Adeno-associated virus vectors

Abstract

This disclosure relates to AAV vectors (e.g., AAV2 vectors). For example, AAV vectors (e.g., AAV2 vectors) comprising an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence, such AAV capsid polypeptides, nucleic acid molecules encoding such vectors, nucleic acid molecules encoding such AAV capsid polypeptides, host cells containing and / or expressing such nucleic acid molecules, and methods and materials for making or using such vectors and / or AAV capsid polypeptides are provided.

Description

Attorney Docket No. 45049-0087W01 / 06839ADENO-ASSOCIATED VIRUS VECTORSCROSS-REFERENCE TO RELATED APPLICATIONThis application claims the benefit of the filing date of U.S. Provisional Patent Application No. 63 / 684,071, filed on August 16, 2024. The contents of U.S. Provisional Patent Application No. 63 / 684,071 are incorporated herein by reference in their entirety.STATEMENT REGARDING FEDERAL FUNDING

[0001] This invention was made with government support under MH120094 awarded by the National Institutes of Health. The government has certain rights in the invention.TECHNICAL FIELD

[0002] This disclosure relates to adeno-associated virus (AAV) vectors. For example, this disclosure provides methods and materials for making and using AAV vectors (e.g, AAV2 vectors) having the ability to deliver nucleic acid to retinal cells.BACKGROUND

[0003] Viral vectors, such as AAV vectors, are efficient vehicles for in vivo nucleic acid delivery, and their use in the clinic is expanding. Improved AAV vectors and AAV production techniques for making effective AAV vector preparations should further expand the use of AAV vectors in the laboratory and clinic.SUMMARY

[0004] The present disclosure provides AAV vectors (e.g., AAV2 vectors). In one aspect, provided herein is an adeno-associated virus (AAV) vector comprising an AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising that of any one of SEQ ID NOs: 5-9. In another aspect, provided herein is an adeno-associated virus (AAV) vector comprising an AAV capsid polypeptide, wherein said capsid polypeptide has an ammo acid sequence comprising that of SEQ ID NO: 5 (HDQVRP).

[0005] In some embodiments, the AAV capsid polypeptide has an amino acid sequence comprising that of any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 581 and 582, between amino acid 582 and 583. between positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 andAttorney Docket No. 45049-0087W01 / 06839587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4. In some embodiments, the AAV capsid polypeptide has an amino acid sequence comprising that of any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1- 4. In some embodiments, the AAV capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9.

[0006] In some embodiments, the AAV vector is an AAV2 vector. In some embodiments, the AAV vector further comprises an exogenous nucleic acid encoding an RNA or a therapeutic polypeptide. In some embodiments, the exogenous nucleic acid encodes an RNA. In some embodiments, the RNA comprises an siRNA or microRNA. In some embodiments, the exogenous nucleic acid encodes a therapeutic polypeptide. In some embodiments, the therapeutic polypeptide is a VEGF polypeptide, a RSI polypeptide, a PRPF31 polypeptide, an ABC A4 polypeptide, a CRB 1 polypeptide, an NPHP5 polypeptide, or an NR2E3 polypeptide.

[0007] In some embodiments, AAV capsid polypeptide has an amino acid sequence comprising SEQ ID NO: 4 except that the amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of SEQ ID NO: 4.

[0008] Also disclosed herein is an AAV capsid polypeptide having an amino acid sequence comprising any one of SEQ ID NOs: 5-9. In some embodiments, the polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 581 and 582, between amino acid 582 and 583, between positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587. between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4. In some embodiments, the polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ IDAttorney Docket No. 45049-0087W01 / 06839NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1- 4. In some embodiments, the AAV capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9. In an embodiment, the polypeptide has an amino acid sequence comprising SEQ ID NO: 4 except that the amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of SEQ ID NO: 4.

[0009] In another aspect, the present disclosure provides one or more nucleic acid molecules encoding an AAV vector. In some embodiments, the one or more nucleic acid molecules encode a capsid polypeptide. In some embodiments, the nucleic acid molecule comprises a DNA.

[0010] In yet another aspect, the present disclosure provides a host cell comprising a nucleic acid molecule. In some embodiments, the host cell comprises a capsid polypeptide. In some embodiments, the host cell comprises a vector. In some embodiments, the host cell expresses any of AAV vectors disclosed herein. In some embodiments, the host cell expresses any of the polypeptides disclosed herein. In some embodiments, the host cell is a retinal cell.

[0011] In another aspect, the present disclosure provides a composition comprising an AAV vector comprising an AAV capsid polypeptide and a pharmaceutically acceptable excipient. The composition can comprise from about 1 x 107to about 1 x 1014of the AAV vector. In some embodiments, the composition can comprise phosphate buffered saline, Hank’s Balanced Salt Solution (e.g., a buffer comprising salts such as sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, magnesium chloride, sodium phosphate, potassium phosphate, sodium bicarbonate, and / or glucose), or Pluronic F68.

[0012] In another aspect, the present disclosure provides a method for delivering an exogenous nucleic acid sequence to a retinal cell of a mammal, wherein said method comprises contacting said retinal cell with an AAV vector comprising an AAV capsid polypeptide and said exogenous nucleic acid sequence, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 5-9, wherein said AAV vector infects said retinal cell, thereby delivering said exogenous nucleic acid sequence to said retinal cell. In some embodiments, the capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4. In some embodiments, the capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 toAttorney Docket No. 45049-0087W01 / 06839590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9. In some embodiments, the mammal is a human. In some embodiments, the human is an infant, a child, or an adult. In some embodiments, the vector is an AAV2 vector. In some embodiments, the exogenous nucleic acid sequence encodes an RNA or a polypeptide. In some embodiments, the exogenous nucleic acid encodes an RNA (e.g., therapeutic). In some embodiments, the RNA is an siRNA or microRNA. In some embodiments, the exogenous nucleic acid encodes a polypeptide (e.g., therapeutic). In some embodiments, the polypeptide is a VEGF polypeptide, a RSI polypeptide, a PRPF31 polypeptide, an ABCA4 polypeptide, a CRB1 polypeptide, an NPHP5 polypeptide, or an NR2E3 polypeptide. In some embodiments, the method comprises intravitreally administering a composition comprising said vector to said mammal, thereby contacting said retinal cell with said vector. In some embodiments, the composition comprises from about 1 x 107to about 1 x 1014of said vector.

[0013] In another aspect, the present disclosure provides a method for treating a retinal condition. In some embodiments, the present disclosure provides a method for treating a retinal condition in a mammal in need thereof, wherein said method comprises contacting a retinal cell of a mammal having said retinal condition with a therapeutically effective amount of an AAV vector comprising an AAV capsid polypeptide and an exogenous nucleic acid sequence, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 5-9, wherein said AAV vector infects said retinal cell and drive expression of said exogenous nucleic acid sequence within said retinal cell, thereby treating said retinal condition. In some embodiments, the contacting comprises a population of retinal cells. In some embodiments, the method comprises contacting the retinal cells of a mammal having the retinal condition with AAV vectors comprising an AAV capsid polypeptide and an exogenous nucleic acid sequence disclosed herein, wherein the AAV vectors infect the retinal cells and drive expression of the exogenous nucleic acid sequence within the retinal cells, thereby treating the retinal condition. In some embodiments, the mammal is a human. In some embodiments, the human is an infant, a child, or an adult. In some embodiments, the retinal condition is selected from the group consisting of cone dystrophy, cone / rod dystrophy, retinitis pigmentosa, macular degeneration, achromatopsia, blue cone monochromacy, and color blindness. In some embodiments, the capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 581 and 582, between amino acid 582 and 583, between positions 583 and 584, between amino acid positions 584 and 585,Attorney Docket No. 45049-0087W01 / 06839 between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4. In some embodiments, the capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4. In some embodiments, the capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9. In some embodiments, the capsid polypeptide has an amino acid sequence comprising SEQ ID NO: 4 except that the amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of SEQ ID NO: 4. In some embodiments, the vector is an AAV2 vector. In some embodiments, the exogenous nucleic acid sequence encodes an RNA or a polypeptide. In some embodiments, the RNA is an siRNA or a microRNA. In some embodiments, the polypeptide is a VEGF polypeptide, a RSI polypeptide, a PRPF31 polypeptide, an ABCA4 polypeptide, a CRB1 polypeptide, an NPHP5 polypeptide, or an NR2E3 polypeptide. In some embodiments, the method comprises intravitreally administering a composition comprising said vector to said mammal, thereby contacting said retinal cell with said vector. In some embodiments, the composition comprises from about 1 x 107to about 1 x 1014of said vectors.

[0014] In another aspect, provided herein is a non-naturally occurring AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 581 and 582, between amino acid positions 582 and 583, between amino acid positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-Li-[6-mer polypeptide] -L2-,Attorney Docket No. 45049-0087W01 / 06839 wherein said Li and said L2 are each, independently, optional amino acid linkers, each having one, two. or three amino acids, and wherein the 6-mer polypeptide has an amino acid sequence of that of SEQ ID NO: 5.

[0015] In another aspect, the present disclosure provides a recombinant AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 581 and 582, between amino acid positions 582 and 583, between amino acid positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-Li-[6-mer polypeptide] -L2-, wherein said Li and said L2 are each, independently, optional amino acid linkers, each having one, two, or three amino acids, and wherein the 6-mer polypeptide has an amino acid sequence of that of SEQ ID NO: 5.

[0016] In another aspect, the present disclosure provides a non-naturally occurring AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-L1-INSERT-L2-, wherein said Li and said L2 are each independently optional amino acid linkers having one, two, or three amino acids, and wherein INSERT represents the amino acid sequence of that of SEQ ID NO: 5.

[0017] In some embodiments, the Li is one amino acid Xi. In some embodiments, Li is two amino acids X2-X1. In some embodiments, Li is three amino acids X3-X2-X1.

[0018] In some embodiments, the Xi is selected from the group of amino acid residues consisting of A, V, I, and L. In some embodiments, Xi is A. In some embodiments, X2 isAttorney Docket No. 45049-0087W01 / 06839 selected from the group of amino acid residues consisting of A, V, I, and L. In some embodiments, X2 is L. In some embodiments, X2-X1 is LA. In some embodiments, X3 is selected from the group of amino acid residues consisting of A, V, I, and L. In some embodiments, Li is absent. In some embodiments, L2 is one amino acid Zi. In some embodiments, L2is two amino acids Z1-Z2. In some embodiments, L2 is three amino acids Z1-Z2-Z3. In some embodiments, Zi is selected from the group of amino acid residues consisting of A, V. I. and L. In some embodiments. Zi is A. In some embodiments, Z2 is selected from the group of amino acid residues consisting of A, V, I, and L. In some embodiments, Z2 is L. In some embodiments, Z1-Z2 is AL. In some embodiments, Z3 is selected from the group of amino acid residues consisting of A, V, I, and L. In some embodiments, L2 is absent.

[0019] In another aspect, the present disclosure provides a non-naturally occurring adeno- associated virus (AAV) vector comprising any AAV capsid polypeptide disclosed herein. In another aspect, the present disclosure provides a recombinant adeno-associated virus (AAV) vector comprising any AAV capsid polypeptide disclosed herein.

[0020] In yet another aspect, the present disclosure provides a viral particle comprising any capsid polypeptide disclosed herein.

[0021] In another aspect, the present disclosure provides a method for administering an exogenous nucleic acid sequence to a mammal in need thereof, wherein said method comprises administering an effective amount of a AAV vector disclosed herein to said mammal, wherein said vector comprising said exogenous nucleic acid sequence. In some embodiments, the mammal is a human. In some embodiments, the human is an infant, a child, or an adult. In some embodiments, administering comprises administering said effective amount to an eye of said mammal. In some embodiments, administering is sufficient to allow for expression of said exogenous nucleic acid sequence in a cell of said mammal. In some embodiments, exogenous nucleic acid sequence encodes a therapeutic polypeptide.

[0022] In another aspect, the present disclosure provides a method of treating a retinal disorder in a patient in need thereof, comprising administering to the patient’s eye an effective amount of an AAV vector, wherein the AAV vector comprises an AAV capsid polypeptide and an exogenous nucleic acid sequence, wherein the AAV capsid polypeptide is represented by Formula A.

[0023] In some cases, an AAV vector (e.g., an AAV2 vector) provided herein having an AAV capsid polypeptide that comprises an amino acid sequence of any one of SEQ ID NOs:Attorney Docket No. 45049-0087W01 / 068395-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can be used in place of the 7m8 AAV2 vector (Dalkara et al., Sei. Transl. Med., 5(189): 189ra76 (2013) and Bennett et al., J. Struct. Biol., 209(2): 107433 (2020)) or in place of the K912 AAV2 vector (Ozttirk et al., eLife, 10:e64175 (2021)) to deliver nucleic acid to retinal cells.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. Although methods and materials similar or equivalent to those described herein can be used to practice any inventio provided herein, suitable methods and methods are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

[0025] The details of one or more embodiments of the disclosure are set forth in the accompanying description below . Other features, objects, and advantages of any invention disclosed herein will be apparent from the description and from the claims.BRIEF DESCRIPTION OF THE DRAWINGS

[0026] The disclosure can be more completely understood with reference to the following.

[0027] FIG. 1A-1D show schematics of AAV vectors that include a wild type AAV2 Rep polypeptide and an indicated AAV2 capsid polypeptide engineered to include an insert sequence (e.g., any one of SEQ ID NOs: 5-9 or a sequence of Formula A) located betw een positions 587 and 588 of SEQ ID NO: 1 (FIG. 1 A), SEQ ID NO: 2 (FIG. IB), SEQ ID NO: 3 (FIG. 1C), or SEQ ID NO: 4 (FIG. ID).

[0028] FIG. 2A-2D show schematics of AAV vectors that include a mutant AAV2 Rep polypeptide (AAV2-M1T-REP) and an indicated AAV2 capsid polypeptide engineered to include an insert sequence (e.g., any one of SEQ ID NOs: 5-9 or a sequence of Formula A) located between positions 587 and 588 of SEQ ID NO: 1 (FIG. 2A), SEQ ID NO: 2 (FIG. 2B), SEQ ID NO: 3 (FIG. 2C), or SEQ ID NO: 4 (FIG. 2D).

[0029] FIGs. 3A-3D show schematics of AAV vectors that include a wild type AAV2 Rep polypeptide and an indicated AAV2 capsid polypeptide engineered to include an insert sequence (e.g., any one of SEQ ID NOs: 5-9 or a sequence of Formula A) as a replacement of amino acid residues at positions 585 to 590 of SEQ ID NO: 1 (FIG. 3A), SEQ ID NO: 2 (FIG. 3B), SEQ ID NO: 3 (FIG. 3C), or SEQ ID NO: 4 (FIG. 3D).Attorney Docket No. 45049-0087W01 / 06839

[0030] FIGs. 4A-4D show schematic of AAV vectors that include a mutant AAV2 Rep polypeptide (AAV2-M1T-REP) and an indicated AAV2 capsid polypeptide engineered to include an insert sequence (e.g., any one of SEQ ID NOs: 5-9 or a sequence of Formula A) as a replacement of amino acid residues at positions 585 to 590 of SEQ ID NO: 1 (FIG. 4A), SEQ ID NO: 2 (FIG. 4B), SEQ ID NO: 3 (FIG. 4C), or SEQ ID NO: 4 (FIG. 4D).

[0031] FIG. 5 contains images showing that right eyes (RE) and left eyes (LE) treated with the indicated variant (SEQ ID NO: 3 with SEQ ID NO: 9 inserted between positions 587 and 588) have higher fluorescent expression than those treated with the 7m8 control or the AAV2 control.DETAILED DESCRIPTION

[0032] The present disclosure provides AAV vectors (e.g., AAV2 vectors). For example, the disclosure provides AAV vectors (e.g, AAV2 vectors) comprising a capsid polypeptide that comprises an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A. Any appropriate AAV vector can be designed to include a capsid polypeptide described herein (e.g, a capsid polypeptide that comprises an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A. For example, AAV2, AAV8, and AAV9 can be designed to include a capsid polypeptide that comprises an amino acid sequence selected from any one of SEQ ID NOs: 5- 9 (or a variant thereof) or according to Formula A.

[0033] Any appropriate AAV capsid polypeptide can be designed to include an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A. For example, AAV2, AAV6, AAV8, and AAV9 capsid polypeptides can be designed to include an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A. In some embodiments, an AAV2 capsid polypeptide can be designed to include an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A. Table 1 provides amino acid sequences of exemplary AAV2 capsid polypeptides that can be designed to include an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A.Table 1: Amino Acid Sequences of Exemplary Capsid Polypeptides. The two bold amino acid residues (“NR ’) in the amino acid sequence denote positions 587 and 588. respectively, and the underlined amino acids (“RGNRQA”) denote positions 585 to 590. Bolded,Attorney Docket No. 45049-0087W01 / 06839 italicized, and underlined amino acid residues in SEQ ID NOs: 2 (E67A), 3 (V708I), and 4 (E67A and V708I) denote amino acid substitutions relative to wild-type.Attorney Docket No. 45049-0087W01 / 06839

[0034] In some embodiments, an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) comprises an amino acid sequence that is at least 95%, at least 96%, at least 97%. at least 98%, or at least 99% identical to the amino acid sequence set forth in Table 1 and can be designed to include an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or Formula A.

[0035] In some embodiments, AAV capsid polypeptide sequences contemplated herein can comprise modifications or mutations in SEQ ID NO: 1. In some embodiments, the AAV capsid polypeptide sequence of the present disclosure comprises a E67A substitution (SEQ ID NO: 2 in Table 1). In some embodiments, the AAV capsid polypeptide sequence of the present disclosure comprises a V708I substitution (SEQ ID NO: 3 in Table 1). In some embodiments, the AAV capsid polypeptide sequence of the present disclosure comprises both E67A and V708I substitutions (SEQ ID NO: 4 in Table 1).

[0036] When designing an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide listed in Table 1) to include an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A, that included amino acid sequence can be located at any appropriate location along the AAV capsid polypeptide (e.g, the AAV2 capsid polypeptide). For example, an amino acid sequence selected from any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A can be located between the naturally- occurring amino acid residues from position 585 to 590 of any one of SEQ ID NOs: 1-4. In some embodiments, the AAV2 capsid polypeptide has an amino acid sequence comprising that of any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 581 and 582, between amino acid 582 and 583, between positions 583 and 584, between amino acid positions 584 and 585, betweenAttorney Docket No. 45049-0087W01 / 06839 amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589. between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4.

[0037] In some embodiments, the present disclosure provides an AAV vector (e.g., AAV2) that comprises an AAV Rep coding sequence (e.g., AAV2 Rep 78 and AAV2 Rep 68 polypeptides) and an AAV Cap coding sequence. In some embodiments, the present disclosure provides an AAV vector (e.g, AAV2) that comprises a variant AAV Rep coding sequence. In some embodiments, the AAV Rep polypeptide has an ACG start codon encoding an AAV2 Rep comprising a substitution of methionine to threonine at position 1 (‘ MIT”). Table 2 provides exemplary AAV Rep polypeptide sequences.Table 2: Exemplary AAV Rep polypeptide sequences.Attorney Docket No. 45049-0087W01 / 06839Attorney Docket No. 45049-0087W01 / 06839Table 3: Exemplary sequences that can be inserted into an AAV capsid.Table 4: Exemplary sequences that can be inserted into an AAV capsid with an exemplary linker.

[0038] The polypeptides listed in Table 3 and Table 4 can be inserted between amino acid positions 581 and 582, between amino acid positions 582 and 583, between amino acid positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588. between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4. In some embodiments, any one of the polypeptides listed in Table 3 and Table 4 is inserted between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4. In some embodiments, any one of the polypeptides listed in Table 3 and Table 4 is inserted between amino acid positions 587 and 588 of SEQ ID NO: 4. In some embodiments, any one of the polypeptides listed in Table 3 and Table 4 replaces the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4.Attorney Docket No. 45049-0087W01 / 06839

[0039] Many of the amino acid sequences set forth in Table 4 start with “LA” and end with “A.” In some embodiments, that “LA” sequence can be a first linker sequence Li, and that “A” amino acid can be a second linker sequence L2, with the Li and L2 linkers each independently being optional amino acid linkers having one, two, or three amino acids according to the following Formula A:-L1-INSERT-L2- . wherein Li and L2 are each independently optional amino acid linkers having one, two, or three amino acids, and wherein INSERT represents the amino acid sequence of any one of SEQ ID NOs: 5-9. For example, in some cases, an AAV vector can be designed to have an AAV capsid polypeptide that comprises an amino acid sequence comprising that of any one of SEQ ID NOs: 5-9.

[0040] In some embodiments, the present disclosure provides a non-naturally occurring AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-L1-INSERT-L2-, wherein said Li and said L2 are each independently optional amino acid linkers having one, two, or three amino acids, and wherein INSERT represents the amino acid sequence of any one of SEQ ID NOs: 5-9.

[0041] In some embodiments, the present disclosure provides a non-naturally occurring AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 581 and 582, between amino acid positions 582 and 583, between amino acid positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-Li-[6-mer polypeptide] -L2-,Attorney Docket No. 45049-0087W01 / 06839 wherein said Li and said L2 are each, independently, optional amino acid linkers, each having one, two. or three amino acids, and wherein the 6-mer polypeptide has an amino acid sequence of any one of SEQ ID NOs: 5-9.

[0042] In some embodiments, the present disclosure provides a recombinant AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 581 and 582, between amino acid positions 582 and 583, between amino acid positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-Li-[6-mer polypeptide]-L2-, wherein said Li and said L2 are each, independently, optional amino acid linkers, each having one, two, or three amino acids, and wherein the 6-mer polypeptide has an amino acid sequence of any one of SEQ ID NOs: 5-9.

[0043] As described herein, an AAV vector can be designed to have an AAV capsid polypeptide that comprises an amino acid sequence of Formula A based on any one of SEQ ID NOs: 5-9. For example, an AAV vector can be designed to have an AAV capsid polypeptide of SEQ ID NO: 1 or an amino acid sequence that is at least 95%. at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 1) that includes an amino acid sequence of Formula A based on any of the amino acid sequences of SEQ ID NOs: 5-9 located between amino acid positions 587 and 588 of SEQ ID NO: 1 (or the appropriate amino acid positions of the alternative sequence).

[0044] In some embodiments, an AAV vector can be designed to have an AAV capsid polypeptide of SEQ ID NO: 2 or an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 2) that includes an amino acid sequence of Formula A based on any of the amino acid sequences of SEQ ID NOs: 5-9 located between amino acid positions 587 and 588 of SEQ ID NO: 2 (or the appropriate amino acid positions of the alternative sequence). In some embodiments, an AAV vector can be designed to have an AAV capsid polypeptide of SEQAttorney Docket No. 45049-0087W01 / 06839ID NO: 3 or an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 3) that includes an amino acid sequence of Formula A based on any of the amino acid sequences of SEQ ID NOs: 5-9 located between amino acid positions 587 and 588 of SEQ ID NO: 3 (or the appropriate amino acid positions of the alternative sequence). In some embodiments, an AAV vector can be designed to have an AAV capsid polypeptide of SEQ ID NO: 4 or an amino acid sequence that is at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 4) that includes an amino acid sequence of Formula A based on any of the amino acid sequences of SEQ ID NOs: 5-9 located between amino acid positions 587 and 588 of SEQ ID NO: 4 (or the appropriate amino acid positions of the alternative sequence).

[0045] In some embodiments, Li, L2, or both Li and L2can be absent. For example, an AAV capsid polypeptide that includes an amino acid sequence of Formula A based on any one of SEQ ID NOs: 5-9 can be included in the absence of the starting “LA” and the ending “A.” In some embodiments, Li can be one amino acid Xi, two amino acids X2-X1, or three amino acids X3-X2-X1. When Xi is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L. When X2 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L. When X3 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L. In some cases, L2 can be one amino acid Zi, two amino acids T -T . or three amino acids Z1-Z2-Z3. When Zi is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L. When Z2 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L. When Z3 is present, it can be an amino acid residue selected from the group consisting of A, V. I, and L. Examples of an Li linkers comprises, without limitation, A, V, I, L, AA, AV, Al, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AV A, AVV, AVI, AVL, AIA, AIV, All, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VW, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI,IAL, IVA, IVV, IVI, IVL. IIA, IIV, III, IIL. ILA, ILV, ILL ILL, LAA, LAV, LAL LAL, LVA, LVV, LVI. LVL, LIA. LIV, LIL LIL, LLA. LLV. LLI, and LLL. Examples of an L2linkers comprises, without limitation, A, V, I, L, AA, AV, Al, AL, VA, VV, VI, VL, I A, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AV A, AVV, AVI, AVL, AIA, AIV, All, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VW, VVI, VVL, VIA, VIV, VII, VIL, VLA. VLV, VLL VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA. IIV, III, IIL,Attorney Docket No. 45049-0087W01 / 06839ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, LIV, LII, LIL, LLA. LLV, LLI, and LLL.

[0046] In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth in any one of SEQ ID NOs: 1-4 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 1-4) with an amino acid sequence of any one of SEQ ID NOs: 5- 9 (or a variant thereof) or according to Formula A between asparagine-587 and arginine-588 (or the appropriate amino acid positions of the alternative sequence). In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth of SEQ ID NO: 1 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 1) with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A between asparagine-587 and arginine-588 (or the appropriate amino acid positions of the alternative sequence). In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth of SEQ ID NO: 2 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 2) with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereol) or according to Formula A between asparagine-587 and arginine-588 (or the appropriate amino acid positions of the alternative sequence). In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth of SEQ ID NO: 3 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 3) with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A between asparagine-587 and arginine-588 (or the appropriate amino acid positions of the alternative sequence). In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth of SEQ ID NO: 4 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 4) with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A between asparagine-587 and arginine-588 (or the appropriate amino acid positions of the alternative sequence).

[0047] In some embodiments, when designing an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) to include an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A, that included amino acid sequence can be used to replace one or more naturally-occurring amino acid residues located at anyAttorney Docket No. 45049-0087W01 / 06839 appropriate location along the AAV capsid polypeptide (e.g., the AAV2 capsid polypeptide). For example, an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) can be used to replace the naturally-occurring amino acid residues at positions 585 to 590 of an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide). For example, an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) can be used to replace the naturally-occurring amino acid residues at positions 585 to 590 of an AAV capsid polypeptide (e.g.. an AAV2 capsid polypeptide). For example, an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) can be used to replace the naturally-occurring amino acid residues at positions 585 to 590 of an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide).

[0048] In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth in any one of SEQ ID NOs: 1-4 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of any one of SEQ ID NOs: 1-4) except that the amino acid residues at positions 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO: 1 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 1) except that the amino acid residues at positions 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO: 2 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 2) except that the amino acid residues at positions 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO: 3 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 3) except that the amino acid residues at positions 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or aAttorney Docket No. 45049-0087W01 / 06839 variant thereof) or according to Formula A based on such a set forth sequence. In some embodiments, an AAV2 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO: 4 (or an amino acid sequence at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence of SEQ ID NO: 4) except that the amino acid residues at positions 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with an amino acid sequence of any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence.

[0049] In some embodiments, an AAV capsid polypeptide (e.g, an AAV2 capsid polypeptide) can be designed to include two or more amino acid sequences set forth in any one of SEQ ID NO: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. For example, an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) can be designed to include two, three, or four amino acid sequences set forth in any one of SEQ ID NO: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence.

[0050] As described herein, an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) can be designed to include an ammo acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. A variant of an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 refers to an amino acid sequence that is identical to that amino acid sequence set forth in any one of SEQ ID NOs: 5-9 except that it has one, two. or three amino acid additions, deletions, substitutions, or combinations thereof. For example, a variant of SEQ ID NO: 5 can be SEQ ID NO: 5 except that it has one, two, or three amino acid additions, deletions, substitutions, or combinations thereof. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs: 5-9, except that it contains one, two, or three amino acid additions. In some embodiments, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs: 5-9, except that it contains one, two, or three amino acid deletions. In some embodiments, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs: 5-9, except that it contains one, two, or three amino acid substitutions. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs: 5-9, except that it contains one amino acid addition, deletion, or substitution. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs: 5-9, except that it contains two amino acid additions, deletions, substitutions, or a combination thereof. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs: 5-9,Attorney Docket No. 45049-0087W01 / 06839 except that it contains three amino acid additions, deletions, substitutions, or a combination thereof.

[0051] In some embodiments, an amino acid substitution present in a variant can be a conservative amino acid substitution. For example, conserv ative amino acid substitutions can be made by substituting one amino acid residue for another amino acid residue having a similar side chain. Families of amino acid residues having similar side chains can include amino acids with basic side chains (e.g.. lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine), and aromatic side chains (e.g.. tyrosine, phenylalanine, tryptophan, histidine).

[0052] In some embodiments, an amino acid substitution present in a variant can be a nonconservative amino acid substitution. Non-conservative amino acid substitutions can be made by substituting one amino acid residue for another amino acid residue having a dissimilar side chain. Examples of non-conservative substitutions include, without limitation, substituting (a) a hydrophilic residue (e.g., serine or threonine) for a hydrophobic residue (e.g., leucine, isoleucine, phenylalanine, valine, or alanine); (b) a cysteine or proline for any other residue; (c) a residue having a basic side chain (e.g., lysine, arginine, or histidine) for a residue having an acidic side chain (e.g., aspartic acid or glutamic acid); and (d) a residue having a bulky side chain (e.g., phenylalanine) for glycine or other residue having a small side chain.

[0053] The percent sequence identity between a particular amino acid sequence and an amino acid sequence referenced by a particular sequence identification number is determined as follows. First, an amino acid sequence is compared to the sequence set forth in a particular sequence identification number using the BLAST 2 Sequences (B12seq) program from the stand-alone version of BLASTZ containing BLASTP version 2.0.14. This stand-alone version of BLASTZ can be obtained from Fish & Richardson’s web site (e.g., www.fr.com / blast / ) or the U.S. government’s National Center for Biotechnology Information web site (www.ncbi.nlm.nih.gov). Instructions explaining how to use the B12seq program can be found in the readme file accompanying BLASTZ. B12seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm. BLASTN is used to compare nucleic acid sequences, while BLASTP is used to compare amino acid sequences. To compare two amino acid sequences, the options of B12seq are set as follows: -i is set to aAttorney Docket No. 45049-0087W01 / 06839 file containing the first amino acid sequence to be compared (e.g., C:\seql.txt); -j is set to a file containing the second amino acid sequence to be compared (e.g., C:\seq2.txt); -p is set to blastp; -o is set to any desired file name (e.g., C:\output.txt); and all other options are left at their default setting. For example, the following command can be used to generate an output file containing a comparison between two amino acid sequences: C:\B12seq -i c:\seql.txt -j c:\seq2.txt -p blastp -o c:\output.txt. If the two compared sequences share homology, then the designated output file will present those regions of homology as aligned sequences. If the two compared sequences do not share homology, then the designated output file will not present aligned sequences. Once aligned, the number of matches is determined by counting the number of positions where an identical amino acid residue is presented in both sequences. A matched position refers to a position in which an identical amino acid residue occurs at the same position in aligned sequences. The percent sequence identity is determined by dividing the number of matches by the length of the sequence set forth in the identified sequence (e.g., SEQ ID NO: 1), followed by multiplying the resulting value by 100. For example, an amino acid sequence that has 725 matches when aligned with the sequence set forth in SEQ ID NO: 1 is 98.6 percent identical to the sequence set forth in SEQ ID NO: 1 (i.e., 725 735 x 100 = 98.6). It is noted that the percent sequence identity value is rounded to the nearest tenth. For example, 78.11, 78.12, 78.13, and 78.14 is rounded down to 78.1, while 78.15, 78.16. 78.17, 78.18, and 78.19 is rounded up to 78.2. It also is noted that the length value will always be an integer.

[0054] Methods for generating an amino acid sequence variant can include site-specific mutagenesis or random mutagenesis (e.g., by PCR) of a nucleic acid encoding an AAV capsid polypeptide. See, for example, Zoller, Curr. Opin. Biotechnol. 3: 348-354 (1992).

[0055] The AAV vectors (e.g.. AAV2 vectors) described herein can be designed to comprise one or more exogenous nucleic acid sequences. For example, an AAV vector (e.g., an AAV2 vector) described herein can be designed to comprise an exogenous nucleic acid sequence that encodes an RNA of interest and / or a polypeptide of interest. An exogenous nucleic acid sequence can be designed to encode any appropriate RNA of interest. Examples of RNAs of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV vector provided herein include, without limitation, siRNAs, RNA components for gene editing, and microRNAs. In some embodiments, an RNA of interest that can be encoded by an exogenous nucleic acid sequence included within an AAV vector provided herein can be SIRNA-027 to treat, e.g., sub-foveal CNVM secondary to age-related macular degeneration (see, e.g., NCT00363714), Cand5 / Bevasiranib to treat, e.g., diabeticAttorney Docket No. 45049-0087W01 / 06839 macular edema (see, e.g., NCT00306904), PF-04523655 to treat, e.g., diabetic macular edema (see, e.g., NCT01445899), QPI-1007 to treat, e.g., optic nerve atrophy in NAION (see, e.g., NCT01064505), Aganirsen to treat, e.g., ischemic CRVO to prevent neovascular glaucoma (see, e.g., NCT02947867), QR-421a to treat, e.g., retinitis pigmentosa / Usher syndrome ty pe 2 (see, e.g., NCT03780257), QR-1123 to treat, e.g., autosomal dominant retinitis pigmentosa (see, e.g., NCT04123626), IONIS-FB-LRX to treat, e.g., geographic atrophy secondary to age-related macular degeneration (see, e.g.. NCT03815825), or Sepofarsen / QR-110 to treat, e.g., Leber’s congenital amaurosis (see, e.g., NCT03913143).

[0056] An exogenous nucleic acid sequence can be designed to encode any appropriate polypeptide of interest. Examples of polypeptides of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV vector provided herein include, without limitation, therapeutic polypeptides, trophic factor polypeptides, gene editing polypeptides (e.g., a Cas9 polypeptide, a TALEN polypeptide, or a zinc finger polypeptide), enzymes, optogenetic tool polypeptides (e.g., a ChR polypeptide, an NhpR polypeptide, or a ReachR polypeptide), antibodies, antibody domains (e.g., VH domains), cytokines, anti-angiogenic polypeptides, and neuroprotective polypeptides. Examples of polypeptides of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV vector provided herein include, without limitation, an ABCA4 polypeptide, a CRB1 polypeptide, an NPHP5 polypeptide, an NR2E3 polypeptide, a PDE6A polypeptide, a PDE6B polypeptide, a PDE6C polypeptide, a PRPF31 polypeptide, a RPE65 polypeptide, a RPGR polypeptide, a RSI polypeptide, a TYR polypeptide, a USH2A polypeptide, a MY07A polypeptide, an REP1 polypeptide, an 0PN1LW polypeptide, an 0PN1MW polypeptide, a CNGA3 polypeptide, a CNGB3 polypeptide, a GUCY2D polypeptide, a GACA1A polypeptide, a GNAT2 polypeptide. a PDE6H polypeptide, a PR0M1 polypeptide, a PRPH2 polypeptide, a CRX polypeptide, an NPHP5 polypeptide, an EYS polypeptide, an ND4 polypeptide, a CLN1-14 polypeptide (e.g., a CLN3 polypeptide, a CLN5 polypeptide, a CLN6 polypeptide, or a CLN8 polypeptide), an NYX polypeptide, a GRM6 polypeptide, a TRPM1 polypeptide, a GPR179 polypeptide, an LRIT3 polypeptide, a glial cell derived neurotrophic factor (GDNF) polypeptide, a brain-derived neurotrophic factor (BDNF) polypeptide, a fibroblast growth factor (FGF) polypeptide, a truncated rod- derived cone viability factor (RdCVF) polypeptide, a full-length rod-derived cone viability factor (RdCVFL) polypeptide, an X-linked inhibitor of apoptosis (XIAP) polypeptide, a soluble fms-related receptor tyrosine kinase 1 (sFLT) polypeptide, a CYP4V2 polypeptide, a palmitoyl protein thioesterase 1 polypeptide, a tripeptidyl peptidase 1 polypeptide, aAttorney Docket No. 45049-0087W01 / 06839DNAJC5 polypeptide, a MFSD8 polypeptide, a cathepsin D polypeptide, a granulin polypeptide, an ATP13A2 polypeptide, a cathepsin F polypeptide, a KCTD7 polypeptide, a “P” gene polypeptide, a TRP1 polypeptide, a MATP (SLC45A2) polypeptide, a SLC24A5 polypeptide, a LRMDA polypeptide, a GPR143 polypeptide, an RPGR-exon 1 -ORF 15 polypeptide, an USH2b polypeptide, an USH1C polypeptide, a CDH23 polypeptide, a PCDH15 polypeptide, a SANS polypeptide, an USH1H polypeptide, a CIB2 polypeptide, an USH1K polypeptide, an ADGRV1 polypeptide, a WHRN polypeptide, a PDZD7 polypeptide, a CLRN1 polypeptide, aHARS polypeptide, an RP2 polypeptide, a FAM161 polypeptide, a DLK polypeptide, a RHO polypeptide, a CHM polypeptide, a BEST1 polypeptide, a RPl polypeptide, an OPA1 polypeptide, a CEP290 polypeptide, aRDH12 polypeptide, a CACNA1F polypeptide, a BBS1 polypeptide, a FAM161A polypeptide, a CERKL polypeptide, a PRPF8 polypeptide, a RP1L1 polypeptide, a SNRNP200 polypeptide, an IMPG2 polypeptide, a CDHR1 polypeptide, an IMPDH1 polypeptide, a CNGB1 polypeptide, a MERTK polypeptide, a KCNV2 polypeptide, an AIPL1 polypeptide, a RPGRIP1 polypeptide, a TULPl polypeptide, a C2ORF71 (aka PCARE) polypeptide, a MAK polypeptide. aTIMP3 polypeptide, a GUCA1A polypeptide, an ALMS1 polypeptide, a BBS10 polypeptide, an IFT140 polypeptide, a CNGA1 polypeptide, aNMNATl polypeptide, a COL2A1 polypeptide, an EFEMP1 polypeptide, a WFS1 polypeptide, a RDH5 polypeptide, a PRPF3 polypeptide, a LRP5 polypeptide, a TOPORS polypeptide, a DHDDS polypeptide, a LCA5 polypeptide, an IQCB1 polypeptide, a RP9 polypeptide, an ATXN7 polypeptide, a BBS2 polypeptide, a SAG RLBP1 polypeptide, a ND6 (MT-ND6) polypeptide, a Cl QTNF5 polypeptide, aVPS13B polypeptide, a KIFll polypeptide, a MT-TLl polypeptide, aKLHL7 polypeptide, an ACO2 polypeptide, a C21orf2 (aka CFAP410) polypeptide, an AHI1 polypeptide, a KIZ polypeptide, a SPATA7 polypeptide, a TTLL5 polypeptide, an HGSNAT polypeptide, aNRL polypeptide, an OAT polypeptide, a FLVCR1 polypeptide, an ABCC6 polypeptide, a LRAT polypeptide, a CEP78 polypeptide, a CDH3 polypeptide, a FZD4 polypeptide, a BBS12 polypeptide, an HKl polypeptide, a PRDM13 polypeptide, an ADAM9 polypeptide, a BBS7 polypeptide, a CABP4 polypeptide, an ABHD12 polypeptide, a COL18Al polypeptide, a MFRP polypeptide, a RIMSl polypeptide, a ROM! polypeptide, a BBS4 polypeptide, an IMPG1 polypeptide, an INPP5E polypeptide, a VC AN polypeptide, a POC1B polypeptide, a RAX2 polypeptide, a TSPAN12 polypeptide, a CACNA2D4 polypeptide, a JAG1 polypeptide, a MKKS polypeptide, a NPHP4 polypeptide, a BBS9 polypeptide, a COL11A1 polypeptide, an ELOVL4 polypeptide, aNDP polypeptide, a NPHP1 polypeptide, a RGR polypeptide, a BBS5 polypeptide, a WDR19 polypeptide, aAttorney Docket No. 45049-0087W01 / 06839C8ORF37 polypeptide, a CTNNA1 polypeptide, a LAMP2 polypeptide, a PEXl polypeptide, a PHYH polypeptide, an ATF6 polypeptide, a PRPS1 polypeptide, a SEMA4A polypeptide, an ARL6 polypeptide, a CNNM4 polypeptide, an OTX2 polypeptide, a PRPF6 polypeptide, a RBP3 polypeptide, aPNPLA6 polypeptide, a SLC24A1 polypeptide, an USH1G polypeptide, a PITPNM3 polypeptide, a TTC8 polypeptide, an ARSG polypeptide, a CWC27 polypeptide, a DRAM2 polypeptide, a PRCD polypeptide, a REEP6 polypeptide, a SSBP1 polypeptide, a LAMA1 polypeptide, a RAB28 polypeptide, a ZNF408 polypeptide, a GNAT1 polypeptide, an IDH3A polypeptide, a PDE6G polypeptide, a PEX6 polypeptide, a TUB polypeptide, a CEP250 polypeptide, a FSCN2 polypeptide, a GRK1 polypeptide, a RBP4 polypeptide, a RD3 polypeptide, an AGBL5 polypeptide, a CAPN5 polypeptide, an IFT172 polypeptide, a KCNJ13 polypeptide, a PAX2 polypeptide, a CC2D2A polypeptide, aHMCNl polypeptide, a MT-ATP6 polypeptide, a RCBTB1 polypeptide, an ARL2BP polypeptide, a CA4 polypeptide, a DFNB31 polypeptide, a GNB3 polypeptide, a MMACHC polypeptide, a PRPF4 polypeptide, a RGS9 polypeptide, an ARHGEF18 polypeptide, a KIAA1549 polypeptide, a MK.S1 polypeptide, a MTTP (not MT-TP) polypeptide, a PLK4 polypeptide, a RPGRIP1L polypeptide, a SDCCAG8 polypeptide, a SRD5A3 polypeptide, a TUBB4B polypeptide, an ADAMTS18 polypeptide, an ARL3 polypeptide, a COL11A2 polypeptide, a MVK polypeptide, a NBAS polypeptide, an OFD 1 polypeptide, a P3H2 polypeptide, a RGS9BP polypeptide, a CSPP1 poly peptide, an ITM2B polypeptide, a PANK2 polypeptide, a PEX7 polypeptide, a P0MGNT1 polypeptide, a SLC4A7 polypeptide, a TMEM231 polypeptide, a TRNTl polypeptide, a TUBGCP6 polypeptide, aZNF513 polypeptide, an AFG3L2 polypeptide, an ARL13B polypeptide, a C5ORF42 (aka CPLANE1) polypeptide, a COL9A1 polypeptide, a CTSD polypeptide, aDTHDl polypeptide, a DYNC2Hl polypeptide, an IFT81 polypeptide, a KIAA0586 polypeptide, a MFN2 polypeptide, a NPHP3 polypeptide, a PCYT1 A polypeptide, a PEX12 polypeptide, a PLA2G5 polypeptide, a POC5 polypeptide, a SCAPER polypeptide, a SLC25A46 polypeptide, a TMEM237 polypeptide, a TRAF3IPl polypeptide, a TTC21B polypeptide, a TUBGCP4 polypeptide, an ADIP0R1 polypeptide, a CEP 164 polypeptide, a CLCC1 polypeptide, a COL9A2 polypeptide, a CTNNB1 polypeptide, a DHX38 polypeptide, a GNPTG polypeptide, a GRN polypeptide, a GUCA1B polypeptide, an IFT27 polypeptide, an IFT74 polypeptide, a KIAA0556 polypeptide, a LRP2 polypeptide, a MAPKAPK3 polypeptide, a MIR204 polypeptide, a MT-ND3 polypeptide, a MT-RNRl polypeptide. a MT-TS2 polypeptide, a ND5 (MT-ND5) polypeptide, aNEK2 polypeptide, an 0PN1SW polypeptide, a PEX13 polypeptide, a PEX2 polypeptide, a RHBDD2 polypeptide, a SAMD11 polypeptide, aAttorney Docket No. 45049-0087W01 / 06839SCLT1 polypeptide, a SLC7A14 polypeptide, a TCTNl polypeptide, a TCTN2 polypeptide, a TLCD3B polypeptide, a TREX1 polypeptide, a TTPA polypeptide, an UNCI 19 polypeptide, a WDPCP polypeptide, an ACBD5 polypeptide, an AHR polypeptide, an ARMC9 polypeptide, an ASRGL1 polypeptide, an AT0H7 polypeptide, a B9Dl polypeptide, a B9D2 polypeptide, a BBIPl polypeptide, a C12ORF65 polypeptide, a C2CD3 polypeptide, a C5AR2 polypeptide, a CCDC188 polypeptide, a CCT2 polypeptide, a CEP104 polypeptide, a CEP 120 polypeptide, a CEP 19 polypeptide, a CEP41 polypeptide, a CISD2 polypeptide, a CLUAP1 polypeptide, a COL9A3 polypeptide, a CRB2 polypeptide, a CTC1 polypeptide, a DACT2 polypeptide, a DDR1 polypeptide, an ENSA polypeptide, an ESPN polypeptide, an EXOSC2 polypeptide, a FBN3 polypeptide, a GDF6 polypeptide, a GPR125 polypeptide, a HKDC1 polypeptide, a HMXl polypeptide, an IDH3B polypeptide, an IFT43 polypeptide, an IFT80 polypeptide, an INVS polypeptide, a KIAA0753 polypeptide, a KIF3B polypeptide, a KIF7 polypeptide, a LRRTM4 polypeptide, a LZTFL1 polypeptide, a MT-ATP8 polypeptide, a MT-COl polypeptide, a MT-CO2 polypeptide, a MT-C03 polypeptide, a MT-CYB polypeptide, a MT-ND2 polypeptide, a MT-ND4L polypeptide, a MT-RNR2 polypeptide, a MT-TA polypeptide, a MT-TC polypeptide, a MT-TD polypeptide, a MT-TE polypeptide, a MT-TF polypeptide, a MT-TG polypeptide, a MT-TH polypeptide, a MT-TI polypeptide, a MT-TK polypeptide, a MT-TL2 polypeptide, a MT-TM polypeptide, a MT-TN polypeptide, a MT-TP (Not MTTP) polypeptide, a MT-TQ polypeptide, a MT-TR polypeptide, a MT-TS1 polypeptide, a MT-TT polypeptide, a MT-TV polypeptide, a MT-TW polypeptide. a MT-TY polypeptide, aNEURODl polypeptide, a PDE6D polypeptide, a PEXl O polypeptide, a PEX11B polypeptide, a PEX14 polypeptide, a PEX16 polypeptide, a PEX19 polypeptide, a PEX26 polypeptide, a PEX3 polypeptide, a PEX5 polypeptide, a PGK1 polypeptide, a PISD polypeptide, a PPP2R3C polypeptide, a PROSl polypeptide, a PSENl polypeptide, a RDHl l polypeptide, a RRM2B polypeptide, a SMARCA4 polypeptide, a SPP2 polypeptide, a TCTN3 polypeptide, a TEADl polypeptide, a TMEM107 polypeptide, a TMEM138 polypeptide, a TMEM216 polypeptide, a TMEM67 polypeptide, a TPPl polypeptide, a TRIM32 polypeptide, a USP45 polypeptide, and a ZNF423 polypeptide. In some cases, a polypeptide of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV vector provided herein can be a MY07A polypeptide, an USH1C polypeptide, a PCDH15 polypeptide, an USH2A polypeptide, or CLRN1 polypeptide to treat, e.g., Usher Syndrome.

[0057] In some embodiments, one or more AAV vectors provided herein can be designed to carry out gene editing within one or more cells (e.g., retinal cells). Such gene editing canAttorney Docket No. 45049-0087W01 / 06839 result in a genomic modification of one or more cells. Examples of such genomic modifications include, without limitation, a targeted insertion of a nucleic acid encoding an RNA and / or polypeptide of interest into one or more cells, a targeted modification (e.g., targeted inactivation or knock-out) of a genomic sequence of one or more cells, and a targeted replacement of nucleic acid (e.g., nucleic acid encoding an RNA, a regulatory nucleic acid sequence, and / or nucleic acid encoding a polypeptide of interest) within one or more cells.

[0058] Any appropriate gene editing components can be engineered into one or more AAV vectors provided herein such that those one or more AAV vectors can be used to deliver the gene editing components to target cells (e.g., one or more retinal cells) within a mammal (e.g. , a human or a non-human primate) in a manner effective to edit the genome of those cells. Typically, the gene editing components include, without limitation, a component that is capable of cleaving genomic nucleic acid at a desired location and an optional donor nucleic acid designed to be inserted into that desired location once it is cleaved. Any appropriate rare-cutting endonuclease can be used to cleave genomic nucleic acid at a desired location. Examples of such rare-cutting endonucleases include, without limitation, meganucleases, transcription activator-like effector (TALE) nucleases (TALENs™; Cellectis, Paris, France), zinc-finger-nucleases (ZFNs), and endonucleases of a clustered regularly interspaced short palindromic repeats (CRISPR)ZCas system (e.g., endonucleases of a CRISPR / Cas 9 system). See, e.g.. Baker, Nature Methods, 9:23-26 (2012); International PCT Patent Application Publication No. WO 2004 / 067736; International PCT Patent Application Publication No. WO 2011 / 072246; U.S. Patent No. 8,586,363; Porteus and Carroll, Nature Biotechnol., 23:967- 973 (2005); Jinek et al., Science. 337:816-821 (2012); Mali et al., Science, 339:823-826 (2013); Li et al., Nature Biotechnology, 31 (8):688-691 (2013); and Makarova et al., Nat. Rev. Microbiol., 9(6):467-477 (2011)).

[0059] In some embodiments, to facilitate gene replacement, two sequences in genomic nucleic acid of a cell (e.g., a retinal cell) - one on either side of a sequence to be removed - can be targeted for endonuclease cleavage. For example, a first target sequence adjacent to the 5‘ end of a sequence to be removed and a second target sequence adjacent to the 3’ end of the sequence to be removed can be targeted by guide RNAs to enable Cas9 cleavage or can be targeted by TALENs designed to specifically recognize those targets. Delivery using one or more AAV vectors provided herein of (a) endonucleases targeted to the genomic DNA and (b) a donor nucleic acid construct can allow cleavage at both genomic targets, removal of theAttorney Docket No. 45049-0087W01 / 06839 sequence between the genomic targets, and insertion of the donor sequence into the location of the deletion.

[0060] An AAV vector (e.g., an AAV2 vector) provided herein can include any appropriate promoter and / or other regulatory’ sequence (e.g., enhancers, transcription initiation sites, translation initiation sites, and termination signals) operably linked an exogenous nucleic acid sequence designed to be expressed. In some cases, a promoter used to drive expression can be a constitutive promotor, a regulatable promotor, a tissue-specific promoter, or a viral promotor. Examples of constitutive promotors that can be used as described herein include, without limitation, chicken 0-actin (CBA) promoters, CAG promoters (e.g., CMV early enhancer / chicken 0 actin), SV40 promoters, cytomegalovirus (CMV) promoters, and E1ALPHA promotors. Examples of regulatable promoters that can be used as described herein include, without limitation, inducible promotors and repressible promotors. Examples of tissue-specific promoters that can be used as described herein include, without limitation, rhodopsin promoters, cone arrestin promoters, and synapsin promoters. Examples of viral promotors that can be used as described herein include, without limitation, adenoviral promotors, vaccinia virus promoters, CMV promoters (e.g.. immediate early CMV promotors), and AAV promoters.

[0061] In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can include a total number of nucleotides up to about 5 kb. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can include a total number of nucleotides that is from about 1 kb to about 5 kb, from about 1 kb to about 4 kb, from about 1 kb to about 3 kb, from about 2 kb to about 5 kb, from about 2 kb to about 4 kb, from about 2 kb to about 3 kb, from about 3 kb to about 5 kb, from about 3 kb to about 4 kb, or from about 4 kb to about 5 kb.

[0062] An AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect retinal cells in vivo and deliver exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability7to infect and drive RNA expression of an exogenous nucleic acid sequence in retinal cells present in a mammal (e.g.. a human or a non-human primate) when compared to wild-type AAV2. In some embodiments, an AAV vector (e.g, an AAV2 vector) provided herein can have the ability toAttorney Docket No. 45049-0087W01 / 06839 drive a level of RNA expression of an exogenous nucleic acid sequence in retinal cells of a mammal (e.g., a human or a non -human primate) that is greater than the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector (e.g., wild-type AAV2) having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 (e.g., a wild-ty pe AAV2 vector) in retinal cells of a control mammal (e.g., a control human or a control non-human primate).

[0063] An AAV vector (e.g., an AAV2 vector) described herein comprising an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) in vivo and deliver exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence (e.g., at high levels). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in at least 2 percent (e.g., at least 2.5 percent, at least 5 percent, at least 7.5 percent, at least 10 percent, or at least 25 percent) of retinal cells of an eye of a mammal (e.g., a human or a non-human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) of a mammal (e.g., a human or a non-human primate) that is greater than (e.g., at least 2 percent greater than, at least 2.5 percent greater than, at least 5 percent greater than, at least 7.5 percent greater than, at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the level of mRNA expression of an exogenous nucleic acid driven by a comparable AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence of SEQ ID NO: 1 (e.g., a wild-type AAV2 vector) in retinal cells of a control mammal (e.g., a control human or a control non-human primate).

[0064] An AAV vector (e.g., an AAV2 vector) described herein comprising an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability' to infect retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) across retinal regions (e.g, across two. three, or four retinal regions) in vivo and deliver exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequenceAttorney Docket No. 45049-0087W01 / 06839(e.g., a high levels). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the retinal cells of a mammal (e.g., a human or a non-human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) in at least two, three, or four different regions of an eye of a mammal (e.g., ahuman or a non-human primate) that is greater than the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 (e.g., a wild-type AAV2 vector) in retinal cells of those regions in a control mammal (e.g., a control human or a control non-human primate).

[0065] An AAV vector (e.g., an AAV2 vector) described herein comprising an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability’ to infect retinal cells of the eye in vivo and deliver exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of retinal cells of the eye of a mammal (e.g.. a human or a non-human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in retinal cells of the eye of a mammal (e.g., a human or a non-human primate) that is greater than (e.g., at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 (e.g., a wild type AAV2 vector) in retinal cells of the eye of a control mammal (e.g., a control human or a control non-human primate).

[0066] An AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence setAttorney Docket No. 45049-0087W01 / 06839 forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect two or more (e.g., two or more, three or more, four or more, five or more, six or more, or seven or more) different retinal cell types within an eye in vivo and deliver exogenous nucleic acid to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid. For example, an AAV vector (e.g., an AAV2 vector) described herein comprising an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect two, three, four, five, six, or seven of the following retinal cell types: retinal ganglion cells, amacrine cells, horizontal cells, bipolar cells, Muller glia cells, photoreceptor cells, and RPE cells. In some embodiments, an AAV vector (e.g.. an AAV2 vector) described herein comprising an AAV capsid polypeptide (e.g, an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereol) or according to Formula A based on such a set forth sequence can have the ability to infect (a) retinal ganglion cells, amacrine cells, and horizontal cells, (b) retinal ganglion cells, amacrine cells, and bipolar cells, (c) retinal ganglion cells, amacrine cells, and Muller glia cells, (d) retinal ganglion cells, amacrine cells, and photoreceptor cells, (e) retinal ganglion cells, amacrine cells, and RPE cells, (f) amacrine cells, horizontal cells, and bipolar cells, (g) amacrine cells, horizontal cells, and Muller glia cells, (h) amacrine cells, horizontal cells, and photoreceptor cells, (i) amacrine cells, horizontal cells, and RPE cells, (j) horizontal cells, bipolar cells, and Muller glia cells, (k) horizontal cells, bipolar cells, and photoreceptor cells, (1) horizontal cells, bipolar cells, and RPE cells, (m) bipolar cells, Muller glia cells, and photoreceptor cells, (n) bipolar cells, Muller glia cells, and RPE cells, or (o) Muller glia cells, photoreceptor cells, and RPE cells. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive mRNA expression of an exogenous nucleic acid in at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the retinal ganglion cells, at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the amacrine cells, at least 2 percent (e g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the horizontal cells, at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the bipolar cells, at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least aboutAttorney Docket No. 45049-0087W01 / 068397.5 percent, at least about 10 percent, or at least about 25 percent) of the Muller glia cells, at least 2 percent (e.g, at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the photoreceptor cells, and / or at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the RPE cells of an eye of a mammal (e.g., a human or a non-human primate) following, for example, an intravitreal administration.

[0067] An AAV vector (e.g., an AAV2 vector) described herein comprising an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect RPE cells in vivo and deliver exogenous nucleic acid sequence to the infected RPE cells such that the infected RPE cells express the exogenous nucleic acid sequence (e.g., at high levels). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of RPE cells of an eye of a mammal (e.g., a human or a non- human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in RPE cells of an eye of a mammal (e.g., a human or a non-human primate) that is greater than (e.g., at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 in RPE cells of an eye of a control mammal (e.g., a control human or a control non-human primate).

[0068] An AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect photoreceptor cells in vivo and deliver exogenous nucleic acid sequence to the infected photoreceptor cells such that the infected photoreceptor cells express the exogenous nucleic acid sequence. For example, an AAV vector (e.g., an AAV2 vector) described herein comprising an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in anyAttorney Docket No. 45049-0087W01 / 06839 one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability- to infect photoreceptor cells in vivo to a greater extent than any other retinal cell type of an eye and deliver exogenous nucleic acid sequence to the infected photoreceptor cells such that the infected photoreceptor cells express the exogenous nucleic acid sequence. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of photoreceptor cells of an eye of a mammal (e.g., a human or a non-human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in photoreceptor cells of an eye of a mammal (e.g., a human or a non-human primate) that is greater than (e.g., at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 in photoreceptor cells of an eye of a control mammal (e.g., a control human or a control non- human primate).

[0069] An AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability- to infect retinal ganglion cells in vivo and deliver exogenous nucleic acid sequence to the infected retinal ganglion cells such that the infected retinal ganglion cells express the exogenous nucleic acid sequence. For example, an AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in anyone of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect retinal ganglion cells in vivo to a greater extent than any other retinal cell type of an eye and deliver exogenous nucleic acid sequence to the infected retinal ganglion cells such that the infected retinal ganglion cells express the exogenous nucleic acid sequence. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25Attorney Docket No. 45049-0087W01 / 06839 percent) of retinal ganglion cells of an eye of a mammal (e.g.. a human or a non-human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in retinal ganglion cells of an eye of a mammal (e.g., a human or a non-human primate) that is greater than (e.g., at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 in retinal ganglion cells of an eye of a control mammal (e.g., a control human or a control non-human primate).

[0070] An AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect bipolar cells of the retina in vivo and deliver exogenous nucleic acid sequence to the infected bipolar cells such that the infected bipolar cells express the exogenous nucleic acid sequence. For example, an AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect bipolar cells of the retina in vivo to a greater extent than any other retinal cell type of an eye and deliver exogenous nucleic acid sequence to the infected bipolar cells such that the infected bipolar cells express the exogenous nucleic acid sequence. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of bipolar cells of the retina of an eye of a mammal (e.g., a human or a non-human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in bipolar cells of the retina of an eye of a mammal (e.g., a human or a non-human primate) that is greater than (e.g., at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 in bipolarAttorney Docket No. 45049-0087W01 / 06839 cells of the retina of an eye of a control mammal (e.g., a control human or a control nonhuman primate).

[0071] An AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have the ability to infect retinal cells in vivo and deliver exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence. In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability' to infect and drive RNA expression of an exogenous nucleic acid sequence in at least 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of retinal cells of an eye of a mammal (e.g., a human or a non-human primate). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in retinal cells of an eye of a mammal (e.g., a human or a non-human primate) that is greater than (e.g., at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the level of RNA expression of an exogenous nucleic acid sequence driven by a control AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence set forth in SEQ ID NO: 1 in retinal cells of an eye of a control mammal (e.g.. a control human or a control non-human primate).

[0072] An AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence can have increased packaging efficiency, the ability to infect cells (e.g., retinal cells) in vivo or in vitro, and the ability to deliver exogenous nucleic acid sequence to the infected cells such that the infected cells express the exogenous nucleic acid sequence (e.g., at high levels). In some embodiments, an AAV vector (e.g., an AAV2 vector) provided herein can have a packaging efficiency that is greater than (e.g.. at least 10 percent greater than, at least 25 percent greater than, at least 50 percent greater than, at least 75 percent greater than, or at least 100 percent greater than) the packaging efficiency of a comparable AAV vector having an AAV capsid polypeptide that comprises the amino acid sequence of SEQ ID NO: 1 (e.g., a wild-type AAV2 vector).Attorney Docket No. 45049-0087W01 / 06839

[0073] Examples of retinal cells that can be infected by an AAV vector (e.g., an AAV2 vector) described herein comprises an AAV capsid polypeptide (e.g.. an AAV2 capsid polypeptide) that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or Formula A include, without limitation, retinal ganglion cells, retinal pigment epithelium cells, photoreceptor cells, bipolar cells, amacrine cells, Muller glia, and horizontal cells.

[0074] The present disclosure also provides compositions comprising one or more AAV vectors provided herein (e.g., one or more AAV2 vectors provided herein). For example, one or more AAV vectors provided herein (e.g., one or more AAV2 vectors provided herein) can be formulated as a pharmaceutical composition for administration to a mammal (e.g., a human or a non-human primate) to treat that mammal. In some embodiments, one or more AAV vectors provided herein (e.g., one or more AAV2 vectors provided herein) can be formulated as a pharmaceutical composition for administration to a mammal (e.g., a human or a non-human primate) to deliver an exogenous nucleic acid sequence to retinal cells for expression within retinal cells. For example, an AAV vector (e g., an AAV2 vector) provided herein can be formulated as a pharmaceutical composition for administration to a mammal (e.g., a human or a non-human primate). In some cases, a pharmaceutical composition provided herein can include a pharmaceutically acceptable carrier such as a buffer, a salt, a surfactant, a sugar, a tonicity modifier, or combinations thereof as, for example, described elsewhere (Gervasi et al., Eur. J. Pharmaceutics and Biopharmaceutics , 131 : 8-24 (2018)). Examples of pharmaceutically acceptable carriers that can be used to make a pharmaceutical composition provided herein include, without limitation, water, lactic acid, citric acid, sodium chloride, sodium citrate, sodium succinate, sodium phosphate, a surfactant (e.g., polysorbate 20, polysorbate 80, or poloxamer 188), dextran 40, or a sugar (e.g., sorbitol, mannitol, sucrose, dextrose, or trehalose), or combinations thereof. For example, a pharmaceutical composition designed to include an AAV vector (e.g., an AAV2 vector) provided herein can be formulated to include a buffer (e.g., an acetate, citrate, histidine, succinate, phosphate, or hydroxymethyl-aminomethane (Tris) buffer), a surfactant (e g., polysorbate 20, polysorbate 80, or poloxamer 188), and a sugar such as sucrose. Other ingredients that can be included within a pharmaceutical composition provided herein include, without limitation, amino acids such as glycine or arginine, antioxidants such as ascorbic acid, methionine, or ethylenediaminetetraacetic acid (EDTA), or combinations thereof.Attorney Docket No. 45049-0087W01 / 06839

[0075] In some embodiments, when a pharmaceutical composition is formulated to include one or more AAV vectors (e.g., one or more AAV2 vectors) provided herein, any appropriate titer of the AAV vectors can be used. For example, a pharmaceutical composition provided herein can be formulated to have AAV vectors (e.g., AAV2 vectors) provided herein at a titer that is greater than 1 x 107viral genomes per mL (vg / mL) (e.g., greater than 1 x 108vg / mL, greater than 1 x 109vg / mL, greater than 1 x 1010vg / mL, greater than 1 x 1011vg / mL, greater than 1 x 1012vg / mL. greater than 1 x 1013vg / mL. or greater than 1 x 1014vg / mL). In some embodiments, a pharmaceutical composition provided herein can be formulated to have AAV vectors (e.g., AAV2 vectors) provided herein at a titer that is from about 1 x 107vg / mL to about 1 x 1014vg / mL (e.g., from about 1 x 107vg / mL to about 1 x 1013vg / mL, from about 1 x 107vg / mL to about 1 x 1012vg / mL, from about 1 x 107vg / mL to about 1 x 1011vg / mL, from about 1 x 107vg / mL to about 1 x 1010vg / mL, from about 1 x 108vg / mL to about 1 x 1014vg / mL, from about 1 x 109vg / mL to about 1 x 1014vg / mL, from about 1 x 1010vg / mL to about 1 x 1014vg / mL, from about 1 x 108vg / mL to about 1 x 1012vg / mL, or from about 1 x 109vg / mL to about 1 x 1011vg / mL). In some embodiments, the titer is vgs / dose, wherein the dose may be administered systemically or directly. In some embodiments, the titer is in vg / eye. In some embodiments, the titer is in vgs / mL.

[0076] A pharmaceutical composition provided herein can be in any appropriate form. For example, a pharmaceutical composition provided herein can be designed to be a liquid, a semi-solid, or a solid. In some embodiments, a pharmaceutical composition provided herein can be a liquid solution (e.g., an injectable and / or infusible solution), a dispersion, a suspension, a tablet, a pill, a powder, a microemulsion, a liposome, or a suppository. In some embodiments, a pharmaceutical composition provided herein can be lyophilized. In some embodiments, a pharmaceutical composition provided herein (e.g., a pharmaceutical composition that includes one or more AAV vectors provided herein such as one or more AAV2 vectors provided herein) can be formulated with a carrier or coating designed to protect against rapid release. For example, a pharmaceutical composition provided herein can be formulated as a controlled release formulation or as a regulated release formulation as described elsewhere (U.S. Patent Application Publication Nos. 2019 / 0241667; 2019 / 0233522; and 2019 / 0233498).

[0077] The present disclosure also provides nucleic acid molecules encoding an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. In some embodiments, a nucleic acid molecule can be designed to encode an AAV capsid polypeptideAttorney Docket No. 45049-0087W01 / 06839 that comprises an amino acid sequence that is encoded by a DNA sequence set forth in any one of CACGACCAGGTGCGGCCC (SEQ ID NO: 18); CACGACCAGGTGCGGCCCGCT (SEQ ID NO: 19); CTAGCACACGACCAGGTGCGGCCC (SEQ ID NO: 20); GCACACGACCAGGTGCGGCCC (SEQ ID NO: 21); and CTAGCACACGACCAGGTGCGGCCCGCT (SEQ ID NO: 17).

[0078] The present disclosure also provides nucleic acid molecules encoding an AAV vector (e.g.. an AAV2 vector) described herein. For example, an isolated nucleic acid molecule can be designed to encode one or more AAV vectors provided herein (e.g., an AAV having an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence). In some embodiments, a nucleic acid molecule can be designed to encode an AAV vector having an AAV capsid polypeptide that comprises an amino acid sequence that is encoded by a DNA sequence set forth in any one of CACGACCAGGTGCGGCCC (SEQ ID NO: 18); CACGACCAGGTGCGGCCCGCT (SEQ ID NO: 19); CTAGCACACGACCAGGTGCGGCCC (SEQ ID NO: 20);GCACACGACCAGGTGCGGCCC (SEQ ID NO: 21); and CTAGCACACGACCAGGTGCGGCCCGCT (SEQ ID NO: 17).

[0079] The present disclosure also provides host cells comprising a nucleic acid molecule provided herein. For example, a host cell can be designed to comprise a nucleic acid molecule encoding an AAV capsid polypeptide described herein and / or a nucleic acid molecule encoding an AAV vector described herein. In some embodiments, a host cell can be designed to comprise a nucleic acid molecule encoding an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. In some embodiments, a host cell can be designed to comprise a nucleic acid molecule encoding an AAV vector having an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. Examples of host cells that can be designed to comprise a nucleic acid molecule encoding an AAV capsid polypeptide described herein and / or a nucleic acid molecule encoding an AAV vector described herein include, without limitation, HEK293T cells (ATCC), 293 AAV cells (Cell Biolabs), NEB 5-alpha cells, TakaraBio Stellar cells, and MegaX cells. Any appropriate method can be used to introduce a nucleic acid molecule provided herein (e.g., a nucleic acid molecule encoding an AAV capsid polypeptide described herein and / or an AAV vector described herein) into a cell. For example, viralAttorney Docket No. 45049-0087W01 / 06839 transfection, electroporation, transient transfection, and gene gun techniques can be used to introduce a nucleic acid molecule provided herein into a cell.

[0080] The present disclosure also provides methods and materials for making an AAV vector (e.g., an AAV2 vector) provided herein. For example, the disclosure provides methods and materials for making AAV vectors (e.g., AAV2 vectors) comprising an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. As described herein, an AAV vector can be constructed to comprise an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. Any appropriate method can be used to construct an AAV vector having an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) provided herein (e.g., a capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence). For example, molecular cloning and AAV vector production techniques such as those described elsewhere can be used to construct and produce an AAV vector having an AAV capsid polypeptide (e.g., an AAV2 capsid polypeptide) provided herein (see, e.g., Sambrook et al.. Molecular Cloning: A Laboratory' Manual, 2nd edition, Cold Spring Harbor Laboratory, NY (1989); Ausubel el al., Current Protocols in Molecular Biology, Green Publishing Associates and John Wiley & Sons, New York, N.Y. (1994); Grieger et al., Nat. Protoc.. 1(3): 1412-28 (2006); and Flannery et al.. Methods Mol. Biol., 935:351 -69 (2013)). In some cases, AAV vectors can be produced in HEK293T cells (ATCC) or 293 AAV cells (Cell Biolabs) using a double or triple transfection method (see, e.g., Grieger et al., Nat. Protoc., 1(3): 1412-28 (2006); and Flannery et al., Methods Mol. Biol.. 935:351-69 (2013)).

[0081] The present disclosure also provides methods and materials for using an AAV vector (e.g., an AAV2 vector) provided herein. For example, the disclosure provides methods and materials for using AAV vectors (e.g., AAV2 vectors) comprising an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence. As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect retinal cells in vivo and to deliver an exogenous nucleic acidAttorney Docket No. 45049-0087W01 / 06839 sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence.

[0082] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) in vivo and to deliver an exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence (e.g., at high levels).

[0083] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) across retinal regions (e.g., across two, three, or four different retinal regions) in vivo and to deliver an exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence (e.g.. at high levels). For example, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) across retinal regions such that the AAV vector infects at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the retinal cells, at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the retinal cells, at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the retinal cells in the vascular arcade region, and / or at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the retinal cells in the periphery region of an eye of a mammal (e.g., a human or a non-human primate).

[0084] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect retinal cells of the eye in vivo and to deliver an exogenousAttorney Docket No. 45049-0087W01 / 06839 nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence.

[0085] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect two or more (e.g., two or more, three or more, four or more, five or more, six or more, or seven or more) different retinal cell types within an eye in vivo and to deliver an exogenous nucleic acid sequence to the infected retinal cells such that the infected retinal cells express the exogenous nucleic acid sequence (e.g., at high levels). For example, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect (a) at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the retinal ganglion cells, (b) at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the amacrine cells, (c) at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the horizontal cells, (d) at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the bipolar cells, (e) at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the Muller glia cells, (f) at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the photoreceptor cells, (g) at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of the RPE cells, all of (a)-(g), or any combination of two, three, four, five, or six of (a)-(g) of an eye of a mammal (e.g.. a human or a non-human primate) following, for example, an intravitreal administration.

[0086] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect RPE cells in vivo and to deliver an exogenous nucleic acid sequence to the infected RPE cells such that the infected RPE cells express the exogenousAttorney Docket No. 45049-0087W01 / 06839 nucleic acid sequence (e.g., at high levels). For example, an AAV vector provided herein (e.g., an AAV vector (e.g.. an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect RPE cells such that the AAV vector infects at least about 2 percent (e.g., at least about 2.5 percent, at least about 5 percent, at least about 7.5 percent, at least about 10 percent, or at least about 25 percent) of RPE cells of an eye of a mammal (e.g, a human or a non-human primate).

[0087] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g, an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect photoreceptor cells in vivo and to deliver an exogenous nucleic acid sequence to the infected photoreceptor cells such that the infected photoreceptor cells express the exogenous nucleic acid sequence. For example, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect photoreceptor cells in vivo to a greater extent than any other retinal cell type of an eye and to deliver an exogenous nucleic acid sequence to the infected photoreceptor cells such that the infected photoreceptor cells express the exogenous nucleic acid sequence to a greater extent than any other retinal cell type of an eye.

[0088] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g, an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect retinal ganglion cells in vivo and to deliver an exogenous nucleic acid sequence to the infected retinal ganglion cells such that the infected retinal ganglion cells express the exogenous nucleic acid sequence. For example, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect retinal ganglion cells in vivo to a greater extent than any other retinal cell type of an eye and to deliver an exogenous nucleic acid sequence to the infected retinal ganglion cells such that the infected retinal ganglion cells express the exogenous nucleic acid sequence to a greater extent than any other retinal cell type of an eye.Attorney Docket No. 45049-0087W01 / 06839

[0089] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect bipolar cells of the retina in vivo and to deliver an exogenous nucleic acid sequence to the infected bipolar cells such that the infected bipolar cells express the exogenous nucleic acid sequence. For example, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect bipolar cells of the retina in vivo to a greater extent than any other retinal cell type of an eye and to deliver an exogenous nucleic acid sequence to the infected bipolar cells such that the infected bipolar cells express the exogenous nucleic acid sequence to a greater extent than any other retinal cell type of an eye.

[0090] As described herein, an AAV vector provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to improve packaging efficiency. In some embodiments, an AAV vector provided herein (e.g., an AAV vector (e g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) can be used to infect cells (e.g., retinal cells) in vivo or in vitro and to deliver an exogenous nucleic acid sequence to the infected cells such that the infected cells express the exogenous nucleic acid sequence (e.g., at high levels).

[0091] In some cases, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 (or a variant thereof) or according to Formula A based on such a set forth sequence) can be used to treat a retinal condition (e.g., a retinal disease). For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects retinal cells (e.g., retinal ganglion cells, photoreceptor cells, and bipolar cells) and (b) drivesAttorney Docket No. 45049-0087W01 / 06839 expression of the delivered exogenous nucleic acid in the infected retinal cells (e.g., at high levels), thereby reducing the severity’ of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that includes an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects retinal cells (e.g., retinal ganglion cells) across at least two, three, or four different retinal regions and (b) drives expression of the delivered exogenous nucleic acid in the infected retinal cells, thereby reducing the severity of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects retinal cells of the eye and (b) drives expression of the delivered exogenous nucleic acid in the infected retinal cells, thereby reducing the severity' of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects two or more (e.g., two or more, three or more, four or more, five or more, six or more, or seven or more) different retinal cell types within an eye and (b) drives expression of the delivered exogenous nucleic acid in the infected retinal cells (e.g., at high levels), thereby reducing the severity of one or more symptoms of the retinal condition and / or slowing the progression of the retinalAttorney Docket No. 45049-0087W01 / 06839 condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects RPE cells and (b) drives expression of the delivered exogenous nucleic acid in the infected RPE cells (e.g., at high levels), thereby reducing the severity' of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g.. an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g.. a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects photoreceptor cells and (b) drives expression of the delivered exogenous nucleic acid in the infected photoreceptor cells, thereby reducing the severity of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects retinal ganglion cells and (b) drives expression of the delivered exogenous nucleic acid in the infected retinal ganglion cells, thereby reducing the severity of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encodingAttorney Docket No. 45049-0087W01 / 06839 an RNA and / or polypeptide capable of treating a retinal condition (e.g.. a retinal disease) can be administered to a mammal (e.g., a human or a non-human primate) having a retinal condition in a manner such that the AAV vector (a) infects bipolar cells of the retina and (b) drives expression of the delivered exogenous nucleic acid in the infected bipolar cells, thereby reducing the severity of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g, an AAV2 vector) provided herein (e.g.. an AAV vector (e.g, an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or anon-human primate) having a retinal condition in a manner such that the AAV vector (a) infects ON-retinal ganglion cells and (b) drives expression of the delivered exogenous nucleic acid in the infected ON-retinal ganglion cells, thereby reducing the severity’ of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g, an AAV2 vector) provided herein (e g., an AAV vector (e g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or anon-human primate) having a retinal condition in a manner such that the AAV vector (a) infects OFF-retinal ganglion cells and (b) drives expression of the delivered exogenous nucleic acid in the infected OFF-retinal ganglion cells, thereby reducing the severity' of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition. For example, an AAV vector (e.g., an AAV2 vector) provided herein (e.g., an AAV vector (e.g., an AAV2 vector) comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence) that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and / or polypeptide capable of treating a retinal condition (e.g., a retinal disease) can be administered to a mammal (e.g., a human or anon-human primate) having a retinal condition in a manner such that the AAV vector (a) infects retinal cells and (b) drives expression of the delivered exogenous nucleic acid in the infected retinal cells (e.g., at high levels), therebyAttorney Docket No. 45049-0087W01 / 06839 reducing the severity of one or more symptoms of the retinal condition and / or slowing the progression of the retinal condition.

[0092] As described herein, an AAV vector (e.g., an AAV2 vector) provided herein can be designed to include and drive expression of an exogenous nucleic acid sequence encoding any appropriate RNA of interest and / or polypeptide of interest. When an AAV vector provided herein is designed to treat a retinal condition (e.g.. a retinal disease), an exogenous nucleic acid sequence that encodes an RNA and / or polypeptide capable of treating the retinal condition can be included within the AAV vector. Examples of RNAs that can be encoded by an exogenous nucleic acid sequence designed to treat a retinal condition (e.g., a retinal disease) and designed to be included within an AAV vector have been provided herein. Examples of polypeptides that can be encoded by an exogenous nucleic acid sequence designed to treat a retinal condition (e.g., a retinal disease) and designed to be included within an AAV vector have been provided herein. In some embodiments, a polypeptide of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV vector provided herein can be a MY07A polypeptide, an USH1C polypeptide, a PCDH15 polypeptide, an USH2A polypeptide, or CLRN1 polypeptide to treat, e.g., Usher Syndrome.

[0093] Any appropriate retinal condition (e.g., a retinal disease) can be treated using an AAV vector (e.g., an AAV2 vector) provided herein (e.g.. an AAV vector comprises an AAV capsid polypeptide that comprises an amino acid sequence set forth in any one of SEQ ID NOs: 5-9 or according to Formula A based on such a set forth sequence and an exogenous nucleic acid sequence encoding a therapeutic RNA and / or polypeptide). Examples of such retinal conditions include, without limitation, Leber congenital amaurosis (LCA), Leber hereditary optic neuropathy (LHON), oculocutaneous albinism type 1 (OCA1). retinitis pigmentosa, rod / cone dystrophy, cone dystrophy, rod dystrophy, Stargardt Disease, Usher syndrome, X-linked retinitis pigmentosa (XLRP), X-linked retinoschisis (XLRS), choroideremia, achromatopsia, blue cone monochromacy, color blindness, glaucoma, optic atrophy, Batten disease, congenital stationary night blindness (CSNB), macular degeneration, CRB 1 -related retinal dystrophy, and foveal cone dystrophy.

[0094] Examples of therapeutic RNAs and polypeptides that can be delivered using an AAV vector provided herein to treat particular retinal conditions are set forth in Tables 5 and 6. Examples of genomic nucleic acids that can be inactivated and / or knocked out to treat particular retinal conditions using one or more AAV vectors provided herein that are designed to deliver gene editing components are set forth in Table 6. Examples of genomicAttorney Docket No. 45049-0087W01 / 06839 nucleic acids of disease causing alleles that can be replaced with healthy alleles to treat particular retinal conditions using one or more AAV vectors provided herein that are designed to deliver gene editing components are set forth in Table 6.Table 5: Examples of therapeutic polypeptide for treating retinal conditionsAttorney Docket No. 45049-0087W01 / 06839Table 6: Examples of polypeptide that can be expressed to treat retinal conditions, examples of polypeptides that can be knocked out to treat retinal conditions, and / orAttorney Docket No. 45049-0087W01 / 06839 examples of polypeptides that can be knocked out and replace with an alternative (e.g., wild-type or non-disease version) to treat retinal conditions.Attorney Docket No. 45049-0087W01 / 06839Attorney Docket No. 45049-0087W01 / 06839Attorney Docket No. 45049-0087W01 / 06839

[0095] In some embodiments, a retinal condition can be treated using an AAV vector provided herein that is designed to express one or more polypeptides having the ability to inhibit vascular angiogenesis. Examples of polypeptides having the ability to inhibit vascular angiogenesis that can be used as described herein include, without limitation, monoclonal anti-VEGF antibody polypeptides, angiostatin polypeptides, siRNA polypeptides, and endostatin polypeptides. In some embodiments, wet AMD can be treated using an AAV vector provided herein that is designed to express a monoclonal anti-VEGF antibody polypeptide, an angiostatin polypeptide, an siRNA, and / or endostatin polypeptide. In some embodiments, diabetic retinopathy can be treated using an AAV vector provided herein that is designed to express a monoclonal anti-VEGF antibody polypeptide, an angiostatin polypeptide, an siRNA, and / or an endostatin polypeptide. In some embodiments, diabetic macular edema can be treated using an AAV vector provided herein that is designed to express a monoclonal anti-VEGF antibody polypeptide, an angiostatin polypeptide, an siRNA, and / or an endostatin polypeptide.

[0096] In some embodiments, a retinal condition can be treated using an AAV vector provided herein that is designed to express one or more polypeptides with neuroprotective capabilities. Examples of polypeptides having the ability to provide neuroprotective activity that can be used as described herein include, without limitation, GDNF polypeptides, CNTF polypeptides, IGF-1 polypeptides, VEGF polypeptides, and BDNF polypeptides. In some embodiments, wet AMD can be treated using an AAV vector provided herein that is designed to express a GDNF polypeptide, a CNTF polypeptide, an IGF-1 polypeptide, a VEGF polypeptide, and / or a BDNF polypeptide. In some embodiments, dry AMD can be treated using an AAV vector provided herein that is designed to express a GDNF polypeptide, a CNTF polypeptide, an IGF-1 polypeptide, a VEGF polypeptide, and / or a BDNF polypeptide. In some embodiments, diabetic retinopathy can be treated using an AAV vector provided herein that is designed to express a GDNF polypeptide, a CNTF polypeptide, an IGF-1 polypeptide, a VEGF polypeptide, and / or a BDNF polypeptide. In some embodiments, diabetic macular edema can be treated using an AAV vector provided herein that is designed to express a GDNF polypeptide, a CNTF polypeptide, an IGF-1 polypeptide, a VEGF polypeptide, and / or a BDNF polypeptide.

[0097] In some embodiments, a retinal condition can be treated using an AAV vector provided herein that is designed to express one or more polypeptides having the ability to provide optogenetic capabilities. Examples of polypeptides having the ability to provide optogenetic capabilities that can be used as described herein include, without limitation, ChRAttorney Docket No. 45049-0087W01 / 06839 polypeptides. ChR2 polypeptides, ArchT polypeptides, NpHR polypeptides, and ChrimsonR polypeptides. In some embodiments, wet AMD can be treated using an AAV vector provided herein that is designed to express a ChR polypeptide, a ChR2 polypeptide, an ArchT polypeptide, a NpHR polypeptide, and / or a ChrimsonR polypeptide. In some embodiments, dry AMD can be treated using an AAV vector provided herein that is designed to express a ChR polypeptide, a ChR2 polypeptide, an ArchT polypeptide, a NpHR polypeptide, and / or a ChrimsonR polypeptide. In some embodiments, diabetic retinopathy can be treated using an AAV vector provided herein that is designed to express a ChR polypeptide, a ChR2 polypeptide, an ArchT polypeptide, a NpHR polypeptide, and / or a ChrimsonR polypeptide. In some embodiments, diabetic macular edema can be treated using an AAV vector provided herein that is designed to express a ChR polypeptide, a ChR2 polypeptide, an ArchT polypeptide, a NpHR polypeptide, and / or a ChrimsonR polypeptide.

[0098] In some embodiments, a retinal condition can be treated using an AAV vector provided herein that is designed to express one or more polypeptides having the ability to inhibit apoptosis. Examples of polypeptides having the ability to inhibit apoptosis that can be used as described herein include, without limitation, XI AP polypeptides, cl API polypeptides, C-IAP2 polypeptides, Livin polypeptides, and Survivin polypeptides. In some embodiments, wet AMD can be treated using an AAV vector provided herein that is designed to express a XIAP polypeptide, a cIAPl polypeptide, a C-IAP2 polypeptide, a Livin polypeptide, and / or a Survivin polypeptide. In some embodiments, diabetic retinopathy can be treated using an AAV vector provided herein that is designed to express a XIAP polypeptide, a cIAPl polypeptide, a C-IAP2 polypeptide, a Livin polypeptide, and / or a Survivin polypeptide. In some embodiments, diabetic macular edema can be treated using an AAV vector provided herein that is designed to express a XIAP polypeptide, a cIAPl polypeptide, a C-IAP2 polypeptide, a Livin polypeptide, and / or a Survivin polypeptide.

[0099] In some embodiments, a retinal condition can be treated using an AAV vector provided herein that is designed to express one or more polypeptides having the ability to inhibit complement. Examples of polypeptides having the ability to inhibit complement that can be used as described herein include, without limitation. Complement Factor I polypeptides. Complement factor H polypeptides, and sCD59 polypeptides. In some cases, wet AMD can be treated using an AAV vector provided herein that is designed to express a Complement Factor I polypeptide, a Complement factor H polypeptide, and / or a sCD59 polypeptide. In some cases, dry AMD can be treated using an AAV vector provided herein that is designed to express a Complement Factor I polypeptide, a Complement factor HAttorney Docket No. 45049-0087W01 / 06839 polypeptide, and / or a sCD59 polypeptide. In some embodiments, diabetic retinopathy can be treated using an AAV vector provided herein that is designed to express a Complement Factor I polypeptide, a Complement factor H polypeptide, and / or a sCD59 polypeptide. In some embodiments, diabetic macular edema can be treated using an AAV vector provided herein that is designed to express a Complement Factor I polypeptide, a Complement factor H polypeptide, and / or a sCD59 polypeptide.

[0100] In some embodiments, a retinal condition can be treated using an AAV vector provided herein that is designed to express one or more polypeptides having the ability to induce survival factors. Examples of polypeptides having the ability to induce survival factors that can be used as described herein include, without limitation, RdCVF polypeptides, RdCVFL polypeptides, HIF-1 polypeptides. IAP family polypeptides, and BCL-2 family polypeptides. In some embodiments, wet AMD can be treated using an AAV vector provided herein that is designed to express a RdCVF polypeptide, a RdCVFL polypeptide, an HIF-1 polypeptide, an IAP family polypeptide, and / or a BCL-2 family polypeptide. In some embodiments, dry AMD can be treated using an AAV vector provided herein that is designed to express a RdCVF polypeptide, a RdCVFL polypeptide, an HIF-1 polypeptide, an IAP family polypeptide, and / or a BCL-2 family polypeptide. In some embodiments, diabetic retinopathy can be treated using an AAV vector provided herein that is designed to express a RdCVF polypeptide, a RdCVFL polypeptide, an HIF-1 polypeptide, an IAP family polypeptide, and / or a BCL-2 family polypeptide. In some embodiments, diabetic macular edema can be treated using an AAV vector provided herein that is designed to express a RdCVF polypeptide, a RdCVFL polypeptide, an HIF-1 polypeptide, an IAP family polypeptide, and / or a BCL-2 family polypeptide.

[0101] A “mammal” as described herein, refers to any animal at risk for, suffering from or diagnosed for a disease or disorder (for example, any disease or disorder associated with a retina) including, but not limited to, primates and humans. In certain embodiments, the mammal is a human. In some embodiments, the human is an infant, a child, or an adult. Any appropriate method can be used to administer an AAV vector provided herein or composition (e.g.. a pharmaceutical composition) provided herein to a mammal (e.g., a human or a nonhuman primate). For example, a composition provided herein (e.g., a pharmaceutical composition containing one or more AAV vectors provided herein) can be administered to a mammal (e.g., a human or a non-human primate) intravitreally, intravenously (e.g., via an intravenous injection or infusion), subcutaneously (e.g.. via a subcutaneous injection), intraperitoneally (e.g., via an intraperitoneal injection), orally, via inhalation, intramuscularlyAttorney Docket No. 45049-0087W01 / 06839(e.g., via intramuscular injection), subretinally, intravitreally, systemically, or suprachoroidally. In some embodiments, the route and / or mode of administration of a composition (e.g., a pharmaceutical composition provided herein) can be adjusted for the mammal being treated.

[0102] In some embodiments, an effective amount of a composition containing an AAV vector provided herein (e.g., a pharmaceutical composition provided herein) to treat a retinal condition can be an amount that reduces the severity of one or more symptoms of the retinal condition and / or slows the progression of the retinal condition without producing significant toxicity7to the mammal. For example, an effective amount of an AAV vector provided herein can be from about 1 x 107viral genomes to about IxlO14viral genomes (e.g., from about 1 x 107viral genomes to about 1 x 1013viral genomes, from about 1 x 107viral genomes to about 1 x 1012viral genomes, from about 1 x 107viral genomes to about 1 x 1011viral genomes, from about 1 x 107viral genomes to about 1 x 1010viral genomes, from about 1 x 108viral genomes to about 1 x 1014viral genomes, from about 1 x 109viral genomes to about 1 x 1014viral genomes, from about 1 x 1010viral genomes to about 1 x 1014viral genomes, from about 1 x 108viral genomes to about 1 x 1012viral genomes, or from about 1 x 109viral genomes to about 1 x 1011viral genomes). In some cases, an effective amount of an AAV vector provided herein can be from about 1 x 1010viral genomes / kg of body weight to about 1 x 1014viral genomes / kg of body weight (e.g., from about 1 x 1010viral genomes / kg of body weight to about 1 x 1013viral genomes / kg of body weight, from about 1 x 1010viral genomes / kg of body weight to about 1 x 1012viral genomes / kg of body weight, from about 1 x 1010viral genomes / kg of body weight to about 1 x 1011viral genomes / kg of body weight). The effective amount can remain constant or can be adjusted as a sliding scale or variable dose depending on the mammal's response to treatment. Various factors can influence the actual effective amount used for a particular application. For example, the severity of a retinal condition, the route of administration, the age and general health condition of the mammal, excipient usage, the possibility of co-usage with other therapeutic or prophylactic treatments such as use of other retinal drugs, and the judgment of the treating physician may require an increase or decrease in the actual effective amount of a composition provided herein (e.g., a pharmaceutical composition containing an AAV vector provided herein) that is administered.

[0103] In some embodiments, an effective frequency of administration of a composition containing an AAV vector provided herein (e.g., a pharmaceutical composition provided herein) can be a frequency that reduces the severity of one or more symptoms of the retinal condition and / or slows the progression of the retinal condition without producing significantAttorney Docket No. 45049-0087W01 / 06839 toxicity to the mammal. Various factors can influence the actual effective frequency used for a particular application. For example, the severity of a retinal condition, the route of administration, the age and general health condition of the mammal, excipient usage, the possibility of co-usage with other therapeutic or prophylactic treatments such as use of other retinal drugs, and the judgment of the treating physician may require an increase or decrease in the actual effective frequency of administration of a composition provided herein.

[0104] In some embodiments, an effective duration of administration of a composition containing an AAV vector provided herein (e.g., a pharmaceutical composition provided herein) can be a duration that reduces the severity7of one or more symptoms of the retinal condition and / or slows the progression of the retinal condition without producing significant toxicity to the mammal. For example, an effective duration of administration of a pharmaceutical composition provided herein can vary from a single time point of administration to several weeks to several months (e.g., 4 to 12 weeks). In some embodiments, the duration can be for as long as the mammal is alive. Multiple factors can influence the actual effective duration used for a particular application. For example, the severity' of a retinal condition, the route of administration, the age and general health condition of the mammal, excipient usage, the possibility of co-usage with other therapeutic or prophylactic treatments such as use of other retinal drugs, and the judgment of the treating physician may require an increase or decrease in the actual effective duration of administration of a composition provided herein (e.g.. a pharmaceutical composition containing an AAV vector provided herein).

[0105] In some embodiments, an effective amount of a composition containing an AAV vector provided herein (e.g., a pharmaceutical composition provided herein) to treat a retinal condition can be administered once or twice to a mammal (e.g.. a human or a non-human primate) to treat that mammal.

[0106] The disclosure will be further described in the following examples, which do not limit the scope of any invention such as those set forth in the claims.EXAMPLESExample 1: Construction of AAV vectors comprising variant capsid polypeptides.

[0107] A low-throughput method was used to create AAV vectors with mutated capsid polypeptides and to screen those created AAV vectors for particular AAV vectors having the ability to exhibit high efficiency and / or specificity for infecting retinal cells. Briefly, a set of AAV variants were cloned and packaged individually, then pooled together. The AAVAttorney Docket No. 45049-0087W01 / 06839 variants were pooled and injected into the eyes of cynomolgus macaques non-human primates (n=3) via intravitreal injection. The AAVs were packaged with a ubiquitous CAG promoter driving expression of a green fluorescent protein (GFP) transgene. Barcodes identifying unique AAV variants were included following the GFP transgene. 30-60 days following injection, single-cell RNA-Seq was used to quantity' the expression of GFP as a metric of the performance of variants in the pool. 7m8 (SEQ ID NO: 16) and AAV2 (SEQ ID NO: 1) were included in the mixture as benchmarking controls in the screen. The performance of each variant was quantified according to the number of cells expressing the transgene. As shown in Table 7, the variant outperformed the engineered 7m8 serotype and AAV2 serotype across retinal cells in all non-human primates.

[0108] SEQ ID NO: 16 denotes VP1 of 7m8MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNGLDKG EPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTS FGGNLGRAVFQAKKR VLEPLGLVEEPVKTAPGKKRPVEHS PVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVPDPQ PLGQPPAAPSGLGTNTMATGSGAPMADNNEGADGVGNSSGNWHCDSTWMGDRVTTTSTRTWA LPTYNNHLYKQI SSQSGASNDNHYFGYSTPWGYFDFNRFHCHFS PRDWQRLINNNWGFRPKR LNFKLFNIQVKEVTQNDGTTT I NNLTSTVQVFTDSEYQLPYVLGSAHQGCLPPFPADVFMV PQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFS YTFEDVPFHSSYAHSQSLDRLM NPLIDQYLYYLSRTNTPSGTTTQSRLQFSQAGASDIRDQSRNWLPGPCYRQQRVSKTSADNN NSEYSWTGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTNVDIEKVM ITDEEEIRTTNPVATEQYGSVSTNLQRGNLALGETTRPARQAATADVNTQGVLPGMVWQDRD VYLQGPIWAKI PHTDGHFHPS PLMGGFGLKHPPPQILIKNTPVPANPSTTFSAAKFAS FITQ YSTGQVSVEIEWELQKENSKRWNPEIQYTSNYNKS INVDFTVDTNGVYSEPRPIGTRYLTRN L (SEQ ID NO: 16)Table 7: Exemplary amino acid sequence of SEQ ID NO: 5 with an exemplary linker as set forth in the insert of SEQ ID NO: 9 was inserted into an AAV capsid polypeptide at position 588. The nucleic acid sequence of SEQ ID NO: 17 encodes the amino acid sequence of SEQ ID NO: 9. Activity level denoting the performance of the exemplary amino acid sequence as compared to 7m8 (SEQ ID NO: 16) and AAV2 (SEQ ID NO: 1) is provided in the “Activity Level” column.Attorney Docket No. 45049-0087W01 / 06839* Each + sign denotes that the modified AAV2 vectors had a performance that increased by 0.51ogl0 in terms of number of retinal cells expressing transgene as compared to wild-type AAV2. “-‘‘indicates the “baseline'’ to which 7m8 and exemplary amino acid sequence were compared.Attorney Docket No. 45049-0087W01 / 06839Example 2: AAV vectors in mouse retina.

[0109] This Example compares the AAV variant and AAV controls, including engineered 7m8 (SEQ ID NO: 16) and wildtype AAV2 (SEQ ID NO: 1), and their ability to transduce retinal cells in the mouse via intravitreal injection based on fluorescent expression (Table 8). The AAV vectors were each packaged separately with CAG-mGL. The packaged variant had titer of 8.81E+12 vg / mL, the packaged 7m8 had titer of 2.61E+12 vg / mL, and packaged AAV2 had titer of 2.99 E+12 vg / mL. Approximately I pL of each virus was injected intravitreally into 3-4 C57BL / 6J mice at age P21. About 1-2 months post-injection, the mouse eyes underwent fundus imaging at low exposure (100). Figure 5 and Table 8 show that the variant has higher fluorescent expression than the 7m8 and AAV2 controls across all right eyes (RE) and left eyes (LE).Table 8* Each + sign indicates stronger fluorescent expression seen in mouse retina and is a qualitative assessment of Figure 5.OTHER EMBODIMENTS

[0110] It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims.

Claims

Attorney Docket No. 45049-0087W01 / 06839WHAT IS CLAIMED IS:1 . An adeno-associated virus (AAV) vector comprising an AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising that of any one of SEQ ID NOs: 5-9.

2. An adeno-associated virus (AAV) vector comprising an AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising that of SEQ ID NO: 5 (HDQVRP).

3. The AAV vector of claim 1 or 2. wherein said AAV capsid polypeptide has an amino acid sequence comprising that of any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 581 and 582, between amino acid 582 and 583, between positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587. between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4.

4. The AAV vector of any one of claims 1-3, wherein said AAV capsid polypeptide has an amino acid sequence comprising that of any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4.

5. The AAV vector of any one of claims 1-4, wherein said AAV capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9.

6. The AAV vector of any one of claims 1-5, wherein said vector is an AAV2 vector.Attorney Docket No. 45049-0087W01 / 068397. The AAV vector of any one of claims 1-6, wherein said vector further comprises an exogenous nucleic acid encoding an RNA or a therapeutic polypeptide.

8. The AAV vector of claim 7, wherein said exogenous nucleic acid encodes an RNA.

9. The AAV vector of claim 8, wherein said RNA is an siRNA or microRNA.

10. The AAV vector of claim 7, wherein said exogenous nucleic acid encodes a therapeutic polypeptide.

11. The AAV vector of claim 10, wherein said therapeutic polypeptide is a VEGF polypeptide, a RSI polypeptide, a PRPF31 polypeptide, an ABCA4 polypeptide, a CRB1 polypeptide, an NPHP5 polypeptide, or an NR2E3 polypeptide.

12. The AAV vector of any one of claims 1-11, wherein AAV capsid polypeptide has an amino acid sequence comprising SEQ ID NO: 4 except that the amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of SEQ ID NO: 4.

13. An AAV capsid polypeptide having an amino acid sequence comprising any one of SEQ ID NOs: 5-9.

14. The AAV capsid polypeptide of claim 13, wherein said polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 581 and 582, between amino acid 582 and 583, between positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4.

15. The polypeptide of claim 13 or 14. wherein said polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence ofAttorney Docket No. 45049-0087W01 / 06839 any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4.

16. The polypeptide of claim 13 or 14, wherein said polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9.

17. The polypeptide of any one of claims 13-16, wherein said polypeptide has an amino acid sequence comprising SEQ ID NO: 4 except that the amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of SEQ ID NO: 4.

18. A nucleic acid molecule encoding the AAV vector of any one of claims 1-12, or the polypeptide of any one of claims 13-17.

19. The nucleic acid molecule of claim 18, wherein said nucleic acid molecule is DNA.

20. A host cell comprising the nucleic acid molecule of claim 18 or 19.

21. The host cell of claim 20, wherein said host cell expresses the AAV vector of any one of claims 1 -12.

22. The host cell of claim 20, wherein said host cell expresses the polypeptide of any one of claims 13-17.

23. A host cell comprising the AAV vector of any one of claims 1-12 or the polypeptide of any one of claims 13-17.

24. The host cell of any one of claims 20-23, wherein said host cell is a retinal cell.

25. A composition comprising the vector of any one of claims 1-12, and a pharmaceutically acceptable excipient.Attorney Docket No. 45049-0087W01 / 0683926. The composition of claim 25, wherein said composition comprises from about 1 x 107vector genomes / mL (vg / mL) to about 1 x 1014vg / mL of said vector.

27. The composition of claim 25 or 26, wherein said pharmaceutically acceptable excipient comprises one or more of: phosphate buffered saline, Hank’s Balanced Salt Solution, and Pluronic F68.

28. A method for delivering an exogenous nucleic acid sequence to a retinal cell of a mammal, wherein said method comprises contacting said retinal cell with an AAV vector comprising an AAV capsid polypeptide and said exogenous nucleic acid sequence, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 5-9, wherein said AAV vector infects said retinal cell, thereby delivering said exogenous nucleic acid sequence to said retinal cell.

29. The method of claim 28, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4.

30. The method of claim 28. wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9.

31. The method of any one of claims 28-30, wherein said mammal is a human.

32. The method of any one of claims 28-31, wherein said vector is an AAV2 vector.

33. The method of any one of claims 28-32. wherein said exogenous nucleic acid sequence encodes an RNA or a polypeptide.

34. The method of claim 33, wherein said exogenous nucleic acid encodes an RNA.

35. The method of claim 34, wherein said RNA is an siRNA or microRNA.Attorney Docket No. 45049-0087W01 / 0683936. The method of claim 33, wherein said exogenous nucleic acid encodes a polypeptide.

37. The method of claim 36, wherein said polypeptide is a VEGF polypeptide, a RSI polypeptide, a PRPF31 polypeptide, an ABCA4 polypeptide, a CRB1 polypeptide, an NPHP5 polypeptide, or an NR2E3 polypeptide.

38. The method of any one of claims 28-37, wherein said method comprises intravitreally administering a composition comprising said vector to said mammal, thereby contacting said retinal cell with said vector.

39. The method of claim 38, wherein said composition comprises from about 1 x 107vector genomes / mL (vg / mL) to about 1 x 1014vg / mL of said vector.

40. A method for treating a retinal condition in a mammal in need thereof, wherein said method comprises contacting a retinal cell of a mammal having said retinal condition with a therapeutically effective amount of an AAV vector comprising an AAV capsid polypeptide and an exogenous nucleic acid sequence, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 5-9, wherein said AAV vector infects said retinal cell and drive expression of said exogenous nucleic acid sequence within said retinal cell, thereby treating said retinal condition.

41. The method of claim 40, wherein said mammal is a human.

42. The method of any one of claims 40-41, wherein said retinal condition is selected from the group consisting of cone dystrophy, cone / rod dystrophy, retinitis pigmentosa, macular degeneration, achromatopsia, blue cone monochromacy, and color blindness.

43. The method of any one of claims 40-42. wherein said polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 581 and 582, between amino acid 582 and 583, between positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589,Attorney Docket No. 45049-0087W01 / 06839 between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between amino acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4.

44. The method of any one of claims 40-43, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that said amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4.

45. The method of any one of claims 40-43, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 except that the amino acids from position 585 to 590 of any one of SEQ ID NOs: 1-4 are replaced with said amino acid sequence of any one of SEQ ID NOs: 5-9.

46. The method of any one of claims 40-45, wherein said capsid polypeptide has an amino acid sequence comprising SEQ ID NO: 4 except that the amino acid sequence of any one of SEQ ID NOs: 5-9 is located between amino acid positions 587 and 588 of SEQ ID NO: 4.

47. The method of any one of claims 40-46. wherein said vector is an AAV2 vector.

48. The method of any one of claims 40-47, wherein said exogenous nucleic acid sequence encodes an RNA.

49. The method of claim 48, wherein said RNA is an siRNA or a microRNA.

50. The method of any one of claims 40-47, wherein said exogenous nucleic acid encodes a polypeptide.

51. The method of claim 50, wherein said polypeptide is a VEGF polypeptide, a RSI polypeptide, a PRPF31 polypeptide, an ABCA4 polypeptide, a CRB1 polypeptide, an NPHP5 polypeptide, or an NR2E3 polypeptide.Attorney Docket No. 45049-0087W01 / 0683952. The method of any one of claims 40-51, wherein said method comprises intravitreally administering a composition comprising said vector to said mammal, thereby contacting said retinal cell with said vector.

53. The method of claim 52, wherein said composition comprises from about 1 x 107vector genomes / mL (vg / mL) to about 1 x 1014vg / mL of said vectors.

54. A non-naturally occurring AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 581 and 582, between amino acid positions 582 and 583, between amino acid positions 583 and 584, between amino acid positions 584 and 585, between amino acid positions 585 and 586, between amino acid positions 586 and 587, between amino acid positions 587 and 588, between amino acid positions 588 and 589, between amino acid positions 589 and 590, between amino acid positions 590 and 591, between amino acid positions 591 and 592, between ammo acid positions 592 and 593, or between amino acid positions 593 and 594 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-Li-[6-mer polypeptide] -L2-, wherein said Li and said L2 are each, independently, optional amino acid linkers, each having one, two. or three amino acids, and wherein the 6-mer polypeptide has an amino acid sequence of that of SEQ ID NO: 5.

55. A non-naturally occurring AAV capsid polypeptide, wherein said capsid polypeptide has an amino acid sequence comprising any one of SEQ ID NOs: 1-4 comprising an amino acid sequence insert of Formula A located between amino acid positions 587 and 588 of any one of SEQ ID NOs: 1-4, wherein said Formula A is:-L1-INSERT-L2-, wherein said Li and said L2 are each independently optional amino acid linkers having one, two. or three amino acids, and wherein INSERT represents the amino acid sequence of that of SEQ ID NO: 5.

56. The capsid polypeptide of claim 54 or 55, wherein said Li is one amino acid Xi.Attorney Docket No. 45049-0087W01 / 0683957. The capsid polypeptide of claim 56, wherein said Xi is selected from the group of amino acid residues consisting of A. V, I. and L.

58. The capsid polypeptide of claim 56, wherein said Xi is A.

59. The capsid polypeptide of claim 54 or 55, wherein said Li is two amino acids X2-X1.

60. The capsid polypeptide of claim 59, wherein said Xi is selected from the group of amino acid residues consisting of A, V, I, and L.

61. The capsid polypeptide of claim 60, wherein said Xi is A.

62. The capsid polypeptide of any one of claims 59-61, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L.

63. The capsid polypeptide of claim 62, wherein said X2 is L.

64. The capsid polypeptide of claim 59, wherein said X2-X1 is LA.

65. The capsid polypeptide of claim 54 or 55, wherein said Li is three amino acids X3-X2-Xi.

66. The capsid polypeptide of claim 65, wherein said Xi is selected from the group of amino acid residues consisting of A. V, I, and L.

67. The capsid polypeptide of claim 66, wherein said Xi is A.

68. The capsid polypeptide of any one of claims 65-67, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L.

69. The capsid polypeptide of claim 68, wherein said X2 is L.

70. The capsid polypeptide of claim 65, wherein said X2-X1 is LA.Attorney Docket No. 45049-0087W01 / 0683971. The capsid polypeptide of any one of claims 65-70, wherein said X3 is selected from the group of amino acid residues consisting of A, V, I, and L.

72. The capsid polypeptide of claim 54 or 55, wherein said Li is absent.

73. The capsid polypeptide of any one of claims 54-72, wherein said L2 is one amino acid Zi.

74. The capsid polypeptide of claim 73, wherein said Zi is selected from the group of amino acid residues consisting of A, V, I, and L.

75. The capsid polypeptide of claim 74, wherein said Zi is A.

76. The capsid polypeptide of any one of claims 54-72, wherein said L2is two amino acids Z1-Z2.

77. The capsid polypeptide of claim 76, wherein said Zi is selected from the group of amino acid residues consisting of A, V, I, and L.

78. The capsid polypeptide of claim 77, wherein said Zi is A.

79. The capsid polypeptide of any one of claims 76-78, wherein said Z2 is selected from the group of amino acid residues consisting of A, V, I, and L.

80. The capsid polypeptide of claim 79, wherein said Z2 is L.

81. The capsid polypeptide of claim 76, wherein said Z1-Z2 is AL.

82. The capsid polypeptide of any one of claims 54-72, wherein said L2 is three amino acids TA-L' i-L' i.

83. The capsid polypeptide of claim 82, wherein said Zi is selected from the group of amino acid residues consisting of A. V, I, and L.Attorney Docket No. 45049-0087W01 / 0683984. The capsid polypeptide of claim 83, wherein said Zi is A.

85. The capsid polypeptide of any one of claims 82-84, wherein said Z2 is selected from the group of amino acid residues consisting of A, V, I, and L.

86. The capsid polypeptide of claim 85, wherein said Z2 is L.

87. The capsid polypeptide of claim 82, wherein said Z1-Z2 is AL.

88. The capsid polypeptide of any one of claims 82-87, wherein said Z3 is selected from the group of amino acid residues consisting of A, V, I, and L.

89. The capsid polypeptide of any one of claims 54-72, wherein said L2 is absent.

90. A non-naturally occurring adeno-associated virus (AAV) vector comprising an AAV capsid polypeptide according to any one of claims 54-89.

91. A viral particle comprising a capsid polypeptide of any one of claims 54-89.

92. A method for administering an exogenous nucleic acid sequence to a mammal in need thereof, wherein said method comprises administering an effective amount of a vector of claim 90 to said mammal, wherein said vector comprising said exogenous nucleic acid sequence.

93. The method of claim 92, wherein said mammal is a human.

94. The method of claim 92 or 93, wherein said administering comprises administering said effective amount to an eye of said mammal.

95. The method of any one of claims 92-94, wherein said administering is sufficient to allow for expression of said exogenous nucleic acid sequence in a cell of said mammal.

96. The method of any one of claims 92-95, wherein said exogenous nucleic acid sequence encodes a therapeutic polypeptide.Attorney Docket No. 45049-0087W01 / 0683997. A method of treating a retinal disorder in a patient in need thereof, comprising administering to the patient’s eye an effective amount of an AAV vector, wherein the AAV vector comprises an AAV capsid polypeptide and an exogenous nucleic acid sequence, wherein the AAV capsid polypeptide is represented by Formula A.

98. A method of treating a retinal disorder in a patient in need thereof, comprising administering to the patient’s eye an effective amount of an AAV vector, wherein the AAV vector comprises an AAV capsid polypeptide and an exogenous nucleic acid sequence, wherein the AAV capsid polypeptide comprises an amino acid sequence insert of Formula A, wherein said Formula A is:-Li -[6-mer polypeptide]-L2-, wherein said Li and said L2 are each, independently, optional amino acid linkers, each having one, two, or three amino acids, and wherein the 6-mer polypeptide has an amino acid sequence of that of SEQ ID NO: 5.

99. A method of treating a retinal disorder in a patient in need thereof, comprising administering to the patient’s eye an effective amount of an AAV vector, wherein the AAV vector comprises an AAV capsid polypeptide of any one of claims 54-89 and an exogenous nucleic acid sequence.

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