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WO2025072698A2 - Aav9 vectors and capsid polypeptides for use in treating cns disorders - Google Patents

Aav9 vectors and capsid polypeptides for use in treating cns disorders Download PDF

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Publication number
WO2025072698A2
WO2025072698A2 PCT/US2024/048892 US2024048892W WO2025072698A2 WO 2025072698 A2 WO2025072698 A2 WO 2025072698A2 US 2024048892 W US2024048892 W US 2024048892W WO 2025072698 A2 WO2025072698 A2 WO 2025072698A2
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polypeptide
amino acid
vector
seq
acid positions
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WO2025072698A3 (en
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Leah Caroline THOMAS BYRNE
Molly E. JOHNSON
William Richard Stauffer
Bilge Esin OZTURK
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University of Pittsburgh
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University of Pittsburgh
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14122—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2750/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssDNA viruses
    • C12N2750/00011—Details
    • C12N2750/14011—Parvoviridae
    • C12N2750/14111—Dependovirus, e.g. adenoassociated viruses
    • C12N2750/14141—Use of virus, viral particle or viral elements as a vector
    • C12N2750/14145—Special targeting system for viral vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2810/00—Vectors comprising a targeting moiety
    • C12N2810/40—Vectors comprising a peptide as targeting moiety, e.g. a synthetic peptide, from undefined source

Definitions

  • Viral vectors such as AAV9 vectors, are efficient vehicles for in vivo nucleic acid delivery, and their use in the clinic is expanding. Improved AAV9 vectors and AAV9 production techniques for making effective AAV9 vector preparations should further expand the use of AAV9 vectors in the laboratory and clinic.
  • SUMMARY OF THE DISCLOSURE provides AAV9 vectors. For example, this document provides AAV9 vectors containing a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • the AAV9 vectors described herein containing a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B can have the ability to infect CNS cells in vivo and deliver exogenous nucleic acid to the infected CNS cells such that the infected CNS cells express the exogenous nucleic acid.
  • This document also provides methods and materials for making and using AAV9 vectors having the ability to deliver nucleic acid to CNS cells.
  • this document provides an adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide having 95% identity or more with SEQ ID NO:1, or a truncated variant thereof, further comprising an amino acid peptide sequence comprising a sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14, wherein the amino acid peptide sequence is located between two existing amino acids of SEQ ID NO:1, e.g., between amino acid positions 588 and 589 (or e.g., between 585 and 590) of SEQ ID NO:1.
  • AAV9 vector comprising an AAV9 capsid polypeptide having 95% identity or more with SEQ ID NO:1, or a truncated variant thereof, further comprising an amino acid peptide sequence comprising a sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14, wherein the amino acid peptide sequence is located between two existing amino acids of SEQ ID NO:1, e.g., between amino
  • Such organisms preferably include, but are not limited to, mammals (e.g., murines, simians, equines, bovines, porcines, primates, canines, felines, and the like), and more preferably include humans.
  • mammals e.g., murines, simians, equines, bovines, porcines, primates, canines, felines, and the like
  • the terms “treat,” “treating,” or “treatment,” and other grammatical equivalents as used in this document include alleviating, abating, ameliorating, or preventing a disease, condition or symptoms, preventing additional symptoms, ameliorating or preventing the underlying metabolic causes of symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition, and are intended to include prophylaxis.
  • the terms further include achieving a therapeutic benefit and/or a prophylactic benefit.
  • therapeutic benefit refers to eradication or amelioration of the underlying disorder being treated.
  • a therapeutic Attorney Docket No.: 45049-0091WO1 / 06521 benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient may still be afflicted with the underlying disorder.
  • the term “effective amount” refers to the amount of a compound (e.g., an AAV9 vector described herein) sufficient to effect beneficial or desired results.
  • an effective amount can be administered in one or more administrations, applications, or dosages and is not intended to be limited to a particular formulation or administration route.
  • the term “combination” in the context of therapies means that two or more different treatments are delivered to the subject during the course of the subject’s affliction with the disorder, such that the effects of the treatments on the patient overlap at a point in time.
  • the delivery of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as “simultaneous” or “concurrent delivery.”
  • the delivery of one treatment ends before the delivery of the other treatment begins.
  • the treatment is more effective because of combined administration.
  • the second treatment is more effective, e.g., an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in the absence of the first treatment, or the analogous situation is seen with the first treatment.
  • delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other.
  • the effect of the two treatments can be partially additive, wholly additive, or greater than additive.
  • the delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered.
  • the term “about” refers to any minimal alteration in the concentration or amount of an agent that does not change the efficacy of the agent in preparation of a formulation and in treatment of a disease or disorder. In certain embodiments, the term “about” may include ⁇ 5%, ⁇ 10%, or ⁇ 15% of a specified numerical value or data point. [0015] Ranges can be expressed in this document as from “about” one particular value, and/or to “about” another particular value. When such a range is expressed, another aspect includes from the one particular value and/or to the other particular value.
  • compositions are described as having, including, containing, incorporating, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is intended that compositions and methods are inclusive or open-ended and do not exclude additional, unrecited components or steps.
  • compositions of the present document that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present document that consist essentially of, or consist of, the recited steps.
  • compositions specifying a percentage are by weight unless otherwise specified. Further, if a variable is not accompanied by a definition, then the previous definition of the variable controls.
  • AAV9 Vectors [0018] The present document provides AAV9 vectors.
  • AAV9 vectors containing a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B can be designed to include a capsid polypeptide described herein (e.g., a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set Attorney Docket No.: 45049-0091WO1 / 06521 forth in Table 1B (or a variant thereof) or Formula B).
  • an AAV9 having an ACG start codon for the AAV9 Rep polypeptides instead of an ATG start codon can be designed to include a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • Any appropriate AAV9 capsid polypeptide can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • an AAV9 capsid polypeptide can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • an AAV9 capsid polypeptide having the following amino acid sequence can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B: MAADGYLPDW LEDNLSEGIR EWWALKPGAP QPKANQQHQD NARGLVLPGY KYLGPGNGLD KGEPVNAADA AALEHDKAYD QQLKAGDNPY LKYNHADAEF QERLKEDTSF GGNLGRAVFQ AKKRLLEPLG LVEEAAKTAP GKKRPVEQSP QEPDSSAGIG KSGAQPAKKR LNFGQTGDTE SVPDPQPIGE PPAAPSGVGS LTMASGGGAP VA
  • an AAV9 capsid polypeptide 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 can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, Attorney Docket No.: 45049-0091WO1 / 06521 e.g., a contemplated AAV9 capside polypeptide is at least 95% identical to SEQ ID NO:1, and/or may include 1, 2, 3, or more amino acid truncations (e.g., on one or both ends), and/or deletions.
  • an AAV9 capsid polypeptide to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, that included amino acid sequence can be located at any appropriate location along the AAV9 capsid polypeptide.
  • an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A such as any one of SEQ ID NOs:2-5 or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B such as any one of SEQ ID NOs:11-14 can be located between the naturally-occurring amino acid residues at positions 588 and 589 of an AAV9 capsid polypeptide or can be located between two naturally-occurring amino acid residues at other positions of e.g., SEQ ID NO:1.
  • an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A such as any one of SEQ ID NOs:2-5 or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B such as any one of SEQ ID NOs:11-14 can be located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of an AAV capsid polypeptide (e.g., an AAV9 capsid polypeptide such as an AAV9 capsid polypeptide comprising the amino acid sequence set forth in SEQ ID NO
  • an AAV9 vector can be designed to have an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • Table 1A Amino acid sequences that can be inserted into an AAV9 capsid polypeptide.
  • Formula A can be as follows: -L1-EGSGRN (SEQ ID NO:2)-L2-, wherein L1 and L2 are each independently optional amino acid linkers having one, two, or three amino acids.
  • L1, L2, or both L1 and L2 can be absent.
  • L1 can be one amino acid X1, two amino acids X2-X1, or three amino acids X3-X2-X1.
  • X1 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • X2 When X2 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • X3 When X3 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • L2 can be one amino acid Z1, two amino acids Z1-Z2, or three amino acids Z1-Z2-Z3.
  • Z1 When Z1 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • Z2 When Z2 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • Z3 When Z3 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • L1 linkers examples include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, LIV, LII, LIL, LLA, LLV, LLI, and LLL.
  • Examples of an Attorney Docket No.: 45049-0091WO1 / 06521 L2 linkers include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, L
  • an AAV9 vector can be designed to have an AAV9 capsid polypeptide of SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) that includes an amino acid sequence insert of Formula B located between amino acid positions 588 and 589 of SEQ ID NO:1 (or the appropriate amino acid positions of the alternative sequence).
  • Formula B can be as follows: -L1- EHQTRP (SEQ ID NO:11)-L2-, wherein L1 and L2 are each independently optional amino acid linkers having one, two, or three amino acids. In some cases, L1, L2, or both L1 and L2 can be absent.
  • L1 can be one amino acid X1, two amino acids X2-X1, or three amino acids X3-X2-X1.
  • X1 can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • X2 When X is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • X3 When X3 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • L2 can be one amino acid Z1, two amino acids Z1-Z2, or three amino acids Z1-Z2-Z3.
  • Z1 is present, it can be an amino acid residue selected from the group consisting of A, V, I, and L.
  • an L1 linkers include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILV
  • L2 linkers examples include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, Attorney Docket No.: 45049-0091WO1 / 06521 ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, LIV
  • an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B inserted between glutamine-588 and alanine-589 (or the appropriate amino acid positions of an alternative sequence).
  • an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with an amino acid sequence set forth in any one of SEQ ID NOs:2-5 (or a variant thereof) inserted between glutamine-588 and alanine-589 (or the appropriate amino acid positions of the alternative sequence).
  • an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with an amino acid sequence set forth in any one of SEQ ID NOs:11-14 (or a variant thereof) inserted between glutamine-588 and alanine-589 (or the appropriate amino acid positions of the alternative sequence).
  • an AAV9 capsid polypeptide when designing an AAV9 capsid polypeptide to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, that included amino acid sequence can be used to replace one or more naturally-occurring amino acid residues located at any appropriate location along the AAV9 capsid polypeptide.
  • an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A such as any one of SEQ ID NOs:2-5 can be used to replace the naturally-occurring amino acid residues at positions 585 to 590 of an AAV9 capsid polypeptide.
  • an amino acid sequence set forth in Table 1B (or a variant thereof) or Attorney Docket No.: 45049-0091WO1 / 06521 Formula B such as any one of SEQ ID NOs:11-14 can be used to replace the naturally-occurring amino acid residues at positions 585 to 590 of an AAV9 capsid polypeptide.
  • an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in 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 set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with the exception that amino acid residues 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with the amino acid sequence set forth in any one of SEQ ID NOs:2-5 (or a variant thereof).
  • an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with the exception that amino acid residues 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with the amino acid sequence set forth in any one of SEQ ID NOs:11-14 (or a variant thereof).
  • an AAV9 capsid polypeptide can be designed to include two or more amino acid sequences set forth in Table 1A (or a variant thereof) or Formula A or two or more amino acid sequences set forth in Table 1B (or a variant thereof) or Formula B.
  • an AAV9 capsid polypeptide can be designed to include two or three amino acid sequences set forth in Table 1A (or a variant thereof) or Formula A.
  • an AAV9 capsid polypeptide can be designed to include two or three amino acid sequences set forth in Table 1B (or a variant thereof) or Formula B.
  • an AAV9 capsid polypeptide can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • a variant of an amino acid sequence set forth in Table 1A refers to an amino acid sequence that is identical to that amino acid sequence Attorney Docket No.: 45049-0091WO1 / 06521 set forth in Table 1A except that it has one, two, or three amino acid additions, deletions, substitutions, or combinations thereof.
  • a variant of SEQ ID NO:2 can be SEQ ID NO:2 except that it has one, two, or three amino acid additions, deletions, substitutions, or combinations thereof.
  • a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:2-5 except that it contains one, two, or three amino acid additions. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:2-5 except that it contains one, two, or three amino acid deletions. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:2-5 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:2-5 except that it contains one amino acid addition, deletion, or substitution.
  • a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:2-5 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:2-5 except that it contains three amino acid additions, deletions, substitutions, or a combination thereof. [0029] A variant of an amino acid sequence set forth in Table 1B refers to an amino acid sequence that is identical to that amino acid sequence set forth in Table 1B except that it has one, two, or three amino acid additions, deletions, substitutions, or combinations thereof.
  • a variant of SEQ ID NO:11 can be SEQ ID NO:11 except that it has one, two, or three amino acid additions, deletions, substitutions, or combinations thereof.
  • a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 except that it contains one, two, or three amino acid additions.
  • a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 except that it contains one, two, or three amino acid deletions.
  • a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 except that it contains one, two, or three amino acid substitutions.
  • a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 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:11-14 except that it contains two amino acid additions, deletions, substitutions, or a combination thereof. In some cases, a variant provided Attorney Docket No.: 45049-0091WO1 / 06521 herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 except that it contains three amino acid additions, deletions, substitutions, or a combination thereof.
  • an amino acid substitution present in a variant can be a conservative amino acid substitution.
  • conservative 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, pheny
  • an amino acid substitution present in a variant can be a non- conservative 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 aresidue 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
  • 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 (Bl2seq) 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).
  • Bl2seq performs a comparison between two sequences using either the BLASTN or BLASTP algorithm.
  • BLASTN is used to compare nucleic acid sequences
  • BLASTP is used to compare amino acid sequences.
  • the following command can be used to generate an output file containing a comparison between two amino acid sequences: C: ⁇ Bl2seq -i c: ⁇ seq1.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.
  • 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 AAV9 capsid polypeptide. See, for example, Zoller, Curr. Opin. Biotechnol.3: 348-354 (1992).
  • the AAV9 vectors described herein can be designed to include one or more exogenous nucleic acid sequences.
  • an AAV9 vector described herein can be designed to include 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.
  • RNAs of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV9 vector provided herein include, without limitation, siRNAs, RNA components for gene editing, and microRNAs.
  • an RNA of interest that can be encoded by an exogenous nucleic acid sequence included within Attorney Docket No.: 45049-0091WO1 / 06521 an AAV9 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., diabetic 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
  • An exogenous nucleic acid sequence can be designed to encode any appropriate polypeptide of interest.
  • polypeptides of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV9 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.
  • therapeutic polypeptides e.g., trophic factor polypeptides, gene editing polypeptides (e.g., a Cas9 polypeptide, a TALEN polypeptide, or
  • polypeptides of interest that can be encoded by an exogenous nucleic acid sequence designed to be included within an AAV9 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 RS1 polypeptide, a TYR polypeptide, a USH2A polypeptide, a MYO7A polypeptide, an REP1 polypeptide, an OPN1LW polypeptide, an OPN1MW polypeptide, a CNGA3 polypeptide, a CNGB3 polypeptide, a GUCY2D polypeptide, a GACA1A polypeptide, a GNAT2 polypeptide, a PDE6
  • one or more AAV9 vectors provided herein can be designed to carry out gene editing within one or more cells (e.g., CNS cells). Such gene editing can 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.
  • 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
  • nucleic acid e.g., nucleic acid encoding
  • any appropriate gene editing components can be engineered into one or more AAV9 vectors provided herein such that those one or more AAV9 vectors can be used to deliver the gene editing components to target cells (e.g., one or more CNS cells) within a mammal (e.g., a human or a non-human primate) in a manner effective to edit the genome of those cells.
  • 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.
  • rare-cutting endonucleases include, without limitation, meganucleases, transcription activator- like effector (TALE) nucleases (TALENsTM; Cellectis, Paris, France), zinc-finger-nucleases (ZFNs), and endonucleases of a clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system (e.g., endonucleases of a CRISPR/Cas 9 system).
  • TALE transcription activator- like effector
  • ZFNs zinc-finger-nucleases
  • CRISPR clustered regularly interspaced short palindromic repeats
  • CRISPR clustered regularly interspaced short palindromic repeats
  • 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)).
  • two sequences in genomic nucleic acid of a cell e.g., a CNS cell
  • 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.
  • An AAV9 vector 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 the sequence between the genomic targets, and insertion of the donor sequence into the location of the deletion.
  • An AAV9 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.
  • 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, SV40 promotors, CMV promotors, 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 promotors that can be used as described herein include, without limitation, rhodopsin promotors, cone arrestin promotors, and synapsin promotors.
  • Examples of viral promotors that can be used as described herein include, without limitation, adenoviral promotors, vaccinia virus promotors, CMV promotors (e.g., immediate early CMV promotors), and AAV promoters.
  • an AAV9 vector provided herein can include a total number of nucleotides up to about 5 kb. In some cases, an AAV9 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.
  • An AAV9 vector described herein containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B can have the ability to Attorney Docket No.: 45049-0091WO1 / 06521 infect CNS cells in vivo and deliver exogenous nucleic acid sequence to the infected CNS cells such that the infected CNS cells express the exogenous nucleic acid sequence.
  • an AAV9 vector provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in more CNS cells of a mammal (e.g., a human or a non-human primate) when compared to wild- type AAV9.
  • a mammal e.g., a human or a non-human primate
  • an AAV9 vector provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in CNS 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 AAV9 vector (e.g., wild-type AAV9) having an AAV9 capsid polypeptide that consists of the amino acid sequence set forth in SEQ ID NO:1 in CNS cells of a control mammal (e.g., a control human or a control non-human primate).
  • a control AAV9 vector e.g., wild-type AAV9 having an AAV9 capsid polypeptide that consists of the amino acid sequence set forth in SEQ ID NO:1 in CNS cells of a control mammal (e.g., a control human or a control non-human primate).
  • Examples of CNS cells that can be infected by an AAV9 vector described herein containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B include, without limitation, CNS cell types such as neurons, glial cells (e.g., astrocytes, oligodendrocytes, ependymal cells, and microglia), choroid plexus cells, and cells related to blood vessels and coverings.
  • This document also provides compositions containing one or more AAV9 vectors provided herein.
  • one or more AAV9 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.
  • a mammal e.g., a human or a non-human primate
  • one or more AAV9 vectors provided 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 CNS cells for expression within CNS cells.
  • an AAV9 vector provided herein can be formulated as a pharmaceutical composition for administration to a mammal (e.g. a human or a non-human primate).
  • 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 (2016)).
  • 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 (2016)).
  • 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, Attorney Docket No.: 45049-0091WO1 / 06521 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.
  • a surfactant e.g., polysorbate 20, polysorbate 80, or poloxamer 188
  • dextran 40 e.g., sorbitol, mannitol, sucrose, dextrose, or trehalose
  • a pharmaceutical composition designed to include an AAV9 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.
  • 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
  • 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.
  • any appropriate titer of the AAV9 vectors can be used.
  • a pharmaceutical composition provided herein can be formulated to have AAV9 vectors provided herein at a titer that is greater than 1x10 7 (e.g., greater than 1 x 10 8 , greater than 1 x 10 9 , greater than 1 x 10 10 , greater than 1 x 10 11 , greater than 1 x 10 12 , greater than 1 x 10 13 , or greater than 1 x 10 14 ).
  • a pharmaceutical composition provided herein can be formulated to have AAV9 vectors (provided herein at a titer that is from about 1x10 7 to about 1x10 14 (e.g., from about 1 x 10 7 to about 1 x 10 13 , from about 1 x 10 7 to about 1 x 10 12 , from about 1 x 10 7 to about 1 x 10 11 , from about 1 x 10 7 to about 1 x 10 10 , from about 1 x 10 8 to about 1 x 10 14 , from about 1 x 10 9 to about 1 x 10 14 , from about 1 x 10 10 to about 1 x 10 14 , from about 1 x 10 8 to about 1 x 10 12 , or from about 1 x 10 9 to about 1 x 10 11 ).
  • AAV9 vectors provided herein at a titer that is from about 1x10 7 to about 1x10 14 (e.g., from about 1 x 10 7 to about 1 x 10 13 , from about 1 x 10
  • a pharmaceutical composition provided herein can be in any appropriate form.
  • a pharmaceutical composition provided herein can be designed to be a liquid, a semi- solid, or a solid.
  • 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.
  • a pharmaceutical composition provided herein can be lyophilized.
  • a pharmaceutical composition provided herein can be formulated with a carrier or coating designed to protect against rapid release.
  • 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).
  • This document also provides nucleic acid molecules encoding an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • a nucleic acid molecule can be designed to encode an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1A (e.g., any one of SEQ ID NOs:6-9).
  • a nucleic acid molecule can be designed to encode an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1B (e.g., any one of SEQ ID NOs:15-18).
  • This document also provides nucleic acid molecules encoding an AAV9 vector described herein.
  • an isolated nucleic acid molecule can be designed to encode one or more AAV9 vectors provided herein (e.g., an AAV9 having an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B).
  • a nucleic acid molecule can be designed to encode an AAV9 vector having an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1A (e.g., any one of SEQ ID NOs:6-9).
  • a nucleic acid molecule can be designed to encode an AAV9 vector having an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1B (e.g., any one of SEQ ID NOs:15-18).
  • This document also provides host cells containing a nucleic acid molecule provided herein.
  • a host cell can be designed to include a nucleic acid molecule encoding an AAV9 capsid polypeptide described herein and/or a nucleic acid molecule encoding an AAV9 vector described herein.
  • a host cell can be designed to include a nucleic acid molecule encoding an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • a host cell can be designed to include a nucleic acid molecule encoding an AAV9 vector having an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • Examples of host cells that can be designed to include a nucleic acid molecule encoding an AAV9 capsid polypeptide described herein and/or a nucleic acid molecule encoding an AAV9 vector described Attorney Docket No.: 45049-0091WO1 / 06521 herein include, without limitation, HEK293T cells (ATCC), 293AAV9 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 AAV9 capsid polypeptide described herein and/or an AAV9 vector described herein) into a cell.
  • This document also provides methods and materials for making an AAV9 provided herein.
  • this document provides methods and materials for making AAV9 vectors containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • an AAV9 vector can be constructed to include an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. Any appropriate method can be used to construct an AAV9 vector having an AAV9 capsid polypeptide provided herein (e.g., a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B).
  • molecular cloning and AAV9 vector production techniques such as those described elsewhere can be used to construct and produce an AAV9 vector having an AAV9 capsid polypeptide provided herein (see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory, NY (1989); Ausubel et 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)).
  • AAV9 vectors can be produced in HEK293T cells (ATCC) or 293AAV9 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)).
  • This document also provides methods and materials for using an AAV9 vector provided herein.
  • this document provides methods and materials for using AAV9 vectors containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B.
  • an AAV9 vector provided herein can be used Attorney Docket No.: 45049-0091WO1 / 06521 to infect CNS cells in vivo and to deliver an exogenous nucleic acid sequence to the infected CNS cells such that the infected CNS cells express the exogenous nucleic acid sequence.
  • an AAV9 vector provided herein e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B
  • a CNS disorder e.g., a CNS disease
  • an AAV9 vector provided herein that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and/or polypeptide capable of treating a CNS disorder (e.g., a CNS disease) can be administered to a mammal (e.g., a human or a non- human primate) having a CNS disorder in a manner such that the AAV9 vector (a) infects CNS cells and (b) drives expression of the delivered exogenous nucleic acid in the infected CNS cells, thereby reducing the severity of one or more symptoms of the CNS disorder and/or slowing the progression of the CNS disorder.
  • a mammal e.g., a human or a non- human primate
  • an AAV9 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.
  • an exogenous nucleic acid sequence that encodes an RNA and/or polypeptide capable of treating the CNS disorder can be included within the AAV9 vector.
  • polypeptides that can be encoded by an exogenous nucleic Attorney Docket No.: 45049-0091WO1 / 06521 acid sequence designed to treat a disorder (e.g, a CNS disorder or other disorder such as an eye disorder) such as disclosed herein and designed to be included within an AAV9 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 RS1 polypeptide, a TYR polypeptide, a USH2A polypeptide, a MYO7A polypeptide, an REP1 polypeptide, an OPN1LW polypeptide, an OPN1MW polypeptide, a CNGA3
  • Any appropriate CNS disorder can be treated using an AAV9 vector provided herein (e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, and an exogenous nucleic acid sequence encoding a therapeutic RNA and/or polypeptide).
  • an AAV9 vector provided herein (e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, and an exogenous nucleic acid sequence encoding a therapeutic RNA and/or polypeptide).
  • CNS conditions include, without limitation, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, restless leg syndrome, amyotrophic lateral sclerosis (ALS), aneurysm, back pain, Bell’s Palsy, birth defects of the brain and spinal cord, brain injury, brain tumor, cerebral palsy, chronic fatigue syndrome, concussion, dementia, disc disease of neck and lower back, dizziness, frontotemporal dementia, Guillain- Barré syndrome, headaches and migraine, Lewy body dementia, mild cognitive impairment, muscular dystrophy, neuralgia, neuropathy, neuromuscular and related diseases, primary Attorney Docket No.: 45049-0091WO1 / 06521 progressive aphasia, psychiatric conditions (e
  • an AAV9 vector provided herein e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, and an exogenous nucleic acid sequence encoding a therapeutic RNA and/or polypeptide
  • PNS peripheral nervous system
  • cisplatin neuropathy diabetic neuropathy, diphtheritic neuropathy, familial amyloid neuropathy, Guillain-Barré syndrome, Lambert-Eaton syndrome, leprosy, neuropathy with IgM1 anti-myelin-associated glycoprotein, pyridoxine neuropathy, or Refsum’s disease.
  • a CNS disorder can be treated using an AAV9 vector provided herein that is designed to express one or more polypeptides having the ability to inhibit vascular angiogenesis.
  • 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.
  • wet AMD can be treated using an AAV9 vector provided herein that is designed to express a monoclonal anti-VEGF antibody polypeptide, an angiostatin polypeptide, an siRNA, and/or endostatin polypeptide.
  • diabetic retinopathy can be treated using an AAV9 vector provided herein that is designed to express a monoclonal anti-VEGF antibody polypeptide, an angiostatin polypeptide, an siRNA, and/or an endostatin polypeptide.
  • diabetic macular edema can be treated using an AAV9 vector provided herein that is designed to express a monoclonal anti-VEGF antibody polypeptide, an angiostatin polypeptide, an siRNA, and/or an endostatin polypeptide.
  • a CNS disorder can be treated using an AAV9 vector provided herein that is designed to express one or more polypeptides with neuroprotective capabilities and/or other CNS delivery capabilities.
  • polypeptides having the ability to provide neuroprotective activity include, without limitation, GDNF polypeptides, CNTF polypeptides, IGF-1 polypeptides, VEGF polypeptides, and BDNF polypeptides.
  • diabetic macular edema can be treated using an AAV9 vector Attorney Docket No.: 45049-0091WO1 / 06521 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.
  • a CNS disorder can be treated using an AAV9 vector provided herein that is designed to express one or more polypeptides having the ability to inhibit apoptosis.
  • polypeptides having the ability to inhibit apoptosis that can be used as described herein include, without limitation, XIAP polypeptides, cIAP1 polypeptides, C-IAP2 polypeptides, Livin polypeptides, and Survivin polypeptides.
  • a CNS disorder can be treated using an AAV9 vector provided herein that is designed to express one or more polypeptides having the ability to inhibit complement.
  • polypeptides having the ability to inhibit complement examples include, without limitation, Complement Factor I polypeptides, Complement factor H polypeptides, and sCD59 polypeptides.
  • a CNS disorder such as contemplated herein can be treated using an AAV9 vector provided herein that is designed to express a Complement Factor I polypeptide, a Complement factor H polypeptide, and/or a sCD59 polypeptide.
  • a CNS disorder can be treated using an AAV9 vector provided herein that is designed to express one or more polypeptides having the ability to induce survival factors.
  • polypeptides having the ability to induce survival factors include, without limitation, RdCVF polypeptides, RdCVFL polypeptides, HIF-1 polypeptides, IAP family polypeptides, and BCL-2 family polypeptides.
  • a CNS disorder such as one or more of Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, restless leg syndrome, amyotrophic lateral sclerosis (ALS), aneurysm, back pain, Bell’s Palsy, birth defects of the brain and spinal cord, brain injury, brain tumor, cerebral palsy, chronic fatigue syndrome, concussion, dementia, disc disease of neck and lower back, dizziness, frontotemporal dementia, Guillain-Barré syndrome, headaches and migraine, Lewy body dementia, mild cognitive impairment, muscular dystrophy, neuralgia, neuropathy, neuromuscular and related diseases, primary progressive aphasia, psychiatric conditions (e.g., severe depression or obsessive- compuls
  • any appropriate method can be used to administer an AAV9 vector provided herein or composition (e.g., a pharmaceutical composition) provided herein to a mammal (e.g., a human or a non-human primate).
  • a composition provided herein e.g., a pharmaceutical composition containing one or more AAV9 vectors provided herein
  • a mammal e.g., a human or a non-human primate
  • intrathecally 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, intramuscularly (e.g., via intramuscular injection), subretinally, intracranially, intravitreally, systemically, or suprachoroidally.
  • an effective amount of a composition containing an AAV9 vector provided herein (e.g., a pharmaceutical composition provided herein) to treat a CNS disorder can be an amount that reduces the severity of one or more symptoms of the CNS disorder and/or slows the progression of the CNS disorder without producing significant toxicity to the mammal.
  • an effective amount of an AAV9 vector provided herein can be from about 1x10 7 viral genomes to about 1x10 14 viral genomes (e.g., from about 1 x 10 7 viral genomes to about 1 x 10 13 viral genomes, from about 1 x 10 7 viral genomes to about 1 x 10 12 viral genomes, from about 1 x 10 7 viral genomes to about 1 x 10 11 viral genomes, from about 1 x 10 7 viral genomes to about 1 x 10 10 viral genomes, from about 1 x 10 8 viral genomes to about 1 x 10 14 viral genomes, from about 1 x 10 9 viral genomes to about 1 x 10 14 viral genomes, from about 1 x 10 10 viral genomes to about 1 x 10 14 viral genomes, from about 1 x 10 8 viral genomes to about 1 x 10 12 viral genomes, or from about 1 x 10 9 viral genomes to about 1 x 10 11 viral genomes).
  • an effective amount of an AAV9 vector provided herein can be from about 1 x 10 10 viral genomes/kg of body weight to about 1 x 10 14 viral genomes/kg of body weight (e.g., from about 1 x 10 10 viral genomes/kg of body weight to about 1 x 10 13 viral genomes/kg of body weight, from about 1 x 10 10 viral genomes/kg of body weight to about 1 x 10 12 viral genomes/kg of body weight, from about 1 x 10 10 viral genomes/kg of body weight to about 1 x 10 11 viral 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.
  • a composition provided herein e.g., a pharmaceutical composition containing an AAV9 vector provided herein
  • an effective frequency of administration of a composition containing an AAV9 vector provided herein can be a frequency that reduces the severity of one or more symptoms of the CNS disorder and/or slows the progression of the CNS disorder without producing significant toxicity to the mammal.
  • a composition containing an AAV9 vector provided herein can be a frequency that reduces the severity of one or more symptoms of the CNS disorder and/or slows the progression of the CNS disorder without producing significant toxicity to the mammal.
  • Various factors can influence the actual effective frequency used for a particular application. For example, the severity of a CNS 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 CNS 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.
  • an effective duration of administration of a composition containing an AAV9 vector provided herein can be a duration that reduces the severity of one or more symptoms of the CNS disorder and/or slows the progression of the CNS disorder without producing significant toxicity to the mammal.
  • 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 cases, 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.
  • the severity of a CNS 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 CNS 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 AAV9 vector provided herein).
  • a composition provided herein e.g., a pharmaceutical composition containing an AAV9 vector provided herein.
  • an effective amount of a composition containing an AAV9 vector provided herein (e.g., a pharmaceutical composition provided herein) to treat a CNS disorder can be administered once or twice to a mammal (e.g., a human or a non-human primate) to treat that mammal.
  • a mammal e.g., a human or a non-human primate
  • CNS disorders such as one or more of Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, restless leg syndrome, amyotrophic lateral sclerosis (ALS), aneurysm, back pain, Bell’s Palsy, birth defects of the brain and spinal cord, brain injury, brain tumor, cerebral palsy, chronic fatigue syndrome, concussion, dementia, disc disease of neck and lower back, dizziness, frontotemporal dementia, Guillain- Barré syndrome, headaches and migraine, Lewy body dementia, mild cognitive impairment, muscular dystrophy, neuralgia, neuropathy, neuromuscular and related diseases, primary progressive aphasia, psychiatric conditions
  • the subject treated in accordance with the methods disclosed herein has not received prior therapy for treating the CNS disorder. In certain embodiments, the subject treated in accordance with the methods disclosed herein has received a prior therapy but continues to experience progression of the CNS disorder despite the prior therapy.
  • EXAMPLES [0067] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.
  • a high-throughput method is used to create AAV9 vectors with mutated capsid polypeptides and to screen those created AAV9 vectors for particular AAV9 vectors having the ability to exhibit high efficiency and/or specificity for infecting CNS cells. See, e.g., ⁇ ztk et al., bioRxiv, 2020.10.01.323196 (2020) and ⁇ ztk et al., eLife, 10:e64175 (2021).
  • AAV9 mutants are created and injected into the CNS of primates (cynolmolgus macaques or rhesus macaques). These libraries are created such that each AAV9 vector in the library contained a unique DNA barcode, which allowed for tracking of a mutated AAV9 capsid polypeptide.
  • successfully packaged AAV9 vectors are polymerase chain reaction (PCR) amplified and repackaged, resulting in a “repack” library.
  • AAV9 vectors are injected into primate retinas, and nucleic acid encoding the AAV9 capsid polypeptides are then amplified from the nuclei of foveal cells, resulting in an “enriched” library.
  • Each iteration of the AAV9 library (e.g., the original library, the repack library, and the enriched library) is injected, e.g., intrathecally into the CNS of primates.
  • the AAV9 vectors compete with each other in vivo. Infection of successful AAV9 vectors can lead to expression of the DNA barcodes.
  • Single cell suspensions are created from isolated CNS fluid, and single cell microfluidic technology (10X Genomics) is used to create cDNA libraries of individual cells. Computational analysis is performed to identify optimal vectors, according to cell specificities, expression levels, and/or other desirable characteristics, based on the presence and quantity of DNA barcodes in transcriptomes from different cells of multiple cell types in parallel.
  • AAV9 capsid polypeptides The performance of AAV9 capsid polypeptides is evaluated on the basis of mRNA transcription levels rather than the presence of DNA, reflecting the ability of the AAV9 vectors to drive expression of the AAV9 vector nucleic acid as opposed to simply having the ability to enter a cell.
  • AAV9 vectors having capsid polypeptides that include SEQ ID NO:5 or 14 located between amino acid residues 588 and 589 of SEQ ID NO:1) can exhibit the ability to infect and drive mRNA expression within CNS cells in a manner similar to that of 7m8 AAV9 vector.
  • the AAV9 vector having a capsid polypeptide that includes SEQ ID NO:5 or 14 can have the ability to exhibit more infectivity of CNS cells than a wild-type AAV9 vector containing an AAV9 capsid polypeptide that consists of SEQ ID NO:1.
  • Attorney Docket No.: 45049-0091WO1 / 06521 [0071] Taken together, this can demonstrate that AAV9 vectors that include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1A (or Formula A) can have the ability to mediate transgene expression in CNS cells.
  • AAV9 vectors that include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1B (or Formula B) can have the ability to mediate transgene expression in CNS cells.
  • Example 2 – Treating a CNS disorder using an AAV9 vector An AAV9 vector is constructed to include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1A (e.g., SEQ ID NO:2 or 5) or Formula A and an exogenous nucleic acid sequence encoding a therapeutic polypeptide.
  • the constructed AAV9 vector is administered intravitreally to a human identified as having a CNS disorder in an amount that is from about 1 x 10 7 to about 1 x 10 14 AAV9 vectors. After the administration, the severity of one or more symptoms of the CNS disorder is reduced and/or the progression of the CNS disorder is slowed.
  • Example 3 – Treating a CNS disorder using an AAV9 vector An AAV9 vector is constructed to include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1B (e.g., SEQ ID NO:11 or 14) or Formula B and an exogenous nucleic acid sequence encoding a therapeutic polypeptide.
  • the constructed AAV9 vector is administered intravitreally to a human identified as having a CNS disorder in an amount that is from about 1 x 10 7 to about 1 x 10 14 AAV9 vectors. After the administration, the severity of one or more symptoms of the CNS disorder is reduced and/or the progression of the CNS disorder is slowed.
  • Example 4 – AAV vectors containing mutated capsids [0073] An in vivo analysis of AAV9 mutated capsids was conducted in mouse CNS via intra cisterna magna injections. Performance was measured by assessing the expression of the fluorophores expressed by each vector using immunohistochemistry (IHC) and/or fluorescent microscopy.
  • IHC immunohistochemistry
  • Fluorophore expression was used as a proxy for capsid efficacy, with higher fluorophore expression indicating higher viral transduction. Fluorophore expression was qualitatively analyzed by scoring relative fluorophore expression in regions of the brain and Attorney Docket No.: 45049-0091WO1 / 06521 spinal cord. As indicated in Table 2, fluorophore expression of both capsids of SEQ ID NO:10 and SEQ ID NO:19 were observed in regions of the brain and spinal cord, indicating their efficacy in viral transduction. Table 2. Qualitative Fluorescence Score (0 to +++) escence. OTHER EMBODIMENTS Embodiment 1.
  • An adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide having 95% identity or more with SEQ ID NO:1, or a truncated variant thereof, further comprising an amino acid peptide sequence comprising a sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14, wherein the amino acid peptide sequence is located between two existing amino acids of SEQ ID NO:1.
  • AAV9 vector comprising an AAV9 capsid polypeptide having 95% identity or more with SEQ ID NO:1, or a truncated variant thereof, further comprising an amino acid peptide sequence comprising a sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14, wherein the amino acid peptide sequence is located between two existing amino acids of SEQ ID NO:1.
  • Embodiment 1 wherein said amino acid peptide sequence is located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of SEQ ID NO:1.
  • Embodiment 3 The vector of Embodiment 1 or 2, wherein the amino acid peptide sequence is SEQ ID NO:2 or 11.
  • Embodiment 4 The vector of Embodiment 1 or 2, wherein the amino acid peptide sequence is SEQ ID NO:5 or 14.
  • Embodiment 5. An adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide of SEQ ID NO:1, wherein the amino acids from position 585 to 590 of SEQ ID NO:1 is replaced with an amino acid peptide sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14.
  • Embodiment 6 The vector of Embodiment 5, wherein the amino acids from position 585 to 590 of SEQ ID NO:1 are replaced with said amino acid sequence of SEQ ID NO:2 or 11.
  • the vector of any one of Embodiments 1-6 wherein said vector infects greater than 2.5 percent of central nervous system (CNS) cells in one or more brain or spinal cord regions including the cortex, hippocampus, midbrain, cerebellum, brainstem, ventral spinal horn, and dorsal spinal horn when a titer of at least 1 x 10 7 of said vector is administered into the CNS of a patient (e.g., via intra cisterna magna (ICM), intraparenchymal (IPa), intraventricular, or intrathecal (IT) administration), or administered intravenously or subcutaneously.
  • ICM intra cisterna magna
  • IPa intraparenchymal
  • IMV intraventricular
  • Thecal intrathecal
  • Embodiment 9 The vector of any one of Embodiments 1-7, wherein said vector comprises an exogenous nucleic acid encoding an RNA or a polypeptide.
  • the vector of Embodiment 8, wherein said exogenous nucleic acid encodes an RNA.
  • Embodiment 10 The vector of Embodiment 9, wherein said RNA is an siRNA or microRNA.
  • Embodiment 11 The vector of Embodiment 8, wherein said exogenous nucleic acid encodes a polypeptide.
  • Embodiment 11 wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide.
  • Embodiment 13 The vector of any one of Embodiments 1-12, wherein said vector expresses more nucleic acid in CNS cells than the level of expression from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1.
  • Embodiment 14 A composition comprising a vector of any one of Embodiments 1-13 and a pharmaceutically acceptable excipient.
  • composition of Embodiment 14 wherein said composition comprises from about 1 x 10 7 to about 1 x 10 14 of said vector.
  • Embodiment 16 The composition of Embodiment 14 or 15, wherein said pharmaceutically acceptable excipient comprises one or more of phosphate buffered saline, Hank’s Balanced Salt Solution, artificial cerebrospinal fluid, and Pluronic F68.
  • a method for delivering an exogenous nucleic acid sequence to a CNS of a mammal comprises: contacting CNS cells of said mammal with an AAV9 vector comprising an AAV9 capsid polypeptide and said exogenous nucleic acid sequence, wherein said capsid polypeptide comprises an amino acid peptide sequence selected from the group consisting of SEQ ID NOs:2- 5 and 11-14; Attorney Docket No.: 45049-0091WO1 / 06521 wherein upon the contacting said AAV9 vector infects said CNS cells and thereby delivering said exogenous nucleic acid sequence to said CNS cells.
  • Embodiment 18 Embodiment 18.
  • said capsid polypeptide comprises the amino acid sequence of SEQ ID NO:1 with the exception that the amino acid peptide sequence is located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of SEQ ID NO:1, or with the exception that the amino acid residues at positions 585 to 590 of SEQ ID NO:1 are replaced with said amino acid peptide sequence.
  • Embodiment 19 The method of Embodiment 18, wherein said amino acid peptide sequence is located between amino acid positions 588 and 589 of SEQ ID NO:1.
  • Embodiment 20 The method of any one of Embodiments 17-19, wherein said amino acid peptide sequence is SEQ ID NO:2 or 11.
  • Embodiment 21 The method of any one of Embodiments 17-20, wherein said mammal is a human.
  • Embodiment 22. The method of any one of Embodiments 17-21, wherein said exogenous nucleic acid sequence encodes an RNA or a polypeptide.
  • Embodiment 23 The method of Embodiment 22, wherein said exogenous nucleic acid encodes said RNA.
  • Embodiment 24 The method of Embodiment 18, wherein said amino acid peptide sequence is located between amino acid positions 588 and 589 of SEQ ID NO:1.
  • Embodiment 20 The method of any one of Embodiments 17-19, wherein said amino acid peptide
  • Embodiment 23 wherein said RNA is an siRNA or microRNA.
  • Embodiment 25 The method of Embodiment 22, wherein said exogenous nucleic acid encodes said polypeptide.
  • Embodiment 26 The method of Embodiment 25, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide.
  • Embodiment 27 Embodiment 27.
  • any one of Embodiments 17-26 wherein said vector expresses more of said exogenous nucleic acid sequence in said CNS cell than the level of expression in a CNS cell from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1; or wherein said vector expresses said exogenous nucleic acid sequence in more CNS cells than the number of CNS cells where said exogenous nucleic acid sequence is expressed from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1; or wherein said vector expresses said exogenous nucleic acid sequence in different CNS cells than the CNS cells where said exogenous nucleic acid sequence is expressed from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1.
  • Embodiment 28 The method of any one of Embodiments 17-27, wherein said method comprises intravenously, subcutaneously, intracranially, and/or intrathecally administering a composition comprising said vector to said mammal, thereby contacting said CNS cell with said vector.
  • Embodiment 29 The method of Embodiment 28, wherein said composition comprises from about 1 x 10 7 to about 1 x 10 14 of said vector, or wherein said composition comprises from about 1 x 10 7 to about 1 x 10 15 of said vector.
  • Embodiment 30 The method of any one of Embodiments 17-27, wherein said method comprises intravenously, subcutaneously, intracranially, and/or intrathecally administering a composition comprising said vector to said mammal, thereby contacting said CNS cell with said vector.
  • a method for treating a CNS disorder in a mammal in need thereof comprises contacting CNS cells of a mammal having said CNS disorder with a therapeutic construct comprising the AAV9 vector of any one of Embodiments 1-13 and an exogenous nucleic acid sequence.
  • Embodiment 31 The method of Embodiment 30, wherein said mammal is a human.
  • Embodiments 30-31 wherein said CNS disorder is selected from the group consisting of Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, and restless leg syndrome.
  • Embodiment 33 The method of any one of Embodiments 30-32, wherein said exogenous nucleic acid sequence encodes an RNA.
  • Embodiment 34 The method of Embodiment 33, wherein said RNA is an siRNA or a microRNA.
  • Embodiment 35 Embodiment 35.
  • Embodiment 36 The method of any one of Embodiments 30-33, wherein said exogenous nucleic acid encodes a polypeptide.
  • Embodiment 36 The method of Embodiment 35, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide.
  • Embodiment 37 Embodiment 37.
  • a non-naturally occurring adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide, wherein said capsid polypeptide comprises at least 95% identity to SEQ ID NO:1, or a truncated variant thereof, and further comprises an amino acid sequence insert of Formula A or Formula B located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 according to SEQ ID NO:1, wherein said Formula A is: -L 1 -EGSGRN (SEQ ID NO:2)-L 2 -, Attorney Docket No.:
  • Embodiment 38 The vector of Embodiment 37, wherein said L 1 is one amino acid X1.
  • Embodiment 39. The vector of Embodiment 38, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 40. The vector of Embodiment 38 or 39, wherein said X1 is A.
  • Embodiment 41. The vector of Embodiment 37, wherein said L 1 is two amino acids X2-X1.
  • Embodiment 42 The vector of Embodiment 41, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 43. The vector of Embodiment 42, wherein said X1 is A.
  • Embodiment 44 The vector of any one of Embodiments 41-43, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 45 The vector of Embodiment 44, wherein said X2 is L.
  • Embodiment 46 The vector of Embodiment 45, wherein said X2-X1 is LA.
  • Embodiment 47 The vector of Embodiment 37, wherein said L1 is three amino acids X3- X2-X1. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 48.
  • Embodiment 47 wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 49 The vector of Embodiment 48, wherein said X1 is A.
  • Embodiment 50 The vector of any one of Embodiments 47-49, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 51 The vector of Embodiment 50, wherein said X2 is L.
  • Embodiment 52 The vector of Embodiment 51, wherein said X2-X1 is LA.
  • Embodiment 53 The vector of Embodiment 53.
  • Embodiment 54 The vector of Embodiment 37, wherein said L1 is absent.
  • Embodiment 55 The vector of any one of Embodiments 37-54, wherein said L2 is one amino acid Z1.
  • Embodiment 56 The vector of Embodiment 55, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 57 The vector of Embodiment 56, wherein said Z1 is A.
  • Embodiment 58 The vector of any one of Embodiments 47-52, wherein said X3 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 59 The vector of any one of Embodiments 37-54, wherein said L2 is two amino acids Z1-Z2.
  • Embodiment 59 The vector of Embodiment 58, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 60.
  • the vector of Embodiment 59, wherein said Z1 is A.
  • Embodiment 61 The vector of any one of Embodiments 58-60, wherein said Z2 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 62 The vector of Embodiment 61, wherein said Z2 is L.
  • Embodiment 63 The vector of Embodiment 62, wherein said Z1-Z2 is AL.
  • Embodiment 64 The vector of any one of Embodiments 37-54, wherein said L2 is three amino acids Z1-Z2-Z3.
  • Embodiment 65 The vector of Embodiment 64, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L, and/or Z2 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 66 The vector of Embodiment 65, wherein said Z1-Z2 is AL.
  • Embodiment 67 The vector of Embodiment 65, wherein said Z1-Z2 is AL.
  • Embodiment 65-66 The vector of any one of Embodiments 65-66, wherein said Z3 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • Embodiment 68 The vector of Embodiment 37, wherein said amino acid sequence insert comprises any one of SEQ ID NOs:2-5 and 11-14.
  • Embodiment 69 The vector of any one of Embodiments 65-66, wherein said Z3 is selected from the group of amino acid residues consisting of A, V, I, and L.
  • a non-naturally occurring AAV9 capsid polypeptide wherein said capsid polypeptide comprises at least 95% identity to SEQ ID NO:1, or a truncated variant thereof, and further comprises an amino acid sequence insert of Formula A or Formula B located 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 Attorney Docket No.: 45049-0091WO1 / 06521 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 according to SEQ ID NO:1, wherein said Formula A is: -L 1 -EGSGRN (SEQ ID NO:2)-L 2 -, wherein said L
  • Embodiment 70 A method for administering an exogenous nucleic acid sequence to a mammal in need thereof, wherein said method comprises administering to the mammal an effective amount of a construct comprising a vector of any one of Embodiments 37-68 and the exogenous nucleic acid sequence.
  • Embodiment 71 A method of treating a CNS disorder in a patient in need thereof, comprising administering to the patient an effective amount of an AAV9 vector comprising an AAV9 capsid polypeptide comprising an amino acid sequence insert represented by Formula A or Formula B and an exogenous nucleic acid sequence.

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Abstract

This document provides AAV9 vectors. For example, AAV9 vectors containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B are provided as are such AAV9 capsid polypeptides, nucleic acid molecules encoding such vectors, nucleic acid molecules encoding such AAV9 capsid polypeptides, host cells containing and/or expressing such nucleic acid molecules, and methods and materials for making or using such vectors and/or AAV9 capsid polypeptides.

Description

Attorney Docket No.: 45049-0091WO1 / 06521 AAV9 VECTORS AND CAPSID POLYPEPTIDES FOR USE IN TREATING CNS DISORDERS CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of U.S. Provisional Application Serial No. 63/540,902, filed September 27, 2023, and the benefit of U.S. Provisional Application Serial No. 63/540,895, filed September 27, 2023. The disclosure of the prior applications are considered part of the disclosure of this application, and are incorporated in its entirety into this application STATEMENT REGARDING FEDERAL FUNDING [0002] This invention was made with government support under MH120094 awarded by the National Institutes of Health. The government has certain rights in the invention. BACKGROUND [0003] Viral vectors, such as AAV9 vectors, are efficient vehicles for in vivo nucleic acid delivery, and their use in the clinic is expanding. Improved AAV9 vectors and AAV9 production techniques for making effective AAV9 vector preparations should further expand the use of AAV9 vectors in the laboratory and clinic. SUMMARY OF THE DISCLOSURE [0004] This document provides AAV9 vectors. For example, this document provides AAV9 vectors containing a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. The AAV9 vectors described herein containing a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B can have the ability to infect CNS cells in vivo and deliver exogenous nucleic acid to the infected CNS cells such that the infected CNS cells express the exogenous nucleic acid. This document also provides methods and materials for making and using AAV9 vectors having the ability to deliver nucleic acid to CNS cells. Attorney Docket No.: 45049-0091WO1 / 06521 [0005] Accordingly, in one aspect, this document provides an adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide having 95% identity or more with SEQ ID NO:1, or a truncated variant thereof, further comprising an amino acid peptide sequence comprising a sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14, wherein the amino acid peptide sequence is located between two existing amino acids of SEQ ID NO:1, e.g., between amino acid positions 588 and 589 (or e.g., between 585 and 590) of SEQ ID NO:1. [0006] Other embodiments and details of the disclosure are presented herein below. DETAILED DESCRIPTION Definitions [0007] To facilitate an understanding of the present document, a number of terms and phrases are defined below. [0008] The terms “a” and “an” as used herein mean “one or more” and include the plural unless the context is inappropriate. [0009] As used herein, the term “pharmaceutical formulation” refers to the combination of an active agent with a carrier, inert or active, making the composition especially suitable for diagnostic or therapeutic use in vivo or ex vivo. [0010] As used herein, the terms “subject” and “patient” refer to an organism to be treated by the methods and compositions described herein. Such organisms preferably include, but are not limited to, mammals (e.g., murines, simians, equines, bovines, porcines, primates, canines, felines, and the like), and more preferably include humans. [0011] The terms “treat,” “treating,” or “treatment,” and other grammatical equivalents as used in this document, include alleviating, abating, ameliorating, or preventing a disease, condition or symptoms, preventing additional symptoms, ameliorating or preventing the underlying metabolic causes of symptoms, inhibiting the disease or condition, e.g., arresting the development of the disease or condition, relieving the disease or condition, causing regression of the disease or condition, relieving a condition caused by the disease or condition, or stopping the symptoms of the disease or condition, and are intended to include prophylaxis. The terms further include achieving a therapeutic benefit and/or a prophylactic benefit. The term “therapeutic benefit” refers to eradication or amelioration of the underlying disorder being treated. Also, a therapeutic Attorney Docket No.: 45049-0091WO1 / 06521 benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient, notwithstanding that the patient may still be afflicted with the underlying disorder. [0012] As used herein, the term “effective amount” refers to the amount of a compound (e.g., an AAV9 vector described herein) sufficient to effect beneficial or desired results. An effective amount can be administered in one or more administrations, applications, or dosages and is not intended to be limited to a particular formulation or administration route. [0013] As used herein, the term “combination” in the context of therapies means that two or more different treatments are delivered to the subject during the course of the subject’s affliction with the disorder, such that the effects of the treatments on the patient overlap at a point in time. In certain embodiments, the delivery of one treatment is still occurring when the delivery of the second begins, so that there is overlap in terms of administration. This is sometimes referred to herein as “simultaneous” or “concurrent delivery.” In other embodiments, the delivery of one treatment ends before the delivery of the other treatment begins. In certain embodiments of either case, the treatment is more effective because of combined administration. For example, the second treatment is more effective, e.g., an equivalent effect is seen with less of the second treatment, or the second treatment reduces symptoms to a greater extent, than would be seen if the second treatment were administered in the absence of the first treatment, or the analogous situation is seen with the first treatment. In certain embodiments, delivery is such that the reduction in a symptom, or other parameter related to the disorder is greater than what would be observed with one treatment delivered in the absence of the other. The effect of the two treatments can be partially additive, wholly additive, or greater than additive. The delivery can be such that an effect of the first treatment delivered is still detectable when the second is delivered. [0014] The term “about” refers to any minimal alteration in the concentration or amount of an agent that does not change the efficacy of the agent in preparation of a formulation and in treatment of a disease or disorder. In certain embodiments, the term “about” may include ±5%, ±10%, or ±15% of a specified numerical value or data point. [0015] Ranges can be expressed in this document as from “about” one particular value, and/or to “about” another particular value. When such a range is expressed, another aspect includes from the one particular value and/or to the other particular value. Similarly, when values are Attorney Docket No.: 45049-0091WO1 / 06521 expressed as approximations, by use of the antecedent “about,” it is understood that the particular value forms another aspect. It is further understood that the endpoints of each of the ranges are significant both in relation to the other endpoint, and independently of the other endpoint. It is also understood that there are a number of values disclosed in this document, and that each value is also disclosed as “about” that particular value in addition to the value itself. It is also understood that throughout the document, data are provided in a number of different formats and that this data represent endpoints and starting points and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point “15” are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units is also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed. [0016] Throughout the description, where compositions are described as having, including, containing, incorporating, or comprising specific components, or where processes and methods are described as having, including, or comprising specific steps, it is intended that compositions and methods are inclusive or open-ended and do not exclude additional, unrecited components or steps. It is contemplated that, additionally, there are compositions of the present document that consist essentially of, or consist of, the recited components, and that there are processes and methods according to the present document that consist essentially of, or consist of, the recited steps. [0017] As a general matter, compositions specifying a percentage are by weight unless otherwise specified. Further, if a variable is not accompanied by a definition, then the previous definition of the variable controls. AAV9 Vectors [0018] The present document provides AAV9 vectors. For example, this document provides AAV9 vectors containing a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. Any appropriate AAV9 vector can be designed to include a capsid polypeptide described herein (e.g., a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set Attorney Docket No.: 45049-0091WO1 / 06521 forth in Table 1B (or a variant thereof) or Formula B). In some cases, an AAV9 having an ACG start codon for the AAV9 Rep polypeptides instead of an ATG start codon (e.g., an AAV9-M1T- REP) can be designed to include a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. [0019] Any appropriate AAV9 capsid polypeptide can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. In some cases, an AAV9 capsid polypeptide can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. In some cases, an AAV9 capsid polypeptide having the following amino acid sequence can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B: MAADGYLPDW LEDNLSEGIR EWWALKPGAP QPKANQQHQD NARGLVLPGY KYLGPGNGLD KGEPVNAADA AALEHDKAYD QQLKAGDNPY LKYNHADAEF QERLKEDTSF GGNLGRAVFQ AKKRLLEPLG LVEEAAKTAP GKKRPVEQSP QEPDSSAGIG KSGAQPAKKR LNFGQTGDTE SVPDPQPIGE PPAAPSGVGS LTMASGGGAP VADNNEGADG VGSSSGNWHC DSQWLGDRVI TTSTRTWALP TYNNHLYKQI SNSTSGGSSN DNAYFGYSTP WGYFDFNRFH CHFSPRDWQR LINNNWGFRP KRLNFKLFNI QVKEVTDNNG VKTIANNLTS TVQVFTDSDY QLPYVLGSAH EGCLPPFPAD VFMIPQYGYL TLNDGSQAVG RSSFYCLEYF PSQMLRTGNN FQFSYEFENV PFHSSYAHSQ SLDRLMNPLI DQYLYYLSKT INGSGQNQQT LKFSVAGPSN MAVQGRNYIP GPSYRQQRVS TTVTQNNNSE FAWPGASSWA LNGRNSLMNP GPAMASHKEG EDRFFPLSGS LIFGKQGTGR DNVDADKVMI TNEEEIKTTN PVATESYGQV ATNHQSAQAQ AQTGWVQNQG ILPGMVWQDR DVYLQGPIWA KIPHTDGNFH PSPLMGGFGM KHPPPQILIK NTPVPADPPT AFNKDKLNSF ITQYSTGQVS VEIEWELQKE NSKRWNPEIQ YTSNYYKSNN VEFAVNTEGV YSEPRPIGTR YLTRNL (SEQ ID NO:1). The two bold amino acid residues are at positions 588 and 589, and the underlined amino acids are at positions 585 to 590. [0020] In some cases, an AAV9 capsid polypeptide 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 can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, Attorney Docket No.: 45049-0091WO1 / 06521 e.g., a contemplated AAV9 capside polypeptide is at least 95% identical to SEQ ID NO:1, and/or may include 1, 2, 3, or more amino acid truncations (e.g., on one or both ends), and/or deletions. [0021] When designing an AAV9 capsid polypeptide to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, that included amino acid sequence can be located at any appropriate location along the AAV9 capsid polypeptide. For example, an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A such as any one of SEQ ID NOs:2-5 or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B such as any one of SEQ ID NOs:11-14 can be located between the naturally-occurring amino acid residues at positions 588 and 589 of an AAV9 capsid polypeptide or can be located between two naturally-occurring amino acid residues at other positions of e.g., SEQ ID NO:1. In some cases, an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A such as any one of SEQ ID NOs:2-5 or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B such as any one of SEQ ID NOs:11-14 can be located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of an AAV capsid polypeptide (e.g., an AAV9 capsid polypeptide such as an AAV9 capsid polypeptide comprising the amino acid sequence set forth in SEQ ID NO:1). [0022] As described herein, an AAV9 vector can be designed to have an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. Table 1A. Amino acid sequences that can be inserted into an AAV9 capsid polypeptide. ID
Figure imgf000007_0001
Attorney Docket No.: 45049-0091WO1 / 06521 LAEGSGRN 4 CTAGCAGAGGGCAGCGGCCGGAAC 8 LAEGSGRNA 5 CTAGCAGAGGGCAGCGGCCGGAACGCT 9 For exa
Figure imgf000008_0001
p , g p p yp p g following amino acid sequence: 1 MAADGYLPDW LEDNLSEGIR EWWALKPGAP QPKANQQHQD NARGLVLPGY 51 KYLGPGNGLD KGEPVNAADA AALEHDKAYD QQLKAGDNPY LKYNHADAEF 101 QERLKEDTSF GGNLGRAVFQ AKKRLLEPLG LVEEAAKTAP GKKRPVEQSP 151 QEPDSSAGIG KSGAQPAKKR LNFGQTGDTE SVPDPQPIGE PPAAPSGVGS 201 LTMASGGGAP VADNNEGADG VGSSSGNWHC DSQWLGDRVI TTSTRTWALP 251 TYNNHLYKQI SNSTSGGSSN DNAYFGYSTP WGYFDFNRFH CHFSPRDWQR 301 LINNNWGFRP KRLNFKLFNI QVKEVTDNNG VKTIANNLTS TVQVFTDSDY 351 QLPYVLGSAH EGCLPPFPAD VFMIPQYGYL TLNDGSQAVG RSSFYCLEYF 401 PSQMLRTGNN FQFSYEFENV PFHSSYAHSQ SLDRLMNPLI DQYLYYLSKT 451 INGSGQNQQT LKFSVAGPSN MAVQGRNYIP GPSYRQQRVS TTVTQNNNSE 501 FAWPGASSWA LNGRNSLMNP GPAMASHKEG EDRFFPLSGS LIFGKQGTGR 551 DNVDADKVMI TNEEEIKTTN PVATESYGQV ATNHQSAQLA EGSGRNAAQA 601 QTGWVQNQGI LPGMVWQDRD VYLQGPIWAK IPHTDGNFHP SPLMGGFGMK 651 HPPPQILIKN TPVPADPPTA FNKDKLNSFI TQYSTGQVSV EIEWELQKEN 701 SKRWNPEIQY TSNYYKSNNV EFAVNTEGVY SEPRPIGTRY LTRNL (SEQ ID NO:10) Table 1B Amino acid sequences that can be inserted into an AAV capsid polypeptide. Amino Acid SEQ ID Nucleic Acid Sequence encoding the Amino Acid SEQ ID Sequence NO: Sequence NO:
Figure imgf000008_0002
o e a p e, a ec o ca e es g e o a e a caps po ypep e a g e following amino acid sequence: 1 MAADGYLPDW LEDNLSEGIR EWWALKPGAP QPKANQQHQD NARGLVLPGY 51 KYLGPGNGLD KGEPVNAADA AALEHDKAYD QQLKAGDNPY LKYNHADAEF 101 QERLKEDTSF GGNLGRAVFQ AKKRLLEPLG LVEEAAKTAP GKKRPVEQSP 151 QEPDSSAGIG KSGAQPAKKR LNFGQTGDTE SVPDPQPIGE PPAAPSGVGS 201 LTMASGGGAP VADNNEGADG VGSSSGNWHC DSQWLGDRVI TTSTRTWALP 251 TYNNHLYKQI SNSTSGGSSN DNAYFGYSTP WGYFDFNRFH CHFSPRDWQR Attorney Docket No.: 45049-0091WO1 / 06521 301 LINNNWGFRP KRLNFKLFNI QVKEVTDNNG VKTIANNLTS TVQVFTDSDY 351 QLPYVLGSAH EGCLPPFPAD VFMIPQYGYL TLNDGSQAVG RSSFYCLEYF 401 PSQMLRTGNN FQFSYEFENV PFHSSYAHSQ SLDRLMNPLI DQYLYYLSKT 451 INGSGQNQQT LKFSVAGPSN MAVQGRNYIP GPSYRQQRVS TTVTQNNNSE 501 FAWPGASSWA LNGRNSLMNP GPAMASHKEG EDRFFPLSGS LIFGKQGTGR 551 DNVDADKVMI TNEEEIKTTN PVATESYGQV ATNHQSAQLA EHQTRPAAQA 601 QTGWVQNQGI LPGMVWQDRD VYLQGPIWAK IPHTDGNFHP SPLMGGFGMK 651 HPPPQILIKN TPVPADPPTA FNKDKLNSFI TQYSTGQVSV EIEWELQKEN 701 SKRWNPEIQY TSNYYKSNNV EFAVNTEGVY SEPRPIGTRY LTRNL (SEQ ID NO:19) [0023] For example, an AAV9 vector can be designed to have an AAV9 capsid polypeptide of SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) that includes an amino acid sequence insert of Formula A located between amino acid positions 588 and 589 of SEQ ID NO:1 (or the appropriate amino acid positions of the alternative sequence). Formula A can be as follows: -L1-EGSGRN (SEQ ID NO:2)-L2-, wherein L1 and L2 are each independently optional amino acid linkers having one, two, or three amino acids. In some cases, L1, L2, or both L1 and L2 can be absent. In some cases, L1 can be one amino acid X1, two amino acids X2-X1, or three amino acids X3-X2-X1. When X1 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 Z1, two amino acids Z1-Z2, or three amino acids Z1-Z2-Z3. When Z1 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 L1 linkers include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, LIV, LII, LIL, LLA, LLV, LLI, and LLL. Examples of an Attorney Docket No.: 45049-0091WO1 / 06521 L2 linkers include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, LIV, LII, LIL, LLA, LLV, LLI, and LLL. For example, an AAV9 vector can be designed to have an AAV9 capsid polypeptide of SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) that includes an amino acid sequence insert of Formula B located between amino acid positions 588 and 589 of SEQ ID NO:1 (or the appropriate amino acid positions of the alternative sequence). Formula B can be as follows: -L1- EHQTRP (SEQ ID NO:11)-L2-, wherein L1 and L2 are each independently optional amino acid linkers having one, two, or three amino acids. In some cases, L1, L2, or both L1 and L2 can be absent. In some cases, L1 can be one amino acid X1, two amino acids X2-X1, or three amino acids X3-X2-X1. When X1 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 Z1, two amino acids Z1-Z2, or three amino acids Z1-Z2-Z3. When Z1 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 L1 linkers include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, LIV, LII, LIL, LLA, LLV, LLI, and LLL. Examples of an L2 linkers include, without limitation, A, V, I, L, AA, AV, AI, AL, VA, VV, VI, VL, IA, IV, II, IL, LA, LV, LI, LL, AAA, AAV, AAI, AAL, AVA, AVV, AVI, AVL, AIA, AIV, AII, AIL, Attorney Docket No.: 45049-0091WO1 / 06521 ALA, ALV, ALI, ALL, VAA, VAV, VAI, VAL, VVA, VVV, VVI, VVL, VIA, VIV, VII, VIL, VLA, VLV, VLI, VLL, IAA, IAV, IAI, IAL, IVA, IVV, IVI, IVL, IIA, IIV, III, IIL, ILA, ILV, ILI, ILL, LAA, LAV, LAI, LAL, LVA, LVV, LVI, LVL, LIA, LIV, LII, LIL, LLA, LLV, LLI, and LLL. [0024] In some cases, an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B inserted between glutamine-588 and alanine-589 (or the appropriate amino acid positions of an alternative sequence). In some cases, an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with an amino acid sequence set forth in any one of SEQ ID NOs:2-5 (or a variant thereof) inserted between glutamine-588 and alanine-589 (or the appropriate amino acid positions of the alternative sequence). In some cases, an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with an amino acid sequence set forth in any one of SEQ ID NOs:11-14 (or a variant thereof) inserted between glutamine-588 and alanine-589 (or the appropriate amino acid positions of the alternative sequence). [0025] In some cases, when designing an AAV9 capsid polypeptide to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, that included amino acid sequence can be used to replace one or more naturally-occurring amino acid residues located at any appropriate location along the AAV9 capsid polypeptide. For example, an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A such as any one of SEQ ID NOs:2-5 can be used to replace the naturally-occurring amino acid residues at positions 585 to 590 of an AAV9 capsid polypeptide. For example, an amino acid sequence set forth in Table 1B (or a variant thereof) or Attorney Docket No.: 45049-0091WO1 / 06521 Formula B such as any one of SEQ ID NOs:11-14 can be used to replace the naturally-occurring amino acid residues at positions 585 to 590 of an AAV9 capsid polypeptide. [0026] In some cases, an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in 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 set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. In some cases, an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with the exception that amino acid residues 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with the amino acid sequence set forth in any one of SEQ ID NOs:2-5 (or a variant thereof). In some cases, an AAV9 capsid polypeptide provided herein can have the sequence set forth in SEQ ID NO:1 (or an alternative sequence that is 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 set forth in SEQ ID NO:1) with the exception that amino acid residues 585 to 590 (or the appropriate amino acid positions of the alternative sequence) are replaced with the amino acid sequence set forth in any one of SEQ ID NOs:11-14 (or a variant thereof). [0027] In some cases, an AAV9 capsid polypeptide can be designed to include two or more amino acid sequences set forth in Table 1A (or a variant thereof) or Formula A or two or more amino acid sequences set forth in Table 1B (or a variant thereof) or Formula B. For example, an AAV9 capsid polypeptide can be designed to include two or three amino acid sequences set forth in Table 1A (or a variant thereof) or Formula A. For example, an AAV9 capsid polypeptide can be designed to include two or three amino acid sequences set forth in Table 1B (or a variant thereof) or Formula B. [0028] As described herein, an AAV9 capsid polypeptide can be designed to include an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. A variant of an amino acid sequence set forth in Table 1A refers to an amino acid sequence that is identical to that amino acid sequence Attorney Docket No.: 45049-0091WO1 / 06521 set forth in Table 1A except that it has one, two, or three amino acid additions, deletions, substitutions, or combinations thereof. For example, a variant of SEQ ID NO:2 can be SEQ ID NO:2 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:2-5 except that it contains one, two, or three amino acid additions. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:2-5 except that it contains one, two, or three amino acid deletions. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:2-5 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:2-5 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:2-5 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:2-5 except that it contains three amino acid additions, deletions, substitutions, or a combination thereof. [0029] A variant of an amino acid sequence set forth in Table 1B refers to an amino acid sequence that is identical to that amino acid sequence set forth in Table 1B except that it has one, two, or three amino acid additions, deletions, substitutions, or combinations thereof. For example, a variant of SEQ ID NO:11 can be SEQ ID NO:11 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:11-14 except that it contains one, two, or three amino acid additions. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 except that it contains one, two, or three amino acid deletions. In some cases, a variant provided herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 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:11-14 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:11-14 except that it contains two amino acid additions, deletions, substitutions, or a combination thereof. In some cases, a variant provided Attorney Docket No.: 45049-0091WO1 / 06521 herein can be the amino acid sequence set forth in any one of SEQ ID NOs:11-14 except that it contains three amino acid additions, deletions, substitutions, or a combination thereof. [0030] In some cases, an amino acid substitution present in a variant can be a conservative amino acid substitution. For example, conservative 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. [0031] In some cases, an amino acid substitution present in a variant can be a non- conservative 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 aresidue 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. [0032] 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 (Bl2seq) 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 Bl2seq program can be found in the readme file accompanying BLASTZ. Bl2seq 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. Attorney Docket No.: 45049-0091WO1 / 06521 [0033] To compare two amino acid sequences, the options of Bl2seq are set as follows: -i is set to a file containing the first amino acid sequence to be compared (e.g., C:\seq1.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:\Bl2seq -i c:\seq1.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 98.5 percent identical to the sequence set forth in SEQ ID NO:1 (i.e., 725 ÷ 736 x 100 = 98.5). 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. [0034] 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 AAV9 capsid polypeptide. See, for example, Zoller, Curr. Opin. Biotechnol.3: 348-354 (1992). [0035] The AAV9 vectors described herein can be designed to include one or more exogenous nucleic acid sequences. For example, an AAV9 vector described herein can be designed to include 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 AAV9 vector provided herein include, without limitation, siRNAs, RNA components for gene editing, and microRNAs. In some cases, an RNA of interest that can be encoded by an exogenous nucleic acid sequence included within Attorney Docket No.: 45049-0091WO1 / 06521 an AAV9 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., diabetic 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 type 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). [0036] 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 AAV9 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 AAV9 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 RS1 polypeptide, a TYR polypeptide, a USH2A polypeptide, a MYO7A polypeptide, an REP1 polypeptide, an OPN1LW polypeptide, an OPN1MW polypeptide, a CNGA3 polypeptide, a CNGB3 polypeptide, a GUCY2D polypeptide, a GACA1A polypeptide, a GNAT2 polypeptide, a PDE6H polypeptide, a PROM1 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 Attorney Docket No.: 45049-0091WO1 / 06521 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, a DNAJC5 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-ORF15 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, a HARS polypeptide, an RP2 polypeptide, a FAM161 polypeptide, a DLK polypeptide, a RHO polypeptide, a CHM polypeptide, a BEST1 polypeptide, a RP1 polypeptide, an OPA1 polypeptide, a CEP290 polypeptide, a RDH12 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 TULP1 polypeptide, a C2ORF71 (aka PCARE) polypeptide, a MAK polypeptide, a TIMP3 polypeptide, a GUCA1A polypeptide, an ALMS1 polypeptide, a BBS10 polypeptide, an IFT140 polypeptide, a CNGA1 polypeptide, a NMNAT1 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 C1QTNF5 polypeptide, a VPS13B polypeptide, a KIF11 polypeptide, a MT-TL1 polypeptide, a KLHL7 polypeptide, an ACO2 polypeptide, a C21orf2 (aka CFAP410) polypeptide, an AHI1 polypeptide, a KIZ polypeptide, a SPATA7 polypeptide, a TTLL5 polypeptide, an HGSNAT polypeptide, a NRL 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 HK1 polypeptide, a PRDM13 polypeptide, an ADAM9 Attorney Docket No.: 45049-0091WO1 / 06521 polypeptide, a BBS7 polypeptide, a CABP4 polypeptide, an ABHD12 polypeptide, a COL18A1 polypeptide, a MFRP polypeptide, a RIMS1 polypeptide, a ROM1 polypeptide, a BBS4 polypeptide, an IMPG1 polypeptide, an INPP5E polypeptide, a VCAN 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, a NDP polypeptide, a NPHP1 polypeptide, a RGR polypeptide, a BBS5 polypeptide, a WDR19 polypeptide, a C8ORF37 polypeptide, a CTNNA1 polypeptide, a LAMP2 polypeptide, a PEX1 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, a PNPLA6 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, a HMCN1 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 MKS1 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 OFD1 polypeptide, a P3H2 polypeptide, a RGS9BP polypeptide, a CSPP1 polypeptide, an ITM2B polypeptide, a PANK2 polypeptide, a PEX7 polypeptide, a POMGNT1 polypeptide, a SLC4A7 polypeptide, a TMEM231 polypeptide, a TRNT1 polypeptide, a TUBGCP6 polypeptide, a ZNF513 polypeptide, an AFG3L2 polypeptide, an ARL13B polypeptide, a C5ORF42 (aka CPLANE1) polypeptide, a COL9A1 polypeptide, a CTSD polypeptide, a DTHD1 polypeptide, a DYNC2H1 polypeptide, an IFT81 polypeptide, a KIAA0586 polypeptide, a MFN2 polypeptide, a NPHP3 polypeptide, a PCYT1A polypeptide, a PEX12 polypeptide, a PLA2G5 polypeptide, a Attorney Docket No.: 45049-0091WO1 / 06521 POC5 polypeptide, a SCAPER polypeptide, a SLC25A46 polypeptide, a TMEM237 polypeptide, a TRAF3IP1 polypeptide, a TTC21B polypeptide, a TUBGCP4 polypeptide, an ADIPOR1 polypeptide, a CEP164 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- RNR1 polypeptide, a MT-TS2 polypeptide, a ND5 (MT-ND5) polypeptide, a NEK2 polypeptide, an OPN1SW polypeptide, a PEX13 polypeptide, a PEX2 polypeptide, a RHBDD2 polypeptide, a SAMD11 polypeptide, a SCLT1 polypeptide, a SLC7A14 polypeptide, a TCTN1 polypeptide, a TCTN2 polypeptide, a TLCD3B polypeptide, a TREX1 polypeptide, a TTPA polypeptide, an UNC119 polypeptide, a WDPCP polypeptide, an ACBD5 polypeptide, an AHR polypeptide, an ARMC9 polypeptide, an ASRGL1 polypeptide, an ATOH7 polypeptide, a B9D1 polypeptide, a B9D2 polypeptide, a BBIP1 polypeptide, a C12ORF65 polypeptide, a C2CD3 polypeptide, a C5AR2 polypeptide, a CCDC188 polypeptide, a CCT2 polypeptide, a CEP104 polypeptide, a CEP120 polypeptide, a CEP19 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 HMX1 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-CO1 polypeptide, a MT-CO2 polypeptide, a MT-CO3 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, a NEUROD1 polypeptide, a PDE6D polypeptide, a PEX10 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 Attorney Docket No.: 45049-0091WO1 / 06521 polypeptide, a PROS1 polypeptide, a PSEN1 polypeptide, a RDH11 polypeptide, a RRM2B polypeptide, a SMARCA4 polypeptide, a SPP2 polypeptide, a TCTN3 polypeptide, a TEAD1 polypeptide, a TMEM107 polypeptide, a TMEM138 polypeptide, a TMEM216 polypeptide, a TMEM67 polypeptide, a TPP1 polypeptide, a TRIM32 polypeptide, a USP45 polypeptide, and a ZNF423 polypeptide. [0037] In some cases, one or more AAV9 vectors provided herein can be designed to carry out gene editing within one or more cells (e.g., CNS cells). Such gene editing can 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. [0038] Any appropriate gene editing components can be engineered into one or more AAV9 vectors provided herein such that those one or more AAV9 vectors can be used to deliver the gene editing components to target cells (e.g., one or more CNS 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)/Cas 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)). Attorney Docket No.: 45049-0091WO1 / 06521 [0039] In some cases, to facilitate gene replacement, two sequences in genomic nucleic acid of a cell (e.g., a CNS 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 AAV9 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 the sequence between the genomic targets, and insertion of the donor sequence into the location of the deletion. [0040] An AAV9 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, SV40 promotors, CMV promotors, 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 promotors that can be used as described herein include, without limitation, rhodopsin promotors, cone arrestin promotors, and synapsin promotors. Examples of viral promotors that can be used as described herein include, without limitation, adenoviral promotors, vaccinia virus promotors, CMV promotors (e.g., immediate early CMV promotors), and AAV promoters. [0041] In some cases, an AAV9 vector provided herein can include a total number of nucleotides up to about 5 kb. In some cases, an AAV9 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. [0042] An AAV9 vector described herein containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B can have the ability to Attorney Docket No.: 45049-0091WO1 / 06521 infect CNS cells in vivo and deliver exogenous nucleic acid sequence to the infected CNS cells such that the infected CNS cells express the exogenous nucleic acid sequence. In some cases, an AAV9 vector provided herein can have the ability to infect and drive RNA expression of an exogenous nucleic acid sequence in more CNS cells of a mammal (e.g., a human or a non-human primate) when compared to wild- type AAV9. In some cases, an AAV9 vector provided herein can have the ability to drive a level of RNA expression of an exogenous nucleic acid sequence in CNS 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 AAV9 vector (e.g., wild-type AAV9) having an AAV9 capsid polypeptide that consists of the amino acid sequence set forth in SEQ ID NO:1 in CNS cells of a control mammal (e.g., a control human or a control non-human primate). [0043] Examples of CNS cells that can be infected by an AAV9 vector described herein containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B include, without limitation, CNS cell types such as neurons, glial cells (e.g., astrocytes, oligodendrocytes, ependymal cells, and microglia), choroid plexus cells, and cells related to blood vessels and coverings. [0044] This document also provides compositions containing one or more AAV9 vectors provided herein. For example, one or more AAV9 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 cases, one or more AAV9 vectors provided 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 CNS cells for expression within CNS cells. For example, an AAV9 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, Attorney Docket No.: 45049-0091WO1 / 06521 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 AAV9 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. [0045] In some cases, when a pharmaceutical composition is formulated to include one or more AAV9 vectors provided herein, any appropriate titer of the AAV9 vectors can be used. For example, a pharmaceutical composition provided herein can be formulated to have AAV9 vectors provided herein at a titer that is greater than 1x107 (e.g., greater than 1 x 108, greater than 1 x 109, greater than 1 x 1010, greater than 1 x 1011, greater than 1 x 1012, greater than 1 x 1013, or greater than 1 x 1014). In some cases, a pharmaceutical composition provided herein can be formulated to have AAV9 vectors (provided herein at a titer that is from about 1x107 to about 1x1014 (e.g., from about 1 x 107 to about 1 x 1013, from about 1 x 107 to about 1 x 1012, from about 1 x 107 to about 1 x 1011, from about 1 x 107 to about 1 x 1010, from about 1 x 108 to about 1 x 1014, from about 1 x 109 to about 1 x 1014, from about 1 x 1010 to about 1 x 1014, from about 1 x 108 to about 1 x 1012, or from about 1 x 109 to about 1 x 1011). [0046] 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 cases, 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 cases, a pharmaceutical composition provided herein can be lyophilized. In some cases, a pharmaceutical composition provided herein (e.g., a pharmaceutical composition that includes one or more AAV9 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). Attorney Docket No.: 45049-0091WO1 / 06521 [0047] This document also provides nucleic acid molecules encoding an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. In some cases, a nucleic acid molecule can be designed to encode an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1A (e.g., any one of SEQ ID NOs:6-9). In some cases, a nucleic acid molecule can be designed to encode an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1B (e.g., any one of SEQ ID NOs:15-18). [0048] This document also provides nucleic acid molecules encoding an AAV9 vector described herein. For example, an isolated nucleic acid molecule can be designed to encode one or more AAV9 vectors provided herein (e.g., an AAV9 having an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B). In some cases, a nucleic acid molecule can be designed to encode an AAV9 vector having an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1A (e.g., any one of SEQ ID NOs:6-9). In some cases, a nucleic acid molecule can be designed to encode an AAV9 vector having an AAV9 capsid polypeptide that includes an amino acid sequence that is encoded by a DNA sequence set forth in Table 1B (e.g., any one of SEQ ID NOs:15-18). [0049] This document also provides host cells containing a nucleic acid molecule provided herein. For example, a host cell can be designed to include a nucleic acid molecule encoding an AAV9 capsid polypeptide described herein and/or a nucleic acid molecule encoding an AAV9 vector described herein. In some cases, a host cell can be designed to include a nucleic acid molecule encoding an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. In some cases, a host cell can be designed to include a nucleic acid molecule encoding an AAV9 vector having an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. Examples of host cells that can be designed to include a nucleic acid molecule encoding an AAV9 capsid polypeptide described herein and/or a nucleic acid molecule encoding an AAV9 vector described Attorney Docket No.: 45049-0091WO1 / 06521 herein include, without limitation, HEK293T cells (ATCC), 293AAV9 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 AAV9 capsid polypeptide described herein and/or an AAV9 vector described herein) into a cell. For example, viral transfection, electroporation, transient transfection, and gene gun techniques can be used to introduce a nucleic acid molecule provided herein into a cell. [0050] This document also provides methods and materials for making an AAV9 provided herein. For example, this document provides methods and materials for making AAV9 vectors containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. As described herein, an AAV9 vector can be constructed to include an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. Any appropriate method can be used to construct an AAV9 vector having an AAV9 capsid polypeptide provided herein (e.g., a capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B). For example, molecular cloning and AAV9 vector production techniques such as those described elsewhere can be used to construct and produce an AAV9 vector having an AAV9 capsid polypeptide provided herein (see, e.g., Sambrook et al., Molecular Cloning: A Laboratory Manual, 2nd edition, Cold Spring Harbor Laboratory, NY (1989); Ausubel et 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, AAV9 vectors can be produced in HEK293T cells (ATCC) or 293AAV9 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)). [0051] This document also provides methods and materials for using an AAV9 vector provided herein. For example, this document provides methods and materials for using AAV9 vectors containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B. As described herein, an AAV9 vector provided herein can be used Attorney Docket No.: 45049-0091WO1 / 06521 to infect CNS cells in vivo and to deliver an exogenous nucleic acid sequence to the infected CNS cells such that the infected CNS cells express the exogenous nucleic acid sequence. [0052] In some cases, an AAV9 vector provided herein (e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B) can be used to treat a CNS disorder (e.g., a CNS disease). For example, an AAV9 vector provided herein that is designed to contain and drive expression of an exogenous nucleic acid sequence encoding an RNA and/or polypeptide capable of treating a CNS disorder (e.g., a CNS disease) can be administered to a mammal (e.g., a human or a non- human primate) having a CNS disorder in a manner such that the AAV9 vector (a) infects CNS cells and (b) drives expression of the delivered exogenous nucleic acid in the infected CNS cells, thereby reducing the severity of one or more symptoms of the CNS disorder and/or slowing the progression of the CNS disorder. [0053] As described herein, an AAV9 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 AAV9 vector provided herein is designed to treat a CNS disorder, an exogenous nucleic acid sequence that encodes an RNA and/or polypeptide capable of treating the CNS disorder can be included within the AAV9 vector. Examples of RNAs that can be encoded by an exogenous nucleic acid sequence designed to treat a disorder (e.g, a CNS disorder or other disorder such as an eye disorder) and designed to be included within an AAV9 vector provided herein include, without limitation, 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., diabetic 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 type 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 toage-related macular degeneration (see, e.g., NCT03815825), and Sepofarsen/QR-110 to treat, e.g., Leber’s congenital amaurosis (see, e.g., NCT03913143). Examples of polypeptides that can be encoded by an exogenous nucleic Attorney Docket No.: 45049-0091WO1 / 06521 acid sequence designed to treat a disorder (e.g, a CNS disorder or other disorder such as an eye disorder) such as disclosed herein and designed to be included within an AAV9 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 RS1 polypeptide, a TYR polypeptide, a USH2A polypeptide, a MYO7A polypeptide, an REP1 polypeptide, an OPN1LW polypeptide, an OPN1MW polypeptide, a CNGA3 polypeptide, a CNGB3 polypeptide, a GUCY2D polypeptide, a GACA1A polypeptide, a GNAT2 polypeptide, a PDE6H polypeptide, a PROM1 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, a DNAJC5 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-ORF15 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, a HARS polypeptide, an RP2 polypeptide, a FAM161 polypeptide, a DLK polypeptide, a RHO polypeptide, a CHM polypeptide, a BEST1 polypeptide, a RP1 polypeptide, an OPA1 polypeptide, a CEP290 polypeptide, a RDH12 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 Attorney Docket No.: 45049-0091WO1 / 06521 AIPL1 polypeptide, a RPGRIP1 polypeptide, a TULP1 polypeptide, a C2ORF71 (aka PCARE) polypeptide, a MAK polypeptide, a TIMP3 polypeptide, a GUCA1A polypeptide, an ALMS1 polypeptide, a BBS10 polypeptide, an IFT140 polypeptide, a CNGA1 polypeptide, a NMNAT1 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 C1QTNF5 polypeptide, a VPS13B polypeptide, a KIF11 polypeptide, a MT-TL1 polypeptide, a KLHL7 polypeptide, an ACO2 polypeptide, a C21orf2 (aka CFAP410) polypeptide, an AHI1 polypeptide, a KIZ polypeptide, a SPATA7 polypeptide, a TTLL5 polypeptide, an HGSNAT polypeptide, a NRL 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 HK1 polypeptide, a PRDM13 polypeptide, an ADAM9 polypeptide, a BBS7 polypeptide, a CABP4 polypeptide, an ABHD12 polypeptide, a COL18A1 polypeptide, a MFRP polypeptide, a RIMS1 polypeptide, a ROM1 polypeptide, a BBS4 polypeptide, an IMPG1 polypeptide, an INPP5E polypeptide, a VCAN 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, a NDP polypeptide, a NPHP1 polypeptide, a RGR polypeptide, a BBS5 polypeptide, a WDR19 polypeptide, a C8ORF37 polypeptide, a CTNNA1 polypeptide, a LAMP2 polypeptide, a PEX1 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, a PNPLA6 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, a HMCN1 polypeptide, a MT-ATP6 polypeptide, a Attorney Docket No.: 45049-0091WO1 / 06521 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 MKS1 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 OFD1 polypeptide, a P3H2 polypeptide, a RGS9BP polypeptide, a CSPP1 polypeptide, an ITM2B polypeptide, a PANK2 polypeptide, a PEX7 polypeptide, a POMGNT1 polypeptide, a SLC4A7 polypeptide, a TMEM231 polypeptide, a TRNT1 polypeptide, a TUBGCP6 polypeptide, a ZNF513 polypeptide, an AFG3L2 polypeptide, an ARL13B polypeptide, a C5ORF42 (aka CPLANE1) polypeptide, a COL9A1 polypeptide, a CTSD polypeptide, a DTHD1 polypeptide, a DYNC2H1 polypeptide, an IFT81 polypeptide, a KIAA0586 polypeptide, a MFN2 polypeptide, a NPHP3 polypeptide, a PCYT1A polypeptide, a PEX12 polypeptide, a PLA2G5 polypeptide, a POC5 polypeptide, a SCAPER polypeptide, a SLC25A46 polypeptide, a TMEM237 polypeptide, a TRAF3IP1 polypeptide, a TTC21B polypeptide, a TUBGCP4 polypeptide, an ADIPOR1 polypeptide, a CEP164 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- RNR1 polypeptide, a MT-TS2 polypeptide, a ND5 (MT-ND5) polypeptide, a NEK2 polypeptide, an OPN1SW polypeptide, a PEX13 polypeptide, a PEX2 polypeptide, a RHBDD2 polypeptide, a SAMD11 polypeptide, a SCLT1 polypeptide, a SLC7A14 polypeptide, a TCTN1 polypeptide, a TCTN2 polypeptide, a TLCD3B polypeptide, a TREX1 polypeptide, a TTPA polypeptide, an UNC119 polypeptide, a WDPCP polypeptide, an ACBD5 polypeptide, an AHR polypeptide, an ARMC9 polypeptide, an ASRGL1 polypeptide, an ATOH7 polypeptide, a B9D1 polypeptide, a B9D2 polypeptide, a BBIP1 polypeptide, a C12ORF65 polypeptide, a C2CD3 polypeptide, a C5AR2 polypeptide, a CCDC188 polypeptide, a CCT2 polypeptide, a CEP104 polypeptide, a CEP120 polypeptide, a CEP19 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 Attorney Docket No.: 45049-0091WO1 / 06521 HKDC1 polypeptide, a HMX1 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-CO1 polypeptide, a MT-CO2 polypeptide, a MT-CO3 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, a NEUROD1 polypeptide, a PDE6D polypeptide, a PEX10 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 PROS1 polypeptide, a PSEN1 polypeptide, a RDH11 polypeptide, a RRM2B polypeptide, a SMARCA4 polypeptide, a SPP2 polypeptide, a TCTN3 polypeptide, a TEAD1 polypeptide, a TMEM107 polypeptide, a TMEM138 polypeptide, a TMEM216 polypeptide, a TMEM67 polypeptide, a TPP1 polypeptide, a TRIM32 polypeptide, a USP45 polypeptide, and a ZNF423 polypeptide. [0054] Any appropriate CNS disorder can be treated using an AAV9 vector provided herein (e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, and an exogenous nucleic acid sequence encoding a therapeutic RNA and/or polypeptide). Examples of such CNS conditions include, without limitation, Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, restless leg syndrome, amyotrophic lateral sclerosis (ALS), aneurysm, back pain, Bell’s Palsy, birth defects of the brain and spinal cord, brain injury, brain tumor, cerebral palsy, chronic fatigue syndrome, concussion, dementia, disc disease of neck and lower back, dizziness, frontotemporal dementia, Guillain- Barré syndrome, headaches and migraine, Lewy body dementia, mild cognitive impairment, muscular dystrophy, neuralgia, neuropathy, neuromuscular and related diseases, primary Attorney Docket No.: 45049-0091WO1 / 06521 progressive aphasia, psychiatric conditions (e.g., severe depression or obsessive-compulsive disorder), scoliosis, seizures, spinal cord injury, spinal deformity and disorders, spine tumor, stroke, sundown syndrome, vascular dementia, and vertigo. In some cases, an AAV9 vector provided herein (e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, and an exogenous nucleic acid sequence encoding a therapeutic RNA and/or polypeptide) can be used to treat a peripheral nervous system (PNS) disorder such as acute motor axonal neuropathy, botulism, Charcot- Marie-Tooth disease types 1A, 1B, and 1X, cisplatin neuropathy, diabetic neuropathy, diphtheritic neuropathy, familial amyloid neuropathy, Guillain-Barré syndrome, Lambert-Eaton syndrome, leprosy, neuropathy with IgM1 anti-myelin-associated glycoprotein, pyridoxine neuropathy, or Refsum’s disease. [0055] In some cases, a CNS disorder can be treated using an AAV9 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 cases, wet AMD can be treated using an AAV9 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 cases, diabetic retinopathy can be treated using an AAV9 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 cases, diabetic macular edema can be treated using an AAV9 vector provided herein that is designed to express a monoclonal anti-VEGF antibody polypeptide, an angiostatin polypeptide, an siRNA, and/or an endostatin polypeptide. [0056] In some cases, a CNS disorder can be treated using an AAV9 vector provided herein that is designed to express one or more polypeptides with neuroprotective capabilities and/or other CNS delivery 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 cases, diabetic macular edema can be treated using an AAV9 vector Attorney Docket No.: 45049-0091WO1 / 06521 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. [0057] In some cases, a CNS disorder can be treated using an AAV9 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, XIAP polypeptides, cIAP1 polypeptides, C-IAP2 polypeptides, Livin polypeptides, and Survivin polypeptides. [0058] In some cases, a CNS disorder can be treated using an AAV9 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, a CNS disorder such as contemplated herein can be treated using an AAV9 vector provided herein that is designed to express a Complement Factor I polypeptide, a Complement factor H polypeptide, and/or a sCD59 polypeptide. [0059] In some cases, a CNS disorder can be treated using an AAV9 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 cases, a CNS disorder such as one or more of Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, restless leg syndrome, amyotrophic lateral sclerosis (ALS), aneurysm, back pain, Bell’s Palsy, birth defects of the brain and spinal cord, brain injury, brain tumor, cerebral palsy, chronic fatigue syndrome, concussion, dementia, disc disease of neck and lower back, dizziness, frontotemporal dementia, Guillain-Barré syndrome, headaches and migraine, Lewy body dementia, mild cognitive impairment, muscular dystrophy, neuralgia, neuropathy, neuromuscular and related diseases, primary progressive aphasia, psychiatric conditions (e.g., severe depression or obsessive- compulsive disorder), scoliosis, seizures, spinal cord injury, spinal deformity and disorders, spine tumor, stroke, sundown syndrome, vascular dementia, and vertigo can be treated using an AAV9 Attorney Docket No.: 45049-0091WO1 / 06521 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. [0060] Any appropriate method can be used to administer an AAV9 vector provided herein or composition (e.g., a pharmaceutical composition) provided herein to a mammal (e.g., a human or a non-human primate). For example, a composition provided herein (e.g., a pharmaceutical composition containing one or more AAV9 vectors provided herein) can be administered to a mammal (e.g., a human or a non-human primate) intrathecally, 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, intramuscularly (e.g., via intramuscular injection), subretinally, intracranially, intravitreally, systemically, or suprachoroidally. In some cases, 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. [0061] In some cases, an effective amount of a composition containing an AAV9 vector provided herein (e.g., a pharmaceutical composition provided herein) to treat a CNS disorder can be an amount that reduces the severity of one or more symptoms of the CNS disorder and/or slows the progression of the CNS disorder without producing significant toxicity to the mammal. For example, an effective amount of an AAV9 vector provided herein can be from about 1x107 viral genomes to about 1x1014 viral genomes (e.g., from about 1 x 107 viral genomes to about 1 x 1013 viral genomes, from about 1 x 107 viral genomes to about 1 x 1012 viral genomes, from about 1 x 107 viral genomes to about 1 x 1011 viral genomes, from about 1 x 107 viral genomes to about 1 x 1010 viral genomes, from about 1 x 108 viral genomes to about 1 x 1014 viral genomes, from about 1 x 109 viral genomes to about 1 x 1014 viral genomes, from about 1 x 1010 viral genomes to about 1 x 1014 viral genomes, from about 1 x 108 viral genomes to about 1 x 1012 viral genomes, or from about 1 x 109 viral genomes to about 1 x 1011 viral genomes). In some cases, an effective amount of an AAV9 vector provided herein can be from about 1 x 1010 viral genomes/kg of body weight to about 1 x 1014 viral genomes/kg of body weight (e.g., from about 1 x 1010 viral genomes/kg of body weight to about 1 x 1013 viral genomes/kg of body weight, from about 1 x 1010 viral genomes/kg of body weight to about 1 x 1012 viral genomes/kg of body weight, from about 1 x 1010 viral genomes/kg of body weight to about 1 x 1011 viral 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 Attorney Docket No.: 45049-0091WO1 / 06521 influence the actual effective amount used for a particular application. For example, the severity of a CNS 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 CNS 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 AAV9 vector provided herein) that is administered. [0062] In some cases, an effective frequency of administration of a composition containing an AAV9 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 CNS disorder and/or slows the progression of the CNS disorder without producing significant toxicity to the mammal. Various factors can influence the actual effective frequency used for a particular application. For example, the severity of a CNS 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 CNS 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. [0063] In some cases, an effective duration of administration of a composition containing an AAV9 vector provided herein (e.g., a pharmaceutical composition provided herein) can be a duration that reduces the severity of one or more symptoms of the CNS disorder and/or slows the progression of the CNS disorder 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 cases, 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 CNS 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 CNS 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 AAV9 vector provided herein). Attorney Docket No.: 45049-0091WO1 / 06521 [0064] In some cases, an effective amount of a composition containing an AAV9 vector provided herein (e.g., a pharmaceutical composition provided herein) to treat a CNS disorder can be administered once or twice to a mammal (e.g., a human or a non-human primate) to treat that mammal. Treatment of CNS disorders [0065] Also contemplated herein are methods of treating patients suffering from a CNS disorder such as one or more of Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, restless leg syndrome, amyotrophic lateral sclerosis (ALS), aneurysm, back pain, Bell’s Palsy, birth defects of the brain and spinal cord, brain injury, brain tumor, cerebral palsy, chronic fatigue syndrome, concussion, dementia, disc disease of neck and lower back, dizziness, frontotemporal dementia, Guillain- Barré syndrome, headaches and migraine, Lewy body dementia, mild cognitive impairment, muscular dystrophy, neuralgia, neuropathy, neuromuscular and related diseases, primary progressive aphasia, psychiatric conditions (e.g., severe depression or obsessive-compulsive disorder), scoliosis, seizures, spinal cord injury, spinal deformity and disorders, spine tumor, stroke, sundown syndrome, vascular dementia, and vertigo by administering an effective amount of therapeutic composition comprising an AAV9 vector provided herein (e.g., an AAV9 vector containing an AAV9 capsid polypeptide that includes an amino acid sequence set forth in Table 1A (or a variant thereof) or Formula A or an amino acid sequence set forth in Table 1B (or a variant thereof) or Formula B, and an exogenous nucleic acid sequence encoding a therapeutic RNA and/or polypeptide). [0066] In certain embodiments, the subject treated in accordance with the methods disclosed herein has not received prior therapy for treating the CNS disorder. In certain embodiments, the subject treated in accordance with the methods disclosed herein has received a prior therapy but continues to experience progression of the CNS disorder despite the prior therapy. EXAMPLES [0067] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims. Attorney Docket No.: 45049-0091WO1 / 06521 Example 1 – Construction of AAV9 vectors containing mutated capsid polypeptides [0068] A high-throughput method is used to create AAV9 vectors with mutated capsid polypeptides and to screen those created AAV9 vectors for particular AAV9 vectors having the ability to exhibit high efficiency and/or specificity for infecting CNS cells. See, e.g., Öztürk et al., bioRxiv, 2020.10.01.323196 (2020) and Öztürk et al., eLife, 10:e64175 (2021). Briefly, highly complex libraries of AAV9 mutants are created and injected into the CNS of primates (cynolmolgus macaques or rhesus macaques). These libraries are created such that each AAV9 vector in the library contained a unique DNA barcode, which allowed for tracking of a mutated AAV9 capsid polypeptide. In one library version, successfully packaged AAV9 vectors are polymerase chain reaction (PCR) amplified and repackaged, resulting in a “repack” library. In another library version, AAV9 vectors are injected into primate retinas, and nucleic acid encoding the AAV9 capsid polypeptides are then amplified from the nuclei of foveal cells, resulting in an “enriched” library. Each iteration of the AAV9 library (e.g., the original library, the repack library, and the enriched library) is injected, e.g., intrathecally into the CNS of primates. [0069] After injection, the AAV9 vectors compete with each other in vivo. Infection of successful AAV9 vectors can lead to expression of the DNA barcodes. Single cell suspensions are created from isolated CNS fluid, and single cell microfluidic technology (10X Genomics) is used to create cDNA libraries of individual cells. Computational analysis is performed to identify optimal vectors, according to cell specificities, expression levels, and/or other desirable characteristics, based on the presence and quantity of DNA barcodes in transcriptomes from different cells of multiple cell types in parallel. The performance of AAV9 capsid polypeptides is evaluated on the basis of mRNA transcription levels rather than the presence of DNA, reflecting the ability of the AAV9 vectors to drive expression of the AAV9 vector nucleic acid as opposed to simply having the ability to enter a cell. [0070] AAV9 vectors having capsid polypeptides that include SEQ ID NO:5 or 14 located between amino acid residues 588 and 589 of SEQ ID NO:1) can exhibit the ability to infect and drive mRNA expression within CNS cells in a manner similar to that of 7m8 AAV9 vector. Thus, the AAV9 vector having a capsid polypeptide that includes SEQ ID NO:5 or 14 can have the ability to exhibit more infectivity of CNS cells than a wild-type AAV9 vector containing an AAV9 capsid polypeptide that consists of SEQ ID NO:1. Attorney Docket No.: 45049-0091WO1 / 06521 [0071] Taken together, this can demonstrate that AAV9 vectors that include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1A (or Formula A) can have the ability to mediate transgene expression in CNS cells. This also can demonstrate that AAV9 vectors that include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1B (or Formula B) can have the ability to mediate transgene expression in CNS cells. Example 2 – Treating a CNS disorder using an AAV9 vector [0072] An AAV9 vector is constructed to include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1A (e.g., SEQ ID NO:2 or 5) or Formula A and an exogenous nucleic acid sequence encoding a therapeutic polypeptide. The constructed AAV9 vector is administered intravitreally to a human identified as having a CNS disorder in an amount that is from about 1 x 107 to about 1 x 1014 AAV9 vectors. After the administration, the severity of one or more symptoms of the CNS disorder is reduced and/or the progression of the CNS disorder is slowed. Example 3 – Treating a CNS disorder using an AAV9 vector [0073] An AAV9 vector is constructed to include an AAV9 capsid polypeptide having an amino acid sequence set forth in Table 1B (e.g., SEQ ID NO:11 or 14) or Formula B and an exogenous nucleic acid sequence encoding a therapeutic polypeptide. The constructed AAV9 vector is administered intravitreally to a human identified as having a CNS disorder in an amount that is from about 1 x 107 to about 1 x 1014 AAV9 vectors. After the administration, the severity of one or more symptoms of the CNS disorder is reduced and/or the progression of the CNS disorder is slowed. Example 4 – AAV vectors containing mutated capsids [0073] An in vivo analysis of AAV9 mutated capsids was conducted in mouse CNS via intra cisterna magna injections. Performance was measured by assessing the expression of the fluorophores expressed by each vector using immunohistochemistry (IHC) and/or fluorescent microscopy. Fluorophore expression was used as a proxy for capsid efficacy, with higher fluorophore expression indicating higher viral transduction. Fluorophore expression was qualitatively analyzed by scoring relative fluorophore expression in regions of the brain and Attorney Docket No.: 45049-0091WO1 / 06521 spinal cord. As indicated in Table 2, fluorophore expression of both capsids of SEQ ID NO:10 and SEQ ID NO:19 were observed in regions of the brain and spinal cord, indicating their efficacy in viral transduction. Table 2. Qualitative Fluorescence Score (0 to +++) escence.
Figure imgf000038_0001
OTHER EMBODIMENTS Embodiment 1. An adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide having 95% identity or more with SEQ ID NO:1, or a truncated variant thereof, further comprising an amino acid peptide sequence comprising a sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14, wherein the amino acid peptide sequence is located between two existing amino acids of SEQ ID NO:1. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 2. The vector of Embodiment 1, wherein said amino acid peptide sequence is located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of SEQ ID NO:1. Embodiment 3. The vector of Embodiment 1 or 2, wherein the amino acid peptide sequence is SEQ ID NO:2 or 11. Embodiment 4. The vector of Embodiment 1 or 2, wherein the amino acid peptide sequence is SEQ ID NO:5 or 14. Embodiment 5. An adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide of SEQ ID NO:1, wherein the amino acids from position 585 to 590 of SEQ ID NO:1 is replaced with an amino acid peptide sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14. Embodiment 6. The vector of Embodiment 5, wherein the amino acids from position 585 to 590 of SEQ ID NO:1 are replaced with said amino acid sequence of SEQ ID NO:2 or 11. Embodiment 7. The vector of any one of Embodiments 1-6, wherein said vector infects greater than 2.5 percent of central nervous system (CNS) cells in one or more brain or spinal cord regions including the cortex, hippocampus, midbrain, cerebellum, brainstem, ventral spinal horn, and dorsal spinal horn when a titer of at least 1 x 107 of said vector is administered into the CNS of a patient (e.g., via intra cisterna magna (ICM), intraparenchymal (IPa), intraventricular, or intrathecal (IT) administration), or administered intravenously or subcutaneously. Embodiment 8. The vector of any one of Embodiments 1-7, wherein said vector comprises an exogenous nucleic acid encoding an RNA or a polypeptide. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 9. The vector of Embodiment 8, wherein said exogenous nucleic acid encodes an RNA. Embodiment 10. The vector of Embodiment 9, wherein said RNA is an siRNA or microRNA. Embodiment 11. The vector of Embodiment 8, wherein said exogenous nucleic acid encodes a polypeptide. Embodiment 12. The vector of Embodiment 11, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide. Embodiment 13. The vector of any one of Embodiments 1-12, wherein said vector expresses more nucleic acid in CNS cells than the level of expression from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1. Embodiment 14. A composition comprising a vector of any one of Embodiments 1-13 and a pharmaceutically acceptable excipient. Embodiment 15. The composition of Embodiment 14, wherein said composition comprises from about 1 x 107 to about 1 x 1014 of said vector. Embodiment 16. The composition of Embodiment 14 or 15, wherein said pharmaceutically acceptable excipient comprises one or more of phosphate buffered saline, Hank’s Balanced Salt Solution, artificial cerebrospinal fluid, and Pluronic F68. Embodiment 17. A method for delivering an exogenous nucleic acid sequence to a CNS of a mammal, wherein said method comprises: contacting CNS cells of said mammal with an AAV9 vector comprising an AAV9 capsid polypeptide and said exogenous nucleic acid sequence, wherein said capsid polypeptide comprises an amino acid peptide sequence selected from the group consisting of SEQ ID NOs:2- 5 and 11-14; Attorney Docket No.: 45049-0091WO1 / 06521 wherein upon the contacting said AAV9 vector infects said CNS cells and thereby delivering said exogenous nucleic acid sequence to said CNS cells. Embodiment 18. The method of Embodiment 17, wherein said capsid polypeptide comprises the amino acid sequence of SEQ ID NO:1 with the exception that the amino acid peptide sequence is located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of SEQ ID NO:1, or with the exception that the amino acid residues at positions 585 to 590 of SEQ ID NO:1 are replaced with said amino acid peptide sequence. Embodiment 19. The method of Embodiment 18, wherein said amino acid peptide sequence is located between amino acid positions 588 and 589 of SEQ ID NO:1. Embodiment 20. The method of any one of Embodiments 17-19, wherein said amino acid peptide sequence is SEQ ID NO:2 or 11. Embodiment 21. The method of any one of Embodiments 17-20, wherein said mammal is a human. Embodiment 22. The method of any one of Embodiments 17-21, wherein said exogenous nucleic acid sequence encodes an RNA or a polypeptide. Embodiment 23. The method of Embodiment 22, wherein said exogenous nucleic acid encodes said RNA. Embodiment 24. The method of Embodiment 23, wherein said RNA is an siRNA or microRNA. Embodiment 25. The method of Embodiment 22, wherein said exogenous nucleic acid encodes said polypeptide. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 26. The method of Embodiment 25, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide. Embodiment 27. The method of any one of Embodiments 17-26, wherein said vector expresses more of said exogenous nucleic acid sequence in said CNS cell than the level of expression in a CNS cell from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1; or wherein said vector expresses said exogenous nucleic acid sequence in more CNS cells than the number of CNS cells where said exogenous nucleic acid sequence is expressed from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1; or wherein said vector expresses said exogenous nucleic acid sequence in different CNS cells than the CNS cells where said exogenous nucleic acid sequence is expressed from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1. Embodiment 28. The method of any one of Embodiments 17-27, wherein said method comprises intravenously, subcutaneously, intracranially, and/or intrathecally administering a composition comprising said vector to said mammal, thereby contacting said CNS cell with said vector. Embodiment 29. The method of Embodiment 28, wherein said composition comprises from about 1 x 107 to about 1 x 1014 of said vector, or wherein said composition comprises from about 1 x 107 to about 1 x 1015 of said vector. Embodiment 30. A method for treating a CNS disorder in a mammal in need thereof, wherein said method comprises contacting CNS cells of a mammal having said CNS disorder with a therapeutic construct comprising the AAV9 vector of any one of Embodiments 1-13 and an exogenous nucleic acid sequence. Embodiment 31. The method of Embodiment 30, wherein said mammal is a human. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 32. The method of any one of Embodiments 30-31, wherein said CNS disorder is selected from the group consisting of Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, and restless leg syndrome. Embodiment 33. The method of any one of Embodiments 30-32, wherein said exogenous nucleic acid sequence encodes an RNA. Embodiment 34. The method of Embodiment 33, wherein said RNA is an siRNA or a microRNA. Embodiment 35. The method of any one of Embodiments 30-33, wherein said exogenous nucleic acid encodes a polypeptide. Embodiment 36. The method of Embodiment 35, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide. Embodiment 37. A non-naturally occurring adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide, wherein said capsid polypeptide comprises at least 95% identity to SEQ ID NO:1, or a truncated variant thereof, and further comprises an amino acid sequence insert of Formula A or Formula B located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 according to SEQ ID NO:1, wherein said Formula A is: -L1-EGSGRN (SEQ ID NO:2)-L2-, Attorney Docket No.: 45049-0091WO1 / 06521 wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids, and wherein said Formula B is: -L1-EHQTRP (SEQ ID NO:11)-L2-, wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids. Embodiment 38. The vector of Embodiment 37, wherein said L1 is one amino acid X1. Embodiment 39. The vector of Embodiment 38, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 40. The vector of Embodiment 38 or 39, wherein said X1 is A. Embodiment 41. The vector of Embodiment 37, wherein said L1 is two amino acids X2-X1. Embodiment 42. The vector of Embodiment 41, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 43. The vector of Embodiment 42, wherein said X1 is A. Embodiment 44. The vector of any one of Embodiments 41-43, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 45. The vector of Embodiment 44, wherein said X2 is L. Embodiment 46. The vector of Embodiment 45, wherein said X2-X1 is LA. Embodiment 47. The vector of Embodiment 37, wherein said L1 is three amino acids X3- X2-X1. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 48. The vector of Embodiment 47, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 49. The vector of Embodiment 48, wherein said X1 is A. Embodiment 50. The vector of any one of Embodiments 47-49, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 51. The vector of Embodiment 50, wherein said X2 is L. Embodiment 52. The vector of Embodiment 51, wherein said X2-X1 is LA. Embodiment 53. The vector of any one of Embodiments 47-52, wherein said X3 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 54. The vector of Embodiment 37, wherein said L1 is absent. Embodiment 55. The vector of any one of Embodiments 37-54, wherein said L2 is one amino acid Z1. Embodiment 56. The vector of Embodiment 55, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 57. The vector of Embodiment 56, wherein said Z1 is A. Embodiment 58. The vector of any one of Embodiments 37-54, wherein said L2 is two amino acids Z1-Z2. Embodiment 59. The vector of Embodiment 58, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L. Attorney Docket No.: 45049-0091WO1 / 06521 Embodiment 60. The vector of Embodiment 59, wherein said Z1 is A. Embodiment 61. The vector of any one of Embodiments 58-60, wherein said Z2 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 62. The vector of Embodiment 61, wherein said Z2 is L. Embodiment 63. The vector of Embodiment 62, wherein said Z1-Z2 is AL. Embodiment 64. The vector of any one of Embodiments 37-54, wherein said L2 is three amino acids Z1-Z2-Z3. Embodiment 65. The vector of Embodiment 64, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L, and/or Z2 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 66. The vector of Embodiment 65, wherein said Z1-Z2 is AL. Embodiment 67. The vector of any one of Embodiments 65-66, wherein said Z3 is selected from the group of amino acid residues consisting of A, V, I, and L. Embodiment 68. The vector of Embodiment 37, wherein said amino acid sequence insert comprises any one of SEQ ID NOs:2-5 and 11-14. Embodiment 69. A non-naturally occurring AAV9 capsid polypeptide, wherein said capsid polypeptide comprises at least 95% identity to SEQ ID NO:1, or a truncated variant thereof, and further comprises an amino acid sequence insert of Formula A or Formula B located 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 Attorney Docket No.: 45049-0091WO1 / 06521 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 according to SEQ ID NO:1, wherein said Formula A is: -L1-EGSGRN (SEQ ID NO:2)-L2-, wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids, and wherein said Formula B is: -L1-EHQTRP (SEQ ID NO:11)-L2-, wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids. Embodiment 70. A method for administering an exogenous nucleic acid sequence to a mammal in need thereof, wherein said method comprises administering to the mammal an effective amount of a construct comprising a vector of any one of Embodiments 37-68 and the exogenous nucleic acid sequence. Embodiment 71. A method of treating a CNS disorder in a patient in need thereof, comprising administering to the patient an effective amount of an AAV9 vector comprising an AAV9 capsid polypeptide comprising an amino acid sequence insert represented by Formula A or Formula B and an exogenous nucleic acid sequence. INCORPORATION BY REFERENCE [0074] The entire disclosure of each of the patent documents and scientific articles referred to herein is incorporated by reference for all purposes. EQUIVALENTS [0075] The disclosure may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The foregoing embodiments are therefore to be Attorney Docket No.: 45049-0091WO1 / 06521 considered in all respects illustrative rather than limiting the disclosure described herein. Various structural elements of the different embodiments and various disclosed method steps may be utilized in various combinations and permutations, and all such variants are to be considered forms of the disclosure. Scope of the disclosure is thus indicated by the appended claims rather than by the foregoing description, and all changes that come within the meaning and range of equivalency of the claims are intended to be embraced therein.

Claims

Attorney Docket No.: 45049-0091WO1 / 06521 WHAT IS CLAIMED IS: 1. An adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide having 95% identity or more with SEQ ID NO:1, or a truncated variant thereof, further comprising an amino acid peptide sequence comprising a sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14, wherein the amino acid peptide sequence is located between two existing amino acids of SEQ ID NO:1. 2. The vector of claim 1, wherein said amino acid peptide sequence is located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of SEQ ID NO:1. 3. The vector of claim 1 or 2, wherein the amino acid peptide sequence is SEQ ID NO:2 or 11. 4. The vector of claim 1 or 2, wherein the amino acid peptide sequence is SEQ ID NO:5 or 14. 5. An adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide of SEQ ID NO:1, wherein the amino acids from position 585 to 590 of SEQ ID NO:1 is replaced with an amino acid peptide sequence selected from the group consisting of SEQ ID NOs:2-5 and 11-14. 6. The vector of claim 5, wherein the amino acids from position 585 to 590 of SEQ ID NO:1 are replaced with said amino acid sequence of SEQ ID NO:2 or 11. 7. The vector of any one of claims 1-6, wherein said vector infects greater than 2.5 percent of central nervous system (CNS) cells when a titer of at least 1 x 107 of said vector is administered into the CNS of a patient, or administered intravenously or subcutaneously. Attorney Docket No.: 45049-0091WO1 / 06521 8. The vector of any one of claims 1-7, wherein said vector comprises an exogenous nucleic acid encoding an RNA or a polypeptide. 9. The vector of claim 8, wherein said exogenous nucleic acid encodes an RNA. 10. The vector of claim 9, wherein said RNA is an siRNA or microRNA. 11. The vector of claim 8, wherein said exogenous nucleic acid encodes a polypeptide. 12. The vector of claim 11, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide. 13. The vector of any one of claims 1-12, wherein said vector expresses more nucleic acid in CNS cells than the level of expression from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1. 14. A composition comprising a vector of any one of claims 1-13 and a pharmaceutically acceptable excipient. 15. The composition of claim 14, wherein said composition comprises from about 1 x 107 to about 1 x 1014 of said vector. 16. The composition of claim 14 or 15, wherein said pharmaceutically acceptable excipient comprises one or more of phosphate buffered saline, Hank’s Balanced Salt Solution, and Pluronic F68. 17. A method for delivering an exogenous nucleic acid sequence to a CNS of a mammal, wherein said method comprises: contacting CNS cells of said mammal with an AAV9 vector comprising an AAV9 capsid polypeptide and said exogenous nucleic acid sequence, wherein said capsid polypeptide comprises an amino acid peptide sequence selected from the group consisting of SEQ ID NOs:2- 5 and 11-14; wherein upon the contacting said AAV9 vector infects said CNS cells and thereby delivering said exogenous nucleic acid sequence to said CNS cells. Attorney Docket No.: 45049-0091WO1 / 06521 18. The method of claim 17, wherein said capsid polypeptide comprises the amino acid sequence of SEQ ID NO:1 with the exception that the amino acid peptide sequence is located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 of SEQ ID NO:1, or with the exception that the amino acid residues at positions 585 to 590 of SEQ ID NO:1 are replaced with said amino acid peptide sequence. 19. The method of claim 18, wherein said amino acid peptide sequence is located between amino acid positions 588 and 589 of SEQ ID NO:1. 20. The method of any one of claims 17-19, wherein said amino acid peptide sequence is SEQ ID NO:2 or 11. 21. The method of any one of claims 17-20, wherein said mammal is a human. 22. The method of any one of claims 17-21, wherein said exogenous nucleic acid sequence encodes an RNA or a polypeptide. 23. The method of claim 22, wherein said exogenous nucleic acid encodes said RNA. 24. The method of claim 23, wherein said RNA is an siRNA or microRNA. 25. The method of claim 22, wherein said exogenous nucleic acid encodes said polypeptide. 26. The method of claim 25, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide. 27. The method of any one of claims 17-26, wherein said vector expresses more of said exogenous nucleic acid sequence in said CNS cell than the level of expression in a CNS cell Attorney Docket No.: 45049-0091WO1 / 06521 from a comparable AAV9 vector comprising a capsid polypeptide consisting of the amino acid sequence set forth in SEQ ID NO:1. 28. The method of any one of claims 17-27, wherein said method comprises intravenously, subcutaneously, intracranially, and/or intrathecally administering a composition comprising said vector to said mammal, thereby contacting said CNS cell with said vector. 29. The method of claim 28, wherein said composition comprises from about 1 x 107 to about 1 x 1014 of said vector. 30. A method for treating a CNS disorder in a mammal in need thereof, wherein said method comprises contacting CNS cells of a mammal having said CNS disorder with a therapeutic construct comprising the AAV9 vector of any one of claims 1-13 and an exogenous nucleic acid sequence. 31. The method of claim 30, wherein said mammal is a human. 32. The method of any one of claims 30-31, wherein said CNS disorder is selected from the group consisting of Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, multiple sclerosis, mucopolysaccharidosis (MPS) IIIB, spinal muscular atrophy (SMA), Pompe disease, GM1 gangliosidosis, phenylketonuria deficiency, dystonia, epilepsy, and restless leg syndrome. 33. The method of any one of claims 30-32, wherein said exogenous nucleic acid sequence encodes an RNA. 34. The method of claim 33, wherein said RNA is an siRNA or a microRNA. 35. The method of any one of claims 30-33, wherein said exogenous nucleic acid encodes a polypeptide. 36. The method of claim 35, wherein said polypeptide is an SMA I polypeptide, a SOD1 polypeptide, an IGF1 polypeptide, a GCaMP6f polypeptide, a Kir4.1 polypeptide, or a GAA polypeptide. Attorney Docket No.: 45049-0091WO1 / 06521 37. A non-naturally occurring adeno-associated virus 9 (AAV9) vector comprising an AAV9 capsid polypeptide, wherein said capsid polypeptide comprises at least 95% identity to SEQ ID NO:1, or a truncated variant thereof, and further comprises an amino acid sequence insert of Formula A or Formula B located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 according to SEQ ID NO:1, wherein said Formula A is: -L1-EGSGRN (SEQ ID NO:2)-L2-, wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids, and wherein said Formula B is: -L1-EHQTRP (SEQ ID NO:11)-L2-, wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids. 38. The vector of claim 37, wherein said L1 is one amino acid X1. 39. The vector of claim 38, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L. 40. The vector of claim 38 or 39, wherein said X1 is A. 41. The vector of claim 37, wherein said L1 is two amino acids X2-X1. Attorney Docket No.: 45049-0091WO1 / 06521 42. The vector of claim 41, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L. 43. The vector of claim 42, wherein said X1 is A. 44. The vector of any one of claims 41-43, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L. 45. The vector of claim 44, wherein said X2 is L. 46. The vector of claim 45, wherein said X2-X1 is LA. 47. The vector of claim 37, wherein said L1 is three amino acids X3-X2-X1. 48. The vector of claim 47, wherein said X1 is selected from the group of amino acid residues consisting of A, V, I, and L. 49. The vector of claim 48, wherein said X1 is A. 50. The vector of any one of claims 47-49, wherein said X2 is selected from the group of amino acid residues consisting of A, V, I, and L. 51. The vector of claim 50, wherein said X2 is L. 52. The vector of claim 51, wherein said X2-X1 is LA. 53. The vector of any one of claims 47-52, wherein said X3 is selected from the group of amino acid residues consisting of A, V, I, and L. 54. The vector of claim 37, wherein said L1 is absent. Attorney Docket No.: 45049-0091WO1 / 06521 55. The vector of any one of claims 37-54, wherein said L2 is one amino acid Z1. 56. The vector of claim 55, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L. 57. The vector of claim 56, wherein said Z1 is A. 58. The vector of any one of claims 37-54, wherein said L2 is two amino acids Z1-Z2. 59. The vector of claim 58, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L. 60. The vector of claim 59, wherein said Z1 is A. 61. The vector of any one of claims 58-60, wherein said Z2 is selected from the group of amino acid residues consisting of A, V, I, and L. 62. The vector of claim 61, wherein said Z2 is L. 63. The vector of claim 62, wherein said Z1-Z2 is AL. 64. The vector of any one of claims 37-54, wherein said L2 is three amino acids Z1-Z2-Z3. 65. The vector of claim 64, wherein said Z1 is selected from the group of amino acid residues consisting of A, V, I, and L, and/or Z2 is selected from the group of amino acid residues consisting of A, V, I, and L. 66. The vector of claim 65, wherein said Z1-Z2 is AL. 67. The vector of any one of claims 65-66, wherein said Z3 is selected from the group of amino acid residues consisting of A, V, I, and L. Attorney Docket No.: 45049-0091WO1 / 06521 68. The vector of claim 37, wherein said amino acid sequence insert comprises any one of SEQ ID NOs:2-5 and 11-14. 69. A non-naturally occurring AAV9 capsid polypeptide, wherein said capsid polypeptide comprises at least 95% identity to SEQ ID NO:1, or a truncated variant thereof, and further comprises an amino acid sequence insert of Formula A or Formula B located 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, between amino acid positions 593 and 594, or between amino acid positions 594 and 595 according to SEQ ID NO:1, wherein said Formula A is: -L1-EGSGRN (SEQ ID NO:2)-L2-, wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids, and wherein said Formula B is: -L1-EHQTRP (SEQ ID NO:11)-L2-, wherein said L1 and said L2 are each independently optional amino acid linkers having one, two, or three amino acids. 70. A method for administering an exogenous nucleic acid sequence to a mammal in need thereof, wherein said method comprises administering to the mammal an effective amount of a construct comprising a vector of any one of claims 37-68 and the exogenous nucleic acid sequence. 71. A method of treating a CNS disorder in a patient in need thereof, comprising administering to the patient an effective amount of an AAV9 vector comprising an AAV9 capsid Attorney Docket No.: 45049-0091WO1 / 06521 polypeptide comprising an amino acid sequence insert represented by Formula A or Formula B and an exogenous nucleic acid sequence.
PCT/US2024/048892 2023-09-27 2024-09-27 Aav9 vectors and capsid polypeptides for use in treating cns disorders Pending WO2025072698A2 (en)

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