeptide represented by UniProt Accession number Q9BXM0 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).462 The method of any one of embodiments 309 to 312, wherein the disease or condition is Distal Muscular Dystrophy, Welander and / to wherein the heterologous nucleic acid sequence encodes TIA1 (e.g., a polypeptide represented by UniProt Accession number P31483 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).463 The method of any one of embodiments 309 to 312, wherein the disease or condition is Distal Muscular Dystrophy, Miyoshi and / to wherein the heterologous nucleic acid sequence encodes DYSF (e.g., a polypeptide represented by UniProt Accession number 075923 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).464 The method of any one of embodiments 309 to 312, wherein the disease or condition is Distal myopathy with anterito tibial onset and / to wherein the heterologous nucleic acid sequence encodes DYSF (e.g., a polypeptide represented by UniProt Accession number 075923 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).465 The method of any one of embodiments 309 to 312, wherein the disease or condition is Duchenne Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes Dystrophin (DMD) (e.g., a polypeptide represented by UniProt Accession number P11532 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).466 The method of any one of embodiments 309 to 312, wherein the disease or condition is Duchenne Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes GALGT2, B4GALNT2 (e.g., a polypeptide represented by UniProt Accession number Q8NHY0 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).467 The method of any one of embodiments 309 to 312, wherein the disease or condition is Dysferlinopathy and / to wherein the heterologous nucleic acid sequence encodes Dysferlin (DYSF) (e.g., a polypeptide represented by UniProt Accession number 075923 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).468 The method of any one of embodiments 309 to 312, wherein the disease or condition is Emery-Dreifuss Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes EMD (e.g., a polypeptide represented by UniProt Accession number P50402 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).469 The method of any one of embodiments 309 to 312, wherein the disease or condition is Emery-Dreifuss Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes SYNE1 (e.g., a polypeptide represented by UniProt Accession number Q8NF91 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).470 The method of any one of embodiments 309 to 312, wherein the disease or condition is Emery-Dreifuss Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes SYNE2 (e.g., a polypeptide represented by UniProt Accession number Q8WXH0 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).471 The method of any one of embodiments 309 to 312, wherein the disease or condition is Emery-Dreifuss Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes TMEM43 (e.g., a polypeptide represented by UniProt Accession number Q9BTV4 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).472 The method of any one of embodiments 309 to 312, wherein the disease or condition is Emery-Dreifuss Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes LMNA (e.g., a polypeptide represented by UniProt Accession number P02545 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).473 The method of any one of embodiments 309 to 312, wherein the disease or condition is Eulenberg Disease (Paramyotonia Congenita) and / to wherein the heterologous nucleic acid sequence encodes SCN4A (e.g., a polypeptide represented by UniProt Accession number P35499 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).474 The method of any one of embodiments 309 to 312, wherein the disease or condition is Facioscapulohumeral Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes SMCHD1 (e.g., a polypeptide represented by UniProt Accession number A6NHR9 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).475 The method of any one of embodiments 309 to 312, wherein the disease or condition is Facioscapulohumeral Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes LRIF1 (e.g., a polypeptide represented by UniProt Accession number Q5T3J3 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).476 The method of any one of embodiments 309 to 312, wherein the disease or condition is Friedreich's ataxia and / to wherein the heterologous nucleic acid sequence encodes Frataxin (FXN)(e.g., a polypeptide represented by UniProt Accession number Q16595 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).477 The method of any one of embodiments 309 to 312, wherein the disease or condition is Fukuyama Congenital Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes FKTN (e.g., a polypeptide represented by UniProt Accession number 075072 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).478 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogen sttoage disease II (Pompe disease or Acid Maltase Deficiency) and / to wherein the heterologous nucleic acid sequence encodes GAA (e.g., a polypeptide represented by UniProt Accession number P10253 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).479 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 10 (Glycogen sttoage disease X) and / to wherein the heterologous nucleic acid sequence encodes PGAM2 (e.g., a polypeptide represented by UniProt Accession number P15259 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).480 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 11 (Glycogen sttoage disease XI) and / to wherein the heterologous nucleic acid sequence encodes LDHA (e.g., a polypeptide represented by UniProt Accession number P00338 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).481 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 2 (Glycogen sttoage disease II to Pompe Disease or Acid maltase deficiency) and / to wherein the heterologous nucleic acid sequence encodes GAA (e.g., a polypeptide represented by UniProt Accession number P10253 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).482 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 3 (Glycogen sttoage disease III) and / to wherein the heterologous nucleic acid sequence encodes AGL (e.g , a polypeptide represented by UniProt Accession number P35573 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).483 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 5 (Glycogen sttoage disease V to McArdle Disease or Myophosphtoylase Deficiency) and / to wherein the heterologous nucleic acid sequence encodes PYGM (e.g., a polypeptide represented by UniProt Accession number P11217 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).484 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 7 (Glycogen sttoage disease VII to Phosphofructokinase Deficiency to TaruiDisease) and / to wherein the heterologous nucleic acid sequence encodes PFKM (e g., a polypeptide represented by UniProt Accession number P08237 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).485 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 9 (Glycogen sttoage disease IX) and / to wherein the heterologous nucleic acid sequence encodes PHKB (e.g., a polypeptide represented by UniProt Accession number Q93100 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).486 The method of any one of embodiments 309 to 312, wherein the disease or condition is Glycogenosis Type 9 (Glycogen sttoage disease IX) and / to wherein the heterologous nucleic acid sequence encodes PHKA2 (e.g., a polypeptide represented by UniProt Accession number P46019 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).487 The method of any one of embodiments 309 to 312, wherein the disease or condition is Hereditary Inclusion-Body Myositis and / to wherein the heterologous nucleic acid sequence encodes GNE (e.g., a polypeptide represented by UniProt Accession number Q9Y223 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).488 The method of any one of embodiments 309 to 312, wherein the disease or condition is Integrin-Deficient Congenital Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes ITGA7 (e g., a polypeptide represented by UniProt Accession number Q 13683 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).489 The method of any one of embodiments 309 to 312, wherein the disease or condition is Kennedy Disease (Spinal-Bulbar Muscular Atrophy) and / to wherein the heterologous nucleic acid sequence encodes Androgen receptto (AR) (e.g., a polypeptide represented by UniProt Accession number P10275 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).490 The method of any one of embodiments 309 to 312, wherein the disease or condition is Kugelberg-Welander Disease and / to wherein the heterologous nucleic acid sequence encodes SMN1 (e.g., a polypeptide represented by UniProt Accession number Q16637 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).491 The method of any one of embodiments 309 to 312, wherein the disease or condition is Lactate dehydrogenase A deficiency and / to wherein the heterologous nucleic acid sequence encodes LDHA (e.g., a polypeptide represented by UniProt Accession number P00338 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).492 The method of any one of embodiments 309 to 312, wherein the disease or condition is Lactate Dehydrogenase B Deficiency and / to wherein the heterologous nucleic acid sequenceencodes LDHB (e.g., a polypeptide represented by UniProt Accession number P07195 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).493 The method of any one of embodiments 309 to 312, wherein the disease or condition is Lambert-Eaton Myasthenic Syndrome and / to wherein the heterologous nucleic acid sequence encodes CACNB2 (e.g., a polypeptide represented by UniProt Accession number Q08289 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).494 The method of any one of embodiments 309 to 312, wherein the disease or condition is Laing Distal Myopathy and / to wherein the heterologous nucleic acid sequence encodes MYH7 (e.g., a polypeptide represented by UniProt Accession number A7E2Y1 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).495 The method of any one of embodiments 309 to 312, wherein the disease or condition is Limb Girdle Muscular Dystrophy Type 20 (LGMD-2C) and / to wherein the heterologous nucleic acid sequence encodes Gamma-sarcoglycan (e.g., a polypeptide represented by UniProt Accession number Q13326 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).496 The method of any one of embodiments 309 to 312, wherein the disease or condition is Limb Girdle Muscular Dystrophy Type 2D (LGMD-2D) and / to wherein the heterologous nucleic acid sequence encodes Alpha-sarcoglycan (e.g., a polypeptide represented by UniProt Accession number Q16586 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto)497 The method of any one of embodiments 309 to 312, wherein the disease or condition is Limb Girdle Muscular Dystrophy Type2E ( LGMD-2E) and / to wherein the heterologous nucleic acid sequence encodes Beta-sarcoglycan (e.g., a polypeptide represented by UniProt Accession number Q16585 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto)498 The method of any one of embodiments 309 to 312, wherein the disease or condition is Limb Girdle Muscular Dystrophy Type 2F (LGMD-2F) and / to wherein the heterologous nucleic acid sequence encodes Delta-sarcoglycan (e.g., a polypeptide represented by UniProt Accession number Q92629 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto)499 The method of any one of embodiments 309 to 312, wherein the disease or condition is Merosin-Deficient Congenital Muscular Dystrophy and / to wherein the heterologous nucleic acid sequence encodes LAMA2 (e.g., a polypeptide represented by UniProt Accession number P24043 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).500 The method of any one of embodiments 309 to 312, wherein the disease or condition is Muscle-Eye-Brain Disease and / to wherein the heterologous nucleic acid sequence encodes POMGNT1 (e.g., a polypeptide represented by UniProt Accession number Q8WZA1 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).501 The method of any one of embodiments 309 to 312, wherein the disease or condition is Mitochondrial Myopathy and / to wherein the heterologous nucleic acid sequence encodes CHCHD10 (e.g., a polypeptide represented by UniProt Accession number Q8WYQ3 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).502 The method of any one of embodiments 309 to 312, wherein the disease or condition is Miyoshi myopathy and / to wherein the heterologous nucleic acid sequence encodes DYSF (e.g., a polypeptide represented by UniProt Accession number 075923 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).503 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myoadenylate Deaminase Deficiency and / to wherein the heterologous nucleic acid sequence encodes AMPD1 (e.g., a polypeptide represented by UniProt Accession number P23109 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).504 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 1 and / to wherein the heterologous nucleic acid sequence encodes DES (e.g., a polypeptide represented by UniProt Accession number P17661 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).505 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 2 and / to wherein the heterologous nucleic acid sequence encodes CRYAB (e.g., a polypeptide represented by UniProt Accession number P02511 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).506 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 3 and / to wherein the heterologous nucleic acid sequence encodes MYOT (e.g., a polypeptide represented by UniProt Accession number Q9UBF9 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).507 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 4 (ZASP related myopathy) and / to wherein the heterologous nucleic acid sequence encodes LDB3, ZASP (e g., a polypeptide represented by UniProt Accession number 075112 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto)508 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 5 and / to wherein the heterologous nucleic acid sequence encodes FLNC(e.g., a polypeptide represented by UniProt Accession number Q14315 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).509 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 6 and / to wherein the heterologous nucleic acid sequence encodes BAG3 (e.g., a polypeptide represented by UniProt Accession number 095817 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).510 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 7 and / to wherein the heterologous nucleic acid sequence encodes KY (e.g., a polypeptide represented by UniProt Accession number Q8NBH2 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).511 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 8 and / to wherein the heterologous nucleic acid sequence encodes PYROXD1 (e.g., a polypeptide represented by UniProt Accession number Q8WU10 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).512 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 9 and / to wherein the heterologous nucleic acid sequence encodes TTN (e g., a polypeptide represented by UniProt Accession number Q8WZ42 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).513 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 10 and / to wherein the heterologous nucleic acid sequence encodes SVIL (e.g., a polypeptide represented by UniProt Accession number 095425 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).514 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 11 and / to wherein the heterologous nucleic acid sequence encodes UNC45B (e.g., a polypeptide represented by UniProt Accession number Q8IWX7 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).515 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myofibrillar Myopathy 12 and / to wherein the heterologous nucleic acid sequence encodes MYL2 (e.g., a polypeptide represented by UniProt Accession number Q99972 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).516 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myotonic dystrophy Type 1 (Steinert Disease) and / to wherein the heterologous nucleic acid sequence encodes Myotonin-protein kinase (DMPK) (e.g., a polypeptide represented by UniProt Accession number Q09013 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).517 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myotonic dystrophy Type 2 and / to wherein the heterologous nucleic acid sequence encodes CNBP (e.g., a polypeptide represented by UniProt Accession number P62633 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).518 The method of any one of embodiments 309 to 312, wherein the disease or condition is Myotubular Myopathy and / to wherein the heterologous nucleic acid sequence encodes MTM1 (e.g., a polypeptide represented by UniProt Accession number Q13496 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).519 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 1 and / to wherein the heterologous nucleic acid sequence encodes TPM3 (e.g., a polypeptide represented by UniProt Accession number P06753 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).520 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 2 and / to wherein the heterologous nucleic acid sequence encodes NEB (e.g., a polypeptide represented by UniProt Accession number P20929 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).521 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 5A, 5B, 5C and / to wherein the heterologous nucleic acid sequence encodes TNNT1 (e.g., a polypeptide represented by UniProt Accession number P13805 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).522 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 3 and / to wherein the heterologous nucleic acid sequence encodes ACTA1 (e.g., a polypeptide represented by UniProt Accession number P68133 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).523 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 6 and / to wherein the heterologous nucleic acid sequence encodes KBTBD13 (e.g., a polypeptide represented by UniProt Accession number C9JR72 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).524 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 4 and / to wherein the heterologous nucleic acid sequence encodes TPM2 (e.g., a polypeptide represented by UniProt Accession number P07951 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).525 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 7 and / to wherein the heterologous nucleic acid sequence encodes CFL2 (e.g.,a polypeptide represented by UniProt Accession number Q9Y281 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).526 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 8 and / to wherein the heterologous nucleic acid sequence encodes KLHL40 (e.g., a polypeptide represented by UniProt Accession number Q2TBA0 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).527 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 9 and / to wherein the heterologous nucleic acid sequence encodes KLHL41 (e.g., a polypeptide represented by UniProt Accession number 060662 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).528 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nemaline Myopathy 10 and / to wherein the heterologous nucleic acid sequence encodes LMOD3 (e.g., a polypeptide represented by UniProt Accession number Q0VAK6 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).529 The method of any one of embodiments 309 to 312, wherein the disease or condition is Nonaka Distal Myopathy and / to wherein the heterologous nucleic acid sequence encodes GNE (e.g., a polypeptide represented by UniProt Accession number Q9Y223 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).530 The method of any one of embodiments 309 to 312, wherein the disease or condition is Oculopharynggeal muscular dystrophy and / to wherein the heterologous nucleic acid sequence encodes PABPN1 (e.g., a polypeptide represented by UniProt Accession number Q86U42 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).531 The method of any one of embodiments 309 to 312, wherein the disease or condition is Ornithine Transcarbamylase deficiency and / to wherein the heterologous nucleic acid sequence encodes OTO (e.g., a polypeptide represented by UniProt Accession number P00480 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).532 The method of any one of embodiments 309 to 312, wherein the disease or condition is Paramyotonia Congenita and / to wherein the heterologous nucleic acid sequence encodes SCN4A (e.g., a polypeptide represented by UniProt Accession number P35499 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).533 The method of any one of embodiments 309 to 312, wherein the disease or condition is Periodic Paralysis, hypokalemic and / to wherein the heterologous nucleic acid sequence encodes CACNA1S (e.g., a polypeptide represented by UniProt Accession number Q13698 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).534 The method of any one of embodiments 309 to 312, wherein the disease or condition is Periodic Paralysis, hyperkalemic and / to wherein the heterologous nucleic acid sequence encodes SCN4A (e.g., a polypeptide represented by UniProt Accession number P35499 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).535 The method of any one of embodiments 309 to 312, wherein the disease or condition is Phosphoglycerate Kinase Deficiency and / to wherein the heterologous nucleic acid sequence encodes PGK1 (e.g., a polypeptide represented by UniProt Accession number P00558 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).536 The method of any one of embodiments 309 to 312, wherein the disease or condition is Polymyositis and / to wherein the heterologous nucleic acid sequence encodes PMSCL2 (e.g., a polypeptide represented by UniProt Accession number Q01780 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).537 The method of any one of embodiments 309 to 312, wherein the disease or condition is Polymyositis and / to wherein the heterologous nucleic acid sequence encodes PMSCL1 (e.g., a polypeptide represented by UniProt Accession number Q06265 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).538 The method of any one of embodiments 309 to 312, wherein the disease or condition is Progressive External Ophthalmoplegia and / to wherein the heterologous nucleic acid sequence encodes POLG (e.g., a polypeptide represented by UniProt Accession number P54098 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).539 The method of any one of embodiments 309 to 312, wherein the disease or condition is Progressive External Ophthalmoplegia and / to wherein the heterologous nucleic acid sequence encodes POLG2 (e.g., a polypeptide represented by UniProt Accession number Q9UHN1 to a polypeptide comprising an amino acid sequence having at least 95% sequence identity thereto).540 The method of any one of embodiments 309 to 312, wherein the di...

Claims

1. WHAT IS CLAIMED IS:1 . A capsid polypeptide comprising the amino acid sequence TRGDYAS (SEQ ID NO:16).

2. The capsid polypeptide of claim 1, wherein at least a portion of the amino acid sequence of SEQ ID NO:16 is in a VR-VIII variable region of the capsid polypeptide3. The capsid polypeptide of claim 1 or 2, wherein the amino acid sequence of SEQ ID NO:16 is C-terminal to a position corresponding to Q585 of the VP1 capsid polypeptide of SEQ ID NOY.

4. The capsid polypeptide of any one of claims 1 to 3, wherein the amino acid sequence of SEQ ID NO:16 is immediately C-terminal to the position corresponding to A587 of the VP1 capsid polypeptide of SEQ ID NO:7.

5. The capsid polypeptide of any one of claims 1 to 4, wherein the amino acid sequence of SEQ ID NO:16 is N-terminal to a position corresponding to Q590 of the VP1 capsid polypeptide of SEQ ID NOY.6 The capsid polypeptide of any one of claims 1 to 5, wherein the amino acid sequence of SEQ ID NO: 16 is immediately N-terminal to the position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.

7. The capsid polypeptide of any one of claims 1 to 6, which comprises an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.

8. The capsid polypeptide of any one of claims 1 to 7, which comprises a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7.

9. The capsid polypeptide of any one of claims 1 to 8, which comprises an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NOY.

10. The capsid polypeptide of any one of claims 1 to 9, which comprises the amino acid sequence TRGDYASI (SEQ ID NO: 18).11 . The capsid polypeptide of any one of claims 1 to 10, which comprises an amino acid sequence having one, two, or three amino acid differences as compared to the amino acid sequence of TRGDYASIAQAQTQ (SEQ ID NO:19).

12. The capsid polypeptide of any one of claims 1 to 10, which comprises the amino acid sequence of SEQ ID NO:19.

13. A capsid polypeptide comprising an amino acid sequence which:(a) comprises the amino acid sequence of SEQ ID NO:16; and(b) has no more than three, two, or one amino acid differences as compared to the amino acid sequence TRGDYASIAQAQTQ (SEQ ID NO:19).

14. The capsid polypeptide of claim 13, wherein at least a portion of the amino acid sequence of SEQ ID NO:16 is in a VR-VIII variable region of the capsid polypeptide15. A capsid polypeptide comprising:(a) the amino acid sequence X1X2RGDX3X4X5X6, where Xi is any amino acid or absent, each of X2-X5 is any amino acid, and Xe is any amino acid or absent; and(b) one or both of:(i) an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7; and(ii) a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7.

16. The capsid polypeptide of claim 15, which comprises an isoleucine at a position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.

17. The capsid polypeptide of claim 15 or 16, which comprises a glutamine at a position corresponding to G594 of the VP1 capsid polypeptide of SEQ ID NO:7.

18. The capsid polypeptide of any one of claims 15 to 17, which comprises an alanine at a position corresponding to 1601 of the VP1 capsid polypeptide of SEQ ID NO:7.

19. The capsid polypeptide of any one of claims 15 to 18, wherein at least a portion of the amino acid sequence X1X2RGDX3X4X5X6 is in a VR-VIII variable region of the capsid polypeptide.

20. A capsid polypeptide comprising the amino acid sequence RGDYSMT (SEQ ID NO:17).21 . The capsid polypeptide of claim 20, wherein at least a portion of the amino acid sequence of SEQ ID NO:17 is in a VR-VIII variable region of the capsid polypeptide22. The capsid polypeptide of claim 20 or 21 , wherein the amino acid sequence of SEQ ID NO:17 is C-terminal to a position corresponding to S586 of the VP1 capsid polypeptide of SEQ ID NOV23. The capsid polypeptide of any one of claims 20 to 22, wherein the amino acid sequence of SEQ ID NO: 17 is immediately C-terminal to the position corresponding to Q588 of the VP1 capsid polypeptide of SEQ ID NO:7.

24. The capsid polypeptide of any one of claims 20 to 23, wherein the amino acid sequence of SEQ ID NO:17 is N-terminal to a position corresponding to A591 of the VPI capsid polypeptide of SEQ ID NO:7.

25. The capsid polypeptide of any one of claims 20 to 24, wherein the amino acid sequence of SEQ ID NO: 17 is immediately N-terminal to the position corresponding to A589 of the VP1 capsid polypeptide of SEQ ID NO:7.

26. The capsid polypeptide of any one of claims 20 to 25, which comprises a histidine at a position corresponding to T582 of the VP1 capsid polypeptide of SEQ ID NO:7.

27. The capsid polypeptide of any one of claims 20 to 26, which comprises the amino acid sequence HNHQSAQRGDYSMT (SEQ ID NO:22).

28. The capsid polypeptide of any one of claims 1 to 27, which comprises an amino acid sequence having at least 90% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof.

29. The capsid polypeptide of any one of claims 1 to 27, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO:7 or the VP2 or VP3 portion thereof.

30. The capsid polypeptide of any one of claims 1 to 27, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.31 . The capsid polypeptide of any one of claims 1 to 27, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.

32. The capsid polypeptide of any one of claims 1 to 27, which comprises an amino acid sequence having at least 95% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.

33. The capsid polypeptide of any one of claims 1 to 27, which comprises an amino acid sequence having at least 99% sequence identity to a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.

34. A capsid polypeptide, which is optionally a capsid polypeptide according to any one of claims 1 to 33, which comprises the amino acid sequence of a VP1 capsid polypeptide of SEQ ID NO: 12 or the VP2 or VP3 portion thereof.

35. A capsid polypeptide, which is optionally a capsid polypeptide according to any one of claims 1 to 33, which comprises the amino acid sequence of a VP1 capsid polypeptide of SEQ ID NO: 14 or the VP2 or VP3 portion thereof.

36. A capsid polypeptide which:(a) comprises the mutations in the mutation set present in the amino acid sequence of SEQ ID NO:12 as compared to the amino acid sequence of SEQ ID NO:7; and(b) comprises an amino acid sequence with an edit distance of 15 or less to the amino acid sequence of SEQ ID NO:12 (or to the VP2 or VP3 portion thereof).

37. The capsid polypeptide of claim 36, comprising an amino acid sequence with an edit distance of 10 or less to the amino acid sequence of SEQ ID NO: 12 (or to the VP2 or VP3 portion thereof)38. A capsid polypeptide which:(a) comprises the mutations in the mutation set present in the amino acid sequence of SEQ ID NO:14 as compared to the amino acid sequence of SEQ ID NO:7; and(b) comprises an amino acid sequence with an edit distance of 15 or less to the amino acid sequence of SEQ ID NO:14 (or to the VP2 or VP3 portion thereof).

39. The capsid polypeptide of claim 38, comprising an amino acid sequence with an edit distance of 8 or less to the amino acid sequence of SEQ ID NO: 14 (or to the VP2 or VP3 portion thereof)40. The capsid polypeptide of any one of claims 1 to 39, whose sequence has an edit distance of (a) 12 or lower, (b) 11 or lower, or (c) 10 or lower to:(a) a VP1 capsid polypeptide of SEQ ID NO: 12;(b) a VP2 capsid polypeptide corresponding to the VP2 portion of SEQ ID NO:12; or(c) a VP3 capsid polypeptide corresponding to the VP3 portion of SEQ ID NO:12.41 . The capsid polypeptide of any one of claims 1 to 39, whose sequence has an edit distance of (a) 12 or lower, (b) 11 or lower, or (c) 10 or lower to:(a) a VP1 capsid polypeptide of SEQ ID NO: 14;(b) a VP2 capsid polypeptide corresponding to the VP2 portion of SEQ ID NO:14; or(c) a VP3 capsid polypeptide corresponding to the VP3 portion of SEQ ID NO:14.

42. The capsid polypeptide of any one of claims 1 to 41 , which is a VP1 capsid polypeptide.

43. A nucleic acid molecule encoding a capsid polypeptide of any one of claims 1 to 42.

44. The nucleic acid molecule of claim 43, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:13 or a fragment thereof (e g., a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof)45. The nucleic acid molecule of claim 43, wherein the nucleic acid molecule comprises a nucleotide sequence having at least 95% sequence identity to the nucleotide sequence of SEQ ID NO:15 or a fragment thereof (e g , a VP1-encoding, a VP2-encoding or a VP3-encoding fragment thereof)46 A virus particle (e g , adeno-associated virus (“AAV”) particle) comprising a capsid polypeptide of any one of claims 1 to 42 or comprising a capsid polypeptide encoded by the nucleic acid molecule of any one of claims 43 to 4547. The virus particle of claim 46, comprising a nucleic acid comprising a payload (e.g., a heterologous transgene) and one or more regulatory elements.

48. The virus particle of claim 46 or 47, wherein the one or more regulatory elements comprise a promoter.

49. The virus particle of claim 48, wherein the promoter is a muscle-specific promoter.50 The virus particle of any one of claims 46 to 49, wherein said virus particle exhibits increased cardiac muscle and / or skeletal muscle biodistribution and / or transduction, e.g , as measured in a mammal, e.g., in NHP, e.g., as described herein, relative to wild-type AAV9 (e.g., a virus particle comprising capsid polypeptides of SEQ ID NO:7 or encoded by SEQ ID NO:8), optionally wherein the increased cardiac and / or skeletal muscle biodistribution is measured using a method described in Section 8.1.51 . An adeno-associated virus particle comprising (a) a capsid polypeptide of any one of claims 1 to 42 or comprising a capsid polypeptide encoded by the nucleic acid molecule of any one of claims 43 to 45 and (b) a nucleic acid comprising a payload (e.g., a heterologous transgene) and one or more regulatory elements.

52. A method of treating a disease or condition in a subject, comprising administering to the subject in an amount effective to treat the disease or condition:(a) a virus particle:(i) comprising the capsid polypeptide of any one of claims 1 to 42 and a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition;(ii) comprising a capsid polypeptide encoded by the nucleic acid molecule of any one of claims 43 to 45 and a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition, or(iii) of any one of claims 46 to 50 comprising a heterologous nucleic acid sequence encoding a therapeutic product suitable for treating the disease or condition; or(b) a composition, e.g., a pharmaceutical composition, comprising the virus particle of (a) and, optionally, a pharmaceutically acceptable carrier.

53. The method of claim 52, wherein the disease or condition is a muscle disease or condition.

54. A method of making a virus (e.g., a dependoparvovirus particle such as an adeno- associated virus (AAV) particle), comprising:(a) providing a cell, cell-free system, or other translation system, comprising the nucleic acid of any one of claims 43 to 45 ; and(b) cultivating the cell, cell-free system, or other translation system, under conditions suitable for the production of the virus particle, thereby making the virus particle.

55. The method of claim 54, wherein the cell, cell-free system, or other translation system comprises a second nucleic acid molecule comprising a payload (e.g , a heterologous nucleic acid sequence encoding a therapeutic product, e.g., as described herein) and at least a portion of said second nucleic acid molecule is packaged in the dependoparvovirus particle.

56. A composition, e.g., a pharmaceutical composition, comprising a virus particle of any one of claims 46 to 50, or a virus particle produced by the method of claims 54 or 55, and a pharmaceutically acceptable carrier or excipient.

57. The capsid polypeptide of any one of claims 1 to 42, the nucleic acid molecule of any one of claims 43 to 45, the virus particle of any one of claims 46 to 50, or the composition of claim 56, for use in treating a disease or condition in a subject.

58. The capsid polypeptide of any one of claims 1 to 42, the nucleic acid molecule of any one of claims 43 to 45, the virus particle of any one of claims 46 to 50, or the composition of claim 56, for use in the manufacture of a medicament for use in treating a disease or condition in a subject.

59. A host cell comprising the nucleic acid of any one of claims 43 to 45.

60. The host cell of claim 59, wherein the host cell comprises a nucleic acid comprising a payload, e.g., a heterologous nucleic acid sequence encoding a therapeutic product, e.g., as described herein.