WO2024064782A2 - Capsides de virus adéno-associé - Google Patents

Capsides de virus adéno-associé Download PDF

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WO2024064782A2
WO2024064782A2 PCT/US2023/074729 US2023074729W WO2024064782A2 WO 2024064782 A2 WO2024064782 A2 WO 2024064782A2 US 2023074729 W US2023074729 W US 2023074729W WO 2024064782 A2 WO2024064782 A2 WO 2024064782A2
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seq
amino acids
acid sequence
amino acid
sequence identity
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Noah DAVIDSOHN
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Rejuvenate Bio
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures
    • C40B40/02—Libraries contained in or displayed by microorganisms, e.g. bacteria or animal cells; Libraries contained in or displayed by vectors, e.g. plasmids; Libraries containing only microorganisms or vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14145—Special targeting system for viral vectors

Definitions

  • Adeno-associated viruses are small, non-enveloped viruses of the genus Dependoviruses within the family Parvoviridae. They package a single-stranded DNA genome that is approximately 4.7kb in length, consisting of rep and cap genes flanked by inverted terminal repeat regions (ITRs). AAVs are an ideal delivery system for gene therapy because AAV infection of cells in culture is non-cytopathic, and natural infection of humans and other animals is silent.
  • the signals directing AAV replication, genome encapsidation and integration are contained within the inverted terminal repeats (ITRs) of the AAV genome, essentially all of the internal 4.7 kb of the AAV genome (encoding the replication and structural capsid proteins, repcap) can be replaced with heterologous nucleic acid sequences, such as a transgene for effecting gene therapy.
  • ITRs inverted terminal repeats
  • Recombinant AAV (rAAV) gene therapy vectors typically consist of a recombinant AAV genome comprising a gene therapy transgene, packaged within an AAV capsid.
  • the AAV capsid which is formed by an assembly of 60 AAV capsid proteins, interacts with the cell surface of target cells and determines the tropism and transduction efficiency of rAAV.
  • tissue-specific genetic diseases it is desirable to have rAAVs that can specifically and efficiently target tissues in need of gene therapy.
  • AAV adeno-associated virus
  • the instant disclosure provides a recombinant adeno- associated virus (AAV) capsid protein comprising: (i) an amino acid sequence having at least 94% sequence identity to amino acids 1-734 of SEQ ID NO: 2, an amino acid sequence having at least 95% sequence identity to amino acids 137-734 of SEQ ID NO: 2, and/or an amino acid sequence having at least 96% sequence identity to amino acids 202-734 of SEQ ID NO: 2: (ii) an amino acid sequence having at least 96% sequence identity to amino acids 1-736 of SEQ ID NO: 4, an amino acid sequence having at least 95% sequence identity to amino acids 138-736 of SEQ ID NO: 4, and/or an amino acid sequence having at least 96% sequence identity to amino acids 204-736 of SEQ ID NO: 4; (iii) an amino acid sequence having at least 94% sequence identity to amino acids 1-734 of SEQ ID NO: 6.
  • AAV adeno- associated virus
  • amino acid sequence having at least 95% sequence identity to amino acids 1-735 of SEQ ID NO: 32 an amino acid sequence having at least 95% sequence identity to amino acids 137-735 of SEQ ID NO: 32, and/or an amino acid sequence having at least 96% sequence identity to amino acids 202-735 of SEQ ID NO: 32;
  • the instant disclosure provides a recombinant AAV capsid protein comprising: (i) an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO: 2. an amino acid sequence having at least 99% sequence identity to amino acids 137-734 of SEQ ID NO: 2, and/or an amino acid sequence having at least 99% sequence identity to amino acids 202-734 of SEQ ID NO: 2; (ii) an amino acid sequence having at least 99% sequence identity to amino acids 1-736 of SEQ ID NO: 4, an amino acid sequence having at least 99% sequence identity to amino acids 138-736 of SEQ ID NO: 4, and/or an amino acid sequence having at least 99% sequence identity to amino acids 204-736 of SEQ ID NO: 4; (iii) an amino acid sequence having at least 99% sequence identity to amino acids 1-734 of SEQ ID NO: 6, an amino acid sequence having at least 99% sequence identity to amino acids 137-734 of SEQ ID NO: 6, and/or an amino acid sequence having at
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 2, amino acids 137-734 of SEQ ID NO: 2, and/or amino acids 202- 734 of SEQ ID NO: 2. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 2.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 4. amino acids 138-736 of SEQ ID NO: 4, and/or amino acids 204-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 4.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 6. amino acids 137-734 of SEQ ID NO: 6, and/or amino acids 202- 734 of SEQ ID NO: 6. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 6.
  • the recombinant AAV capsid protein comprises amino acids 1-731 of SEQ ID NO: 8. amino acids 134-731 of SEQ ID NO: 8, and/or amino acids 199- 731 of SEQ ID NO: 8. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 10.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 12, amino acids 137-736 of SEQ ID NO: 12, and/or amino acids 203- 736 of SEQ ID NO: 12. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 12.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 14, amino acids 138-736 of SEQ ID NO: 14, and/or amino acids 204- 736 of SEQ ID NO: 14. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 14.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 16, amino acids 137-734 of SEQ ID NO: 16, and/or amino acids 202- 734 of SEQ ID NO: 16. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 16.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 18, amino acids 138-735 of SEQ ID NO: 18, and/or amino acids 203-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 18.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 20, amino acids 137-736 of SEQ ID NO: 20, and/or amino acids 202-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 20.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 22, amino acids 137-736 of SEQ ID NO: 22, and/or amino acids 203- 736 of SEQ ID NO: 22. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 22.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 24, amino acids 138-736 of SEQ ID NO: 24, and/or amino acids 204- 736 of SEQ ID NO: 24. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 24.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 26, amino acids 137-735 of SEQ ID NO: 26, and/or amino acids 202- 735 of SEQ ID NO: 26. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 26.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 28, amino acids 137-734 of SEQ ID NO: 28, and/or amino acids 202- 734 of SEQ ID NO: 28. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 28.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 30, amino acids 137-734 of SEQ ID NO: 30, and/or amino acids 202-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 30.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 32, amino acids 137-735 of SEQ ID NO: 32, and/or amino acids 202-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 32.
  • the recombinant AAV capsid protein comprises amino acids 1 -735 of SEQ ID NO: 34, amino acids 137-735 of SEQ ID NO: 34, and/or amino acids 202- 735 of SEQ ID NO: 34.
  • the recombinant AAV capsid protein comprises SEQ ID NO: 34.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 36, amino acids 138-735 of SEQ ID NO: 36, and/or amino acids 203- 735 of SEQ ID NO: 36. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 36.
  • the recombinant AAV capsid protein comprises amino acids 1-731 of SEQ ID NO: 38, amino acids 134-731 of SEQ ID NO: 38, and/or amino acids 199- 731 of SEQ ID NO: 38. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 38.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 40, amino acids 137-735 of SEQ ID NO: 40, and/or amino acids 203-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 40.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 42, amino acids 138-736 of SEQ ID NO: 42, and/or amino acids 204-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 42.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 44, amino acids 137-736 of SEQ ID NO: 44, and/or amino acids 202- 736 of SEQ ID NO: 44. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 44.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 46, amino acids 137-736 of SEQ ID NO: 46, and/or amino acids 203- 736 of SEQ ID NO: 46. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 46.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 48, amino acids 137-734 of SEQ ID NO: 48, and/or amino acids 202- 734 of SEQ ID NO: 48. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 48.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 50, amino acids 137-734 of SEQ ID NO: 50, and/or amino acids 202- 734 of SEQ ID NO: 50. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 50.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 52, amino acids 137-734 of SEQ ID NO: 52, and/or amino acids 202-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 52.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 54, amino acids 138-735 of SEQ ID NO: 54, and/or amino acids 203-
  • the recombinant AAV capsid protein comprises SEQ ID NO: 54.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 56, amino acids 137-735 of SEQ ID NO: 56, and/or amino acids 202- 735 of SEQ ID NO: 56. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 56.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 58, amino acids 137-735 of SEQ ID NO: 58, and/or amino acids 202- 735 of SEQ ID NO: 58. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 58.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 60, amino acids 137-735 of SEQ ID NO: 60, and/or amino acids 202- 735 of SEQ ID NO: 60. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 60.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 62, amino acids 137-735 of SEQ ID NO: 62, and/or amino acids 202- 735 of SEQ ID NO: 62. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 62.
  • the recombinant AAV capsid protein of the present disclosure is a variant of a clade A, clade B, clade C, clade D, clade E, clade F, clade G, clade H, clade I, AAVgo. 1, AAV3, AAV4, AAV10, AAV11, AAV12, rh.32, rh32.33, rh.33, rh.34, BAAV, or AAV5 capsid protein.
  • the instant disclosure provides an isolated polynucleotide encoding a recombinant AAV capsid protein as described in the instant disclosure.
  • the isolated polynucleotide has at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, 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 with the nucleic acid sequence set forth in any one of SEQ ID NOs: 1, 3. 5, 7, 9. 11.
  • the isolated polynucleotide comprises the nucleic acid sequence set forth in any one of SEQ ID NOs: 1, 3, 5, 7, 9, 11. 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, and 61.
  • the instant disclosure provides a vector comprising an isolated polynucleotide as described in the instant disclosure.
  • the vector is a plasmid or a viral vector.
  • the viral vector is a retrovirus vector, a herpes virus vector, a baculovirus vector, or an adenovirus vector.
  • the vector is an expression vector.
  • the instant disclosure provides a recombinant cell comprising a polynucleotide as described in the instant disclosure or a vector as described in the instant disclosure.
  • the instant disclosure provides a method of producing a recombinant AAV capsid protein, the method comprising culturing a recombinant cell as described in the instant disclosure under conditions where the polynucleotide is expressed, and the capsid protein is produced.
  • the instant disclosure provides a recombinant adeno-associated virus (rAAV) comprising: (a) a capsid comprising one or more of the recombinant AAV capsid proteins of the instant disclosure; and (b) an rAAV genome.
  • the rAAV genome comprises a transgene.
  • the transgene encodes a polypeptide.
  • the transgene encodes a miRNA, shRNA, siRNA, antisense RNA, gRNA, antagomir, miRNA sponge, RNA aptazyme, RNA aptamer, IncRNA. ribozyme or mRNA.
  • the transgene is operably linked to a transcriptional regulatory element.
  • the instant disclosure provides a method for transducing a cell, the method comprising contacting the cell with a rAAV of the instant disclosure under conditions whereby the cell is transduced.
  • the instant disclosure provides a method for delivering a transgene in a cell, the method comprising contacting the cell with a rAAV of the instant disclosure under conditions whereby the cell is transduced and the transgene is expressed.
  • the cell is in a subject and the rAAV is administered to the subject.
  • the cell is a muscle cell and/or a liver cell.
  • the rAAV is administered to the subject intravenously, intraperitoneally, subcutaneously, intramuscularly, intrathecally, or intradermally.
  • the subject is selected from the group consisting of a human subject, a canine subject, a feline subject, and an equine subject.
  • the subject is a human subject.
  • the instant disclosure provides a recombinant AAV comprising: (a) a capsid comprising one or more of the recombinant AAV capsid proteins of the instant disclosure; and (b) an rAAV genome, for use in medicine, for use as therapy, or for use as a medicament.
  • the instant disclosure provides a packaging system for preparation of an rAAV, wherein the packaging system comprises: (a) a first nucleotide sequence encoding one or more AAV Rep proteins; (b) a second nucleotide sequence encoding a recombinant AAV capsid protein of the instant disclosure; and (c) a third nucleotide sequence comprising an rAAV genome sequence.
  • the packaging system comprises a first vector compnsing the first nucleotide sequence and the second nucleotide sequence, and a second vector comprising the third nucleotide sequence.
  • the packaging system of the instant disclosure further comprises a fourth nucleotide sequence comprising one or more helper virus genes.
  • the fourth nucleotide sequence is comprised within a third vector.
  • the fourth nucleotide sequence comprises one or more genes from a virus selected from the group consisting of adenovirus, herpesvirus, vaccinia virus, and cytomegalovirus (CMV).
  • CMV cytomegalovirus
  • the first vector, second vector, and/or the third vector of the packaging system of the instant disclosure is a plasmid.
  • the instant disclosure provides a method for recombinant preparation of an rAAV, the method comprising introducing the packaging system as described in the instant disclosure into a cell under conditions whereby the rAAV is produced.
  • FIG. 1 shows the expression level of FGF21 (FIG. 1A) or sTGFbR2 (FIG. IB) in plasma samples obtained from dogs intravenously administered 5 x 10 12 vg/kg of vector genomes comprising FGF21 and sTGFbR2 coding sequences packaged in CL01, CL03, CL11, CL13, CL27, or AAV8 (control) capsids.
  • FIG. 1A shows the expression of FGF21 as assessed by ELISA
  • FIG. IB shows the expression of sTGFbR2 as assessed by Western blot.
  • AAV capsid proteins recombinant adeno-associated virus (AAV) capsid proteins, compositions (e.g., rAAV) comprising the capsid proteins, nucleic acids encoding the capsid proteins, and methods of making and using the capsid proteins.
  • AAV capsid proteins described herein have been isolated from screening a shuffled capsid library in vivo for AAVs that are enriched in the liver and/or muscle.
  • AAV is a standard abbreviation for adeno-associated virus.
  • rAAV recombinant adeno-associated virus
  • capsid gene refers to a nucleic acid sequence that encodes a capsid protein.
  • the capsid protein may be VP1, VP2, or VP3.
  • VP1, VP2, and/or VP3 capsid proteins assemble into a capsid that surrounds the rAAV genome.
  • the term “rep gene” refers to the nucleic acid sequences that encode the non-structural proteins (e.g., rep78, rep68, rep52, and rep40) required for the replication and production of an AAV.
  • rAAV genome refers to a nucleic acid molecule (e.g., DNA and/or RNA) comprising the genome sequence of an rAAV.
  • a nucleic acid molecule e.g., DNA and/or RNA
  • the rAAV genome can be in the sense or antisense orientation relative to the direction of transcription of the transgene.
  • an “isolated polynucleotide” refers to a polynucleotide that has been separated from one or more nucleic acid molecules present in the natural source of the polynucleotide.
  • the “percentage identity” between two nucleotide sequences or between two amino acid sequences is calculated by multiplying the number of matches between the pair of aligned sequences by 100, and dividing by the length of the aligned region, including internal gaps. Identity scoring only counts perfect matches, and does not consider the degree of similarity of amino acids to one another. Note that only internal gaps are included in the length, not gaps at the sequence ends.
  • a “vector” refers to a nucleic acid molecule that is a vehicle for introducing anucleic acid molecule (e.g., a polynucleotide disclosed herein) into a cell.
  • an “expression vector” refers to a vector comprising transcriptional regulatory elements operably linked to a gene of interest (e.g., a polynucleotide disclosed herein) that facilitate the expression of the gene of interest in a cell and/or a cell free expression system.
  • a gene of interest e.g., a polynucleotide disclosed herein
  • transgene refers to a non-AAV nucleic acid sequence that encodes a polypeptide ( ⁇ ?.g., an antibody or scFv) or non-coding RNA ( ⁇ ?.g., an miRNA, shRNA, siRNA, antisense RNA, gRNA, antagomir, miRNA sponge, RNA aptazyme, or RNA aptamer).
  • a polypeptide ⁇ ?.g., an antibody or scFv
  • non-coding RNA ⁇ ?.g., an miRNA, shRNA, siRNA, antisense RNA, gRNA, antagomir, miRNA sponge, RNA aptazyme, or RNA aptamer.
  • transcriptional regulatory element refers to a cis-acting nucleotide sequence, for example, a DNA sequence, that regulates (e.g., controls, increases, or reduces) transcription of an operably linked nucleotide sequence by an RNA polymerase to form an RNA molecule.
  • a TRE may comprise one or more promoter elements and/or enhancer elements.
  • promoter and enhancer elements in a gene may be close in location, and the term “promoter” may refer to a sequence comprising a promoter element and an enhancer element. Thus, the term “promoter” does not exclude an enhancer element in the sequence.
  • the promoter and enhancer elements do not need to be derived from the same gene or species, and the sequence of each promoter or enhancer element may be either identical or substantially identical to the corresponding endogenous sequence in the genome.
  • operably linked is used to describe the connection between a TRE and a polynucleotide sequence (e.g., a transgene disclosed herein) to be transcribed.
  • gene expression is placed under the control of a TRE comprising one or more promoter and/or enhancer elements.
  • the transgene is “operably linked” to the TRE if the transcription of the transgene is controlled or influenced by the TRE.
  • the promoter and enhancer elements of the TRE may be in any orientation and/or distance from the trans gene, as long as the desired transcriptional activity is obtained.
  • the TRE is upstream from the transgene.
  • the term “effective amount” in the context of the administration of an AAV to a subject refers to the amount of the AAV that achieves a desired prophylactic or therapeutic effect.
  • the present disclosure provides a recombinant adeno-associated virus (AAV) capsid protein.
  • AAV capsid proteins described herein have been isolated from screening a shuffled capsid library in vivo for AAVs that are enriched in the liver and/or muscle.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 2.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%. at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 2.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 2. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 2. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 2. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 2.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 4.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 138-736 of SEQ ID NO: 4.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 4.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 4. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-736 of SEQ ID NO: 4. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 204-736 of SEQ ID NO: 4. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 4.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 6.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 6.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 6.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 6. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 6. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 6. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 6.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%. at least 99%, or 100% identity to amino acids 1-731 of SEQ ID NO: 8.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity’ to amino acids 134-731 of SEQ ID NO: 8.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 199-731 of SEQ ID NO: 8.
  • the recombinant AAV capsid protein comprises amino acids 1-731 of SEQ ID NO: 8. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 134-731 of SEQ ID NO: 8. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 199-731 of SEQ ID NO: 8. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 8.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 10.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-736 of SEQ ID NO: 10.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 10.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 10. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-736 of SEQ ID NO: 10. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-736 of SEQ ID NO: 10. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 10.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 12.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 12.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 12.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 12. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-736 of SEQ ID NO: 12. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-736 of SEQ ID NO: 12. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 12.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 1-736 of SEQ ID NO: 14.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 138-736 of SEQ ID NO: 14.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%. at least 87%. at least 88%.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 14. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-736 of SEQ ID NO: 14. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 204-736 of SEQ ID NO: 14. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 14.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 16.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 137-734 of SEQ ID NO: 16.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 16. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 16. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 16. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 16.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 18.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 18.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 18. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-735 of SEQ ID NO: 18. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-735 of SEQ ID NO: 18. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 18.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 20.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-736 of SEQ ID NO: 20.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 20. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-736 of SEQ ID NO: 20. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-736 of SEQ ID NO: 20. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 20.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 22.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 22.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 22.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 22. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-736 of SEQ ID NO: 22. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-736 of SEQ ID NO: 22. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 22.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 24.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 24.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 24. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-736 of SEQ ID NO: 24. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 204-736 of SEQ ID NO: 24. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 24.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 26.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 26.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 26.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 26. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 26. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-735 of SEQ ID NO: 26. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 26.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 28.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 28.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 28. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 28. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 28. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 28.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 30.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 30.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 30.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 30. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 30. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 30. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 30.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 32.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 32.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 32. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 32. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-735 of SEQ ID NO: 32. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 32.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 34.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 34.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 34.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 34. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 34. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-735 of SEQ ID NO: 34. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 34.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 36.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 36.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 36. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-735 of SEQ ID NO: 36. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-735 of SEQ ID NO: 36. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 36.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-731 of SEQ ID NO: 38.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 134-731 of SEQ ID NO: 38.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 199-731 of SEQ ID NO: 38.
  • the recombinant AAV capsid protein comprises amino acids 1-731 of SEQ ID NO: 38. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 134-731 of SEQ ID NO: 38. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 199-731 of SEQ ID NO: 38. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 38.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 40.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 40.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 40. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 40. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-735 of SEQ ID NO: 40. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 40.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 42.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 138-736 of SEQ ID NO: 42.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 42.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 42. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-736 of SEQ ID NO: 42. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 204-736 of SEQ ID NO: 42. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 42.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 44.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-736 of SEQ ID NO: 44.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 44. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-736 of SEQ ID NO: 44. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-736 of SEQ ID NO: 44. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 44.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 46.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 46.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 46.
  • the recombinant AAV capsid protein comprises amino acids 1-736 of SEQ ID NO: 46. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-736 of SEQ ID NO: 46. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-736 of SEQ ID NO: 46. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 46.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 48.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 48.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 48. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 48. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 48. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 48.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 50.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 50.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 50.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 50. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 50. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 50. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 50.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 52.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 52.
  • the recombinant AAV capsid protein comprises amino acids 1-734 of SEQ ID NO: 52. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-734 of SEQ ID NO: 52. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-734 of SEQ ID NO: 52. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 52.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 54.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 138-735 of SEQ ID NO: 54.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 54.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 54. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 138-735 of SEQ ID NO: 54. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 203-735 of SEQ ID NO: 54. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 54.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 56.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 56.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 56. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 56. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-735 of SEQ ID NO: 56. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 56.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 58.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 58.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 58.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 58. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 58. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-735 of SEQ ID NO: 58. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 58.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 60.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 60.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 60. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 60. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-735 of SEQ ID NO: 60. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 60.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 62.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 62.
  • the recombinant AAV capsid protein comprises an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 62.
  • the recombinant AAV capsid protein comprises amino acids 1-735 of SEQ ID NO: 62. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 137-735 of SEQ ID NO: 62. In certain embodiments, the recombinant AAV capsid protein comprises amino acids 202-735 of SEQ ID NO: 62. In certain embodiments, the recombinant AAV capsid protein comprises SEQ ID NO: 62.
  • the recombinant AAV capsid protein is a variant of a clade A, clade B, clade C, clade D, clade E, clade F, clade G. clade H, clade I, AAVgo. 1, AAV3, AAV4, AAV10, AAV11, AAV12. rh.32, rh32.33. rh.33, rh.34, BAAV. or AAV5 capsid protein.
  • the present disclosure provides an isolated polynucleotide encoding one or more recombinant AAV capsid proteins as disclosed herein.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 2.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 2.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 2. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 734 of SEQ ID NO: 2. In certain embodiments, the isolated polynucleotide encodes amino acids 202-734 of SEQ ID NO: 2. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 2.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 4.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-736 of SEQ ID NO: 4.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 4. In certain embodiments, the isolated polynucleotide encodes amino acids 138- 736 of SEQ ID NO: 4. In certain embodiments, the isolated polynucleotide encodes amino acids 204-736 of SEQ ID NO: 4. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 4.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 6.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 6.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%. at least 86%, at least 87%, at least 88%. at least 89%.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 6. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 734 of SEQ ID NO: 6. In certain embodiments, the isolated polynucleotide encodes amino acids 202-734 of SEQ ID NO: 6. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 6.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to amino acids 1-731 of SEQ ID NO: 8.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%. at least 89%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%. at least 91%. at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 199-731 of SEQ ID NO: 8.
  • the isolated polynucleotide encodes amino acids 1-731 of SEQ ID NO: 8. In certain embodiments, the isolated polynucleotide encodes amino acids 134- 731 of SEQ ID NO: 8. In certain embodiments, the isolated polynucleotide encodes amino acids 199-731 of SEQ ID NO: 8. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 8.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 10.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%. at least 87%, at least 88%, at least 89%, at least 90%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%. at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 10.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 10. In certain embodiments, the isolated polynucleotide encodes amino acids 138- 736 of SEQ ID NO: 10. In certain embodiments, the isolated polynucleotide encodes amino acids 203-736 of SEQ ID NO: 10. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 10.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%. at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1 -736 of SEQ ID NO: 12.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 12.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 12. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 736 of SEQ ID NO: 12. In certain embodiments, the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 12.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 14.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 14.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 14. In certain embodiments, the isolated polynucleotide encodes amino acids 138- 736 of SEQ ID NO: 14. In certain embodiments, the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 14.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 16.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 16.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 16.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 16. In certain embodiments, the isolated polynucleotide encodes amino acids 137-
  • the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 16.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 18.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-735 of SEQ ID NO: 18.
  • the isolated polynucleotide encodes an ammo acid sequence having at least 85%.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 18. In certain embodiments, the isolated polynucleotide encodes amino acids 138-
  • the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 18.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 20.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 20.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 20. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 736 of SEQ ID NO: 20. In certain embodiments, the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 20.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 22.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 22.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%. at least 86%, at least 87%, at least 88%.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 22. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 736 of SEQ ID NO: 22. In certain embodiments, the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 22.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 1-736 of SEQ ID NO: 24.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%. at least 87%, at least 88%, at least 89%, at least 90%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%. at least 89%. at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 24.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 24. In certain embodiments, the isolated polynucleotide encodes amino acids 138- 736 of SEQ ID NO: 24. In certain embodiments, the isolated polynucleotide encodes amino acids 204-736 of SEQ ID NO: 24. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 24.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 1-735 of SEQ ID NO: 26.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%. at least 87%, at least 88%, at least 89%, at least 90%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%. at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 26.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 26. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 735 of SEQ ID NO: 26. In certain embodiments, the isolated polynucleotide encodes amino acids 202-735 of SEQ ID NO: 26. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 26.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 28.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 28.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 28. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 734 of SEQ ID NO: 28. In certain embodiments, the isolated polynucleotide encodes amino acids 202-734 of SEQ ID NO: 28. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 28.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 30.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 30.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 30. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 734 of SEQ ID NO: 30. In certain embodiments, the isolated polynucleotide encodes amino acids 202-734 of SEQ ID NO: 30. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 30.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 32.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 32.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 32.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 32. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 735 of SEQ ID NO: 32. In certain embodiments, the isolated polynucleotide encodes amino acids 202-735 of SEQ ID NO: 32. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 32.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 34.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 34.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 34. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 735 of SEQ ID NO: 34. In certain embodiments, the isolated polynucleotide encodes amino acids 202-735 of SEQ ID NO: 34. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 34.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 36.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-735 of SEQ ID NO: 36.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 36.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 36. In certain embodiments, the isolated polynucleotide encodes amino acids 138- 735 of SEQ ID NO: 36. In certain embodiments, the isolated polynucleotide encodes amino acids 203-735 of SEQ ID NO: 36. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 36.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-731 of SEQ ID NO: 38.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 199-731 of SEQ ID NO: 38.
  • the isolated polynucleotide encodes amino acids 1-731 of SEQ ID NO: 38. In certain embodiments, the isolated polynucleotide encodes amino acids 134- 731 of SEQ ID NO: 38. In certain embodiments, the isolated polynucleotide encodes amino acids 199-731 of SEQ ID NO: 38. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 38.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 40.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 40.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 40.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 40. In certain embodiments, the isolated polynucleotide encodes amino acids 137-
  • the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 40.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 42.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 42.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 42. In certain embodiments, the isolated polynucleotide encodes amino acids 138-
  • the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 42.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 44.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 44.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-736 of SEQ ID NO: 44.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 44. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 736 of SEQ ID NO: 44. In certain embodiments, the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 44.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 46.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 46.
  • the isolated polynucleotide encodes amino acids 1-736 of SEQ ID NO: 46. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 736 of SEQ ID NO: 46. In certain embodiments, the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 46.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 48.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 48.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 48.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 48. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 734 of SEQ ID NO: 48. In certain embodiments, the isolated polynucleotide encodes amino acids 202-734 of SEQ ID NO: 48. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 48.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 50.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 50.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 50. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 734 of SEQ ID NO: 50. In certain embodiments, the isolated polynucleotide encodes amino acids 202-734 of SEQ ID NO: 50. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 50.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 52.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 52.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 52.
  • the isolated polynucleotide encodes amino acids 1-734 of SEQ ID NO: 52. In certain embodiments, the isolated polynucleotide encodes amino acids 137-
  • the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 52.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 54.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 54.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 54. In certain embodiments, the isolated polynucleotide encodes amino acids 138-
  • the isolated polynucleotide encodes amino acids
  • the isolated polynucleotide encodes SEQ ID NO: 54.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 56.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 56.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 56.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 56. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 735 of SEQ ID NO: 56. In certain embodiments, the isolated polynucleotide encodes amino acids 202-735 of SEQ ID NO: 56. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 56.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 58.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 58.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 58. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 735 of SEQ ID NO: 58. In certain embodiments, the isolated polynucleotide encodes amino acids 202-735 of SEQ ID NO: 58. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 58.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 60.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 60.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 60.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 60. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 735 of SEQ ID NO: 60. In certain embodiments, the isolated polynucleotide encodes amino acids 202-735 of SEQ ID NO: 60. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 60.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 62.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the isolated polynucleotide encodes an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 62.
  • the isolated polynucleotide encodes amino acids 1-735 of SEQ ID NO: 62. In certain embodiments, the isolated polynucleotide encodes amino acids 137- 735 of SEQ ID NO: 62. In certain embodiments, the isolated polynucleotide encodes amino acids 202-735 of SEQ ID NO: 62. In certain embodiments, the isolated polynucleotide encodes SEQ ID NO: 62.
  • the present disclosure provides an isolated polynucleotide having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%. at least 97%, at least 98%, at least 99%, or 100% sequence identity with the nucleic acid sequence set forth in any one of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, and 61.
  • the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 1. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 3. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 5. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 7. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 9. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 11.
  • the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 13. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 15. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 17. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 19. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 21. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 23.
  • the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 25. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 27. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 29. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 31. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 33. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 35.
  • the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 37. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 39. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 41. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 43. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 45. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 47.
  • the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 49. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 51. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 53. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 55. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 57. In certain embodiments, the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 59.
  • the isolated polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO: 61. [00195] In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 1 . In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 3. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 5. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 7.
  • the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 9. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 11. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 13. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 15. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 17. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 19.
  • the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 21. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 23. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 25. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 27. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 29.
  • the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 31. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 33. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 35. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 37. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 39.
  • the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 41. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 43. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 45. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 47. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 49.
  • the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 51. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 53. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 55. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 57. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 59. In certain embodiments, the isolated polynucleotide consists of the nucleic acid sequence set forth in SEQ ID NO: 61.
  • the polynucleotide is optimized, e.g, by codon/RNA optimization, replacement with heterologous signal sequences, and/or elimination of mRNA instability elements.
  • Methods to generate optimized polynucleotides for recombinant expression by introducing codon changes and/or eliminating inhibitory regions in the mRNA can be carried out by adapting the optimization methods described in, e.g.. U.S. Patent Nos. 5,965.726; 6,174,666; 6,291,664; 6,414,132; and 6,794,498, accordingly, all of which are herein incorporated by reference in their entireties.
  • potential splice sites and instability elements within the RNA can be mutated without altering the amino acids encoded by the nucleic acid sequences to increase stability of the RNA for recombinant expression.
  • the alterations utilize the degeneracy of the genetic code, e.g, using an alternative codon for an identical amino acid.
  • Such methods can increase expression of the encoded capsid protein relative to the expression of the capsid encoded by polynucleotides that have not been optimized.
  • the present disclosure provides a vector comprising a polynucleotide disclosed herein.
  • Suitable vectors include, without limitation, plasmids, viruses, cosmids, artificial chromosomes, linear DNA, and mRNA.
  • the vector is a plasmid or a viral vector.
  • the vector is a retrovirus vector, a herpes virus vector, a baculovirus vector, or an adenovirus vector.
  • the vector is an expression vector.
  • Vectors e.g. , expression vectors
  • the instant disclosure provides a recombinant cell comprising a polynucleotide or a vector (e.g., an expression vector) disclosed herein.
  • the instant disclosure provides a method of producing an AAV capsid protein, the method comprising culturing the recombinant cell under conditions whereby the polynucleotide is expressed and the capsid is produced.
  • Suitable vectors include, without limitation, plasmids, minimal vectors (e.g.. minicircles, NanoplasmidsTM, doggybones, MIDGE vectors, and the like), viruses, cosmids, artificial chromosomes, linear DNA, and mRNA.
  • the first nucleic acid vector and/or the second nucleic acid vector is a DNA plasmid or a DNA minimal vector. Any DNA plasmid or DNA minimal vector that can accommodate the necessary vector elements can be used for the first nucleic acid vector and the second nucleic acid vector.
  • Suitable DNA minimal vectors include, without limitation, linear covalently closed DNA (e.g., ministring DNA), linear covalently closed dumbbell shaped DNA (e.g, doggybone DNA, dumbbell DNA), minicircles, NanoplasmidsTM, minimalistic immunologically defined gene expression (MIDGE) vectors, and others known to those of skill in the art.
  • DNA minimal vectors and their methods of production are described in, e.g., U.S. Patent Application Nos. 20100233814, 20120282283, 20130216562, 20150218565, 20150218586. 20160008488, 20160215296, 20160355827, 20190185924, 20200277624, and 20210010021, all of which are herein incorporated by reference in their entireties.
  • a variety of host cells and expression vector systems can be utilized to express the capsid proteins described herein.
  • Such expression systems represent vehicles by which the coding sequences of interest can be produced and subsequently purified, but also represent cells which can, when transformed or transfected with the appropriate nucleotide coding sequences, express a capsid protein described herein in situ.
  • These include but are not limited to microorganisms such as bacteria (e.g., E. coli and B.
  • subtilis transformed with, e.g., recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing capsid protein coding sequences; yeast (e.g, Saccharomyces Pichia) transformed with, e.g., recombinant yeast expression vectors containing capsid protein coding sequences; insect cell systems infected with, e.g., recombinant virus expression vectors (e.g., baculovirus) containing capsid protein coding sequences; plant cell systems (e.g...
  • green algae such as Chlamydomonas reinhardtii
  • recombinant virus expression vectors e.g, cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV
  • recombinant plasmid expression vectors e.g., recombinant plasmid expression vectors
  • COS e.g., COS1 or COS
  • CHO BHK, MDCK, HEK 293, NS0, PER.C6, VERO, CRL7O3O, HsS78Bst, HeLa, and NIH 3T3, HEK-293T, HepG2, SP210, Rl. l, B-W, L-M, BSC1, BSC40, YB/20 and BMT10 cells
  • COS e.g., COS1 or COS
  • CHO BHK, MDCK, HEK 293, NS0, PER.C6, VERO, CRL7O3O, HsS78Bst, HeLa, and NIH 3T3, HEK-293T, HepG2, SP210, Rl. l, B-W, L-M, BSC1, BSC40, YB/20 and BMT10 cells
  • Rl. l B-W, L-M, BSC1, BSC40, YB/20 and BMT10
  • recombinant expression constructs containing promoters derived from the genome of mammalian cells e.g., metallothionein promoter
  • mammalian viruses e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter
  • cells for expressing the capsid proteins described herein are human cells, e.g., human cell lines.
  • a mammalian expression vector is pOptiVECTM or pcDNA3.3.
  • bacterial cells such as Escherichia coli, or eukaryotic cells (e.g., mammalian cells) are used for the expression of a capsid protein.
  • mammalian cells such as CHO or HEK293 cells, in conjunction with a vector such as the major intermediate early gene promoter element from human cytomegalovirus is an effective expression system for capsid proteins disclosed herein.
  • a number of expression vectors can be advantageously selected depending upon the use intended for the capsid protein being expressed. For example, when a large quantity of a capsid protein is to be produced, vectors which direct the expression of high levels of fusion protein products that are readily purified can be desirable. Such vectors include, but are not limited to, the E.
  • coll expression vector pUR278 (Ruether U & Mueller-Hill B (1983) EMBO J 2: 1791-1794), in which the capsid protein coding sequence can be ligated individually into the vector in frame with the lac Z coding region so that a fusion protein is produced; plN vectors (Inouye S & Inouye M (1985) Nuc Acids Res 13: 3101-3109; Van Heeke G & Schuster SM (1989) J Biol Chem 24: 5503-5509); and the like, all of which are herein incorporated by reference in their entireties.
  • pGEX vectors can also be used to express foreign polypeptides as fusion proteins with glutathione 5-transferase (GST).
  • fusion proteins are soluble and can easily be purified from lysed cells by adsorption and binding to matrix glutathione agarose beads followed by elution in the presence of free glutathione.
  • the pGEX vectors are designed to include thrombin or factor Xa protease cleavage sites so that the cloned target gene product can be released from the GST moiety.
  • Autographa californica nuclear polyhedrosis virus (AcNPV), for example, can be used as a vector to express foreign genes.
  • the virus grows in Spodoptera frugiperda cells.
  • the capsid protein coding sequence can be cloned individually into non-essential regions (for example the polyhedrin gene) of the virus and placed under control of an AcNPV promoter (for example the polyhedrin promoter).
  • a number of viral-based expression systems can be utilized.
  • the capsid protein coding sequence of interest can be ligated to an adenovirus transcription/translation control complex, e.g., the late promoter and tripartite leader sequence. This chimeric gene can then be inserted in the adenovirus genome by in vitro or in vivo recombination.
  • Insertion in a non-essential region of the viral genome will result in a recombinant virus that is viable and capable of expressing the capsid protein molecule in infected hosts (see, e.g., Logan J & Shenk T (1984) PNAS 81(12): 3655-9, which is herein incorporated by reference in its entirety ).
  • Specific initiation signals can also be required for efficient translation of inserted capsid protein coding sequences. These signals include the ATG initiation codon and adjacent sequences. Furthermore, the initiation codon must be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert.
  • exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic.
  • the efficiency of expression can be enhanced by the inclusion of appropriate transcription enhancer elements, transcription terminators, etc. (see, e.g. , Bitter G et al. (1987) Methods Enzymol. 153: 516-544, which is herein incorporated by reference in its entirety).
  • a host cell strain can be chosen which modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific fashion desired. Such modifications (e.g., glycosylation) and processing (e.g. cleavage) of protein products can be important for the function of the protein.
  • Different host cells have characteristic and specific mechanisms for the post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the foreign protein expressed.
  • eukaryotic host cells which possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product can be used.
  • Such mammalian host cells include but are not limited to CHO, VERO, BHK, Hela, MDCK, HEK 293, NIH 3T3, W138, BT483, Hs578T, HTB2, BT2O and T47D, NSO (a murine myeloma cell line that does not endogenously produce any immunoglobulin chains), CRL7O3O, COS (e.g, COS1 or COS), PER.C6, VERO, HsS78Bst, HEK-293T, HepG2, SP210, Rl. l, B-W, L-M, BSC1, BSC40, YB/20, BMT10 and HsS78Bst cells.
  • stable expression cells can be generated.
  • cell lines which stably express a capsid protein described herein can be engineered.
  • host cells can be transformed with a polynucleotide (e.g., DNA or RNA) controlled by appropriate transcriptional regulatory elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker.
  • a polynucleotide e.g., DNA or RNA
  • appropriate transcriptional regulatory elements e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.
  • engineered cells can be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media.
  • the selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci which in turn can be cloned and expanded into cell lines.
  • This method can advantageously be used to engineer cell lines which express a capsid protein described herein or a fragment thereof.
  • a number of selection systems can be used, including but not limited to the herpes simplex virus thymidine kinase (Wigler M et al. (1977) Cell 11(1): 223-32); hypoxanthineguanine phosphoribosyltransferase (Szybalska EH & Szybalski W (1962) PNAS 48(12): 2026-2034); and adenine phosphoribosyltransferase (Lowy I et al. (1980) Cell 22(3): 817-23) genes in tk-, hgprt- or aprt-cells, respectively, all of which are herein incorporated by reference in their entireties.
  • antimetabolite resistance can be used as the basis of selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler M et al. (1980) PNAS 77(6): 3567-70; O’Hare K et al.
  • the present disclosure provides an rAAV composition comprising a capsid comprising a recombinant AAV capsid protein disclosed herein.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 2.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 2.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 2.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 4.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-736 of SEQ ID NO: 4.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity' to amino acids 204-736 of SEQ ID NO: 4.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 6.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 6.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 6.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identity to amino acids 1-731 of SEQ ID NO: 8.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 134-731 of SEQ ID NO: 8.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 199-731 of SEQ ID NO: 8.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 10.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-736 of SEQ ID NO: 10.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 10.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 12.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 12.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 12.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 14.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 138-736 of SEQ ID NO: 14.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%. at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 14.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%. at least 90%, at least 91%, at least 92%. at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 1 .
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 16.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 16.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 18.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-735 of SEQ ID NO: 18.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 18.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 20.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 20.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 202-736 of SEQ ID NO: 20.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 22.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 22.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 203-736 of SEQ ID NO: 22.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 24.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 138-736 of SEQ ID NO: 24.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 24.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 26.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 26.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%. at least 93%. at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 26.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 28.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 28.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 28.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 30.
  • the rAAV composition comprises a capsid compnsing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 30.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 30.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 32.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 32.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 32.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 34.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 34.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%. at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 34.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 36.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 138-735 of SEQ ID NO: 36.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 36.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-731 of SEQ ID NO: 38.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 134-731 of SEQ ID NO: 38.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 199-731 of SEQ ID NO: 38.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 40.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 40.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-735 of SEQ ID NO: 40.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 42.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 204-736 of SEQ ID NO: 42.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 44.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity- to amino acids 137-736 of SEQ ID NO: 44.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-736 of SEQ ID NO: 44.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-736 of SEQ ID NO: 46.
  • the rAAV composition comprises a capsid compnsing an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-736 of SEQ ID NO: 46.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 203-736 of SEQ ID NO: 46.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 48.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity- to amino acids 137-734 of SEQ ID NO: 48.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 48.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%. at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 50.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%. at least 92%. at least 93%.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%. at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 50.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-734 of SEQ ID NO: 52.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-734 of SEQ ID NO: 52.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-734 of SEQ ID NO: 52.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 54.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 138-735 of SEQ ID NO: 54.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity 7 to amino acids 203-735 of SEQ ID NO: 54.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 56.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91 %, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 56.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 56.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 58.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%. at least 96%. at least 97%. at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 58.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%. at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 60.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 60.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 202-735 of SEQ ID NO: 60.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 1-735 of SEQ ID NO: 62.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to amino acids 137-735 of SEQ ID NO: 62.
  • the rAAV composition comprises a capsid comprising an amino acid sequence having at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity' to amino acids 202-735 of SEQ ID NO: 62.
  • the present disclosure provides an rAAV comprising a capsid compnsing one or more recombinant capsid proteins disclosed herein; and an rAAV genome.
  • the rAAVs disclosed herein generally comprise a recombinant genome (e.g., an rAAV genome) packaged within the capsid.
  • the rAAV genome can be of any ty pe that is capable of being packages within an AAV capsid disclosed herein.
  • the rAAV genome is a single-stranded DNA genome.
  • the rAAV genome is a self- complementary 7 genome, for example as described in US7790154, which is hereby incorporated by reference in its entirety.
  • the rAAV genome comprises a trans gene.
  • the transgene encodes a polypeptide.
  • the transgene encodes a therapeutic protein.
  • the transgene encodes an antibody or a fragment thereof (e g. , a Fab, scFv, or full-length antibody).
  • the transgene encodes an scFv, nanobody, or VHH.
  • the transgene encodes a non-coding RNA.
  • the transgene comprises one or more sequences encoding an RNA molecule.
  • RNA molecules include, without limitation, miRNA, shRNA, siRNA, antisense RNA, gRNA, antagomirs, miRNA sponges, RNA aptazymes, RNA aptamers, mRNA, IncRNAs, ribozymes, and synthetic RNAs known in the art.
  • the transgene encodes one or more polypeptides, or a fragment thereof.
  • Such transgenes can comprise the complete coding sequence of a polypeptide, or only a fragment of a coding sequence of a polypeptide.
  • the transgene encodes a polypeptide that is useful to treat a disease or disorder in a subject.
  • Suitable polypeptides include, without limitation, (3-globin, hemoglobin, tissue plasminogen activator, and coagulation factors; colony stimulating factors (CSF); interleukins, such as IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, etc.; growth factors, such as keratinocyte growth factor (KGF).
  • CSF colony stimulating factors
  • interleukins such as IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, etc.
  • growth factors such as keratinocyte growth factor (KGF).
  • SCF stem cell factor
  • FGF fibroblast growth factor
  • HGF hepatocyte growth factor
  • IGFs insulin-like growth factors
  • BMP bone morphogenetic protein
  • EGF epidermal growth factor
  • GDF-9 growth differentiation factor-9
  • HDGF hepatoma derived growth factor
  • GDF-8 nerve growth factor
  • soluble receptors such as soluble TNF-a receptors, soluble interleukin receptors (e.g., soluble IL-1 receptors and soluble type II IL-1 receptors), soluble y/A T cell receptors, ligand-binding fragments of a soluble receptor, and the like; enzymes, such as a-glucosidase, imiglucerase, P- glucocerebrosidase, and alglucerase; enzyme activators, such as tissue plasminogen activator; chemokines, such as IP-10, monokine induced by interferon-gamma (Mig), Groa/IL-8, RANTES, MIP-la, MIP-ip, MCP-1, PF-4, and the
  • VEGF121, VEGF165, VEGF-C, VEGF-2 glioma-derived growth factor, angiogenin, angiogenin-2; and the like; anti- angiogenic agents, such as a soluble VEGF receptor; protein vaccine; neuroactive peptides, such as nerve grow th factor (NGF), bradykinin, cholecystokinin, gastrin, secretin, oxytocin, gonadotropin-releasing hormone, beta-endorphin, enkephalin, substance P, somatostatin, prolactin, galanin, growth hormone-releasing hormone, bombesin, dynorphin, warfarin, neurotensin, motilin, thyrotropin, neuropeptide Y, luteinizing hormone, calcitonin, insulin, glucagons, vasopressin, angiotensin II, thyrotropin-releasing hormone, vasoactive intestinal peptide, a sleep
  • the transgene encodes a protein that may be defective in one or more lysosomal storage diseases.
  • suitable proteins include, without limitation, a-sialidase, cathepsin A, a-mannosidase, P-mannosidase, glycosyl asparaginase, a-fucosidase, a-N- acetylglucosaminidase, P-galactosidase, P-hexosaminidase a-subunit, P-hexosaminidase P- subunit, GM2 activator protein, glucocerebrosidase, Saposin C, Arylsulfatase A, Saposin B, formyl-glycine generating enzyme, P-galactosylceramidase, a-galactosidase A, iduronate sulfatase, a-iduronidase, hepara
  • the transgene encodes an antibody or a fragment thereof (e.g., a Fab, scFv. or full-length antibody).
  • Suitable antibodies include, without limitation, muromonab-cd3, efalizumab, tositumomab, daclizumab, nebacumab, catumaxomab, edrecolomab, abciximab, rituximab, basiliximab, palivizumab, infliximab, trastuzumab, adalimumab, ibritumomab tiuxetan, omalizumab, cetuximab, bevacizumab, natalizumab, panitumumab, ranibizumab, eculizumab, certolizumab, ustekinumab, canakinumab, golimumab, ofatum
  • tocilizumab denosumab. belimumab, ipilimumab. brentuximab vedotin, pertuzumab, raxibacumab, obinutuzumab, alemtuzumab, siltuximab, ramucirumab, vedolizumab, blinatumomab, nivolumab, pembrolizumab, idarucizumab, necitumumab, dinutuximab, secukinumab, mepolizumab, alirocumab, evolocumab, daratumumab, elotuzumab, ixekizumab, reslizumab.
  • olaratumab bezlotoxumab, atezolizumab, obiltoxaximab, inotuzumab ozogamicin, brodalumab, guselkumab, dupilumab, sarilumab, avelumab, ocrehzumab, emicizumab, benralizumab, gemtuzumab ozogamicin, durvalumab.
  • burosumab erenumab, galcanezumab, lanadelumab, mogamulizumab, tildrakizumab, cemiplimab, fremanezumab, ravulizumab, emapalumab, ibalizumab, moxetumomab, caplacizumab, romosozumab, risankizumab, polatuzumab. eptinezumab, leronlimab, sacituzumab. brolucizumab, isatuximab. and teprotumumab.
  • the transgene encodes a nuclease.
  • Suitable nucleases include, without limitation, zinc fingers nucleases (ZFN) (see, e.g., Porteus, and Baltimore (2003) Science 300: 763; Miller et al. (2007) Nat. Biotechnol. 25:778-785; Sander et al. (2011) Nature Methods 8:67-69; and Wood et al. (2011) Science 333:307, each of which is hereby incorporated by reference in its entirety); transcription activator-like effectors nucleases (TALEN) (see, e.g., Wood et al. (2011) Science 333:307; Boch et al.
  • ZFN zinc fingers nucleases
  • TALEN transcription activator-like effectors nucleases
  • the transgene encodes an RNA-guided nuclease.
  • Suitable RNA-guided nucleases include, without limitation, Class I and Class II clustered regularly interspaced short palindromic repeats (CRISPR)-associated nucleases.
  • Class I is divided into types I, III, and IV, and includes, without limitation, type I (Cas3), type I-A (Cas8a, Cas5), type I-B (Cas8b), type I-C (Cas8c), type 1-D (CaslOd), type I-E (Csel, Cse2), type I-F (Csyl, Csy2, Csy3), type I-U (GSU0054), type III (CaslO), type III-A (Csm2), type III-B (Cmr5), type III-C (CsxlO or Csxl 1), type III-D (CsxlO), and type IV (Csfl).
  • type I Cas3
  • type I-A Cas8a, Cas5
  • type I-B Cas8b
  • type I-C Cas8c
  • type 1-D CaslOd
  • type I-E Csel, Cs
  • Class II is divided into types II, V, and VI, and includes, without limitation, type II (Cas9), type 1I-A (Csn2), type Il-B (Cas4), type V (Cpfl, C2cl, C2c3), and type VI (Casl3a, Casl3b, Casl3c).
  • RNA-guided nucleases also include naturally-occurring Class II CRISPR nucleases such as Cas9 (Type II) or Casl2a/Cpfl (Type V), as well as other nucleases derived or obtained therefrom.
  • Exemplary Cas9 nucleases that may be used in the present invention include, but are not limited to, S', pyogenes Cas9 (SpCas9), S’, aureus Cas9 (SaCas9), N. meningitidis Cas9 (NmCas9), C. jejuni Cas9 (CjCas9), and Geobacillus Cas9 (GeoCas9).
  • the transgene encodes reporter sequences, which upon expression produce a detectable signal.
  • reporter sequences include, without limitation, DNA sequences encoding P-lactamase, P -galactosidase (LacZ), alkaline phosphatase, thymidine kinase, green fluorescent protein (GFP), red fluorescent protein (RFP), chloramphenicol acetyltransferase (CAT), luciferase, membrane bound proteins including, for example, CD2, CD4, CD8, the influenza hemagglutinin protein, and others well known in the art, to which high affinity antibodies directed thereto exist or can be produced by conventional means, and fusion proteins comprising a membrane bound protein appropriately fused to an antigen tag domain from, among others, hemagglutinin or Myc.
  • the rAAV genome comprises a transcriptional regulatory' element (TRE) operably linked to the transgene, to control expression of an RNA or polypeptide encoded by the transgene.
  • the TRE comprises a constitutive promoter.
  • the TRE can be active in any mammalian cell e.g., any human cell).
  • the TRE is active in a broad range of human cells.
  • Such TREs may comprise constitutive promoter and/or enhancer elements including any of those described herein, and any of those known to one of skill in the art.
  • the TRE comprises an inducible promoter.
  • the TRE may be a tissue-specific TRE, i.e., it is active in specific tissue(s) and/or organ(s).
  • a tissue-specific TRE comprises one or more tissue-specific promoter and/or enhancer elements, and optionally one or more constitutive promoter and/or enhancer elements.
  • tissue-specific promoter and/or enhancer elements can be isolated from genes specifically expressed in the tissue by methods well known in the art.
  • Suitable promoters include, e.g., cytomegalovirus promoter (CMV) (Stinski et al. (1985) Journal of Virology 55(2): 431-441); CMV early enhancer/chicken [3-actin (CBA) promoter/rabbit P-globin intron (CAG) (Miyazaki et al. (1989) Gene 79(2): 269-277); CB SB (Jacobson et al. (2006) Molecular Therapy 13(6): 1074-1084); human elongation factor la promoter (EFla) (Kim et al.
  • CMV cytomegalovirus promoter
  • CBA CMV early enhancer/chicken [3-actin (CBA) promoter/rabbit P-globin intron
  • CB SB Jacobson et al. (2006) Molecular Therapy 13(6): 1074-1084
  • EFla human elongation factor la promoter
  • the TRE comprises a cytomegalovirus (CMV) promoter/enhancer, an SV40 promoter, a chicken beta actin (CBA) promoter, an smCBA promoter, a human elongation factor 1 alpha (EFla) promoter, a minute virus of mouse (MVM) intron which comprises transcription factor binding sites, a human phosphoglycerate kinase (PGK1) promoter, a human ubiquitin C (Ubc) promoter, a human beta actin promoter, a human neuron-specific enolase (ENO2) promoter, a human beta-glucuronidase (GUSB) promoter, a rabbit beta-globin element, a human calmodulin 1 (CALM1) promoter, a human ApoE/C-I hepatic control region (HCR1).
  • CMV cytomegalovirus
  • CBA chicken beta actin
  • smCBA smCBA promoter
  • EFla e
  • a human al -antitrypsin (hAAT) promoter an extended HCR1, a HS-CRM8 element of an hAAT promoter, a human transthyretin (TTR) promoter, and/or a human Methyl-CpG Binding Protein 2 (MeCP2) promoter.
  • hAAT human al -antitrypsin
  • TTR human transthyretin
  • MeCP2 Methyl-CpG Binding Protein 2
  • a transfer genome may comprise a TRE comprising a CMV enhancer, a CBA promoter, and the splice acceptor from exon 3 of the rabbit beta-globin gene, collectively called a CAG promoter.
  • a transfer genome may comprise a TRE comprising a hybrid of CMV enhancer and CBA promoter followed by a splice donor and splice acceptor, collectively called a C ASI promoter region.
  • a transfer genome may comprise a TRE comprising a HCR1 and hAAT promoter (also referred to as an LP1 promoter).
  • the TRE is liver-specific.
  • Exemplary liver-specific TREs may comprise one or more elements from, without limitation, the ApoA-I promoter, the ApoA-II promoter, the ApoA-IV promoter, the ApoB promoter, the ApoC-I promoter, the ApoC-II promoter, the ApoC-III promoter, the ApoE promoter, the albumin promoter, the a-fetoprotein promoter, the phosphoenolpyruvate carboxykinase 1 (PCK1) promoter, the phosphoenol pyruvate carboxykinase 2 (PCK2) promoter, the transthyretin (TTR) promoter, the a-antitrypsin (AAT or SERPINA1) promoter, the TK (thymidine kinase) promoter, the hemopexin promoter, the alcohol dehydrogenase 6 promoter, the cholesterol 7alpha-hydroxylase promoter, the factor
  • the TRE is muscle specific.
  • Exemplary 7 muscle-specific TREs may comprise one or more elements from, without limitation, the human skeletal muscle a- actin (EISA) promoter, the muscle creatine kinase (MCK) promoter, the MHCK7 promoter, the dMCK promoter, the tMCK promoter, the CK.6 promoter, the CK8 promoter, the CK8e promoter, the human desmin (DES) promoter or variant thereof, the cardiac troponin T (cTnT) promoter, the myosin light-chain (MLC2v) promoter, the human a-myosin heavy chain gene (aMHC) promoter, the MLC promoter, the human troponin I (TNNI1) promoter, the AUSEx3 promoter, the SPcA5- 12 promoter, the SP-301 promoter, the MH promoter, and the Sk-CRM4/DES promoter.
  • EISA human skeletal muscle a- actin
  • MCK7
  • the native promoter for the transgene may be used.
  • the native promoter may be preferred when it is desired that expression of the transgene should mimic the native expression.
  • the native promoter may be used when expression of the transgene must be regulated temporally or developmentally, or in a tissue-specific manner, or in response to specific transcriptional stimuli.
  • other native expression control elements such as enhancer elements, polyadenylation sites or Kozak consensus sequences may also be used to mimic the native expression.
  • compositions compnsing an AAV as disclosed herein together with a pharmaceutically acceptable excipient, adjuvant, diluent, vehicle or carrier, or a combination thereof.
  • a '‘pharmaceutically acceptable carrier’ includes any material which, when combined with an active ingredient of a composition, allows the ingredient to retain biological activity and without causing disruptive physiological reactions, such as an unintended immune reaction.
  • Pharmaceutically acceptable carriers include water, phosphate buffered saline, emulsions such as oil/water emulsion, and wetting agents. Compositions comprising such carriers are formulated by well-known conventional methods such as those set forth in Remington’s Pharmaceutical Sciences, current ed., Mack Publishing Co., Easton Pa.
  • the instant disclosure provides methods for transducing a cell.
  • the methods generally comprise contacting the cell with an rAAV disclosed herein under conditions whereby the cell is transduced.
  • the rAAV disclosed herein can be used to transduce cells in vitro, in vivo and ex vivo.
  • the instant disclosure provides methods for delivering a transgene into a cell.
  • the methods generally comprise comprising contacting the cell with an rAAV disclosed herein under conditions whereby the cell is transduced and the transgene is expressed.
  • the rAAV disclosed herein can comprise a transgene under the control of a TRE.
  • the instant disclosure provides methods for expressing a transgene in a cell, the method generally comprising contacting the cell with such an rAAV under conditions whereby the cell is transduced and the transgene is expressed.
  • the transgene can encode a polypeptide and/or an RNA molecule, as described herein.
  • the instant disclosure provides methods for producing a polypeptide and/or an RNA molecule in a cell, the method generally comprising contacting the cell with such an rAAV under conditions whereby the cell is transduced and the polypeptide and/or an RNA molecule is produced.
  • the cell is in a subject and the rAAV is administered to the subject.
  • the rAAV disclosed herein can be administered to a subject by all routes suitable for an rAAV, including, without limitation, intravenously, intraperitoneally, subcutaneously, intramuscularly, intrathecally, or intradermally.
  • the subject is a member of any mammalian or nonmammalian species. Suitable subjects include, without limitation, humans, non-human primates, canines, felines, ungulates (e.g., equine, bovine, swine (e.g., pig)), avians, rodents (e.g., rats, mice), and other subjects.
  • the subject is canine.
  • the subject is human.
  • the invention provides an rAAV as disclosed herein for use in medicine. In another aspect, the invention provides an rAAV as disclosed herein for use as therapy. In another aspect, the invention provides an rAAV as disclosed herein for use as a medicament.
  • the instant disclosure provides packaging systems for recombinant preparation of a recombinant adeno-associated virus (rAAV) disclosed herein.
  • packaging systems generally comprise: a first nucleotide encoding one or more AAV Rep proteins; a second nucleotide encoding a capsid protein of any of the AAVs as disclosed herein; and a third nucleotide sequence comprising any of the rAAV genome sequences as disclosed herein, wherein the packaging system is operative in a cell for enclosing the transfer genome in the capsid to form the AAV.
  • the packaging system comprises a first vector comprising the first nucleotide sequence encoding the one or more AAV Rep proteins and the second nucleotide sequence encoding the AAV capsid protein, and a second vector comprising the third nucleotide sequence comprising the rAAV genome.
  • a “vector” refers to a nucleic acid molecule that is a vehicle for introducing nucleic acids into a cell (e.g., a plasmid, a virus, a cosmid, an artificial chromosome, etc.).
  • the packaging system further comprises a fourth nucleotide sequence comprising one or more helper virus genes.
  • the fourth nucleotide sequence comprises adenoviral E2, E4, and VA genes.
  • the packaging system further comprises a third vector (e.g. , a helper virus vector), comprising the fourth nucleotide sequence.
  • the third vector may be an independent third vector, integral with the first vector, or integral with the second vector.
  • the packaging system comprises a first vector comprising the first nucleotide sequence encoding one or more AAV Rep proteins, the second nucleotide sequence encoding one or more recombinant AAV capsid protein as disclosed herein, and the third nucleotide sequence comprising any of the rAAV genome sequences as disclosed herein, wherein the packaging system is operative in a cell for enclosing the transfer genome in the capsid to form the AAV.
  • the packaging system further comprises a fourth nucleotide sequence comprising one or more helper virus genes.
  • the fourth nucleotide sequence comprises adenoviral E2, E4, and VA genes.
  • the packaging system further comprises a second vector (e.g. , a helper virus vector), comprising the fourth nucleotide sequence.
  • the second vector may be an independent second vector, integral with the first vector.
  • AAV Rep protein can be employed in the packaging systems disclosed herein.
  • the Rep nucleotide sequence encodes an AAV2 Rep protein.
  • Suitable AAV2 Rep proteins may include, without limitation, Rep 78/68 or Rep 68/52.
  • the helper virus is selected from the group consisting of adenovirus, herpes virus (including herpes simplex virus (HSV)), poxvirus (such as vaccinia virus), cytomegalovirus (CMV), and baculovirus.
  • the adenovirus genome comprises one or more adenovirus RNA genes selected from the group consisting of El, E2, E4 and VA.
  • the adenovirus genome comprises one or more adenovirus RNA genes selected from the group consisting of E2, E4, and VA.
  • the HSV genome comprises one or more of HSV genes selected from the group consisting of UL5/8/52, ICPO, ICP4, ICP22, and UL30/UL42.
  • the vectors e.g.. first, second, and/or third vectors
  • the vectors are contained within one or more plasmids.
  • the first, second, and/or third vectors are contained within one or more recombinant helper viruses.
  • the first vector and the third vector are contained within a recombinant helper virus.
  • the second vector and the third vector are contained within a recombinant helper virus.
  • the disclosure provides a method for recombinant preparation of an AAV as described herein, wherein the method comprises transfecting or transducing a cell with a packaging system as described herein under conditions operative for enclosing the rAAV genome in the capsid to form the rAAV as described herein.
  • Exemplary’ methods for recombinant preparation of an rAAV include transient transfection (e.g. , with one or more transfection plasmids containing a first, and a second, and optionally a third vector as described herein), viral infection (e.g.
  • helper viruses such as a adenovirus, poxvirus (such as vaccinia virus), herpes virus (including HSV, cytomegalovirus, or baculovirus, containing a first, and a second, and optionally a third vector as described herein)), and stable producer cell line transfection or infection (e.g, with a stable producer cell, such as a mammalian or insect cell, containing a Rep nucleotide sequence encoding one or more AAV Rep proteins and/or a Cap nucleotide sequence encoding one or more capsid proteins as described herein, and with a transfer genome as described herein being delivered in the form of a plasmid or a recombinant helper virus).
  • a stable producer cell such as a mammalian or insect cell, containing a Rep nucleotide sequence encoding one or more AAV Rep proteins and/or a Cap nucleotide sequence encoding one or more capsid proteins as described herein,
  • the instant disclosure provides a packaging system for preparation of an rAAV, wherein the packaging system comprises: a first nucleotide sequence encoding one or more AAV Rep proteins; a second nucleotide sequence encoding a capsid protein of any one of the AAVs described herein; a third nucleotide sequence comprising an rAAV genome sequence of any one of the AAVs described herein; and optionally a fourth nucleotide sequence comprising one or more helper virus genes (e.g, adenoviral E2, E4, and VA genes).
  • helper virus genes e.g, adenoviral E2, E4, and VA genes.
  • Example 1 AAV Capsid Library Generation and Liver and/or Muscle Enrichment
  • a shuffled capsid library was generated using capsid sequences of AAV1 through AAV14, AAVrh8, and AAVrhlO as inputs. Briefly, nucleic acid molecules encoding each AAV capsid were digested to create fragments of approximately 200-400 base pairs in length, gel extracted, mixed, and assembled via PCR. The assembled shuffled capsids were then digested and ligated into a vector for subsequent virus generation. Virus was produced using standard triple plasmid transfection techniques. Virus was then collected and harvested via precipitation with PEG 8000.
  • Non-functional capsids were removed via ultracentrifugation in an iodixanol gradient, and buffer exchange of the 40% iodixanol fraction. See. e.g., Zolotukhin et al. (1999) Gene Therapy 6(6): 973-985. Nucleic acid encoding functional capsids were then extracted from the virus preparation using PCR, and the process of viral production was repeated with the functional viral capsid nucleic acids.
  • an rAAV vector comprising a first coding sequence encoding canine FGF21 and second coding sequence encoding a soluble form of canine transforming growth factor-beta receptor 2 (sTGFbR2; see, e.g., Davidsohn et al. (2019) Proc. Natl. Acad. Sci. USA 116 (47): 23505-2351 1) was packaged into CL01, CL03, CL11, CL13, CL27, or AAV8 capsids. A total of 30 young adult (1- to 3-year old) naive male and female Beagle dogs were intravenously administered the CL01, CL03, CL11.

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Abstract

L'invention concerne des protéines de capside de virus adéno-associé (AAV) recombinant, des compositions ( par exemple, rAAV) comprenant les protéines de capside, des acides nucléiques codant pour les protéines de capside, et des procédés de fabrication et d'utilisation des protéines de capside.
PCT/US2023/074729 2022-09-21 2023-09-21 Capsides de virus adéno-associé WO2024064782A2 (fr)

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