Frataxin expression constructs having engineered promoters and methods of use thereof

Abstract

To provide an adeno-associated virus (AAV) vector genome for altering, e.g., enhancing, expression of frataxin (FXN) in vitro and / or in vivo, including an engineered promoter.SOLUTION: Provided is an adeno-associated virus (AAV) vector genome comprising a 5' inverted terminal repeat (ITR), an engineered promoter, a payload region, and a 3' ITR, wherein the payload region encodes a frataxin (FXN) protein.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to frataxin-based compositions and methods for enhancing expression of frataxin (FXN), whether in vitro or in vivo, at least in part through the development of novel engineered promoters. Such frataxin-based compositions may be delivered in adeno-associated virus (AAV) vectors. In other embodiments, frataxin-based compositions, e.g., AAV-frataxin compositions, are used to treat subjects in need thereof, e.g., human subjects diagnosed with Friedreich's ataxia or other neurological conditions resulting from deficiencies in frataxin amount and / or function, or as research tools in the study of diseases or conditions in cellular or animal models of such diseases or conditions. [Background technology]

[0002] Friedreich's ataxia (FA), first described by German physician Nikolas Friedreich in the 1860s, is an autosomal recessive genetic disorder that causes progressive damage to the nervous system. See, e.g., "Friedreich's ataxia," IEEE Transactions on Neuroscience, Vol. 1, No. 1, pp. 111-114, the contents of which are incorporated herein by reference in their entirety. Onset usually occurs during adolescence, usually by age 25. See, e.g., "Friedreich's ataxia," IEEE Transactions on Neuroscience, Vol. 1, No. 1, pp. 111-114, the contents of which are incorporated herein by reference in their entirety. FA typically results from degeneration of nerve tissue in the spinal cord due to reduced expression of the mitochondrial protein frataxin (FXN) in sensory neurons that direct muscle movement in the arms and legs (through connections with the cerebellum). See, e.g., "Friedreich's ataxia," IEEE Transactions on Neuroscience, Vol. 1, No. 1, pp. 111-114, the contents of which are incorporated herein by reference in their entirety. The spinal cord thins, and peripheral nerve cells lose some of their myelin sheath, the insulation that covers some nerve cells and helps transmit nerve impulses. Early symptoms of FA include poor coordination, e.g., gait disturbance, poor balance, leg weakness, decreased gait, incoordination, dysarthria, nystagmus, paresthesia, kyphoscoliosis, and foot deformities. See Non-Patent Document 1. FA is also associated with scoliosis, heart disease, and diabetes. The disease typically progresses to the point where a wheelchair is required for mobility. The incidence of FA in Caucasian populations ranges from approximately 1 in 20,000 to approximately 1 in 50,000, with an estimated carrier frequency of approximately 1 in 120 in European populations. See Non-Patent Document 4; Non-Patent Document 2, the contents of each of which are incorporated herein by reference in their entirety.

[0003] An intronic GAA triplet repeat expansion in the FXN gene is the genetic cause of reduced expression of frataxin, which leads to FA. See Non-Patent Document 1. Over time, this deficiency causes the symptoms described above, as well as frequent fatigue due to effects on cellular metabolism.

[0004] Sclerosis and degeneration are most frequent in the dorsal root ganglia, spinocerebellar tracts, lateral corticospinal tracts, and dorsal columns. See, J. Neuropsychiatry, 1999, 10, 1111-1115, the contents of which are incorporated herein by reference in their entirety.

[0005] Progressive destruction of the dorsal root ganglia leads to thinning of the dorsal roots, degeneration of the dorsal columns, transsynaptic atrophy of neurons in Clarke's columns and dorsal spinocerebellar fibers, atrophy of the gracile and cuneate nuclei, and neuropathy of sensory nerves. See Non-Patent Document 3, the contents of which are incorporated herein by reference in their entirety. Lesions in the dentate nucleus consist of progressive and selective atrophy of large glutamatergic neurons and gamma-aminobutyric acid (GABA)-containing cortico-nuclear synaptic terminals. Small GABAergic neurons in the dentato-olivary tract and their projection fibers survive. Atrophy of Betz cells and the corticospinal tract constitutes secondary lesions. Currently, there is no effective treatment for FA, and patients are often simply monitored for symptom management.

[0006] As a result, there remains a long-felt need in the art to develop pharmaceutical compositions and methods for the treatment of FXN-related disorders, as well as to ameliorate the deficiency of this protein in patients suffering from FA. Adeno-associated virus (AAV) has emerged as one of the most widely studied and utilized viral particles for the delivery of therapeutically effective polypeptides to mammalian cells. See, for example, Non-Patent Document 6 and Non-Patent Document 7 (the contents of each of which are incorporated herein by reference in their entirety). Therefore, this modality is well suited for development toward the treatment of FA and the delivery of frataxin and frataxin-related proteins and peptides. [Prior art documents] [Non-patent literature]

[0007] [Non-Patent Document 1] Parkinson et al., Journal of Neurochemistry, 2013, 126(Suppl.1), 103-117 [Non-patent document 2] Campuzano, et al., Science, 271.5254 (Mar. 8, 1996): 1423 [Non-patent document 3] Koeppen, Arnulf; J Neurol Sci., 2011, April 15; 303(1-2):1-12 [Non-patent document 4] Nageshwaran and Festenstein, Frontiers in Neurology, Vol. 6, Art. 262 (2015) [Non-Patent Document 5] Sandi et al., Frontiers in Genetics, Vol. 5, Art. 165 (June 2014) [Non-patent document 6] Tratschin et al., Mol. Cell Biol., 5(11):3251-3260 (1985) [Non-Patent Document 7] Grimm et al., Hum. Gene Ther., 10(15):2445-2450 (1999) Summary of the Invention

[0008] In some aspects, the present disclosure provides an AAV viral genome comprising at least one inverted terminal repeat (ITR) and a payload region, wherein the payload region encodes a frataxin protein. In some embodiments, the AAV viral genome comprises a 5' ITR, an engineered promoter, a payload region, and a 3' ITR. The encoded frataxin protein can be human (Homo sapiens) frataxin, cynomolgus monkey (Macaca fascicularis) frataxin, or a 3' ITR. The frataxin may be (Macaca fascicularis) frataxin or (Macaca mulatta) frataxin, a synthetic (non-naturally occurring) frataxin, or a derivative thereof, e.g., a variant that retains one or more functions of wild-type frataxin. In some embodiments, the frataxin protein may be at least partially humanized.

[0009] The engineered promoter of the AAV viral genome may be derived from the cytomegalovirus (CMV) promoter, the chicken beta-actin (CBA) promoter, or the frataxin (FXN) promoter. In some embodiments, the engineered promoter is a promoter variant or derivative of the parent promoter sequence.

[0010] In some embodiments, the engineered promoter is derived from a CMV promoter. In some embodiments, the engineered promoter is derived from a CBA promoter. In some embodiments, the engineered promoter is derived from the FXN promoter.

[0011] The engineered promoter of the AAV viral genome described herein may comprise a sequence given by any of SEQ ID NOs: 1734-1777. In some embodiments, the engineered promoter comprises a sequence having at least 90% sequence identity to any of SEQ ID NOs: 1734-1777. In some embodiments, the engineered promoter comprises a sequence having at least 95% sequence identity to any of SEQ ID NOs: 1734-1777. In some embodiments, the engineered promoter comprises a sequence having at least 99% sequence identity to any of SEQ ID NOs: 1734-1777. In some embodiments, the engineered promoter may consist of any of SEQ ID NOs: 1734-1777. In some embodiments, the engineered promoter is derived from a CMV promoter and may comprise a sequence given by any of SEQ ID NOs: 1743-1751, 1767, and 1772-1774. In some embodiments, the engineered promoter comprises SEQ ID NO: 1777. In some embodiments, the engineered promoter is derived from the CBA promoter and may include a sequence given by any of SEQ ID NOs: 1734-1742, 1760-1766, 1768, and 1775-1776. In some embodiments, the engineered promoter is derived from the FXN promoter and may include a sequence given by any of SEQ ID NOs: 1752-1759 and 1769-1770.

[0012] In some embodiments, the engineered promoter comprises a sequence given by SEQ ID NO: 1738. In some embodiments, the engineered promoter comprises a sequence having at least 90% sequence identity to SEQ ID NO: 1738. In some embodiments, the engineered promoter comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1738. In some embodiments, the engineered promoter comprises a sequence having at least 99% sequence identity to SEQ ID NO: 1738. In some embodiments, the engineered promoter consists of SEQ ID NO: 1738.

[0013] In some embodiments, the engineered promoter comprises a sequence given by SEQ ID NO: 1740. In some embodiments, the engineered promoter comprises a sequence having at least 90% sequence identity to SEQ ID NO: 1740. In some embodiments, the engineered promoter comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1740. In some embodiments, the engineered promoter comprises a sequence having at least 99% sequence identity to SEQ ID NO: 1740. In some embodiments, the engineered promoter consists of SEQ ID NO: 1740.

[0014] In some embodiments, the engineered promoter comprises a sequence given by SEQ ID NO: 1742. In some embodiments, the engineered promoter comprises a sequence having at least 90% sequence identity to SEQ ID NO: 1742. In some embodiments, the engineered promoter comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1742. In some embodiments, the engineered promoter comprises a sequence having at least 99% sequence identity to SEQ ID NO: 1742. In some embodiments, the engineered promoter consists of SEQ ID NO: 1742.

[0015] In some embodiments, the engineered promoter comprises a sequence given by SEQ ID NO: 1750. In some embodiments, the engineered promoter comprises a sequence having at least 90% sequence identity to SEQ ID NO: 1750. In some embodiments, the engineered promoter comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1750. In some embodiments, the engineered promoter comprises a sequence having at least 99% sequence identity to SEQ ID NO: 1750. In some embodiments, the engineered promoter consists of SEQ ID NO: 1750.

[0016] In some embodiments, the engineered promoter comprises a sequence given by SEQ ID NO: 1756. In some embodiments, the engineered promoter comprises a sequence having at least 90% sequence identity to SEQ ID NO: 1756. In some embodiments, the engineered promoter comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1756. In some embodiments, the engineered promoter comprises a sequence having at least 99% sequence identity to SEQ ID NO: 1756. In some embodiments, the engineered promoter consists of SEQ ID NO: 1756.

[0017] The engineered promoters described herein may have a length of 50 to 1400 nucleotides (nt). In some embodiments, the engineered promoter is derived from a CMV promoter and is 50 to 700 nt in length. In some embodiments, the engineered promoter is derived from a CMV promoter and is 109 nt in length. In some embodiments, the engineered promoter is derived from a CBA promoter and is 100 to 700 nt in length. In some embodiments, the engineered promoter is derived from a CBA promoter and is 100 to 400 nt in length. In some embodiments, the engineered promoter is derived from a CBA promoter and is 100 nt in length. In some embodiments, the engineered promoter is derived from a CBA promoter and is 200 to 350 nt in length. In some embodiments, the engineered promoter is derived from a CBA promoter and is 260 nt in length. In some embodiments, the engineered promoter is derived from a CBA promoter and is 332 nt in length. In some embodiments, the engineered promoter is derived from the FXN promoter and is 200-1400 nt in length. In some embodiments, the engineered promoter is 950-1150 nt in length. In some embodiments, the engineered promoter is derived from the FXN promoter and is 1060 nt in length.

[0018] In some embodiments, the engineered promoter comprises an enhancer region. The engineered promoter and the payload region encoding frataxin may be incorporated into the AAV viral genome.

[0019] In some embodiments, the AAV viral genome comprises, in addition to the engineered promoter and payload region, a 5'ITR, an enhancer, an intron, at least one miR binding site (e.g., 1, 2, or 3 miR binding sites), a polyA sequence, a filler sequence, and a 3'ITR. In some embodiments, the AAV viral genome comprises multiple miR binding sites (an "miR binding site series") that may appear consecutively or separated by one or more nucleotides. In some embodiments, the 5'ITR and / or 3'ITR are AAV2 ITRs.

[0020] In some embodiments, the viral genome comprises at least one ITR sequence. In some embodiments, the ITR may be an AAV2 ITR. In some embodiments, the 5' ITR may be an AAV2 ITR. In some embodiments, the 3' ITR may be an AAV2 ITR. In some embodiments, the 5' and / or 3' ITR may be 141 nt in length. In some embodiments, the 5' ITR comprises a sequence at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1811. In some embodiments, the 3' ITR comprises a sequence at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1812.

[0021] In some embodiments, the sequence from ITR to ITR comprises an intron / exon region. In some embodiments, the intron / exon region may be an enhancer sequence. As a non-limiting example, the enhancer sequence may comprise two or more subcomponents, such as, but not limited to, an iE1 exon (e.g., exon 1), an iE1 intron (e.g., intron 1), a human beta-globin intron (e.g., intron 2), and / or a human beta-globin exon (e.g., exon 3), or fragments thereof. In some embodiments, the intron / exon region comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to a sequence set forth in any of SEQ ID NOs: 1815-1821. In some embodiments, the enhancer comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to a sequence set forth in any of SEQ ID NOs: 1815-1821. In some embodiments, the enhancer comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1777. In some embodiments, the intron / exon region comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to one or more human beta-globin sequences, e.g., SEQ ID NOs: 1820 and / or 1821. In some embodiments, the intron may comprise a sequence given by any of SEQ ID NOs: 1815-1821. In some embodiments, the intron has a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1816. In some embodiments, the intron may consist of SEQ ID NO: 1816.

[0022] In some embodiments, the miR binding site series comprises at least one miR122 binding site sequence. In some embodiments, at least one miR122 binding site comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1827. In some embodiments, at least one miR122 binding site consists of SEQ ID NO: 1827. In some embodiments, the AAV vector genome comprises three copies of miR122 binding sites, e.g., three copies of SEQ ID NO: 1827 or a variant thereof having at least 90% sequence identity. In some embodiments, the miR binding site series may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1826. In some embodiments, the miR binding site series may consist of SEQ ID NO: 1826.

[0023] In some embodiments, the polyA sequence is a human growth hormone (hGH) polyA sequence. In some embodiments, the viral genome comprises an hGH polyA sequence that is at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1828. In some embodiments, the polyA sequence consists of SEQ ID NO: 1828.

[0024] In some embodiments, the AAV viral genome further comprises a filler sequence, e.g., an albumin filler sequence. In some embodiments, the filler sequence may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to a sequence provided by any of SEQ ID NOs: 1829-1842. In some embodiments, the filler sequence may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1838. In some embodiments, the filler sequence may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1839. In some embodiments, the filler sequence may consist of SEQ ID NO: 1839. In some embodiments, the filler sequence may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1840. In some embodiments, the filler sequence may consist of SEQ ID NO: 1840. In some embodiments, the filler sequence may comprise a sequence that is at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1841. In some embodiments, the filler sequence may consist of SEQ ID NO: 1841.

[0025] In some embodiments, the AAV viral genome may comprise a sequence given by any of SEQ ID NOs: 1778-1810. In some embodiments, the AAV viral genome comprises a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any of SEQ ID NOs: 1778-1810. In some embodiments, the AAV viral genome comprises a sequence having 80-85%, 80-90%, 80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or 90-100% sequence identity to any of SEQ ID NOs: 1778-1810. An AAV viral genome in which the encoded frataxin is a frataxin from a Cynomolgus monkey (Cynomolgus monkey) species may comprise a sequence given by any of SEQ ID NOs: 1778-1795. In some embodiments, the AAV viral genome comprises a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any of SEQ ID NOs: 1778-1795. In some embodiments, the AAV viral genome comprises a sequence having 80-85%, 80-90%, 80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or 90-100% sequence identity to any of SEQ ID NOs: 1778-1795. An AAV viral genome in which the encoded frataxin is human frataxin may comprise a sequence given by any of SEQ ID NOs: 1796-1810. In some embodiments, the AAV viral genome comprises a sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to any of SEQ ID NOs: 1796-1810. In some embodiments, the AAV viral genome comprises a sequence having 80-85%, 80-90%, 80-95%, 80-99%, 80-100%, 90-95%, 90-99%, or 90-100% sequence identity to any of SEQ ID NOs: 1796-1810.

[0026] In some embodiments, the AAV viral genome may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1797. In some embodiments, the AAV viral genome may consist of SEQ ID NO: 1797. In some embodiments, the AAV viral genome may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1801. In some embodiments, the AAV viral genome may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1808. In some embodiments, the AAV viral genome may comprise a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1809. In some embodiments, the AAV viral genome may consist of SEQ ID NO: 1809.

[0027] In some embodiments, the payload region of an AAV vector genome encoding frataxin comprises a nucleic acid sequence at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% identical to the sequence provided by any of SEQ ID NOs: 1822-1824. In some embodiments, the payload region of an AAV vector genome encoding frataxin comprises a nucleic acid sequence provided by any of SEQ ID NOs: 1822-1824. In some embodiments, the nucleic acid sequence encoding frataxin comprises SEQ ID NO: 1822. In some embodiments, the nucleic acid sequence encoding frataxin comprises SEQ ID NO: 1823. In some embodiments, the nucleic acid sequence encoding frataxin comprises SEQ ID NO: 1824. In some embodiments, the nucleic acid sequence encoding frataxin comprises a fragment of SEQ ID NO: 1728, 1729, or 1730, or a variant thereof having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity thereto. In some embodiments, the nucleic acid sequence encoding frataxin comprises a fragment of SEQ ID NO: 1728. In some embodiments, the nucleic acid sequence encoding frataxin comprises nucleotides 221-853 of SEQ ID NO:1728.

[0028] In some embodiments, the payload region of an AAV vector genome comprises a nucleic acid sequence encoding a frataxin polypeptide having at least 80%, at least 85%, at least 90%, at least 95%, or at least 99% sequence identity to SEQ ID NO: 1725, 1726, or 1727. In some embodiments, the payload region of an AAV vector genome comprises a nucleic acid sequence encoding a frataxin polypeptide of SEQ ID NO: 1725, 1726, or 1727. In some embodiments, the AAV vector genome comprises a nucleic acid sequence encoding a frataxin polypeptide comprising SEQ ID NO: 1725. In some embodiments, the payload region of an AAV vector genome comprises a nucleic acid sequence encoding a frataxin polypeptide having at least 80%, 85%, 90%, 95%, or 99% sequence identity to SEQ ID NO: 1731, 1732, or 1733. In some embodiments, the payload region of an AAV vector genome comprises a nucleic acid sequence encoding a frataxin polypeptide of SEQ ID NO: 1731, 1732, or 1733.

[0029] The viral genome containing the engineered promoter or promoter variant may be incorporated into an AAV particle, which comprises the viral genome and a capsid. In some embodiments, the capsid comprises a sequence set forth in Table 1 or selected from the group consisting of SEQ ID NOs: 1-1724. Non-limiting examples of capsids include AAV9, AAV9 K449R, AAVPHP.B, AAVPHP.N, VOY101 (having the amino acid sequence of SEQ ID NO: 1 and / or having the nucleic acid sequence of SEQ ID NO: 1722), and / or VOY201 (having the amino acid sequence of SEQ ID NO: 1724 and / or having the nucleic acid sequence of SEQ ID NO: 1723). In some embodiments, the capsid is encoded by a nucleic acid sequence selected from SEQ ID NOs: 4, 135, 1722, and 1723. In some embodiments, the capsid may have the amino acid sequence given by any of SEQ ID NOs: 1, 2, 3, 9, 136, or 1724. In some embodiments, the capsid comprises the amino acid sequence given by SEQ ID NO: 136. In some embodiments, the capsid comprises the amino acid sequence encoded by the nucleic acid sequence given by SEQ ID NO: 135. In some embodiments, the capsid comprises the amino acid sequence given by SEQ ID NO: 9. In some embodiments, the capsid comprises the amino acid sequence given by SEQ ID NO: 3. In some embodiments, the capsid comprises the amino acid sequence encoded by the nucleic acid sequence given by SEQ ID NO: 4. In some embodiments, the capsid comprises the amino acid sequence given by SEQ ID NO: 2. In some embodiments, the capsid comprises the amino acid sequence given by SEQ ID NO: 1. In some embodiments, the capsid comprises the amino acid sequence encoded by the nucleic acid sequence given by SEQ ID NO: 1722. In some embodiments, the capsid comprises the amino acid sequence encoded by the nucleic acid sequence given by SEQ ID NO: 1723. In some embodiments, the capsid comprises the amino acid sequence given by SEQ ID NO: 1724.

[0030] In some embodiments, the AAV particles described herein can be used in pharmaceutical compositions. In some embodiments, the pharmaceutical composition comprises sodium chloride, sodium phosphate, potassium chloride, potassium phosphate, and poloxamer 188. In some embodiments, the pharmaceutical composition comprises 192 mM sodium chloride, 10 mM sodium phosphate, 2.7 mM potassium chloride, 2 mM potassium phosphate, and 0.001% poloxamer 188 (v / v). In some embodiments, the sodium phosphate of the composition is dibasic. In some embodiments, the potassium phosphate of the composition is monobasic. In some embodiments, the pH of the pharmaceutical composition is 7.3 to 7.7. In some embodiments, the pH of the pharmaceutical composition is 7.4.

[0031] In some embodiments, the AAV particles may comprise a vector genome given by SEQ ID NO: 1797 and a VOY101 capsid. In some embodiments, a pharmaceutical composition comprising an AAV particle comprising a vector genome given by SEQ ID NO: 1797 and a VOY101 capsid comprises sodium chloride, sodium phosphate, potassium chloride, potassium phosphate, and poloxamer 188; optionally, the pharmaceutical composition comprises 192 mM sodium chloride, 10 mM sodium phosphate, 2.7 mM potassium chloride, 2 mM potassium phosphate, and 0.001% poloxamer 188 (v / v), and the pH of the composition is 7.4.

[0032] In some embodiments, the AAV particles may comprise a vector genome provided by SEQ ID NO: 1801 and a VOY101 capsid. In some embodiments, a pharmaceutical composition comprising an AAV particle comprising a vector genome provided by SEQ ID NO: 1801 and a VOY101 capsid comprises sodium chloride, sodium phosphate, potassium chloride, potassium phosphate, and poloxamer 188; optionally, the pharmaceutical composition comprises 192 mM sodium chloride, 10 mM sodium phosphate, 2.7 mM potassium chloride, 2 mM potassium phosphate, and 0.001% poloxamer 188 (v / v), and the pH of the composition is 7.4.

[0033] In some embodiments, the AAV particles may comprise a vector genome given by SEQ ID NO: 1808 and a VOY101 capsid. In some embodiments, a pharmaceutical composition comprising an AAV particle comprising a vector genome given by SEQ ID NO: 1808 and a VOY101 capsid comprises sodium chloride, sodium phosphate, potassium chloride, potassium phosphate, and poloxamer 188; optionally, the pharmaceutical composition comprises 192 mM sodium chloride, 10 mM sodium phosphate, 2.7 mM potassium chloride, 2 mM potassium phosphate, and 0.001% poloxamer 188 (v / v), and the pH of the composition is 7.4.

[0034] In some embodiments, the AAV particles may comprise a vector genome given by SEQ ID NO: 1809 and a VOY101 capsid. In some embodiments, a pharmaceutical composition comprising an AAV particle comprising a vector genome given by SEQ ID NO: 1809 and a VOY101 capsid comprises sodium chloride, sodium phosphate, potassium chloride, potassium phosphate, and poloxamer 188; optionally, the pharmaceutical composition comprises 192 mM sodium chloride, 10 mM sodium phosphate, 2.7 mM potassium chloride, 2 mM potassium phosphate, and 0.001% poloxamer 188 (v / v), and the pH of the composition is 7.4.

[0035] The pharmaceutical compositions and / or AAV particles of the present disclosure can be used to treat neurological or neuromuscular disorders, such as, but not limited to, Friedreich's ataxia.

[0036] In some embodiments, the AAV particles of the present disclosure are used to treat a disorder or condition associated with decreased frataxin expression or protein levels. In some embodiments, the disorder or condition associated with decreased frataxin expression or protein levels is a neurological or neuromuscular disorder. In some embodiments, the disorder or condition associated with decreased frataxin protein levels is FA or frataxin deficiency. In some embodiments, administration of AAV particles can result in increased frataxin expression in target cells to a level that is 0.5-3x (e.g., 0.5-1x, 1-1.5x, 1.5-2x, 2-2.5x, 2.5-3x) of frataxin expression in equivalent target cells of a normal subject not suffering from a disorder associated with decreased frataxin levels. In some embodiments, administration of AAV particles can result in frataxin expression in target cells of approximately 5.5-32.8 ng / mg protein.

[0037] Details of various aspects or embodiments of the present disclosure are set forth below. Other features, objects, and advantages of the present disclosure will be apparent from the description and claims. In the description, the singular also includes the plural unless the context clearly dictates otherwise. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this disclosure belongs. In case of conflict, the description in this specification shall prevail.

[0038] The foregoing and other objects, features, and advantages will be apparent from the following description of specific embodiments presented herein, as illustrated in the accompanying drawings, in which: The drawings are not necessarily to scale, emphasis instead being placed upon illustrating the principles of the various embodiments described herein. [Brief explanation of the drawings]

[0039] [Figure 1A]FIG. 1 shows graphs depicting the quantification results of frataxin expression levels (ng / mg) by ELISA and AAV biodistribution (VG / DC) by quantitative PCR in cardiac tissue. [Figure 1B] FIG. 1 shows graphs depicting the quantification results of frataxin expression levels (ng / mg) by ELISA and AAV biodistribution (VG / DC) by quantitative PCR in cerebellar tissue. [Figure 1C] FIG. 1 shows graphs depicting the quantification results of frataxin expression levels (ng / mg) by ELISA and AAV biodistribution (VG / DC) by quantitative PCR in dorsal root ganglia (DRG). [Figure 1D] FIG. 1 shows graphs depicting the quantification results of frataxin expression levels (ng / mg) by ELISA and AAV biodistribution (VG / DC) by quantitative PCR in liver tissue. [Figure 2] FIG. 10 shows a graph depicting quantification results for striatal cFXN protein levels by ELISA for certain promoter constructs of the present disclosure. [Figure 3A] FIG. 1 shows a graph illustrating the quantification results of frataxin expression levels (ng / mg) in lumbar DRG tissues by ELISA. [Figure 3B] FIG. 1 shows a graph showing the quantification results of frataxin expression levels (ng / mg) in cerebellar tissue by ELISA. [Figure 4] FIG. 10 shows a graph depicting electromyography (H-wave intensity) measurements in Pvalb cKO animals treated intravenously with VOY101-CMV-D7-hFXN or VOY101-CBA-D8-hFXN AAV particles compared to Pvalb cKO and wild-type (WT) mice. [Figure 5]FIG. 10 shows a graph depicting behavioral analysis via the notched-bar test in Pvalb cKO mice treated intravenously with VOY101-CMV-D7-hFXN or VOY101-CBA-D8-hFXN AAV particles compared to Pvalb cKO mice and wild-type (WT) mice. [Figure 6A] FIG. 1 shows a graph depicting quantification results for frataxin expression levels (ng / mg) by ELISA for certain DRG tissues of the present disclosure. [Figure 6B] A diagram showing an enlarged view of the quantification results in Figure 6A for hFXN13 (CBA.D4) having sequence number 1808, hFXN14 (CBA.D6) having sequence number 1809, and hFXN2 (CBA.D8) having sequence number 1797. [Figure 6C] FIG. 1 shows a graph depicting quantification results for frataxin expression levels (ng / mg) by ELISA for certain ventricular tissues of the present disclosure. [Figure 6D] A diagram showing an enlarged view of the quantification results in Figure 6C for hFXN13 (CBA.D4) having sequence number 1808, hFXN14 (CBA.D6) having sequence number 1809, and hFXN2 (CBA.D8) having sequence number 1797. [Figure 7] FIG. 1 shows a graph depicting the quantification results of frataxin expression levels (FXN:luciferase ratio) by luciferase expression for promoter constructs of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0040] I. Composition Adeno-associated virus (AAV) vectors Parvoviridae viruses are small, non-enveloped, icosahedral-capsid viruses characterized by a single-stranded DNA genome. Parvoviridae viruses consist of two subfamilies: the Parvovirinae, which infect vertebrates, and the Densovirinae, which infect invertebrates. This virus family can be useful as a biological tool due to its relatively simple structure, which can be manipulated using standard molecular biology techniques. The viral genome can be modified to contain the minimum components required to assemble a functional recombinant virus or virus particle carrying or engineered to express or deliver a desired nucleic acid construct or payload, such as a transgene, a polypeptide-encoding polynucleotide, or FXN, and deliver it to a target cell, tissue, or organism. In some embodiments, the target cell is a CNS cell. In some embodiments, the target tissue is a CNS tissue.

[0041] Parvoviruses and other members of the Parvoviridae family are reviewed in Kenneth I. Berns, "The Parvoviridae: The Viruses and Their Replication," Chapter 69 of FIELDS VIROLOGY (3rd ed., 1996), the contents of which are incorporated herein by reference in their entirety.

[0042] The Parvoviridae family includes the genus Dependovirus, which comprises the adeno-associated viruses (AAV) that are capable of replication in vertebrate hosts, including but not limited to, humans, primates, bovine, canine, equine, and ovine species.

[0043] Adeno-associated virus (AAV) is a single-stranded, non-enveloped DNA virus with a genome approximately 5,000 nucleotides long. It is a dependent parvovirus (similar to other parvoviruses) containing two open reading frames encoding replication proteins (Rep) and capsid structural proteins (Cap). These open reading frames are flanked by two inverted terminal repeat (ITR) sequences that serve as origins of replication for the viral genome. The wild-type AAV viral genome contains nucleotide sequences for two open reading frames: one for four nonstructural Rep proteins (Rep78, Rep68, Rep52, and Rep40 encoded by the Rep genes) and one for three capsid or structural proteins (VP1, VP2, and VP3 encoded by the capsid or Cap genes). While Rep proteins are important for replication and packaging, capsid proteins assemble to create the protein shell of AAV, i.e., the AAV capsid. Alternative splicing and alternative initiation codons and promoters result in the generation of four different Rep proteins from a single open reading frame and the generation of three capsid proteins from a single open reading frame. Depending on the AAV serotype, such as, but not limited to, AAV9 / hu.14 (SEQ ID NO: 123 of U.S. Pat. No. 7,906,111, the contents of which are incorporated herein by reference in their entirety), VP1 refers to amino acids 1-736, VP2 refers to amino acids 138-736, and VP3 refers to amino acids 203-736. In other words, VP1 is the full-length capsid sequence, while VP2 and VP3 are shorter components of the whole. As a result, changes in sequence in the VP3 region will also be changes relative to VP1 and VP2, but because VP3 is the shortest of the three, the percent difference compared to the parent sequence will be highest in VP3. Although described herein in terms of amino acid sequences, the nucleic acid sequences encoding these proteins can also be described similarly.Together, the three capsid proteins assemble to create the AAV capsid protein. Without wishing to be bound by theory, the AAV capsid protein typically comprises a 1:1:10 molar ratio of VP1:VP2:VP3. As used herein, "AAV serotype" is primarily defined by the AAV capsid. In some cases, the ITRs are also specifically described by the AAV serotype (e.g., AAV2 / 9).

[0044] AAV vectors typically require a co-helper (e.g., adenovirus) to undergo productive infection in infected cells. In the absence of such helper functions, AAV virions essentially enter host cells but do not integrate into the cellular genome. As used herein, the term "AAV vector" or "AAV particle" includes a capsid and a viral genome containing a polynucleotide payload. As used herein, "payload" or "payload region" refers to one or more polynucleotides or polynucleotide regions encoded by or within the viral genome, or such polynucleotides or polynucleotide regions, such as transgenes, polynucleotides encoding polypeptides or multi-polypeptides, e.g., FXN.

[0045] AAV vectors have been investigated for delivery due to several unique characteristics. Non-limiting examples of these characteristics include: (i) the ability to infect both dividing and non-dividing cells; (ii) a wide host range for infection, including human cells; (iii) wild-type AAV has not been associated with any disease and has been shown not to replicate in infected cells; (iv) the lack of a cell-mediated immune response to the vector; and (v) the tendency not to integrate into host chromosomes, reducing the potential for long-term genetic alteration. Furthermore, infection with AAV vectors has minimal impact on altering patterns of cellular gene expression (Stilwell and Samulski et al., Biotechniques, 2003, 34, 148 (the contents of which are incorporated herein by reference in their entirety)).

[0046] Typically, the AAV vector for FXN delivery can be a replication-deficient recombinant viral vector, because it lacks the sequence encoding functional Rep and Cap proteins in the viral genome.In some cases, the defective AAV vector can lack most or all coding sequences, and essentially only contains one or two AAV ITR sequences and payload sequences.In certain embodiments, the viral genome encodes FXN.For example, the viral genome encodes human FXN.

[0047] In one embodiment, the AAV particles of the present disclosure can be introduced into mammalian cells. AAV vectors can be modified to enhance the efficiency of delivery, and such modified AAV vectors of the present disclosure can be packaged efficiently and used to successfully infect target cells at high frequency and with minimal toxicity.

[0048] In other embodiments, the AAV particles of the present disclosure can be used to deliver FXN to the central nervous system (see, e.g., U.S. Patent No. 6,180,613, the contents of which are incorporated herein by reference in their entirety).

[0049] AAV serotype The AAV particles of the present disclosure may comprise or be derived from any natural or recombinant AAV serotype. According to the present disclosure, the AAV particles may be any of the following: VOY101, VOY201, AAV9, AAV9 K449R, AAVPHP.B (PHP.B), AAVPHP.A (PHP.A), AAVG2B-26, AAVG2B-13, AAVTH1.1-32, AAVTH1.1-35, AAVPHP.B2 (PHP.B2), AAVPHP.B3 (PHP.B3), AAVPHP.N / PHP.B-DGT, AAVPHP.B-EST, AAVPHP.B-GGT, AAVPHP.B-ATP, AAVPHP.B-ATT-T. , AAVPHP.B-DGT-T, AAVPHP.B-GGT-T, AAVPHP.B-SGS, AAVPHP.B-AQP, AAVPHP.B-QQP, AAVPHP.B-SNP(3), AAVPHP.B- SNP, AAVPHP.B-QGT, AAVPHP.B-NQT, AAVPHP.B-EGS, AAVPHP.B-SGN, AAVPHP.B-EGT, AAVPHP.B-DST, AAVPHP.B-DST, A AVPHP.B-STP, AAVPHP.B-PQP, AAVPHP.B-SQP, AAVPHP.B-QLP, AAVPHP.B-TMP, AAVPHP.B-TTP, AAVPHP.S / G2A12, AAV G2A15 / G2A3(G2A3), AAVG2B4(G2B4), AAVG2B5(G2B5), PHP.S, AAV1, AAV2, AAV2G9, AAV3, AAV3a, AAV3b, AAV3-3, AAV 4, AAV4-4, AAV5, AAV6, AAV6.1, AAV6.2, AAV6.1.2, AAV7, AAV7.2, AAV8, AAV9, AAV9.11, AAV9.13, AAV9.16, AAV9.24 , AAV9.45, AAV9.47, AAV9.61, AAV9.68, AAV9.84, AAV9.9, AAV10, AAV11, AAV12, AAV16.3, AAV24.1, AAV27.3, AAV42.12、AAV42-1b、AAV42-2、AAV42-3a、AAV42-3b、AAV42-4、AAV42-5a、AAV42-5b、AAV42-6b、AAV42-8、AAV42-10、AAV42-11、AAV42-12、AAV42-13、AAV42-1 5、AAV42-aa、AAV43-1、AAV43-12、AAV43-20、AAV43-21、AAV43-23、AAV43-25、AAV43-5、AAV44.1、AAV44.2、AAV44.5、AAV223.1、AAV223.2、AAV223.4、AA V223.5、AAV223.6、AAV223.7、AAV1-7 / rh.48、AAV1-8 / rh.49、AAV2-15 / rh.62、AAV2-3 / rh.61、AAV2-4 / rh.50、AAV2-5 / rh.51、AAV3.1 / rh.6、AAV3.1 / rh.61 .9、AAV3-9 / rh.52、AAV3-11 / rh.53、AAV4-8 / r11.64、AAV4-9 / rh.54、AAV4-19 / rh.55、AAV5-3 / rh.57、AAV5-22 / rh.58、AAV7.3 / hu.7、AAV16.8 / hu.10 AV16.12 / hu.11、AAV29.3 / bb.1、AAV29.5 / bb.2、AAV106.1 / hu.37、AAV114.3 / hu.40、AAV127.2 / hu.41、AAV127.5 / hu.42、AAV128.3 / hu.44、AAV130.4 / hu.48、AAV145.1 / hu.53、AAV145.5 / hu.54、AAV145.6 / hu.55、AAV161.10 / hu.60、AAV161.6 / hu.61、AAV33.12 / hu.17、AAV33.4 / hu.15、AAV33.8 / hu.16、A AV52 / hu.19、AAV52.1 / hu.20、AAV58.2 / hu.25、AAVA3.3、AAVA3.4、AAVA3.5、AAVA3.7、AAVC1、AAVC2、AAVC5、AAV-DJ、AAV-DJ8、AAV3、AAV5、AAV2 rh.72、AAVhu.8、AAVrh.68、AAVrh.70、AAVpi.1、AAVpi.3、AAVpi.2、AAVrh.60、AAVrh.44、AAVrh.65、AAVrh.55、AAVrh.47、AAVrh.69、AAVrh.45、AAVrh.59、AAVhu.12、AAVH6、AAVLK03、AAVH-1 / hu.1、AAVH-5 / hu.3、AAVLG-10 / rh.40、AAVLG-4 / rh.38、AAVLG-9 / hu.39、AAV N721-8 / rh.43、AAVCh.5、AAVCh.5R1、AAVcy.2、AAVcy.3、AAVcy.4、AAVcy.5、AAVCy.5R1、AAVCy.5R2、AAVCy.5R3、AAVC y.5R4、AAVcy.6、AAVhu.1、AAVhu.2、AAVhu.3、AAVhu.4、AAVhu.5、AAVhu.6、AAVhu.7、AAVhu.9、AAVhu.10、AAVhu.11、 AAVhu.13、AAVhu.15、AAVhu.16、AAVhu.17、AAVhu.18、AAVhu.20、AAVhu.21、AAVhu.22、AAVhu.23.2、AAVhu.24、AAVhu .25、AAVhu.27、AAVhu.28、AAVhu.29、AAVhu.29R、AAVhu.31、AAVhu.32、AAVhu.34、AAVhu.35、AAVhu.37、AAVhu.39、A AVhu.40、AAVhu.41、AAVhu.42、AAVhu.43、AAVhu.44、AAVhu.44R1、AAVhu.44R2、AAVhu.44R3、AAVhu.45、AAVhu.46、AA Vhu.47、AAVhu.48、AAVhu.48R1、AAVhu.48R2、AAVhu.48R3、AAVhu.49、AAVhu.51、AAVhu.52、AAVhu.54、AAVhu.55、AA Vhu.56、AAVhu.57、AAVhu.58、AAVhu.60、AAVhu.61、AAVhu.63、AAVhu.64、AAVhu.66、AAVhu.67、AAVhu.14 / 9、AAVhu.t 19、AAVrh.2、AAVrh.2R、AAVrh.8、AAVrh.8R、AAVrh.10、AAVrh.12、AAVrh.13、AAVrh.13R、AAVrh.14、AAVrh.17、AAVrh.18、AAVrh.19、AA Vrh.20、AAVrh.21、AAVrh.22、AAVrh.23、AAVrh.24、AAVrh.25、AAVrh.31、AAVrh.32、AAVrh.33、AAVrh.34、AAVrh.35、AAVrh.36、AAVrh.37、AAVrh.37R2、AAVrh.38、AAVrh.39、AAVrh.40、AAVrh.46、AAVrh.48、AAVrh.48.1、AAVrh.48.1.2、AAVrh.48.2、AAVrh.49、AAVrh.51、AAVrh.52、AAVrh. rh.53、AAVrh.54、AAVrh.56、AAVrh.57、AAVrh.58、AAVrh.61、AAVrh.64、AAVrh.64R1、AAVrh.64R2、AAVrh.67、AAVrh.73、AAVrh.74、AAVrh.8R、AAVrh.8R. A586R mutation、AAVrh8R R533A mutation、AAAV、BAAV、ヤギAAV、ウシAAV、AAVhE1.1、AAVhEr1.5、AAVhEr1.14、AAVhEr1.8、AAVhEr1.16、AAVhEr1.18、AAVhEr1.35、AAVhEr1.7、AAVhEr1.36、AAV hEr2.29、AAVhEr2.4、AAVhEr2.16、AAVhEr2.30、AAVhEr2.31、AAVhEr2.36、AAVhER1.23、AAVhEr3.1、AAV2.5T、AAV-PAEC、AAV-LK01、AAV-LK02、AAV-LK03、AAV-LK03 LK04、AAV-LK05、AAV-LK06、AAV-LK07、AAV-LK08、AAV-LK09、AAV-LK10、AAV-LK11、AAV-LK12、AAV-LK13、AAV-LK14、AAV-LK15、AAV-LK16、AAV-LK17、AAV-LK18 、AAV-LK19、AAV-PAEC2、AAV-PAEC4、AAV-PAEC6、AAV-PAEC7、AAV-PAEC8、AAV-PAEC11、AAV-PAEC12、AAV-2-pre-miRNA-101、AAV-8h、AAV-8b、AAV-h、AAV-b、AAV SM 10-2、AAVシャッフル100-1、AAVシャッフル100-3、AAVシャッフル100-7、AAVシャッフル10-6、AAVシャッフル10-8、AAV SM 10-1、AAV SM 10-8、AAV SM 100-3 100-10、BNP61 AAV、BNP62 AAV、BNP63 AAV、AAVrh.50、AAVrh.43、AAVrh.62、AAVrh.48、AAVhu.19、AAVhu.11、AAVhu.53、AAV4-8 / rh.64、AAVLG-9 / hu.39、AAV54.5 / hu.23、AAV54.2 / hu.22、AAV54.7 / hu.24、AAV54.1 / hu.21、AAV54.4R / hu.27、AAV46.2 / hu.28、AAV46.6 / hu.29、AAV128.1 / hu.43、トゥルータップAAV(ttAAV)、UPENN AAV 10、ジャパニーズAAV10 serotype、AAV CBr-7.1、AAV CBr-7.10、AAV CBr-7.2、AAV CBr-7.3、AAV CBr-7.4、AAV CBr-7.5、AAV CBr-7.7、AAV CBr-7.8、AAV CBr-B7.3、AAV CBr-B7.4、AAV CBr-E1、AAV CBr-E2、AAV CBr-E3、AAV. CBr-E4、AAV CBr-E5、AAV CBr-e5、AAV CBr-E6、AAV CBr-E7、AAV CBr-E8、AAV CHt-1、AAV CHt-2、AAV CHt-3、AAV CHt-6.1、AAV CHt-6.10、AAV CHt-6.5、AAV CHt-6.6、AAV CHt-6.7、AAV CHt-6.8、AAV CHt-P1、AAV CHt-P2、AAV CHt-P5、AAV CHt-P6、AAV CHt-P8、AAV CHt-P9、AAV CKd-1、AAV CKd-10、AAV CKd-2、AAV CKd-3、AAV CKd-4、AAV CKd-6、AAV CKd-7、AAV CKd-8、AAV CKd-B1、AAV CKd-B2、AAV CKd-B3、AAV CKd-B4、AAV CKd-B5、AAV CKd-B6、AAV CKd-B7、AAV CKd-B8、AAV CKd-H1、AAV CKd-H2、AAV CKd-H3、AAV CKd-H4、AAV CKd-H5、AAV CKd-H6、AAV CKd-N3、AAV CKd-N4、AAV CKd-N9、AAV CLg-F1、AAV CLg-F2、AAV CLg-F3、AAV CLg-F4、AAV CLg-F5、AAV CLg-F6、AAV CLg-F7、AAV CLg-F8、AAV CLv-1、AAV CLv1-1、AAV Clv1-10、AAV CLv1-2、AAV CLv-12、AAV CLv1-3、AAV CLv-13、AAV CLv1-4、AAV Clv1-7、AAV Clv1-8、AAV Clv1-9、AAV CLv-2、AAV CLv-3、AAV CLv-4、AAV CLv-6、AAV CLv-8、AAV CLv-D1、AAV CLv-D2、AAV CLv-D3、AAV CLv-D4、AAV CLv-D5、AAV CLv-D6、AAV CLv-D7、AAV CLv-D8、AAV CLv-E1、AAV CLv-K1、AAV CLv-K3、A AV CLv-K6、AAV CLv-L4、AAV CLv-L5、AAV CLv-L6、AAV CLv-M1、AAV CLv-M11、AAV CLv-M2、AAV CLv-M5, AAV CLv-M6, AAV CLv-M7, AAV CLv-M8, AAV CLv-M9, AAV CLv-R1, AAV CLv-R2, AAV CLv-R3, AAV CLv-R4, AAV CLv-R5, AAV CLv-R6, AAV CLv-R7, AAV CLv-R8, AAV CLv-R9. CSp-8.6, AAV CSp-8.7, AAV CSp-8.8, AAV CSp-8.9, AAV The serotype may utilize, be based on, or comprise a peptide selected from CSp-9, AAV.hu.48R3, AAV.VR-355, AAV3B, AAV4, AAV5, AAVF1 / HSC1, AAVF11 / HSC11, AAVF12 / HSC12, AAVF13 / HSC13, AAVF14 / HSC14, AAVF15 / HSC15, AAVF16 / HSC16, AAVF17 / HSC17, AAVF2 / HSC2, AAVF3 / HSC3, AAVF4 / HSC4, AAVF5 / HSC5, AAVF6 / HSC6, AAVF7 / HSC7, AAVF8 / HSC8, and / or AAVF9 / HSC9, and variants or hybrids / chimeras / combinations thereof.

[0050] In some embodiments, the AAV serotypes used in the compositions disclosed herein include, but are not limited to, AAV1 (SEQ ID NOs: 6 and 64 in U.S. Patent Application Publication No. 20030138772), AAV2 (SEQ ID NOs: 7 and 70 in U.S. Patent Application Publication No. 20030138772), AAV3 (SEQ ID NOs: 8 and 71 in U.S. Patent Application Publication No. 20030138772), AAV4 (SEQ ID NO: 63 in U.S. Patent Application Publication No. 20030138772), AAV5 (SEQ ID NO: 114 in U.S. Patent Application Publication No. 20030138772), AAV6 (SEQ ID NO: 65 in U.S. Patent Application Publication No. 20030138772), AAV7 (SEQ ID NO: 76 in U.S. Patent Application Publication No. 20030138772), AAV8 (SEQ ID NO: 77 in U.S. Patent Application Publication No. 20030138772), AAV9 (SEQ ID NO: 90 in U.S. Patent Application Publication No. 20030138772), AAV10 (SEQ ID NO: 102 in U.S. Patent Application Publication No. 20030138772), AAV11 (SEQ ID NO: 113 in U.S. Patent Application Publication No. 20030138772), AAV12 (SEQ ID NO: 114 in U.S. Patent Application Publication No. 20030138772), AAV13 (SEQ ID NO: 115 in U.S. Patent Application Publication No. 20030138772), AAV14 (SEQ ID NO No. 30138772), AAV8 (SEQ ID NOs: 4 and 95 of U.S. Patent Application Publication No. 20030138772), AAV9 (SEQ ID NOs: 5 and 100 of U.S. Patent Application Publication No. 20030138772), AAV10 (SEQ ID NO: 117 of U.S. Patent Application Publication No. 20030138772), AAV11 (SEQ ID NO: 118 of U.S. Patent Application Publication No. 20030138772), AAV12 (SEQ ID NO: 119 of U.S. Patent Application Publication No. 20030138772), AAVrh10 (amino acids 1 to 738 of SEQ ID NO: 81 of U.S. Patent Application Publication No. 20030138772), AAV16.3 (U.S. Patent Application Publication No. 20030138772 SEQ ID NO: 10), AAV29.3 / bb.1 (US Patent Application Publication No. 20030138772, SEQ ID NO: 11), AAV29.4 (US Patent Application Publication No. 20030138772, SEQ ID NO: 12), AAV29.5 / bb.2 (US Patent Application Publication No. 20030138772, SEQ ID NO: 13), AAV1.3 (US Patent Application Publication No. 20030138772, SEQ ID NO: 14), AAV13.3 (US Patent Application Publication No. 20030138772, SEQ ID NO: 15), AAV24.1 (US Patent Application Publication No. 20030138772, SEQ ID NO: 16), AAV27.3 (US Patent Application Publication No. 20030138772, SEQ ID NO: 17), AAV7.2 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 18), AAVC1 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 19), AAVC3 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 20), AAVC5 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 21), AAVF1 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 22), AAVF3 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 23), AAVF5 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 24), AAVH6 (U.S. Patent Application Publication No. 20030138772, SEQ ID NO: 25), SEQ ID NO: 25), AAVH2 (US Patent Application Publication No. 20030138772 SEQ ID NO: 26), AAV42-8 (US Patent Application Publication No. 20030138772 SEQ ID NO: 27), AAV42-15 (US Patent Application Publication No. 20030138772 SEQ ID NO: 28), AAV42-5b (US Patent Application Publication No. 20030138772 SEQ ID NO: 29), AAV42-1b (US Patent Application Publication No. 20030138772 SEQ ID NO: 30), AAV42-13 (US Patent Application Publication No. 20030138772 SEQ ID NO: 31), AAV42-3a (US Patent Application Publication No. 20030138772 SEQ ID NO: 32), AAV42-5b (US Patent Application Publication No. 20030138772 SEQ ID NO: 33), AAV42-1b (US Patent Application Publication No. 20030138772 SEQ ID NO: 34), AAV42-1c (US Patent Application Publication No. 20030138772 SEQ ID NO: 35), AAV42-3d (US Patent Application Publication No. 20030138772 SEQ ID NO: 36), AAV42-3e (US Patent Application Publication No. 20030138772 SEQ ID NO: 37), AAV42-3f (US Patent Application Publication No. 20030138772 SEQ ID NO: 38), AAV42-3g (US Patent Application Publication No. 20030138772 SEQ ID NO: 39), AAV42-4g (US Patent Application Publication No. 2 SEQ ID NO: 32), AAV42-4 (US Patent Application Publication No. 20030138772). SEQ ID NO: 33), AAV42-5a (US Patent Application Publication No. 20030138772 SEQ ID NO: 34), AAV42-10 (US Patent Application Publication No. 20030138772 SEQ ID NO: 35), AAV42-3b (US Patent Application Publication No. 20030138772 SEQ ID NO: 36), AAV42-11 (US Patent Application Publication No. 20030138772 SEQ ID NO: 37), AAV42-6b (US Patent Application Publication No. 20030138772 SEQ ID NO: 38), AAV43-1 (US Patent Application Publication No. 20030138772 SEQ ID NO: 39), AAV43-5 (US Patent Application Publication No. 20030138772 SEQ ID NO: 39), AAV43-6b (US Patent Application Publication No. 20030138772 SEQ ID NO: 39), AAV43-5 (US Patent Application Publication No. 20030138772 SEQ ID NO: 40), AAV42-5b (US Patent Application Publication No. 20030138772 SEQ ID NO: 41), AAV42-5c (US Patent Application Publication No. 20030138772 SEQ ID NO: 42), AAV42-5d (US Patent Application Publication No. 20030138772 SEQ ID NO: 42), AAV42-5e (US Patent Application Publication No. 20030138772 SEQ ID NO: 42), AAV42-5f (US Patent Application Publication No. 20030138772 SEQ ID NO: 43), AAV42-5g (US Patent Application Publication No. SEQ ID NO: 40), AAV43-12 (US Patent Application Publication No. 20030138772 SEQ ID NO: 41), AAV43-20 (US Patent Application Publication No. 20030138772 SEQ ID NO: 42), AAV43-21 (US Patent Application Publication No. 20030138772 SEQ ID NO: 43), AAV43-23 (US Patent Application Publication No. 20030138772 SEQ ID NO: 44), AAV43-25 (US Patent Application Publication No. 20030138772 SEQ ID NO: 45), AAV44.1 (US Patent Application Publication No. 20030138772 SEQ ID NO: 46), AAV44.5 (US Patent Application Publication No. 20030138772 SEQ ID NO: 47), AAV223.1 (US Patent Application Publication No. 20030138772 SEQ ID NO: 48), AAV223.2 (US Patent Application Publication No. 20030138772 SEQ ID NO: 49), AAV223.4 (US Patent Application Publication No. 20030138772 SEQ ID NO: 50), AAV223.5 (US Patent Application Publication No. 20030138772 SEQ ID NO: 51), AAV223.6 (US Patent Application Publication No. 20030138772 SEQ ID NO: 52), AAV223.7 (US Patent Application Publication No. 20030138772 SEQ ID NO: 53), AAVA3.4 (US Patent Application Publication No. 20030138772 SEQ ID NO: 54), SEQ ID NO: 54), AAVA3.5 (US Patent Application Publication No. 20030138772 SEQ ID NO: 55), AAVA3.7 (US Patent Application Publication No. 20030138772 SEQ ID NO: 56), AAVA3.3 (US Patent Application Publication No. 20030138772 AAV42.12 (US Patent Application Publication No. 20030138772 SEQ ID NO: 57), AAV42.12 (US Patent Application Publication No. 20030138772 SEQ ID NO: 58), AAV44.2 (US Patent Application Publication No. 20030138772 SEQ ID NO: 59), AAV42-2 (US Patent Application Publication No. 20030138772 SEQ ID NO: 9), or a variant or hybrid / chimeric / combination thereof.

[0051] In some embodiments, the AAV serotype may be, but is not limited to, AAV2 (SEQ ID NOs: 7 and 23 of US Patent Application Publication No. 20150159173), rh20 (SEQ ID NO: 1 of US Patent Application Publication No. 20150159173), rh32 / 33 (SEQ ID NO: 2 of US Patent Application Publication No. 20150159173), rh39 (SEQ ID NOs: 3, 20, and 36 of US Patent Application Publication No. 20150159173), rh46 (SEQ ID NOs: 4 and 22 of US Patent Application Publication No. 20150159173), rh73 (SEQ ID NOs: 5 and 6 of US Patent Application Publication No. 20150159173), or AAV2 (SEQ ID NOs: 6 and 7 of US Patent Application Publication No. 20150159173). US Patent Application Publication No. 20150159173, SEQ ID NO: 5), rh74 (US Patent Application Publication No. 20150159173, SEQ ID NO: 6), AAV6.1 (US Patent Application Publication No. 20150159173, SEQ ID NO: 29), rh.8 (US Patent Application Publication No. 20150159173, SEQ ID NO: 41), rh.48.1 (US Patent Application Publication No. 20150159173, SEQ ID NO: 44), hu.44 (US Patent Application Publication No. 20150159173, SEQ ID NO: 45), hu.29 (US Patent Application Publication No. 20150159173, SEQ ID NO: 6 ...7), hu.49 (US Patent Application Publication No. 20150159173, SEQ ID NO: 68), hu.50 (US Patent Application Publication No. 20150159173, SEQ ID NO: 69), hu.60 (US Patent Application Publication No. 20150159173, SEQ ID NO: 70), hu.51 (US Patent No. 20150159173), hu.48 (SEQ ID NO: 38 in U.S. Patent Application Publication No. 20150159173), rh54 (SEQ ID NO: 49 in U.S. Patent Application Publication No. 20150159173), AAV2 (SEQ ID NO: 7 in U.S. Patent Application Publication No. 20150159173), cy.5 (SEQ ID NOs: 8 and 24 in U.S. Patent Application Publication No. 20150159173), rh.10 (SEQ ID NOs: 9 and 25 in U.S. Patent Application Publication No. 20150159173), rh.13 (SEQ ID NO: 10 in U.S. Patent Application Publication No. 20150159173), rh.14 (SEQ ID NO: 11 in U.S. Patent Application Publication No. 20150159173), rh.15 (SEQ ID NO: 12 in U.S. Patent Application Publication No. 20150159173), rh.16 (SEQ ID NO: 13 in U.S. Patent Application Publication No. 20150159173), rh.17 (SEQ ID NO: 14 in U.S. Patent Application Publication No. 20150159173), rh.18 (SEQ ID NO: 15 in U.S. Patent Application Publication No. 20150159173), rh.19 (SEQ ID NO: 16 in U.S. Patent Application Publication No. 20150159173), rh.20 (SEQ ID NO: 17 in U.S. Patent Application Publication No. 20150159173), rh.21 (SEQ ID NO: No. 159173), AAV1 (SEQ ID NOs: 11 and 27 in U.S. Patent Application Publication No. 20150159173), AAV3 (SEQ ID NOs: 12 and 28 in U.S. Patent Application Publication No. 20150159173), AAV6 (SEQ ID NOs: 13 and 29 in U.S. Patent Application Publication No. 20150159173), AAV7 (SEQ ID NOs: 14 and 30 in U.S. Patent Application Publication No. 20150159173), AAV8 (SEQ ID NOs: 15 and 31 in U.S. Patent Application Publication No. 20150159173), hu.13 (SEQ ID NOS: 16 and 32 in U.S. Patent Application Publication No. 20150159173), hu.26 (SEQ ID NOS: 17 and 33 in U.S. Patent Application Publication No. 20150159173), hu.37 (SEQ ID NOS: 18 and 34 in U.S. Patent Application Publication No. 20150159173), hu.53 (SEQ ID NOS: 19 and 35 in U.S. Patent Application Publication No. 20150159173), rh.43 (SEQ ID NOS: 10 and 11 in U.S. Patent Application Publication No. 20150159173), rh.54 (SEQ ID NOS: 11 and 12 in U.S. Patent Application Publication No. 20150159173), rh.63 (SEQ ID NOS: 12 and 13 in U.S. Patent Application Publication No. 20150159173), rh.73 (SEQ ID NOS: 13 and 14 in U.S. Patent Application Publication No. 20150159173), rh.83 (SEQ ID NOS: 14 and 15 in U.S. Patent Application Publication No. 20150159173), rh.93 (SEQ ID NOS: 15 and 16 in U.S. Patent Application Publication No. 20150159173), rh.10 (SEQ ID NOS: 16 and 17 in U.S. Patent Application Publication No. 20150159173), rh.11 (SEQ ID NOS: 17 and 18 in U.S. Patent Application Publication No. 20150159173), rh.12 (SEQ ID 21 and 37 in U.S. Patent Application Publication No. 20150159173), rh.2 (SEQ ID NO: 39 in U.S. Patent Application Publication No. 20150159173), rh.37 (SEQ ID NO: 40 in U.S. Patent Application Publication No. 20150159173), rh.64 (SEQ ID NO: 43 in U.S. Patent Application Publication No. 20150159173), rh.48 (SEQ ID NO: 44 in U.S. Patent Application Publication No. 20150159173), ch.5 (SEQ ID NO: 50 in U.S. Patent Application Publication No. 20150159173), No. 46), rh.67 (SEQ ID NO: 47 of U.S. Patent Application Publication No. 20150159173), rh.58 (SEQ ID NO: 48 of U.S. Patent Application Publication No. 20150159173), or variants thereof, such as, but not limited to, Cy5R1, Cy5R2, Cy5R3, Cy5R4, rh.13R, rh.37R2, rh.2R, rh.8R, rh.48.1, rh.48.2, rh.48.1.2, hu.44R1, hu.44R2, hu. The nucleic acid sequence may be or include a sequence as described in U.S. Patent Application Publication No. 20150159173 (the contents of which are incorporated by reference herein in their entirety), such as hu.44R3, hu.29R, ch.5R1, rh64R1, rh64R2, AAV6.2, AAV6.1, AAV6.12, hu.48R1, hu.48R2, or hu.48R3, or a variant or hybrid / chimeric / combination thereof.

[0052] In some embodiments, the AAV serotype may be or may include a sequence described in U.S. Pat. No. 7,198,951 (the contents of which are incorporated herein by reference in their entirety), such as, but not limited to, AAV9 (SEQ ID NOs: 1-3 of U.S. Pat. No. 7,198,951), AAV2 (SEQ ID NO: 4 of U.S. Pat. No. 7,198,951), AAV1 (SEQ ID NO: 5 of U.S. Pat. No. 7,198,951), AAV3 (SEQ ID NO: 6 of U.S. Pat. No. 7,198,951), or AAV8 (SEQ ID NO: 7 of U.S. Pat. No. 7,198,951), or a variant or hybrid / chimera / combination thereof.

[0053] In some embodiments, the AAV serotype may be an AAV9 sequence as described by N. Pulicherla et al. (Molecular Therapy, Vol. 19, No. 6, pp. 1070-1078, 2011, the contents of which are incorporated herein by reference in their entirety), such as, but not limited to, AAV9.9, AAV9.11, AAV9.13, AAV9.16, AAV9.24, AAV9.45, AAV9.47, AAV9.61, AAV9.68, or AAV9.84, or a variant thereof.

[0054] In some embodiments, the AAV serotype may be or may include a sequence described in U.S. Pat. No. 6,156,303 (the contents of which are incorporated herein by reference in their entirety), such as, but not limited to, AAV3B (SEQ ID NOs: 1 and 10 of U.S. Pat. No. 6,156,303), AAV6 (SEQ ID NOs: 2, 7, and 11 of U.S. Pat. No. 6,156,303), AAV2 (SEQ ID NOs: 3 and 8 of U.S. Pat. No. 6,156,303), AAV3A (SEQ ID NOs: 4 and 9 of U.S. Pat. No. 6,156,303), or a derivative or variant or hybrid / chimera / combination thereof.

[0055] In some embodiments, the AAV serotype may be or may include a sequence as described in U.S. Patent Application Publication No. 20140359799 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, AAV8 (SEQ ID NO: 1 of U.S. Patent Application Publication No. 20140359799), AAVDJ (SEQ ID NOs: 2 and 3 of U.S. Patent Application Publication No. 20140359799), or variants thereof.

[0056] In some embodiments, the serotype is selected from the group described by Grimm et al. (Journal of The AAV-DJ sequence may be AAVDJ or a variant thereof, such as AAVDJ8 (or AAV-DJ8), as described in Virology, Vol. 82, No. 12, pp. 5887-5911, 2008, incorporated herein by reference in its entirety. The amino acid sequence of AAVDJ8 may contain two or more mutations effective to remove the heparin-binding domain (HBD). As a non-limiting example, the AAV-DJ sequence set forth as SEQ ID NO: 1 in U.S. Pat. No. 7,588,772 (the contents of which are incorporated herein by reference in their entirety) may contain two mutations: (1) R587Q (arginine (R; Arg) at amino acid 587 is changed to glutamine (Q; Gln)) and (2) R590T (arginine (R; Arg) at amino acid 590 is changed to threonine (T; Thr)). As another non-limiting example, the AAV-DJ sequence described in U.S. Pat. No. 7,588,772 can contain three mutations: (1) K406R (lysine (K; Lys) at amino acid 406 is changed to arginine (R; Arg)), (2) R587Q (arginine (R; Arg) at amino acid 587 is changed to glutamine (Q; Gln)), and (3) R590T (arginine (R; Arg) at amino acid 590 is changed to threonine (T; Thr)).

[0057] In some embodiments, the AAV serotype may be or include the sequence of AAV4 as described in WO1998011244 (the contents of which are incorporated by reference in their entirety), including, but not limited to, AAV4 (SEQ ID NOs: 1-20 of WO1998011244).

[0058] In some embodiments, the AAV serotype may be or may include a mutation in the AAV2 sequence that gives rise to AAV2G9 as described in WO2014144229 and incorporated by reference herein in its entirety.

[0059] In some embodiments, the AAV serotype includes, but is not limited to, AAV3-3 (SEQ ID NO: 217 in WO2005033321), AAV1 (SEQ ID NOs: 219 and 202 in WO2005033321), AAV106.1 / hu.37 (SEQ ID NO: 10 in WO2005033321), AAV114.3 / hu.40 (SEQ ID NO: 11 in WO2005033321), AAV127.2 / hu.41 (SEQ ID NO: 6 and 7 in WO2005033321), AAV127.2 / hu.41 (SEQ ID NO: 7 and 8 in WO2005033321), AAV127.2 / hu.41 (SEQ ID NO: 8 and 9 in WO2005033321), AAV127.2 / hu.41 (SEQ ID NO: 9 and 10 in WO2005033321), AAV127.2 / hu.41 (SEQ ID NO: 10 and 11 in WO2005033321), AAV127.2 / hu.41 (SEQ ID NO: 1 ... and 8), AAV128.3 / hu.44 (SEQ ID NO: 81 in WO 2005033321), AAV130.4 / hu.48 (SEQ ID NO: 78 in WO 2005033321), AAV145.1 / hu.53 (SEQ ID NOs: 176 and 177 in WO 2005033321), AAV145.6 / hu.56 (SEQ ID NOs: 168 and 192 in WO 2005033321), AAV16.12 / hu.11 (SEQ ID NO: 153 in WO 2005033321), and 57), AAV16.8 / hu.10 (SEQ ID NOs: 156 and 56 in WO 2005033321), AAV161.10 / hu.60 (SEQ ID NO: 170 in WO 2005033321), AAV161.6 / hu.61 (SEQ ID NO: 174 in WO 2005033321), AAV1-7 / rh.48 (SEQ ID NO: 32 in WO 2005033321), AAV1-8 / rh.49 (SEQ ID NOs: 103 and 25 in WO 2005033321), AAV2 (SEQ ID NOs: 211 and 221 in WO 2005033321), AAV2-15 / rh.62 (SEQ ID NOs: 33 and 114 in WO 2005033321), AAV2-3 / rh.61 (SEQ ID NO: 21 in WO 2005033321), AAV2-4 / rh.50 (SEQ ID NOs: 23 and 108 in WO 2005033321), AAV2-5 / rh.51 (SEQ ID NOs: 104 and 22 in WO 2005033321), AAV3.1 / hu.AAV3.1 / hu.6 (SEQ ID NOs: 5 and 84 in WO 2005033321), AAV3.1 / hu.9 (SEQ ID NOs: 155 and 58 in WO 2005033321), AAV3-11 / rh.53 (SEQ ID NOs: 186 and 176 in WO 2005033321), AAV3-3 (SEQ ID NO: 200 in WO 2005033321), AAV33.12 / hu.17 (SEQ ID NO: 4 in WO 2005033321), AAV33.4 / hu.15 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.16 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.17 (SEQ ID NO: 4 in WO 2005033321), AAV3.1 / hu.18 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.19 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.20 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.21 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.22 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.23 (SEQ ID NO: 5 in WO 2005033321), AAV3.1 / hu.24 (SEQ ID NO: 5 in WO 20050333 3321), AAV33.8 / hu.16 (SEQ ID NO: 51 in WO 2005033321), AAV3-9 / rh.52 (SEQ ID NOs: 96 and 18 in WO 2005033321), AAV4-19 / rh.55 (SEQ ID NO: 117 in WO 2005033321), AAV4-4 (SEQ ID NOs: 201 and 218 in WO 2005033321), AAV4-9 / rh.54 (SEQ ID NO: 116 ... ), AAV5 (SEQ ID NOs: 199 and 216 in WO 2005033321), AAV52.1 / hu.20 (SEQ ID NO: 63 in WO 2005033321), AAV52 / hu.19 (SEQ ID NO: 133 in WO 2005033321), AAV5-22 / rh.58 (SEQ ID NO: 27 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 105 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 110 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 111 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 112 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 113 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 114 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 115 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 116 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 117 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 118 in WO 2005033321), AAV5-3 / rh.57 (SEQ ID NO: 119 in 33321), AAV58.2 / hu.25 (SEQ ID NO: 49 in WO 2005033321), AAV6 (SEQ ID NOs: 203 and 220 in WO 2005033321), AAV7 (SEQ ID NOs: 222 and 213 in WO 2005033321), AAV7.3 / hu.7 (SEQ ID NO: 55 in WO 2005033321), AAV8 (SEQ ID NOs: 223 and 214 in WO 2005033321), AAVH-1 / hu.AAVhu.1 (SEQ ID NO: 46 in WO 2005033321), AAVH-5 / hu.3 (SEQ ID NO: 44 in WO 2005033321), AAVhu.1 (SEQ ID NO: 144 in WO 2005033321), AAVhu.10 (SEQ ID NO: 156 in WO 2005033321), AAVhu.11 (SEQ ID NO: 153 in WO 2005033321), AAVhu.12 (SEQ ID NO: 156 in WO 2005033321), AAVhu.13 (SEQ ID NO: 153 in WO 2005033321), AAVhu.14 (SEQ ID NO: 144 in WO 2005033321), AAVhu.15 (SEQ ID NO: 153 in WO 2005033321), AAVhu.16 (SEQ ID NO: 156 in WO 2005033321), AAVhu.17 (SEQ ID NO: 153 in WO 2005033321), AAVhu.18 (SEQ ID NO: 153 in WO 2005033321), AAVhu.19 (SEQ ID NO: 153 in WO 2005033321), AAVhu.20 (SEQ ID NO: 153 in WO 2005033321), AAVhu.21 (SEQ ID NO: 153 in WO 2005033321), AAVhu.22 (SEQ ID NO: 153 in WO 2005033321), AAVhu.23 (SEQ ID NO: 153 SEQ ID NO: 59), AAVhu.13 (SEQ ID NO: 129 in WO 2005033321), AAVhu.14 / AAV9 (SEQ ID NOs: 123 and 3 in WO 2005033321), AAVhu.15 (SEQ ID NO: 147 in WO 2005033321), AAVhu.16 (SEQ ID NO: 148 in WO 2005033321), AAVhu.17 ( AAVhu.18 (SEQ ID NO: 83 in WO 2005033321), AAVhu.19 (SEQ ID NO: 133 in WO 2005033321), AAVhu.2 (SEQ ID NO: 143 in WO 2005033321), AAVhu.20 (SEQ ID NO: 1 in WO 2005033321), AAVhu.30 (SEQ ID NO: 1 in WO 2005033321), AAVhu.40 (SEQ ID NO: 1 in WO 2005033321), AAVhu.50 (SEQ ID NO: 1 in WO 2005033321), AAVhu.60 (SEQ ID NO: 1 in WO 2005033321), AAVhu.70 (SEQ ID NO: 1 in WO 2005033321), AAVhu.80 (SEQ ID NO: 1 in WO 2005033321), AAVhu.90 (SEQ ID NO: 1 in WO 2005033321), AAVhu.10 (SEQ ID NO: 1 in WO 2005033321), AAVhu.11 (SEQ ID NO: 1 in WO 2005033321), AAVhu.12 (SEQ ID NO: 1 in WO 2005033321), AAVhu.13 (SEQ ID NO: 1 in WO 2005033321), AAVhu.14 (SEQ ID NO: 1 in WO 2005033321), AAVhu.15 (SEQ ID NO: 1 in WO 2005033321), AAVhu.16 (SEQ ID NO: 1 34), AAVhu.21 (SEQ ID NO: 135 in WO 2005033321), AAVhu.22 (SEQ ID NO: 138 in WO 2005033321), AAVhu.23.2 (SEQ ID NO: 137 in WO 2005033321), AAVhu.24 (SEQ ID NO: 136 in WO 2005033321), AAVhu.25 (SEQ ID NO: 137 in WO 2005033321), AAVhu.26 (SEQ ID NO: 138 in WO 2005033321), AAVhu.27 (SEQ ID NO: 139 in WO 2005033321), AAVhu.28 (SEQ ID NO: 140 in WO 2005033321), AAVhu.29 (SEQ ID NO: 141 in WO 2005033321), AAVhu.29 (SEQ ID NO: 142 in WO 2005033321), AAVhu.20 (SEQ ID NO: 143 in WO 2005033321), AAVhu.21 (SEQ ID NO: 135 in WO 2005033321), AAVhu.22 (SEQ ID NO: 138 in WO 2005033321), AAVhu.23.2 (SEQ ID NO: 144 in WO 2005033321), AAVhu.24 (SEQ ID NO: 145 in WO 2005033321), AAVhu.25 (SEQ ID NO: 146 in WO 2005033321), AAVhu.26 ( 3321), AAVhu.27 (SEQ ID NO: 140 in WO 2005033321), AAVhu.29 (SEQ ID NO: 132 in WO 2005033321), AAVhu.3 (SEQ ID NO: 145 in WO 2005033321), AAVhu.31 (SEQ ID NO: 121 in WO 2005033321), AAVhu.32 (SEQ ID NO: 122 in WO 2005033321), AAVhu.34 (SEQ ID NO: 125 in WO 2005033321), AAVhu.35 (SEQ ID NO: 164 in WO 2005033321), AAVhu.37 (SEQ ID NO: 88 in WO 2005033321), AAVhu.39 (SEQ ID NO: 102 in WO 2005033321), AAVhu.4 (SEQ ID NO: 141 in WO 2005033321), AAVhu. AAVhu.40 (SEQ ID NO: 87 in WO 2005033321), AAVhu.41 (SEQ ID NO: 91 in WO 2005033321), AAVhu.42 (SEQ ID NO: 85 in WO 2005033321), AAVhu.43 (SEQ ID NO: 160 in WO 2005033321), AAVhu.44 (SEQ ID NO: 144 in WO 2005033321), AAVhu.45 (SEQ ID NO: 127 in WO 2005033321), AAVhu.46 (SEQ ID NO: 159 in WO 2005033321), AAVhu.47 (SEQ ID NO: 128 in WO 2005033321), AAVhu.48 (SEQ ID NO: 157 in WO 2005033321), AAVhu.49 (SEQ ID NO: 189 in WO 2005033321), AAVhu.51 (SEQ ID NO: 190 in WO 2005033321), AAVhu.52 (SEQ ID NO: 191 in WO 2005033321), AAVhu.5 3 (SEQ ID NO: 186 in WO 2005033321), AAVhu.54 (SEQ ID NO: 188 in WO 2005033321), AAVhu.55 (SEQ ID NO: 187 in WO 2005033321), AAVhu.56 (SEQ ID NO: 192 in WO 2005033321), AAVhu.57 (SEQ ID NO: 193 in WO 2005033321), AAVhu.58 (SEQ ID NO: 194 in WO 2005033321), AAVhu.AAVhu.6 (SEQ ID NO: 84 in WO 2005033321), AAVhu.60 (SEQ ID NO: 184 in WO 2005033321), AAVhu.61 (SEQ ID NO: 185 in WO 2005033321), AAVhu.63 (SEQ ID NO: 195 in WO 2005033321), AAVhu.64 (WO 2005 AAVhu.66 (SEQ ID NO: 197 in WO 2005033321), AAVhu.67 (SEQ ID NO: 198 in WO 2005033321), AAVhu.7 (SEQ ID NO: 150 in WO 2005033321), AAVhu.8 (SEQ ID NO: 151 in WO 2005033321), AAVhu.9 (SEQ ID NO: 152 in WO 2005033321), AAVhu.10 (SEQ ID NO: 153 in WO 2005033321), AAVhu.11 (SEQ ID NO: 154 in WO 2005033321), AAVhu.12 (SEQ ID NO: 155 in WO 2005033321), AAVhu.13 (SEQ ID SEQ ID NO: 12), AAVhu.9 (SEQ ID NO: 155 in WO 2005033321), AAVLG-10 / rh.40 (SEQ ID NO: 14 in WO 2005033321), AAVLG-4 / rh.38 (SEQ ID NO: 86 in WO 2005033321), AAVLG-4 / rh.38 (SEQ ID NO: 7 in WO 2005033321), AAVN721-8 / rh.43 (SEQ ID NO: 163 in WO 2005033321), AAVN721-8 / rh.43 (SEQ ID NO: 43 in WO 2005033321), AAVpi.1 (WO 2005033321 SEQ ID NO: 28), AAVpi.2 (WO 2005033321 SEQ ID NO: 30), AAVpi.3 (WO 2005033321 SEQ ID NO: 29), AAVrh.38 (WO 2005033321 SEQ ID NO: 86), AAVrh.40 (WO 2005033321 SEQ ID NO: 92), AAVrh.43 (WO 2005033321 SEQ ID NO: 163), AAVrh.44 (WO 2005033321 SEQ ID NO: 34), AAVrh.45 (WO 2005033321 SEQ ID NO: 41), AAVrh.47 (WO 2005033321 SEQ ID NO: 38), AAVrh.AAVrh.48 (SEQ ID NO: 115 in WO 2005033321), AAVrh.49 (SEQ ID NO: 103 in WO 2005033321), AAVrh.50 (SEQ ID NO: 108 in WO 2005033321), AAVrh.51 (SEQ ID NO: 104 in WO 2005033321), AAVrh.52 (SEQ ID NO: 105 in WO 2005033321), AAVrh.53 (SEQ ID NO: 106 in WO 2005033321), AAVrh.54 (SEQ ID NO: 107 in WO 2005033321), AAVrh.55 (SEQ ID NO: 109 in WO 2005033321), AAVrh.56 (SEQ ID NO: 110 in WO 2005033321), AAVrh.57 (SEQ ID NO: 111 in WO 2005033321), AAVrh.58 (SEQ ID NO: 112 in WO 2005033321), AAVrh.59 (SEQ ID NO: 113 in WO 2005033321), AAVrh.60 (SEQ ID NO: 114 in WO 2005033321), AAVrh.61 (SEQ ID NO: 115 in WO 2005033321), AAVrh.62 (SEQ ID NO: 116 in WO 2005033321), AAVrh.63 (SEQ ID NO: 117 in WO 2005033321), AAVrh.64 (SEQ ID NO: 11 321), AAVrh.53 (SEQ ID NO: 97 in WO 2005033321), AAVrh.55 (SEQ ID NO: 37 in WO 2005033321), AAVrh.56 (SEQ ID NO: 152 in WO 2005033321), AAVrh.57 (SEQ ID NO: 105 in WO 2005033321), AAVrh.58 (SEQ ID NO: 106 in WO 2005033321), AAVrh.59 (SEQ ID NO: 42 in WO 2005033321), AAVrh.60 (SEQ ID NO: 106 in WO 2005033321), AAVrh.61 (SEQ ID NO: 106 in WO 2005033321), AAVrh.62 (SEQ ID NO: 106 in WO 2005033321), AAVrh.63 (SEQ ID NO: 106 in WO 2005033321), AAVrh.64 (SEQ ID NO: 106 in WO 2005033321), AAVrh.65 (SEQ ID NO: 106 in WO 2005033321), AAVrh.66 (SEQ ID NO: 106 in WO 2005033321), AAVrh.67 (SEQ ID NO: 106 in WO 2005033321), AAVrh.68 (SEQ ID NO: 106 in WO 2005033321), AAVrh.69 (SEQ ID NO: 106 in WO 2005033321), AAVrh.69 (SEQ ID NO: SEQ ID NO: 31), AAVrh.61 (SEQ ID NO: 107 in WO 2005033321), AAVrh.62 (SEQ ID NO: 114 in WO 2005033321), AAVrh.64 (SEQ ID NO: 99 in WO 2005033321), AAVrh.65 (SEQ ID NO: 35 in WO 2005033321), AAVrh.68 (SEQ ID NO: 16 in WO 2005033321), AAVrh.69 (SEQ ID NO: 39 in WO 2005033321), AAVrh.70 (SEQ ID NO: 20 in WO 2005033321), AAVrh.72 (SEQ ID NO: 106 in WO 2005033321), AAVrh.73 (SEQ ID NO: 107 in WO 2005033321), AAVrh.74 (SEQ ID NO: 106 in WO 2005033321), AAVrh.75 (SEQ ID NO: 106 in WO 2005033321), AAVrh.76 (SEQ ID NO: 106 in WO 2005033321), AAVrh.77 (SEQ ID NO: 106 in WO 2005033321), AAVrh.78 (SEQ ID NO: 106 in WO 2005033321), AAVrh.79 (SEQ ID NO: 106 in WO 2005033321), AAVrh.80 (SEQ ID NO: 106 in WO 2005033321), AAVrh.81 (SEQ ID NO and AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, or AAVrh14, such as SEQ ID NO:9, or a variant thereof, including but not limited to AAVcy.2, AAVcy.3, AAVcy.4, AAVcy.5, AAVcy.6, AAVrh.12, AAVrh.17, AAVrh.18, AAVrh.19, AAVrh.21, AAVrh.22, AAVrh.23, AAVrh.24, AAVrh.25, AAVrh.25 / 42 15, AAVrh.31, AAVrh.32, AAVrh.33, AAVrh.34, AAVrh.35, AAVrh.36, AAVrh.37, or AAVrh14, as described in WO2005033321 (the contents of which are incorporated by reference in their entirety).Non-limiting examples of variants include SEQ ID NOs: 13, 15, 17, 19, 24, 36, 40, 45, 47, 48, 51, 52, 53, 54, 60, 61, 62, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 79, 80, 82, 89, 90, 93, 94, 95, 98, 100, 101, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 0, 124, 126, 131, 139, 142, 151, 154, 158, 161, 162, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 202, 204, 205, 206, 207, 208, 209, 210, 211, 212, 215, 219, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235 or 236.

[0060] In some embodiments, the AAV serotype may be or may include a sequence as described in WO2015168666 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, AAVrh8R (SEQ ID NO: 9 of WO2015168666), AAVrh8R A586R mutant (SEQ ID NO: 10 of WO2015168666), AAVrh8R R533A mutant (SEQ ID NO: 11 of WO2015168666), or variants thereof.

[0061] In some embodiments, the AAV serotype may be, but is not limited to, AAVhE1.1 (SEQ ID NO: 44 of U.S. Pat. No. 9,233,131), AAVhEr1.5 (SEQ ID NO: 45 of U.S. Pat. No. 9,233,131), AAVhER1.14 (SEQ ID NO: 46 of U.S. Pat. No. 9,233,131), AAVhEr1.8 (SEQ ID NO: 47 of U.S. Pat. No. 9,233,131), AAVhEr1.16 (U.S. Pat. No. 9,233,131), AAVhEr1.18 (SEQ ID NO: 48 of U.S. Pat. No. 9,233,131), AAVhEr1.19 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.20 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.21 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.22 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.23 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.24 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.25 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.26 (S No. 31), AAVhEr1.18 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr1.35 (SEQ ID NO: 50 of U.S. Pat. No. 9,233,131), AAVhEr1.7 (SEQ ID NO: 51 of U.S. Pat. No. 9,233,131), AAVhEr1.36 (SEQ ID NO: 52 of U.S. Pat. No. 9,233,131), AAVhEr2.29 (SEQ ID NO: 53 of U.S. Pat. No. 9,233,131), AAVhEr3.18 (SEQ ID NO: 49 of U.S. Pat. No. 9,233,131), AAVhEr3.35 (SEQ ID NO: 50 of U.S. Pat. No. 9,233,131), AAVhEr3.7 (SEQ ID NO: 51 of U.S. Pat. No. 9,233,131), AAVhEr3.36 (SEQ ID NO: 52 of U.S. Pat. No. 9,233,131), AAVhEr4.29 (SEQ ID NO: 53 of U.S. Pat. No. 9,233,131), AAVhEr5.29 (SEQ ID NO: 53 of U.S. Pat. No. 9,233,131), AAVhEr6.29 (SEQ ID NO: 53 of U.S. Pat. No. 9,233,131), AAVhEr7.29 (SEQ ID NO: 53 of U.S. Pat. No. 9,233,131), AAVhEr8.29 (SEQ ID 53), AAVhEr2.4 (SEQ ID NO: 54 in U.S. Pat. No. 9,233,131), AAVhEr2.16 (SEQ ID NO: 55 in U.S. Pat. No. 9,233,131), AAVhEr2.30 (SEQ ID NO: 56 in U.S. Pat. No. 9,233,131), AAVhEr2.31 (SEQ ID NO: 58 in U.S. Pat. No. 9,233,131), AAVhEr2.36 (SEQ ID NO: 57 in U.S. Pat. No. 9,233,131), AAVhEr1 The sequence may be or include a sequence as described in U.S. Patent No. 9,233,131 (the contents of which are incorporated by reference in their entirety herein), such as AAVhEr3.1 (SEQ ID NO: 59 of U.S. Patent No. 9,233,131), AAV2.5T (SEQ ID NO: 42 of U.S. Patent No. 9,233,131), or a variant thereof.

[0062] In some embodiments, the AAV serotype may be, but is not limited to, AAV-PAEC (SEQ ID NO: 1 in US Patent Application Publication No. 20150376607), AAV-LK01 (SEQ ID NO: 2 in US Patent Application Publication No. 20150376607), AAV-LK02 (SEQ ID NO: 3 in US Patent Application Publication No. 20150376607), AAV-LK03 (SEQ ID NO: 4 in US Patent Application Publication No. 20150376607), AAV-LK04 (SEQ ID NO: 5 in US Patent Application Publication No. 20150376607), AAV-LK05 (US Patent Application Publication No. AAV-LK06 (SEQ ID NO: 6 in U.S. Patent Application Publication No. 20150376607), AAV-LK07 (SEQ ID NO: 8 in U.S. Patent Application Publication No. 20150376607), AAV-LK08 (SEQ ID NO: 9 in U.S. Patent Application Publication No. 20150376607), AAV-LK09 (SEQ ID NO: 10 in U.S. Patent Application Publication No. 20150376607), AAV-LK10 (SEQ ID NO: 11 in U.S. Patent Application Publication No. 20150376607), AAV-LK11 (U.S. Patent Application Publication No. 20150376607), AAV-LK12 (SEQ ID NO: 13 in U.S. Patent Application Publication No. 20150376607), AAV-LK13 (SEQ ID NO: 14 in U.S. Patent Application Publication No. 20150376607), AAV-LK14 (SEQ ID NO: 15 in U.S. Patent Application Publication No. 20150376607), AAV-LK15 (SEQ ID NO: 16 in U.S. Patent Application Publication No. 20150376607), AAV-LK16 (SEQ ID NO: 17 in U.S. Patent Application Publication No. 20150376607), AAV-LK17 (SEQ ID NO: 18 in U.S. Patent Application Publication No. 20150376607), AAV-LK18 (SEQ ID NO: 19 in U.S. Patent Application Publication No. 20150376607), AAV-LK19 (SEQ ID NO: 20 AAV-LK12 (SEQ ID NO: 12 in U.S. Patent Application Publication No. 20150376607), AAV-LK12 (SEQ ID NO: 13 in U.S. Patent Application Publication No. 20150376607), AAV-LK13 (SEQ ID NO: 14 in U.S. Patent Application Publication No. 20150376607), AAV-LK14 (SEQ ID NO: 15 in U.S. Patent Application Publication No. 20150376607), AAV-LK15 (SEQ ID NO: 16 in U.S. Patent Application Publication No. 20150376607), AAV-LK16 (SEQ ID NO: 17 in U.S. Patent Application Publication No. 20150376607), AAV-LK1 7 (SEQ ID NO: 18 in U.S. Patent Application Publication No. 20150376607), AAV-LK18 (SEQ ID NO: 19 in U.S. Patent Application Publication No. 20150376607), AAV-LK19 (SEQ ID NO: 20 in U.S. Patent Application Publication No. 20150376607), AAV-PAEC2 (SEQ ID NO: 21 in U.S. Patent Application Publication No. 20150376607), AAV-PAEC4 (SEQ ID NO: 22 in U.S. Patent Application Publication No. 20150376607), AAV-PAEC6 (SEQ ID NO: 23 in U.S. Patent Application Publication No. 20150376607),The sequence may be or may include a sequence as described in U.S. Patent Application Publication No. 20150376607 (the contents of which are incorporated by reference in their entirety), such as AAV-PAEC7 (SEQ ID NO: 24 of U.S. Patent Application Publication No. 20150376607), AAV-PAEC8 (SEQ ID NO: 25 of U.S. Patent Application Publication No. 20150376607), AAV-PAEC11 (SEQ ID NO: 26 of U.S. Patent Application Publication No. 20150376607), AAV-PAEC12 (SEQ ID NO: 27 of U.S. Patent Application Publication No. 20150376607), or a variant thereof.

[0063] In some embodiments, the AAV serotype may be or include a sequence as described in U.S. Pat. No. 9,163,261 (the contents of which are incorporated by reference in their entirety) such as, but not limited to, AAV-2-pre-miRNA-101 (SEQ ID NO: 1 of U.S. Pat. No. 9,163,261), or a variant thereof.

[0064] In some embodiments, the AAV serotype may be or have a sequence as described in U.S. Patent Application Publication No. 20150376240 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, AAV-8h (SEQ ID NO: 6 of U.S. Patent Application Publication No. 20150376240), AAV-8b (SEQ ID NO: 5 of U.S. Patent Application Publication No. 20150376240), AAV-h (SEQ ID NO: 2 of U.S. Patent Application Publication No. 20150376240), AAV-b (SEQ ID NO: 1 of U.S. Patent Application Publication No. 20150376240), or variants thereof.

[0065] In some embodiments, the AAV serotype includes, but is not limited to, AAV SM 10-2 (US Patent Application Publication No. 20160017295 SEQ ID NO: 22), AAV Shuffle 100-1 (US Patent Application Publication No. 20160017295 SEQ ID NO: 23), AAV Shuffle 100-3 (US Patent Application Publication No. 20160017295 SEQ ID NO: 24), AAV Shuffle 100-7 (US Patent Application Publication No. 20160017295 SEQ ID NO: 25), AAV Shuffle 100-8 (US Patent Application Publication No. 20160017295 SEQ ID NO: 26), AAV Shuffle 100-9 (US Patent Application Publication No. 20160017295 SEQ ID NO: 27), AAV Shuffle 100-1 (US Patent Application Publication No. 20160017295 SEQ ID NO: 28), AAV Shuffle 100-2 (US Patent Application Publication No. 20160017295 SEQ ID NO: 29), AAV Shuffle 100-3 (US Patent Application Publication No. 20160017295 SEQ ID NO: 30), AAV Shuffle 100-4 (US Patent Application Publication No. 20160017295 SEQ ID NO: 31), AAV Shuffle 100-5 (US Patent Application Publication No. 20160017295 SEQ ID NO: 32), AAV Shuffle 100-6 (US Patent Application Publication No. 20160017295 SEQ ID NO: 33), AAV Shuffle 100-7 (US Patent Application Publication No. 20160017295 SEQ ID NO: 34 shuffle10-2 (SEQ ID NO: 34 in U.S. Patent Application Publication No. 20160017295), AAV shuffle10-6 (SEQ ID NO: 35 in U.S. Patent Application Publication No. 20160017295), AAV shuffle10-8 (SEQ ID NO: 36 in U.S. Patent Application Publication No. 20160017295), AAV shuffle100-2 (SEQ ID NO: 37 in U.S. Patent Application Publication No. 20160017295), AAV The AAV may be or have a sequence as described in U.S. Patent Application Publication No. 20160017295 (the contents of which are incorporated by reference in their entirety), such as SM 10-1 (SEQ ID NO: 38 of U.S. Patent Application Publication No. 20160017295), AAV SM 10-8 (SEQ ID NO: 39 of U.S. Patent Application Publication No. 20160017295), AAV SM 100-3 (SEQ ID NO: 40 of U.S. Patent Application Publication No. 20160017295), AAV SM 100-10 (SEQ ID NO: 41 of U.S. Patent Application Publication No. 20160017295), or a variant thereof.

[0066] In some embodiments, the AAV serotype may be or may include a sequence as described in U.S. Patent Application Publication No. 20150238550 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, BNP61 AAV (SEQ ID NO: 1 of U.S. Patent Application Publication No. 20150238550), BNP62 AAV (SEQ ID NO: 3 of U.S. Patent Application Publication No. 20150238550), BNP63 AAV (SEQ ID NO: 4 of U.S. Patent Application Publication No. 20150238550), or a variant thereof.

[0067] In some embodiments, the AAV serotype includes, but is not limited to, AAVrh.50 (US Patent Application Publication No. 20150315612, SEQ ID NO: 108), AAVrh.43 (US Patent Application Publication No. 20150315612, SEQ ID NO: 163), AAVrh.62 (US Patent Application Publication No. 20150315612, SEQ ID NO: 114), AAVrh.48 (US Patent Application Publication No. 20150315612, SEQ ID NO: 115), AAVhu.19 (US Patent Application Publication No. 20150315612, SEQ ID NO: 116), AAVrh.50 (US Patent Application Publication No. 20150315612, SEQ ID NO: 117), AAVrh.51 (US Patent Application Publication No. 20150315612, SEQ ID NO: 118), AAVrh.52 (US Patent Application Publication No. 20150315612, SEQ ID NO: 119), AAVrh.53 (US Patent Application Publication No. 20150315612, SEQ ID NO: 120), AAVrh.54 (US Patent Application Publication No. 20150315612, SEQ ID NO: 121), AAVrh.55 (US Patent Application Publication No. 20150315612, SEQ ID NO: 122), AAVrh.56 (US Patent Application Publication No. 20150315612, SEQ ID NO: 123), AAVrh.57 (US Patent Application Publication No. 20150315612, SEQ ID NO: 124), AAVrh.58 (US Patent Application Publication No. 2015031561 AAVhu.11 (SEQ ID NO: 153 in U.S. Patent Application Publication No. 20150315612), AAVhu.53 (SEQ ID NO: 186 in U.S. Patent Application Publication No. 20150315612), AAV4-8 / rh.64 (SEQ ID NO: 15 in U.S. Patent Application Publication No. 20150315612), AAVLG-9 / hu.39 (SEQ ID NO: 24 in U.S. Patent Application Publication No. 20150315612), AAV54.5 / hu.23 (U.S. Patent Application Publication No. 20150315612), ...hu.53 (SEQ ID NO: 186 in U.S. Patent Application Publication No. 20150315612), AAV No. 315612), AAV54.2 / hu.22 (SEQ ID NO: 67 of U.S. Patent Application Publication No. 20150315612), AAV54.7 / hu.24 (SEQ ID NO: 66 of U.S. Patent Application Publication No. 20150315612), AAV54.1 / hu.21 (SEQ ID NO: 65 of U.S. Patent Application Publication No. 20150315612), AAV54.4R / hu.27 (SEQ ID NO: 64 of U.S. Patent Application Publication No. 20150315612), AAV46.2 / hu.28 (U.S. Patent Application Publication No. 20150 ...4R / hu.27 (SEQ ID NO: 64 of U.S. Patent Application Publication No. 20150315612), AAV46.2 / hu.28 (U.S. Patent Application Publication No. 20150315612), AAV54.4R / hu The AAV may be or may include a sequence as described in U.S. Patent Application Publication No. 20150315612 (the contents of which are incorporated by reference in their entirety), such as AAV46.6 / hu.29 (U.S. Patent Application Publication No. 20150315612 SEQ ID NO: 68), AAV46.6 / hu.29 (U.S. Patent Application Publication No. 20150315612 SEQ ID NO: 69), AAV128.1 / hu.43 (U.S. Patent Application Publication No. 20150315612 SEQ ID NO: 80), or a variant thereof.

[0068] In some embodiments, the AAV serotype may be or may include a sequence as described in WO2015121501 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, truetype AAV (ttAAV) (SEQ ID NO: 2 of WO2015121501), "Eupen AAV10" (SEQ ID NO: 8 of WO2015121501), "Japanese AAV10" (SEQ ID NO: 9 of WO2015121501), or variants thereof.

[0069] According to the present disclosure, the selection or use of AAV capsid serotypes can be from various species. In one embodiment, the AAV can be avian AAV (AAAV). The AAAV serotype can be or have a sequence as described in U.S. Patent No. 9,238,800 (the contents of which are incorporated herein by reference in their entirety), such as, but not limited to, AAAV (SEQ ID NO: 1, 2, 4, 6, 8, 10, 12, or 14 of U.S. Patent No. 9,238,800), or a variant thereof.

[0070] In one embodiment, the AAV may be bovine AAV (BAAV). BAAV serotypes may be or have sequences as described in U.S. Patent No. 9,193,769 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, BAAV (SEQ ID NOS: 1 and 6 of U.S. Patent No. 9,193,769), or variants thereof. BAAV serotypes may be or have sequences as described in U.S. Patent No. 7,427,396 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, BAAV (SEQ ID NOS: 5 and 6 of U.S. Patent No. 7,427,396), or variants thereof.

[0071] In some embodiments, the AAV may be a caprine AAV. The caprine AAV serotype may be or have a sequence as described in U.S. Patent No. 7,427,396 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, a caprine AAV (SEQ ID NO: 3 of U.S. Patent No. 7,427,396), or a variant thereof.

[0072] In some embodiments, AAV can be engineered as a hybrid AAV from two or more parental serotypes.In one embodiment, this AAV can be AAV2G9, which comprises sequences from AAV2 and AAV9.The AAV2G9 AAV serotype can be or have the sequence described in US Patent Publication No. 20160017005 (the contents of which are incorporated herein by reference in their entirety).

[0073] In one embodiment, the AAV may be a serotype generated by an AAV9 capsid library having mutations at amino acids 390-627 (VP1 numbering) as described by Pulicherla et al. (Molecular Therapy, Vol. 19, No. 6, pp. 1070-1078, 2011, the contents of which are incorporated by reference in their entirety herein). Serotypes and corresponding nucleotide and amino acid substitutions include, but are not limited to, AAV9.1 (G1594C; D532H), AAV6.2 (T1418A and T1436X; V473D and I479K), AAV9.3 (T1238A; F413Y), AAV9.4 (T1250C and A1617T; F417S), AAV9.5 (A1235G, A1314T, A1642G, C1760T; Q412R, T548A, A5 87V), AAV9.6(T1231A;F411I), AAV9.9(G1203A, G1785T;W595C), AAV9.10(A1500G, T1676C;M559T), AAV9.11(A1425T, A1702C, A1769T; T568P, Q590L), AAV9.13 (A1369C, A1720T; N457H, T574S), AAV9.14 (T1340A, T1362C, T1560C, G1713A; L 447H), AAV9.16(A1775T;Q592L), AAV9.24(T1507C, T1521G;W503R), AAV9.26(A1337G, A1769C;Y446C, Q590P), AAV9.3 3(A1667C;D556A), AAV9.34(A1534G, C1794T;N512D), AAV9.35(A1289T, T1450A, C1494T, A1515T, C1794A, G1816A;Q430 L, Y484N, N98K, V606I), AAV9.40 (A1694T, E565V), AAV9.41 (A1348T, T1362C; T450S), AAV9.44 (A1684C, A1701T, A1737G) ;N562H, K567N), AAV9.45 (A1492T, C1804T; N498Y, L602F), AAV9.46 (G1441C, T1525C, T1549G; G481R, W509R, L517V), 9.47 (G1241A, G1358A, A1669G, C1745T; S414N, G453D, K557E, T582I), AAV9.48 (C1445T, A1736T; P482L, Q579L) , AAV9.50(A1638T, C1683T, T1805A; Q546H, L602H), AAV9.53(G1301A, A1405C, C1664T, G1811T; R134Q, S469R, A555V, G604V), AAV9.54 (C1531A, T1609A; L511I, L537M), AAV9.55 (T1605A; F535L), AAV9.58 (C1475T, C1579A ;T492I, H527N), AAV.59(T1336C;Y446H), AAV9.61(A1493T;N498I), AAV9.64(C1531A, A1617T;L511I), AAV9. 65 (C1335T, T1530C, C1568A; A523D), AAV9.68 (C1510A; P504T), AAV9.80 (G1441A; G481R), AAV9.83 (C1402A, A 1500T;P468T, E500D), AAV9.87(T1464C, T1468C;S490P), AAV9.90(A1196T;Y399F), AAV9.91(T1316G, A1583T , C1782G, T1806C; L439R, K528I), AAV9.93 (A1273G, A1421G, A1638C, C1712T, G1732A, A1744T, A1832T; S425G, Q474R, Q546H, P571L, G578R, T582S, D611V), AAV9.94 (A1675T; M559L), or AAV9.95 (T1605A; F535L).

[0074] In some embodiments, the AAV serotypes include, but are not limited to, AAVF1 / HSC1 (SEQ ID NOs: 2 and 20 in WO2016049230), AAVF2 / HSC2 (SEQ ID NOs: 3 and 21 in WO2016049230), AAVF3 / HSC3 (SEQ ID NOs: 5 and 22 in WO2016049230), AAVF4 / HSC4 (SEQ ID NOs: 6 and 23 in WO2016049230), AAVF5 / HSC5 (SEQ ID NOs: 11 and 25 in WO 2016049230), AAVF6 / HSC6 (SEQ ID NOs: 7 and 24 in WO 2016049230), AAVF7 / HSC7 (SEQ ID NOs: 8 and 27 in WO 2016049230), AAVF8 / HSC8 (SEQ ID NOs: 9 and 28 in WO 2016049230), AAVF9 / HSC9 (SEQ ID NOs: 10 and 29 in WO 2016049230), A AVF11 / HSC11 (SEQ ID NOs: 4 and 26 in WO 2016049230), AAVF12 / HSC12 (SEQ ID NOs: 12 and 30 in WO 2016049230), AAVF13 / HSC13 (SEQ ID NOs: 14 and 31 in WO 2016049230), AAVF14 / HSC14 (SEQ ID NOs: 15 and 32 in WO 2016049230), AAVF15 / HSC15 (SEQ ID NOs: 16 and 33 in WO 2016049230), The AAVF16 / HSC16 sequence may be or may include a sequence as described in WO2016049230 (the contents of which are incorporated by reference in their entirety), such as AAVF16 / HSC16 (SEQ ID NOs: 16 and 33 of WO2016049230), AAVF16 / HSC16 (SEQ ID NOs: 17 and 34 of WO2016049230), AAVF17 / HSC17 (SEQ ID NOs: 13 and 35 of WO2016049230), or a variant or derivative thereof.

[0075] In some embodiments, the AAV serotypes include, but are not limited to, AAV CBr-El (SEQ ID NOs: 13 and 87 in U.S. Pat. No. 8,734,809), AAV CBr-E2 (SEQ ID NOs: 14 and 88 in U.S. Pat. No. 8,734,809), AAV CBr-E3 (SEQ ID NOs: 15 and 89 in U.S. Pat. No. 8,734,809), AAV CBr-E4 (SEQ ID NOs: 16 and 90 in U.S. Pat. No. 8,734,809), AAV CBr-E5 (SEQ ID NOs: 17 and 91 in U.S. Pat. No. 8,734,809), AAV CBr-e5 (SEQ ID NOs: 18 and 92 in U.S. Pat. No. 8,734,809), AAV CBr-E6 (SEQ ID NOs: 19 and 93 in U.S. Pat. No. 8,734,809), AAV CBr-E7 (SEQ ID NOs: 19 and 94 in U.S. Pat. No. 8,734,809), AAV CBr-E8 (SEQ ID NOs: 20 and 21 in U.S. Pat. No. 8,734,809), AAV CBr-E9 (SEQ ID NOs: 21 and 22 in U.S. Pat. No. 8,734,809), AAV CBr-E10 (SEQ ID NOs: 22 and 23 in U.S. Pat. No. 8,734,809), AAV CBr-E11 (SEQ ID NOs: 23 and 24 in U.S. Pat. No. 8,734,80 CBr-E7 (SEQ ID NOs: 20 and 94 in U.S. Patent No. 8,734,809), AAV CBr-E8 (SEQ ID NOs: 21 and 95 in U.S. Patent No. 8,734,809), AAV CLv-D1 (SEQ ID NOs: 22 and 96 in U.S. Patent No. 8,734,809), AAV CLv-D2 (SEQ ID NOs: 23 and 97 in U.S. Patent No. 8,734,809), AAV CLv-D3 (SEQ ID NOs: 24 and 98 in U.S. Patent No. 8,734,809), AAV CLv-D4 (SEQ ID NOs: 25 and 99 in U.S. Patent No. 8,734,809), AAV CLv-D5 (SEQ ID NOs: 26 and 100 in U.S. Patent No. 8,734,809), AAV CLv-D6 (SEQ ID NOs: 27 and 101 in U.S. Patent No. 8,734,809), AAV CLv-D7 (SEQ ID NOs: 28 and 102 in U.S. Patent No. 8,734,809), AAV CLv-D8 (SEQ ID NOs: 29 and 103 in U.S. Patent No. 8,734,809), AAV CLv-E1 (SEQ ID NOs: 13 and 87 in U.S. Patent No. 8,734,809), AAV CLv-R1 (SEQ ID NOs: 30 and 104 in U.S. Patent No. 8,734,809), AAV CLv-R2 (SEQ ID NOs: 31 and 105 in U.S. Patent No. 8,734,809), AAV CLv-R3 (SEQ ID NOs: 32 and 106 in U.S. Patent No. 8,734,809), AAV CLv-R4 (SEQ ID NOs: 33 and 107 in U.S. Patent No. 8,734,809), AAV CLv-R5 (SEQ ID NOs: 34 and 108 in U.S. Patent No. 8,734,809), AAV CLv-R6 (SEQ ID NOs: 35 and 109 in U.S. Patent No. 8,734,809), AAV CLv-R7 (SEQ ID NOs: 36 and 110 in U.S. Patent No. 8,734,809), AAV CLv-R8 (SEQ ID NOs: 37 and 111 in U.S. Patent No. 8,734,809), AAV CLv-R9 (SEQ ID NOs: 38 and 112 in U.S. Patent No. 8,734,809), AAV CLg-F1 (SEQ ID NOs: 39 and 113 in U.S. Patent No. 8,734,809), AAV CLg-F2 (SEQ ID NOs: 40 and 114 in U.S. Patent No. 8,734,809), AAV CLg-F3 (SEQ ID NOs: 41 and 115 in U.S. Patent No. 8,734,809), AAV CLg-F4 (SEQ ID NOs: 42 and 116 in U.S. Patent No. 8,734,809), AAVCLg-F5 (SEQ ID NOs: 43 and 117 in U.S. Patent No. 8,734,809), AAV CLg-F6 (SEQ ID NOs: 43 and 117 in U.S. Patent No. 8,734,809), AAV CLg-F7 (SEQ ID NOs: 44 and 118 in U.S. Patent No. 8,734,809), AAV CLg-F8 (SEQ ID NOs: 43 and 117 in U.S. Patent No. 8,734,809), AAV CSp-1 (SEQ ID NOs: 45 and 119 in U.S. Patent No. 8,734,809), AAV CSp-10 (SEQ ID NOs: 46 and 120 in U.S. Patent No. 8,734,809), AAV CSp-11 (SEQ ID NOs: 47 and 121 in U.S. Patent No. 8,734,809), AAV CSp-2 (SEQ ID NOs: 48 and 122 in U.S. Patent No. 8,734,809), AAV CSp-3 (SEQ ID NOs: 49 and 123 in U.S. Patent No. 8,734,809), AAV CSp-4 (SEQ ID NOs: 50 and 124 in U.S. Patent No. 8,734,809), AAV CSp-6 (SEQ ID NOs: 51 and 125 in U.S. Patent No. 8,734,809), AAV CSp-7 (SEQ ID NOs: 52 and 126 in U.S. Patent No. 8,734,809), AAV CSp-8 (SEQ ID NOs: 53 and 127 in U.S. Patent No. 8,734,809), AAV CSp-9 (SEQ ID NOs: 54 and 128 in U.S. Patent No. 8,734,809), AAV CHt-2 (SEQ ID NOs: 55 and 129 in U.S. Patent No. 8,734,809), AAV CHt-3 (SEQ ID NOs: 56 and 130 in U.S. Patent No. 8,734,809), AAV CKd-1 (SEQ ID NOs: 57 and 131 in U.S. Patent No. 8,734,809), AAV CKd-10 (SEQ ID NOs: 58 and 132 in U.S. Patent No. 8,734,809), AAV CKd-2 (SEQ ID NOs: 59 and 133 in U.S. Patent No. 8,734,809), AAV CKd-3 (SEQ ID NOs: 60 and 134 in U.S. Patent No. 8,734,809), AAV CKd-4 (SEQ ID NOs: 61 and 135 in U.S. Patent No. 8,734,809), AAV CKd-6 (SEQ ID NOs: 62 and 136 in U.S. Patent No. 8,734,809), AAV CKd-7 (SEQ ID NOs: 63 and 137 in U.S. Patent No. 8,734,809), AAV CKd-8 (SEQ ID NOs: 64 and 138 in U.S. Patent No. 8,734,809), AAVCLv-1 (SEQ ID NOs: 35 and 139 in U.S. Patent No. 8,734,809), AAV CLv-12 (SEQ ID NOs: 66 and 140 in U.S. Patent No. 8,734,809), AAV CLv-13 (SEQ ID NOs: 67 and 141 in U.S. Patent No. 8,734,809), AAV CLv-2 (SEQ ID NOs: 68 and 142 in U.S. Patent No. 8,734,809), AAV CLv-3 (SEQ ID NOs: 69 and 143 in U.S. Patent No. 8,734,809), AAV CLv-4 (SEQ ID NOs: 70 and 144 in U.S. Patent No. 8,734,809), AAV CLv-6 (SEQ ID NOs: 71 and 145 in U.S. Patent No. 8,734,809), AAV CLv-8 (SEQ ID NOs: 72 and 146 in U.S. Patent No. 8,734,809), AAV CKd-B1 (SEQ ID NOs: 73 and 147 in U.S. Patent No. 8,734,809), AAV CKd-B2 (SEQ ID NOs: 74 and 148 in U.S. Patent No. 8,734,809), AAV CKd-B3 (SEQ ID NOs: 75 and 149 in U.S. Patent No. 8,734,809), AAV CKd-B4 (SEQ ID NOs: 76 and 150 in U.S. Patent No. 8,734,809), AAV CKd-B5 (SEQ ID NOs: 77 and 151 in U.S. Patent No. 8,734,809), AAV CKd-B6 (SEQ ID NOs: 78 and 152 in U.S. Patent No. 8,734,809), AAV CKd-B7 (SEQ ID NOs: 79 and 153 in U.S. Patent No. 8,734,809), AAV CKd-B8 (SEQ ID NOs: 80 and 154 in U.S. Patent No. 8,734,809), AAV CKd-H1 (SEQ ID NOs: 81 and 155 in U.S. Patent No. 8,734,809), AAV CKd-H2 (SEQ ID NOs: 82 and 156 in U.S. Patent No. 8,734,809), AAV CKd-H3 (SEQ ID NOs: 83 and 157 in U.S. Patent No. 8,734,809), AAV CKd-H4 (SEQ ID NOs: 84 and 158 in U.S. Patent No. 8,734,809), AAV CKd-H5 (SEQ ID NOs: 85 and 159 in U.S. Patent No. 8,734,809), AAV CKd-H6 (SEQ ID NOs: 77 and 151 in U.S. Patent No. 8,734,809), AAV CHt-1 (SEQ ID NOs: 86 and 160 in U.S. Patent No. 8,734,809), AAV CLv1-1 (SEQ ID NO: 171 in U.S. Patent No. 8,734,809), AAV CLv1-2 (SEQ ID NO: 172 in U.S. Patent No. 8,734,809), AAV CLv1-3 (SEQ ID NO: 173 in U.S. Patent No. 8,734,809), AAV CLv1-4 (SEQ ID NO: 174 in U.S. Patent No. 8,734,809), AAV Clv1-7 (SEQ ID NO: 175 in U.S. Patent No. 8,734,809), AAV Clv1-8 (SEQ ID NO: 176 in U.S. Patent No. 8,734,809), AAV Clv1-9 (SEQ ID NO: 177 in U.S. Patent No. 8,734,809), AAVIt may be or include a sequence as described in U.S. Pat. No. 8,734,809 (the contents of which are incorporated by reference in their entirety), such as Clv1-10 (SEQ ID NO: 178 of U.S. Pat. No. 8,734,809), AAV.VR-355 (SEQ ID NO: 181 of U.S. Pat. No. 8,734,809), AAV.hu.48R3 (SEQ ID NO: 183 of U.S. Pat. No. 8,734,809), or a variant or derivative thereof.

[0076] In some embodiments, the AAV serotypes include, but are not limited to, AAV CHt-P2 (SEQ ID NOs: 1 and 51 in WO2016065001), AAV CHt-P5 (SEQ ID NOs: 2 and 52 in WO2016065001), AAV CHt-P9 (SEQ ID NOs: 3 and 53 in WO2016065001), AAV CBr-7.1 (SEQ ID NOs: 4 and 54 in WO2016065001), AAV CBr-7.2 (SEQ ID NOs: 5 and 55 in WO2016065001), AAV CBr-7.3 (SEQ ID NOs: 6 and 56 in WO2016065001), AAV CBr-7.4 (SEQ ID NOs: 7 and 57 in WO2016065001), AAV CBr-7.5 (SEQ ID NOs: 8 and 58 in WO 2016065001), AAV CBr-7.7 (SEQ ID NOs: 9 and 59 in WO 2016065001), AAV CBr-7.8 (SEQ ID NOs: 10 and 60 in WO 2016065001), AAV CBr-7.10 (SEQ ID NOs: 11 and 61 in WO 2016065001), AAV CKd-N3 (SEQ ID NOs: 12 and 62 in WO 2016065001), AAV CKd-N4 (SEQ ID NOs: 13 and 63 in WO 2016065001), AAV CKd-N9 (SEQ ID NOs: 14 and 64 in WO 2016065001), AAV CLv-L4 (SEQ ID NOs: 15 and 65 in WO 2016065001), AAV CLv-L5 (SEQ ID NOs: 16 and 66 in WO 2016065001), AAV CLv-L6 (SEQ ID NOs: 17 and 67 in WO 2016065001), AAV CLv-K1 (SEQ ID NOs: 18 and 68 in WO 2016065001), AAV CLv-K3 (SEQ ID NOs: 19 and 69 in WO 2016065001), AAV CLv-K6 (SEQ ID NOs: 20 and 70 in WO 2016065001), AAV CLv-M1 (SEQ ID NOs: 21 and 71 in WO 2016065001), AAV CLv-M11 (SEQ ID NOs: 22 and 72 in WO 2016065001), AAV CLv-M2 (SEQ ID NOs: 23 and 73 in WO 2016065001), AAV CLv-M5 (SEQ ID NOs: 24 and 74 in WO 2016065001), AAV CLv-M6 (SEQ ID NOs: 25 and 75 in WO 2016065001), AAV CLv-M7 (SEQ ID NOs: 26 and 76 in WO 2016065001), AAV CLv-M8 (SEQ ID NOs: 27 and 77 in WO 2016065001), AAV CLv-M9 (SEQ ID NOs: 28 and 78 in WO 2016065001), AAV CHt-P1 (SEQ ID NOs: 29 and 79 in WO 2016065001), AAV CHt-P6 (SEQ ID NOs: 30 and 80 in WO 2016065001), AAV CHt-P8 (SEQ ID NOs: 31 and 81 in WO 2016065001), and AAV CHt-6.1 (SEQ ID NOs: 32 and 82 in International Publication No. 2016065001), AAV CHt-6.10 (SEQ ID NOs: 33 and 83 in International Publication No. 2016065001), AAV CHt-6.5 (SEQ ID NOs: 34 and 84 in International Publication No. 2016065001), AAV CHt-6.6 (SEQ ID NOs: 35 and 85 in International Publication No. 2016065001), AAV CHt-6.7 (SEQ ID NOs: 36 and 86 in International Publication No. 2016065001), AAV CHt-6.8 (SEQ ID NOs: 37 and 87 in International Publication No. 2016065001), AAV CSp-8.10 (SEQ ID NOs: 38 and 88 in International Publication No. 2016065001), AAV CSp-8.2 (SEQ ID NOs: 39 and 89 in WO 2016065001), AAV CSp-8.4 (SEQ ID NOs: 40 and 90 in WO 2016065001), AAV CSp-8.5 (SEQ ID NOs: 41 and 91 in WO 2016065001), AAV CSp-8.6 (SEQ ID NOs: 42 and 92 in WO 2016065001), AAV CSp-8.7 (SEQ ID NOs: 43 and 93 in WO 2016065001), AAV CSp-8.8 (SEQ ID NOs: 44 and 94 in WO 2016065001), AAV CSp-8.9 (SEQ ID NOs: 45 and 95 in WO 2016065001), AAV The sequences may be or include sequences as described in WO2016065001 (the contents of which are incorporated by reference in their entirety), such as CBr-B7.3 (SEQ ID NOs: 46 and 96 in WO2016065001), AAV CBr-B7.4 (SEQ ID NOs: 47 and 97 in WO2016065001), AAV3B (SEQ ID NOs: 48 and 98 in WO2016065001), AAV4 (SEQ ID NOs: 49 and 99 in WO2016065001), AAV5 (SEQ ID NOs: 50 and 100 in WO2016065001), or variants or derivatives thereof.

[0077] In some embodiments, the AAV particles may be or comprise a serotype selected from any of those found in Table 1. In some embodiments, the AAV particles may comprise a sequence, fragment, or variant of any of the sequences in Table 1.

[0078] In some embodiments, the AAV particles may be encoded by a sequence, fragment, or variant of any of the sequences in Table 1. In the DNA and RNA sequences cited and / or described herein, the single letter symbols have the following explanations: A is adenine; C is cytosine; G is guanine; T is thymine; U is uracil; W is a weak base such as adenine or thymine; S is a strong nucleotide such as cytosine and guanine; M is an amino nucleotide such as adenine and cytosine; K is a keto nucleotide such as guanine and thymine; R is the purines adenine and guanine; Y is the pyrimidines cytosine and thymine; B is any base other than A (e.g., cytosine, guanine, and thymine); D is any base other than C (e.g., adenine, guanine, and thymine); H is any base other than G (e.g., adenine, cytosine, and thymine); V is any base other than T (e.g., adenine, cytosine, and guanine); N is any nucleotide (not a gap); and Z is zero.

[0079] In any of the amino acid sequences cited and / or described herein, the single letter symbols have the following explanation: G (Gly) is glycine; A (Ala) is alanine; L (Leu) is leucine; M (Met) is methionine; F (Phe) is phenylalanine; W (Trp) is tryptophan; K (Lys) is lysine; Q (Gln) is glutamine; E (Glu) is glutamic acid; S (Ser) is serine; P (Pro) is proline; V (Val) is valine; and I (Ile) is isoform. leucine; C (Cys) is cysteine; Y (Tyr) is tyrosine; H (His) is histidine; R (Arg) is arginine; N (Asn) is asparagine; D (Asp) is aspartic acid; T (Thr) is threonine; B (Asx) is aspartic acid or asparagine; J (Xle) is leucine or isoleucine; O (Pyl) is pyrrolysine; U (Sec) is selenocysteine; X (Xaa) is any amino acid; and Z (Glx) is glutamine or glutamic acid.

[0080] [Table 1-1]

[0081] [Table 1-2]

[0082] [Table 1-3]

[0083] [Table 1-4]

[0084] [Table 1-5]

[0085] Table 1-6

[0086] Table 1-7

[0087] Table 1-8

[0088] Table 1-9

[0089] Table 1-10

[0090] Table 1-11

[0091] Table 1-12

[0092] Table 1-13

[0093] Table 1-14

[0094] Table 1-15

[0095] Table 1-16

[0096] Table 1-17

[0097] Table 1-18

[0098] Table 1-19

[0099] Table 1-20

[0100] Table 1-21

[0101] Table 1-22

[0102] Table 1-23

[0103] Table 1-24

[0104] Table 1-25

[0105] Table 1-26

[0106] [Table 1-27]

[0107] [Table 1-28]

[0108] [Table 1-29]

[0109] [Table 1-30]

[0110] [Table 1-31]

[0111] [Table 1-32]

[0112] [Table 1-33]

[0113] [Table 1-34]

[0114] The contents of each of the patents, applications and / or publications listed in Table 1 are incorporated herein by reference in their entirety. In some embodiments, the AAV serotype includes, but is not limited to, AAV9 (SEQ ID NOs: 2 and 11 in WO2015038958, or SEQ ID NOs: 135 and 136 herein), PHP.B (SEQ ID NOs: 8 and 9 in WO2015038958, SEQ ID NOs: 3 and 4 herein), G2B-13 (SEQ ID NO: 12 in WO2015038958, SEQ ID NO: 5 herein), G2B-26 (SEQ ID NO: 13 in WO2015038958, SEQ ID NO: 14 herein), G2B-36 (SEQ ID NO: 15 in WO2015038958, SEQ ID NO: 16 herein), G2B-46 (SEQ ID NO: 17 in WO2015038958, SEQ ID NO: 18 herein), G2B-56 (SEQ ID NO: 19 in WO2015038958, SEQ ID NO: 20 herein), G2B-66 (SEQ ID NO: 21 in WO2015038958, SEQ ID NO: 22 herein), G2B-76 (SEQ ID NO: 23 in WO2015038958, SEQ ID NO: 24 herein), G2B-86 (SEQ ID NO: 25 in WO2015038958, SEQ ID NO: 26 herein), G2B-96 (SEQ ID NO: 26 in WO2015038958, SEQ ID NO: 27 herein), G2B-106 (SEQ ID NO: 28 in WO2015038958, SEQ ID NO: 29 herein), G2B-116 (SEQ ID The targeting peptides may be or may include sequences described in WO2015038958 (the contents of which are incorporated herein by reference in their entirety), such as TH1.1-32 (SEQ ID NO: 13 in WO2015038958, SEQ ID NO: 3 herein), TH1.1-32 (SEQ ID NO: 14 in WO2015038958, SEQ ID NO: 6 herein), TH1.1-35 (SEQ ID NO: 15 in WO2015038958, SEQ ID NO: 7 herein), or variants thereof. Additionally, any of the "targeting peptides" or "amino acid inserts" (used interchangeably herein to mean sequences that may be inserted into an AAV capsid sequence to facilitate delivery to CNS tissue) described in WO2015038958 may be inserted into any parent AAV serotype, such as, but not limited to, AAV9 (SEQ ID NO: 135 for the DNA sequence and SEQ ID NO: 136 for the amino acid sequence). In some embodiments, the amino acid insert is inserted at amino acids 586 to 592 of the parent AAV (e.g., AAV9). In some embodiments, the amino acid insert is inserted at amino acids 588 to 589 of the parent AAV sequence.The amino acid inserts may include, but are not limited to, the following amino acid sequences: TLAVPFK (SEQ ID NO: 1 in WO2015038958; SEQ ID NO: 1260 herein), KFPVALT (SEQ ID NO: 3 in WO2015038958; SEQ ID NO: 1261 herein), LAVPFK (SEQ ID NO: 31 in WO2015038958; SEQ ID NO: 1262 herein), AVPFK (SEQ ID NO: 32 in WO2015038958; SEQ ID NO: 1263 herein), VPFK (SEQ ID NO: 33 in WO 2015038958; SEQ ID NO: 1264 herein), TLAVPF (SEQ ID NO: 34 in WO 2015038958; SEQ ID NO: 1265 herein), TLAVP (SEQ ID NO: 35 in WO 2015038958; SEQ ID NO: 1266 herein), TLAV (SEQ ID NO: 36 in WO 2015038958; SEQ ID NO: 1267 herein) , SVSKPFL (SEQ ID NO: 28 in WO 2015038958; SEQ ID NO: 1268 herein), FTLTTPK (SEQ ID NO: 29 in WO 2015038958; SEQ ID NO: 1269 herein), MNATKNV (SEQ ID NO: 30 in WO 2015038958; SEQ ID NO: 1270 herein), QSSQTPR (SEQ ID NO: 54 in WO 2015038958; SEQ ID NO: 1271 herein), ILGTGTS (SEQ ID NO: 55 in WO2015038958; SEQ ID NO: 1272 herein), TRTNPEA (SEQ ID NO: 56 in WO2015038958; SEQ ID NO: 1273 herein), NGGTSSS (SEQ ID NO: 58 in WO2015038958; SEQ ID NO: 1274 herein), or YTLSQGW (SEQ ID NO: 60 in WO2015038958; SEQ ID NO: 1275 herein).Non-limiting examples of nucleotide sequences that may encode amino acid inserts include, but are not limited to, the following: AAGTTTCCTGTGGGCGTTGACT (for SEQ ID NO: 3 in WO2015038958; SEQ ID NO: 1276 herein), ACTTTGGCGGTGCCTTTTAAG (SEQ ID NOs: 24 and 49 in WO2015038958; SEQ ID NO: 1277 herein), AGTGTGAGTAAGCCTTTTTTG (SEQ ID NO: 25 in WO2015038958; SEQ ID NO: 1278 herein), TTTACGTTGACGACGCCTAAG (SEQ ID NO: 26 in WO2015038958; SEQ ID NO: 1279 herein), ATGAATGCTACGAAGAATGTG (SEQ ID NO: 3 in WO2015038958; SEQ ID NO: 1279 herein), ACTTTGGCGGTGCCTTTTAAG (for SEQ ID NO: 24 and 49 in WO2015038958; SEQ ID NO: 1277 herein), ACTTTGGCGGTGCCTTTTAAG (for SEQ ID NO: 25 in WO2015038958; SEQ ID NO: 1278 herein), TTTACGTTGACGACGCCTAAG (for SEQ ID NO: 26 in WO2015038958; SEQ ID NO: 1279 herein), ATGAATGCTACGAAGAATGTG (for SEQ ID NO: 3 in WO2015038958; SEQ ID NO: 1279 herein), ACTTTGGCGGTGCCTTTTAAG (for SEQ ID NO: 24 and 49 in WO2015038958; SEQ ID NO: 1279 herein), ACTTT 27; SEQ ID NO: 1280 herein), CAGTCGTCGCAGACGCCTAGG (SEQ ID NO: 48 in WO2015038958; SEQ ID NO: 1281 herein), ATTCTGGGGACTGGTACTTCG (SEQ ID NOs: 50 and 52 in WO2015038958; SEQ ID NO: 1282 herein), ACGCGGACTAATCCTGAGGCT (SEQ ID NO: 51 in WO2015038958; SEQ ID NO: 1283 herein), AATGGGGGGACTAGTAGTTCT (SEQ ID NO: 53 in WO2015038958; SEQ ID NO: 1284 herein), or TATACTTTGTCGCAGGGTTGG (SEQ ID NO: 59 in WO2015038958; SEQ ID NO: 1285 herein).

[0115] In some embodiments, the AAV serotype may be or may include a sequence as described in WO2017100671 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, AAV9 K449R (SEQ ID NO: 45 in WO2017100671, SEQ ID NO: 9 herein), PHP.N (SEQ ID NO: 46 in WO2017100671, SEQ ID NO: 2 herein), PHP.S (SEQ ID NO: 47 in WO2017100671, SEQ ID NO: 8 herein), or a variant thereof. Additionally, any of the targeting peptides or amino acid inserts described in WO2017100671 may be inserted into any parent AAV serotype, such as, but not limited to, AAV9 (SEQ ID NO: 9 or SEQ ID NO: 136). In some embodiments, the amino acid insert is inserted at amino acids 586-592 of the parent AAV (e.g., AAV9). In some embodiments, the amino acid insert is inserted at amino acids 588-589 of the parent AAV sequence. The amino acid insert may include, but is not limited to, the following amino acid sequences: AQTLAVPFKAQ (SEQ ID NO: 1 in WO2017100671; SEQ ID NO: 1286 herein), AQSVSKPFLAQ (SEQ ID NO: 2 in WO2017100671; SEQ ID NO: 1287 herein), AQFTLTTPKAQ (SEQ ID NO: 3 in the Sequence Listing of WO2017100671; SEQ ID NO: 1288 herein), DGTLAVPFKAQ (SEQ ID NO: 4 in the Sequence Listing of WO2017100671; SEQ ID NO: 1289 herein), DGTLAVPFKAQ (SEQ ID NO: 5 in the Sequence Listing of WO2017100671; SEQ ID NO: 1289 herein), DGTLAVPFKAQ (SEQ ID NO: 6 in the Sequence Listing of WO2017100671; SEQ ID NO: 1289 herein), DGTLAVPFKAQ (SEQ ID NO: 7 in the Sequence Listing of WO2017100671; SEQ ID NO: 1289 herein), DGTLAVPFKAQ (SEQ ID NO: 8 in the Sequence Listing of WO2017100671; SEQ ID NO: 1289 herein), DGTLAVPFKAQ (SEQ ID NO: 9 in the Sequence Listing of WO2017100671; SEQ ID NO: 1289 herein), DGTLAVPFKAQ (SEQ ID NO: 10 in the Sequence Listing of WO2017100671; SEQ ID NO: 1289 herein), DGTLAVPFKAQ (SEQ ID NO: SEQ ID NO: 4 in the Table; SEQ ID NO: 1289 herein), ESTLAVPFKAQ (SEQ ID NO: 5 in WO 2017100671; SEQ ID NO: 1290 herein), GGTLAVPFKAQ (SEQ ID NO: 6 in WO 2017100671; SEQ ID NO: 1291 herein), AQTLATPFKAQ (SEQ ID NOs: 7 and 33 in WO 2017100671; SEQ ID NO: 1292 herein), ATTLATPFKAQ (SEQ ID NO: 8 in WO 2017100671;SEQ ID NO: 1293 herein), DGTLATPFKAQ (SEQ ID NO: 9 in WO 2017100671; SEQ ID NO: 1294 herein), GGTLATPFKAQ (SEQ ID NO: 10 in WO 2017100671; SEQ ID NO: 1295 herein), SGSLAVPFKAQ (SEQ ID NO: 11 in WO 2017100671; SEQ ID NO: 1296 herein), AQTLAQPFKAQ (SEQ ID NO: 12 in WO 2017100671; SEQ ID NO: 12 SEQ ID NO: 1297 herein), AQTLQQPFKAQ (SEQ ID NO: 13 in WO 2017100671; SEQ ID NO: 1298 herein), AQTLSNPFKAQ (SEQ ID NO: 14 in WO 2017100671; SEQ ID NO: 1299 herein), AQTLAVPFSNP (SEQ ID NO: 15 in WO 2017100671; SEQ ID NO: 1300 herein), QGTLAVPFKAQ (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1301 herein), AQTLQQPFKAQ (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1302 herein), AQTLAVPFSNP (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1303 herein), AQTLAVPFSNP (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1304 herein), AQTLAVPFSNP (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1305 herein), AQTLAVPFSNP (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1306 herein), AQTLAVPFSNP (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1307 herein), AQTLAVPFSNP (SEQ ID NO: 16 in WO 2017100671; SEQ ID NO: 1308 herein), AQTLAVPFSNP (SEQ ID NO: 16 in WO 2017 SEQ ID NO: 1301 in WO 2017100671; SEQ ID NO: 1302 herein), NQTLAVPFKAQ (SEQ ID NO: 17 in WO 2017100671; SEQ ID NO: 1302 herein), EGSLAVPFKAQ (SEQ ID NO: 18 in WO 2017100671; SEQ ID NO: 1303 herein), SGNLAVPFKAQ (SEQ ID NO: 19 in WO 2017100671; SEQ ID NO: 1304 herein), EGTLAVPFKAQ (SEQ ID NO: 20 in WO 2017100671; SEQ ID NO: 1305 herein), SEQ ID NO: 1305 in WO 2017100671), DSTLAVPFKAQ (SEQ ID NO: 21 in Table 1 of WO 2017100671; SEQ ID NO: 1306 herein), AVTLAVPFKAQ (SEQ ID NO: 22 in WO 2017100671; SEQ ID NO: 1307 herein), AQTLSTPFKAQ (SEQ ID NO: 23 in WO 2017100671; SEQ ID NO: 1308 herein), AQTLPQPFKAQ (SEQ ID NOs: 24 and 32 in WO 2017100671;SEQ ID NO: 1309 herein), AQTLSQPFKAQ (SEQ ID NO: 25 in WO 2017100671; SEQ ID NO: 1310 herein), AQTLQLPFKAQ (SEQ ID NO: 26 in WO 2017100671; SEQ ID NO: 1311 herein), AQTLTMPFKAQ (SEQ ID NOs: 27 and 34 in WO 2017100671 and SEQ ID NO: 35 in the Sequence Listing of WO 2017100671; SEQ ID NO: 1312 herein), AQTLT TPFKAQ (SEQ ID NO: 28 in WO 2017100671; SEQ ID NO: 1313 herein), AQYTLSQGWAQ (SEQ ID NO: 29 in WO 2017100671; SEQ ID NO: 1314 herein), AQMNATKNVAQ (SEQ ID NO: 30 in WO 2017100671; SEQ ID NO: 1315 herein), AQVSGGHHSAQ (SEQ ID NO: 31 in WO 2017100671; SEQ ID NO: 1316 herein), AQTLTAPFKA Q (SEQ ID NO: 35 in Table 1 of WO 2017100671; SEQ ID NO: 1317 herein), AQTLSKPFKAQ (SEQ ID NO: 36 in WO 2017100671; SEQ ID NO: 1318 herein), QAVRTSL (SEQ ID NO: 37 in WO 2017100671; SEQ ID NO: 1319 herein), YTLSQGW (SEQ ID NO: 38 in WO 2017100671; SEQ ID NO: 1275 herein), LAKERLS (WO 2017100 SEQ ID NO: 39 in WO 2017100671; SEQ ID NO: 1320 herein), TLAVPFK (SEQ ID NO: 40 in the Sequence Listing of WO 2017100671; SEQ ID NO: 1260 herein), SVSKPFL (SEQ ID NO: 41 in WO 2017100671; SEQ ID NO: 1268 herein), FTLTTPK (SEQ ID NO: 42 in WO 2017100671; SEQ ID NO: 1269 herein), MNSTKNV (SEQ ID NO: 43 in WO 2017100671;SEQ ID NO: 1321 herein), VSGGHHS (SEQ ID NO: 44 in WO2017100671; SEQ ID NO: 1322 herein), SAQTLAVPFKAQAQ (SEQ ID NO: 48 in WO2017100671; SEQ ID NO: 1323 herein), SXXXLAVPFKAQAQ (SEQ ID NO: 49 in WO2017100671, where X may be any amino acid; SEQ ID NO: 1324 herein), SAQXXXVPFKAQAQ (WO2017100671 No. 50 in WO 2017100671, where X may be any amino acid; SEQ ID NO: 1325 herein), SAQTLXXXFKAQAQ (SEQ ID NO: 51 in WO 2017100671, where X may be any amino acid; SEQ ID NO: 1326 herein), SAQTLAVXXXAQAQ (SEQ ID NO: 52 in WO 2017100671, where X may be any amino acid; SEQ ID NO: 1327 herein), SAQTLAVPFXXXAQ (SEQ ID NO: 53 in WO 2017100671, where X may be any amino acid; SEQ ID NO: 1328 herein), SAQTLAVXXXAQAQ (SEQ ID NO: 54 in WO 2017100671, where X may be any amino acid; SEQ ID NO: 1329 herein), SAQTLAVPFXXXAQ (SEQ ID NO: 55 in WO 2017100671, where X may be any amino acid; SEQ ID NO: 1330 herein), SAQTLAVPFXXXAQ (SEQ ID NO: 56 in WO 2017100671, where X may be any amino acid; SEQ ID NO: 1331 SEQ ID NO: 53 of WO 2017100671, where X may be any amino acid; SEQ ID NO: 1328 herein), TNHQSAQ (SEQ ID NO: 65 of WO 2017100671; SEQ ID NO: 1329 herein), AQAQTGW (SEQ ID NO: 66 of WO 2017100671; SEQ ID NO: 1330 herein), DGTLATPFK (SEQ ID NO: 67 of WO 2017100671; SEQ ID NO: 1331 herein), DGTLATPFKXX (SEQ ID NO: 67 of WO 2017100671; SEQ ID NO: 1332 herein), SEQ ID NO: 68 of Lett, where X may be any amino acid; SEQ ID NO: 1332 herein), LAVPFKAQ (SEQ ID NO: 80 of WO2017100671; SEQ ID NO: 1333 herein), VPFKAQ (SEQ ID NO: 81 of WO2017100671; SEQ ID NO: 1334 herein), FKAQ (SEQ ID NO: 82 of WO2017100671; SEQ ID NO: 1335 herein), AQTLAV (SEQ ID NO: 83 of WO2017100671;SEQ ID NO: 1336 herein), AQTLAVPF (SEQ ID NO: 84 in WO 2017100671; SEQ ID NO: 1337 herein), QAVR (SEQ ID NO: 85 in WO 2017100671; SEQ ID NO: 1338 herein), AVRT (SEQ ID NO: 86 in WO 2017100671; SEQ ID NO: 1339 herein), VRTS (SEQ ID NO: 87 in WO 2017100671; SEQ ID NO: 1340 herein), RTSL (SEQ ID NO: 88 in WO 2017100671; SEQ ID NO: 1341 herein), It may be any of QAVRT (SEQ ID NO: 89 in WO 2017100671; SEQ ID NO: 1342 herein), AVRTS (SEQ ID NO: 90 in WO 2017100671; SEQ ID NO: 1343 herein), VRTSL (SEQ ID NO: 91 in WO 2017100671; SEQ ID NO: 1344 herein), QAVRTS (SEQ ID NO: 92 in WO 2017100671; SEQ ID NO: 1345 herein), or AVRTSL (SEQ ID NO: 93 in WO 2017100671; SEQ ID NO: 1346 herein);

[0116] Non-limiting examples of nucleotide sequences that may encode the amino acid insert include the following: GATGGGACTTTGGCGGTGCCTTTTAAGGCACAG (SEQ ID NO: 54 in WO2017100671; SEQ ID NO: 1347 herein), GATGGGACGTTGGCGGTGCCTTTTAAGGCACAG (SEQ ID NO: 55 in WO2017100671; SEQ ID NO: 1348 herein), CAGGCGGTTAGGACGTCTTTG (SEQ ID NO: 56 in WO2017100671; SEQ ID NO: 1349 herein), CAGGTCTTCACGGACTCAGACTATCAG (SEQ ID NOs: 57 and 78 in WO2017100671; SEQ ID NO: 1350 herein), CAAGTAAAACCTCTACAAATGTGGTAAAATCG (SEQ ID NO: 58 in WO2017100671; SEQ ID NO: 1351 herein), ACT CATCGACCAATACTTGTACTATCTCTCTAGAAC (SEQ ID NO: 59 in WO 2017100671; SEQ ID NO: 1352 herein), GGAAGTATTCCTTGGTTTTGAACCCA (SEQ ID NO: 60 in WO 2017100671; SEQ ID NO: 1353 herein), GGTCGCGGTTCTTGTTTGTGGAT (SEQ ID NO: 61 in WO 2017100671; SEQ ID NO: 1354 herein), SEQ ID NO: 1354), CGACCTTGAAGCGCATGAACTCCT (SEQ ID NO: 62 in WO2017100671; SEQ ID NO: 1355 herein), GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCMNNMNNMNNMNNMNNMNNMNNTTGGGCACTCTGGTGGTTTGTC (SEQ ID NO: 63 in WO2017100671, where N can be A, C, T, or G;SEQ ID NO: 1356 herein), GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCMNNMNNMNNAAAAGGCACCGCCAAAGTTTG (SEQ ID NO: 69 in WO2017100671, where N can be A, C, T, or G; SEQ ID NO: 1357 herein), GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCMNNMNNMNNCACCGCCAAAGTTTGGGCACT (WO2017100671 SEQ ID NO: 70 in WO2017100671, where N may be A, C, T, or G; SEQ ID NO: 1358 herein), GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGCCTGTGCCTTAAAMNNMNNMNNCAAAGTTTGGGCACTCTGGTGG (SEQ ID NO: 71 in WO2017100671, where N may be A, C, T, or G; SEQ ID NO: 1359 herein), GTATTCCTTGGTTTTGAACCCAACCGGTCTGCGC CTGTGCCTTAAAAGGCACMNNMNNMNNTTGGGCACTCTGGTGGTTTGTG (SEQ ID NO: 72 in WO2017100671, where N can be A, C, T, or G; SEQ ID NO: 1360 herein), ACTTTGGCGGTGCCTTTTAAG (SEQ ID NO: 74 in WO2017100671; SEQ ID NO: 1277 herein), AGTGTGAGTAAGCCTTTTTTG (SEQ ID NO: 1360 in WO2017100671, where N can be A, C, T, or G; SEQ ID NO: 1360 herein), 75; SEQ ID NO: 1278 herein), TTTACGTTGACGACGCCTAAG (SEQ ID NO: 76 in WO2017100671; SEQ ID NO: 1279 herein), TATACTTTGTCGCAGGGTTGG (SEQ ID NO: 77 in WO2017100671; SEQ ID NO: 1285 herein), or CTTGCGAAGGAGCGGCTTTCG (SEQ ID NO: 79 in WO2017100671; SEQ ID NO: 1361 herein).

[0117] In some embodiments, the AAV serotype may include, but is not limited to, AAV1 (SEQ ID NO: 181 in U.S. Pat. No. 9,624,274), AAV6 (SEQ ID NO: 182 in U.S. Pat. No. 9,624,274), AAV2 (SEQ ID NO: 183 in U.S. Pat. No. 9,624,274), AAV3b (SEQ ID NO: 184 in U.S. Pat. No. 9,624,274), AAV7 (SEQ ID NO: 185 in U.S. Pat. No. 9,624,274), AAV8 (SEQ ID NO: 186 in U.S. Pat. No. 9,624,274), AAV10 (SEQ ID NO: 187 in U.S. Pat. No. 9,624,274), AAV4 (SEQ ID NO: 188 in U.S. Pat. No. 9,624,274), AAV11 (SEQ ID NO: 189 in U.S. Pat. No. 9,624,274), bAAV (SEQ ID NO: 190 in U.S. Pat. No. 9,624,274), AAV5 (U.S. Pat. No. 9,624,274), AAV6 (SEQ ID NO: 182 in U.S. Pat. No. 9,624,274), AAV7 (SEQ ID NO: 185 in U.S. Pat. No. 9,624,274), AAV8 (SEQ ID NO: 186 in U.S. Pat. No. 9,624,274), AAV10 (SEQ ID NO: 187 in U.S. Pat. No. 9,624,274), AAV No. 9624274 (the contents of which are incorporated by reference herein in their entirety), such as GPV (SEQ ID NO: 191 in U.S. Pat. No. 9624274; SEQ ID NO: 992 herein), B19 (SEQ ID NO: 193 in U.S. Pat. No. 9624274; SEQ ID NO: 993 herein), MVM (SEQ ID NO: 194 in U.S. Pat. No. 9624274; SEQ ID NO: 994 herein), FPV (SEQ ID NO: 195 in U.S. Pat. No. 9624274; SEQ ID NO: 995 herein), CPV (SEQ ID NO: 196 in U.S. Pat. No. 9624274; SEQ ID NO: 996 herein), or variants thereof. Additionally, any of the structural protein inserts described in U.S. Patent No. 9,624,274 may be inserted into, but not limited to, I-453 and I-587 of any parent AAV serotype, including, but not limited to, AAV2 (U.S. Patent No. 9,624,274 SEQ ID NO: 183). Amino acid inserts may include, but are not limited to, the following amino acid sequences: VNLTWSRASG (U.S. Patent No. 9,624,274 SEQ ID NO: 50; SEQ ID NO: 1362 herein), EFCINHRGYWVCGD (U.S. Patent No. 9,624,274 SEQ ID NO: 55;No. 9,624,274; SEQ ID NO: 1363 herein), EDGQVMDVDLS (SEQ ID NO: 85 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1364 herein), EKQRNGTLT (SEQ ID NO: 86 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1365 herein), TYQCRVTHPHLPRALMR (SEQ ID NO: 87 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1366 herein), RHSTTQPRKTKGSG (SEQ ID NO: 88 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1366 herein). 367), DSNPRGVSAYLSR (SEQ ID NO: 89 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1368 herein), TITCLWDLAPSK (SEQ ID NO: 90 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1369 herein), KTKGSGFFVF (SEQ ID NO: 91 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1370 herein), THPHLPRALMRS (SEQ ID NO: 92 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1371 herein), GETYQCRVTHPHL PRALMRSTTK (SEQ ID NO: 93 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1372 herein), LPRALMRS (SEQ ID NO: 94 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1373 herein), INHRGYWV (SEQ ID NO: 95 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1374 herein), CDAGSVRTNAPD (SEQ ID NO: 60 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1375 herein), AKAVSNLTESRSESLQS (U.S. Pat. No. 9,624,274; SEQ ID NO: 1376 herein), and AKAVSNLTESRSESLQS (U.S. Pat. No. 9,624,274; SEQ ID NO: 1377 herein). 274 , SEQ ID NO: 96; SEQ ID NO: 1376 herein), SLTGDEFKKVLET (U.S. Pat. No. 9,624,274 , SEQ ID NO: 97; SEQ ID NO: 1377 herein), REAVAYRFEED (U.S. Pat. No. 9,624,274 , SEQ ID NO: 98; SEQ ID NO: 1378 herein), INPEIITLDG (U.S. Pat. No. 9,624,274 , SEQ ID NO: 99; SEQ ID NO: 1379 herein), DISVTGAPVITATYL (U.S. Pat. No. 9,624,274 , SEQ ID NO: 100;No. 9,624,274; SEQ ID NO: 1380 herein), DISVTGAPVITA (SEQ ID NO: 101 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1381 herein), PKTVSNLTESSSESVQS (SEQ ID NO: 102 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1382 herein), SLMGDEFKAVLET (SEQ ID NO: 103 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1383 herein), QHSVAYTFEED (SEQ ID NO: 104 in U.S. Pat. No. 9,624,274; SEQ ID NO: 13 84), INPEIITRDG (SEQ ID NO: 105 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1385 herein), DISLTGDPVITASYL (SEQ ID NO: 106 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1386 herein), DISLTGDPVITA (SEQ ID NO: 107 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1387 herein), DQSIDFEIDSA (SEQ ID NO: 108 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1388 herein), KNVSEDLPLPTFSPT LLGDS (SEQ ID NO: 109 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1389 herein), KNVSEDLPLPT (SEQ ID NO: 110 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1390 herein), CDSGRVRTDAPD (SEQ ID NO: 111 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1391 herein), FPEHLLVDFLQSLS (SEQ ID NO: 112 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1392 herein), DAEFRHDSG (SEQ ID NO: 113 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1393 herein), No. 9,624,274; SEQ ID NO: 1393 herein), HYAAAQWDFGNTMCQL (SEQ ID NO: 113 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1394 herein), YAAQWDFGNTMCQ (SEQ ID NO: 114 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1395 herein), RSQKEGLHYT (SEQ ID NO: 115 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1396 herein), SSRTPSDKPVAHWANPQAE (SEQ ID NO: 116 in U.S. Pat. No. 9,624,274;No. 9,624,274; SEQ ID NO: 1397 herein), SRTPSDKPVAHWANP (SEQ ID NO: 117 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1398 herein), SSRTPSDKP (SEQ ID NO: 118 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1399 herein), NADGNVDYHMNSVP (SEQ ID NO: 119 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1400 herein), DGNVDYHMNSV (U.S. Pat. No. 9,624,274 No. 9,624,274; SEQ ID NO: 1401 herein), RSFKEFLQSSLRALRQ (SEQ ID NO: 121 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1402 herein), FKEFLQSSLRA (SEQ ID NO: 122 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1403 herein), or QMWAPQWGPD (SEQ ID NO: 123 in U.S. Pat. No. 9,624,274; SEQ ID NO: 1404 herein);

[0118] In some embodiments, the AAV serotype may be or have a sequence as described in U.S. Pat. No. 9,475,845, the contents of which are incorporated by reference in their entirety, including, but not limited to, an AAV capsid protein comprising one or more amino acid modifications at amino acid positions 585-590 of the native AAV2 capsid protein. Further modifications may be made to the following amino acid sequences, including, but not limited to, RGNRQA (SEQ ID NO: 3 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1405 herein), SSSTDP (SEQ ID NO: 4 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1406 herein), SSNTAP (SEQ ID NO: 5 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1407 herein), SNSNLP (SEQ ID NO: 6 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1408 herein), SSTTAP (SEQ ID NO: 7 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1409 herein), AANTAA (SEQ ID NO: 8 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1410 herein), QQNTAP (SEQ ID NO: 9 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1411 herein), SAQAQA (SEQ ID NO: 10 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1412 herein), SAQAQA (SEQ ID NO: 11 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1413 herein), SAQAQA (SEQ ID NO: 12 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1414 herein), SAQAQA (SEQ ID NO: 13 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1415 herein), No. 9,475,845; SEQ ID NO: 1412 herein), QANTGP (SEQ ID NO: 11 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1413 herein), NATTAP (SEQ ID NO: 12 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1414 herein), SSTAGP (SEQ ID NOs: 13 and 20 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1415 herein), QQNTAA (SEQ ID NO: 14 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1416 herein), PSTAGP (SEQ ID NO: 15 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1417 herein), NQNTAP (SEQ ID NO: 16 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1418 herein), QAANAP (SEQ ID NO: 17 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1419 herein), SIVGLP (SEQ ID NO: 18 in U.S. Pat. No. 9,475,845;No. 9,475,845; SEQ ID NO: 1420 herein), AASTAA (SEQ ID NOs: 19 and 27 of U.S. Pat. No. 9,475,845; SEQ ID NO: 1421 herein), SQNTTA (SEQ ID NO: 21 of U.S. Pat. No. 9,475,845; SEQ ID NO: 1422 herein), QQDTAP (SEQ ID NO: 22 of U.S. Pat. No. 9,475,845; SEQ ID NO: 1423 herein), QTNTGP (SEQ ID NO: 23 of U.S. Pat. No. 9,475,845; SEQ ID NO: 1424 herein), QTNGAP (SEQ ID NO: 24 of U.S. Pat. No. 9,475,845; SEQ ID NO: 1425 herein), QQNAAP (SEQ ID NO: 25 of U.S. Pat. No. 9,475,845; SEQ ID NO: 1426 herein), or AANTQA (SEQ ID NO: 26 of U.S. Pat. No. 9,475,845; SEQ ID NO: 1427 herein). In some embodiments, the amino acid modification is a substitution at amino acid positions 262-265 of the native AAV2 capsid protein, or at the corresponding positions of a capsid protein of another AAV that has a targeting sequence, including, but not limited to, the following amino acid sequences: NGRAHA (SEQ ID NO: 38 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1428 herein), QPEHSST (SEQ ID NOs: 39 and 50 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1429 herein), VNTANST (SEQ ID NO: 40 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1430 herein), HGPMQKS (SEQ ID NO: 41 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1431 herein), PHKPPLA (SEQ ID NO: 143 ... SEQ ID NO: 42 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1432 herein), IKNNEMW (SEQ ID NO: 43 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1433 herein), RNLDTPM (SEQ ID NO: 44 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1434 herein), VDSHRQS (SEQ ID NO: 45 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1435 herein), YDSKTKT (SEQ ID NO: 46 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1436 herein), SQLPHQK (SEQ ID NO: 47 in U.S. Pat. No. 9,475,845;No. 9,475,845; SEQ ID NO: 1437 herein), STMQQNT (SEQ ID NO: 48 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1438 herein), TERYMTQ (SEQ ID NO: 49 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1439 herein), DASLSTS (SEQ ID NO: 51 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1440 herein), DLPNKKT (SEQ ID NO: 52 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1441 herein), DLTAARL (U.S. Pat. No. 9,475,845; SEQ ID NO: 1445 herein), No. 9,475,845; SEQ ID NO: 1442 herein), EPHQFNY (SEQ ID NO: 54 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1443 herein), EPQSNHT (SEQ ID NO: 55 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1444 herein), MSSWPSQ (SEQ ID NO: 56 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1445 herein), NPKHNAT (SEQ ID NO: 57 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1446 herein), PDGMRTT (U.S. Pat. No. 9,475,845; SEQ ID NO: 1446 herein), SEQ ID NO: 58 in U.S. Patent No. 9475845; SEQ ID NO: 1447 herein), PNNNKTT (SEQ ID NO: 59 in U.S. Patent No. 9475845; SEQ ID NO: 1448 herein), QSTTHDS (SEQ ID NO: 60 in U.S. Patent No. 9475845; SEQ ID NO: 1449 herein), TGSKQKQ (SEQ ID NO: 61 in U.S. Patent No. 9475845; SEQ ID NO: 1450 herein), SLKHQAL (SEQ ID NO: 62 in U.S. Patent No. 9475845; SEQ ID NO: 1451 herein) , SPIDGEQ (SEQ ID NO: 63 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1452 herein), WIFPWIQL (SEQ ID NOs: 64 and 112 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1453 herein), CDCRGDCFC (SEQ ID NO: 65 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1454 herein), CNGRC (SEQ ID NO: 66 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1455 herein), CPRECES (SEQ ID NO: 67 in U.S. Pat. No. 9,475,845;SEQ ID NO: 1456 herein), CTTHWGFTLC (SEQ ID NOs: 68 and 123 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1457 herein), CGRRAGGSC (SEQ ID NO: 69 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1458 herein), CKGGRAKDC (SEQ ID NO: 70 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1459 herein), CVPELGHEC (SEQ ID NOs: 71 and 115 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1460 herein), CR RETAWAK (SEQ ID NO: 72 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1461 herein), VSWFSHRYSPFAVS (SEQ ID NO: 73 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1462 herein), GYRDGYAGPILYN (SEQ ID NO: 74 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1463 herein), XXXYXXX (SEQ ID NO: 75 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1464 herein), YXNW (SEQ ID NO: 76 in U.S. Pat. No. 9,475,845 ; SEQ ID NO: 1465 herein), RPLPPLP (SEQ ID NO: 77 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1466 herein), APPLPPR (SEQ ID NO: 78 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1467 herein), DVFYPYPYASGS (SEQ ID NO: 79 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1468 herein), MYWYPY (SEQ ID NO: 80 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1469 herein), DITWDQLWDLMK ​​(U.S. Pat. No. No. 9,475,845; SEQ ID NO: 1470 herein), CWDDXWLC (SEQ ID NO: 82 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1471 herein), EWCEYLGGYLRCYA (SEQ ID NO: 83 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1472 herein), YXCXXGPXTWXCXP (SEQ ID NO: 84 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1473 herein), IEGPTLRQWLAARA (SEQ ID NO: 85 in U.S. Pat. No. 9,475,845;SEQ ID NO: 1474 herein), LWXXX (SEQ ID NO: 86 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1475 herein), XFXXYLW (SEQ ID NO: 87 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1476 herein), SSIISHFRWGLCD (SEQ ID NO: 88 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1477 herein), MSRPACPPNDKYE (SEQ ID NO: 89 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1478 herein), CLRSGRGC (U.S. Pat. No. 9475,845; SEQ ID NO: 1479 herein), No. 5845; SEQ ID NO: 1479 herein), CHWMFSPWC (SEQ ID NO: 91 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1480 herein), WXXF (SEQ ID NO: 92 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1481 herein), CSSRLDAC (SEQ ID NO: 93 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1482 herein), CLPVASC (SEQ ID NO: 94 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1483 herein), CGFECVRQCPER C (SEQ ID NO: 95 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1484 herein), CVALCREACGEGC (SEQ ID NO: 96 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1485 herein), SWCEPGWCR (SEQ ID NO: 97 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1486 herein), YSGKWGW (SEQ ID NO: 98 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1487 herein), GLSGGRS (SEQ ID NO: 99 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1488 herein), No. 1488), LMLPRAD (SEQ ID NO: 100 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1489 herein), CSCFRDVCC (SEQ ID NO: 101 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1490 herein), CRDVVSVIC (SEQ ID NO: 102 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1491 herein), MARSGL (SEQ ID NO: 103 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1492 herein), MARAKE (SEQ ID NO: 104 in U.S. Pat. No. 9,475,845;SEQ ID NO: 1493 herein), MSRTMS (SEQ ID NO: 105 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1494 herein), KCCYSL (SEQ ID NO: 106 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1495 herein), MYWGDSHWLQYWYE (SEQ ID NO: 107 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1496 herein), MQLPLAT (SEQ ID NO: 108 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1497 herein), EWLS (SEQ ID NO: 109 in U.S. Pat. No. 9,475,845; SEQ ID NO: 149 herein; 8), SNEW (SEQ ID NO: 110 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1499 herein), TNYL (SEQ ID NO: 111 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1500 herein), WDLAWMFRLPVG (SEQ ID NO: 113 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1501 herein), CTVALPGGYVRVC (SEQ ID NO: 114 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1502 herein), CVAYCIEHHCWTC (U.S. Pat. No. 9,475,845 No. 9,475,845; SEQ ID NO: 1503 herein), CVFAHNYDYLVC (SEQ ID NO: 117 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1504 herein), CVFTSNYAFC (SEQ ID NO: 118 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1505 herein), VHSPNKK (SEQ ID NO: 119 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1506 herein), CRGDGWC (SEQ ID NO: 120 in U.S. Pat. No. 9,475,845; SEQ ID NO: 150 7), XRGCDX (SEQ ID NO: 121 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1508 herein), PXXX (SEQ ID NO: 122 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1509 herein), SGKGPRQITAL (SEQ ID NO: 124 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1510 herein), AAAAAAAAAXXXXX (SEQ ID NO: 125 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1511 herein), VYMSPF (SEQ ID NO: 126 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1512 herein), VYMSPF (SEQ ID NO: 127 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1513 herein), VYMSPF (SEQ ID NO: 128 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1514 herein), VYMSPF (SEQ ID NO: 129 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1515 herein), VYMSPF (SEQ ID NO: 129 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1516 herein), VYMSPF (SEQ ID NO: 129 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1517 herein), VYMSPF (SEQ ID NO: 129 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1518 SEQ ID NO: 126; SEQ ID NO: 1512 herein), ATWLPPR (SEQ ID NO: 127 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1513 herein), HTMYYHHYQHHL (SEQ ID NO: 128 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1514 herein), SEVGCRAGPLQWLCEKYFG (SEQ ID NO: 129 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1515 herein), CGLLPVGRPDRNVWRWLC (SEQ ID NO: 130 in U.S. Pat. No. 9,475,845;No. 9,475,845; SEQ ID NO: 1516 herein), CKGQCDRFKGLPWEC (SEQ ID NO: 131 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1517 herein), SGRSA (SEQ ID NO: 132 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1518 herein), WGFP (SEQ ID NO: 133 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1519 herein), AEPMPHSLNFSQYLWYT (SEQ ID NO: 134 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1520 herein), WAYXSP (U.S. Pat. No. 9,475,845; SEQ ID NO: 1521 herein), No. 475845; SEQ ID NO: 1521 herein), IELLQAR (U.S. Pat. No. 9,475,845; SEQ ID NO: 1522 herein), AYTKCSRQWRTCMTTH (U.S. Pat. No. 9,475,845; SEQ ID NO: 137 herein), PQNSKIPGPTFLDPH (U.S. Pat. No. 9,475,845; SEQ ID NO: 1524 herein), SMEPALPDWWWKMFK (U.S. Pat. No. 9,475,845; SEQ ID NO: 139 herein), No. 9,475,845; SEQ ID NO: 1526 herein), TACHQHVRMVRP (SEQ ID NO: 141 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1527 herein), VPWMEPAYQRFL (SEQ ID NO: 142 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1528 herein), DPRATPGS (SEQ ID NO: 143 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1529 herein), FRPNRA QDYNTN (SEQ ID NO: 144 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1530 herein), CTKNSYLMC (SEQ ID NO: 145 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1531 herein), CXXTXXXGXGC (SEQ ID NO: 146 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1532 herein), CPIEDRPMC (SEQ ID NO: 147 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1533 herein), HEWSYLAPYPWF (SEQ ID NO: 148 in U.S. Pat. No. 9,475,845;No. 9,475,845; SEQ ID NO: 1534 herein), MCPKHPLGC (SEQ ID NO: 149 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1535 herein), RMWPSSTVNLSAGRR (SEQ ID NO: 150 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1536 herein), SAKTAVSQRVWLPSHRGGEP (SEQ ID NO: 151 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1537 herein), KSREHVNNSACPSKRITAAL (SEQ ID NO: 152 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1538 herein), EGFR (SEQ ID NO: 153 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1539 herein), AGLGVR (SEQ ID NO: 154 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1540 herein), GTRQGHTMRLGVSDG (U.S. Pat. No. No. 9,475,845; SEQ ID NO: 1541 herein), IAGLAATPGWSHWLAL (SEQ ID NO: 156 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1542 herein), SMSIARL (SEQ ID NO: 157 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1543 herein), HTFEPGV (SEQ ID NO: 158 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1544 herein), NTSLKRISNKRIRRK (SEQ ID NO: 159 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1545 herein), LRIKRKRRKRKKTRK (SEQ ID NO: 160 in U.S. Pat. No. 9,475,845; SEQ ID NO: 1546 herein), GGG, GFS, LWS, EGG, LLV, LSP, LBS, AGG, GRR, GGH, or GTV;

[0119] In some embodiments, the AAV serotype may be or have a sequence as described in U.S. Patent Application Publication No. 20160369298 (the contents of which are incorporated by reference in their entirety), including, but not limited to, a site-directed mutant capsid protein of AAV2 (SEQ ID NO: 97 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1547 herein), or a variant thereof, wherein the specific mutation site is at least one site selected from sites R447, G453, S578, N587, N587+1, S662 of VP1 or a fragment thereof.

[0120] Additionally, any of the mutant sequences described in US Patent Application Publication No. 20160369298 may be mutated using any of the following sequences, including, but not limited to, SDSGASN (SEQ ID NO: 1 and SEQ ID NO: 231 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1548 herein), SPSGASN (SEQ ID NO: 2 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1549 herein), SHSGASN (SEQ ID NO: 3 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1550 herein), SR SGASN (SEQ ID NO: 4 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1551 herein), SKSGASN (SEQ ID NO: 5 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1552 herein), SNSGASN (SEQ ID NO: 6 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1553 herein), SGSGASN (SEQ ID NO: 7 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1554 herein), SASGASN (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1555 herein), SEQ ID NOs: 8, 175, and 221 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1555 herein), SESGTSN (SEQ ID NO: 9 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1556 herein), STTGGSN (SEQ ID NO: 10 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1557 herein), SSAGSTN (SEQ ID NO: 11 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1558 herein), NNDSQA (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1559 herein), and NNDSQA (U.S. Patent Application Publication No. 20160369298). 69298; SEQ ID NO:1559 herein), NNRNQA (SEQ ID NO:13 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO:1560 herein), NNNKQA (SEQ ID NO:14 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO:1561 herein), NAKRQA (SEQ ID NO:15 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO:1562 herein), NDEHQA (SEQ ID NO:16 in U.S. Patent Application Publication No. 20160369298;SEQ ID NO: 1563 herein), NTSQKA (SEQ ID NO: 17 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1564 herein), YYLSRTNTPSGTDTQSRLVFSQAGA (SEQ ID NO: 18 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1565 herein), YYLSRTNTDSGTETQSGLDFSQAGA (SEQ ID NO: 19 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1566 herein), YYLSRTNTESGTPTQ SALEFSQAGA (SEQ ID NO: 20 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1567 herein), YYLSRTNTHSGTHTQSPLHFSQAGA (SEQ ID NO: 21 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1568 herein), YYLSRTNTSSGTITISHLIFSQAGA (SEQ ID NO: 22 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1569 herein), YYLSRTNTRSGIMTKSSLMFSQAGA (U ... YYLSRTNTKSGRKTLSNLSFSQAGA (SEQ ID NO: 23 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1570 herein), YYLSRTNTKSGRKTLSNLSFSQAGA (SEQ ID NO: 24 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1571 herein), YYLSRTNDGSGPVTPSKLRFSQRGA (SEQ ID NO: 25 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1572 herein), YYLSRTNAASGHATHSDLKFSQPGA (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1573 herein), YYLSRTNTKSGRKTLSNLSFSQAGA (SEQ ID NO: 24 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1574 herein), YYLSRTNDGSGPVTPSKLRFSQRGA (SEQ ID NO: 25 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1575 herein), YYLSRTNAASGHATHSDLKFSQPGA (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1576 herein), YYLSRTNTKSGRKTLSNLSFSQAGA (SEQ ID NO: 25 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1577 herein), YYLSRTNDGSGPVTPSKLRFSQRGA (SEQ ID NO: 25 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1578 herein), Y 298; SEQ ID NO: 1573 herein), YYLSRTNGQAGSLTMSELGFSQVGA (SEQ ID NO: 27 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1574 herein), YYLSRTNSTGGNQTTSQLLFSQLSA (SEQ ID NO: 28 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1575 herein), YFLSRTNNNTGLNTNSTLNFSQGRA (SEQ ID NO: 29 in U.S. Patent Application Publication No. 20160369298;SEQ ID NO: 1576 herein), SKTGADNNNSEYSWTG (SEQ ID NO: 30 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1577 herein), SKTDADNNNSEYSWTG (SEQ ID NO: 31 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1578 herein), SKTEADNNNSEYSWTG (SEQ ID NO: 32 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1579 herein), SKTPADNNNSEYSWT G (SEQ ID NO: 33 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1580 herein), SKTHADNNNSEYSWTG (SEQ ID NO: 34 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1581 herein), SKTQADNNNSEYSWTG (SEQ ID NO: 35 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1582 herein), SKTIADNNNSEYSWTG (SEQ ID NO: 36 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1583 herein), SKTMADNNNSEYSWTG (SEQ ID NO: 37 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1584 herein), SKTRADNNNSEYSWTG (SEQ ID NO: 38 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1585 herein), SKTNADNNNSEYSWTG (SEQ ID NO: 39 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1586 herein), SKTVGRNNNSEYSWT G (SEQ ID NO: 40 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1587 herein), SKTADRNNNSEYSWTG (SEQ ID NO: 41 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1588 herein), SKKLSQNNNSKYSWQG (SEQ ID NO: 42 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1589 herein), SKPTTGNNNSDYSWPG (SEQ ID NO: 43 in U.S. Patent Application Publication No. 20160369298;SEQ ID NO: 1590 herein), STQKNENNNSNYSWPG (SEQ ID NO: 44 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1591 herein), HKDDEGKF (SEQ ID NO: 45 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1592 herein), HKDDNRKF (SEQ ID NO: 46 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1593 herein), HKDDTNKF (SEQ ID NO: 46 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1594 herein), SEQ ID NO: 47 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1594 herein), HEDSDKNF (SEQ ID NO: 48 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1595 herein), HRDGADSF (SEQ ID NO: 49 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1596 herein), HGDNKSRF (SEQ ID NO: 50 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1597 herein), KQGSEKTNVDFEEV (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1598 ... KQGSEKTNVDSEEV (SEQ ID NO: 51 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1598 herein), KQGSEKTNVDSEEV (SEQ ID NO: 52 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1599 herein), KQGSEKTNVDVEEV (SEQ ID NO: 53 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1600 herein), KQGSDKTNVDDAGV (SEQ ID NO: 54 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1601 herein), KQGSSK TNVDPREV (SEQ ID NO: 55 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1602 herein), KQGSRKTNVDHKQV (SEQ ID NO: 56 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1603 herein), KQGSKGGNVDTNRV (SEQ ID NO: 57 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1604 herein), KQGSGEANVDNGDV (SEQ ID NO: 58 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1604 herein), KQGSGEANVDNGDV (SEQ ID NO: 59 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1605 herein), KQGSRKTNVDHKQV (SEQ ID NO: 56 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1606 herein), KQGSKGGNVDTNRV (SEQ ID NO: 57 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1606 herein), KQGSGEANVDNGDV (SEQ ID NO: 59 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1605 ...SEQ ID NO: 1605 herein), KQDAAADNIDYDHV (SEQ ID NO: 59 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1606 herein), KQSGTRSNAAASSV (SEQ ID NO: 60 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1607 herein), KENTNTNDTELTNV (SEQ ID NO: 61 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1608 herein), QRGNNVAATADVNT (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1609 herein), No. 69298; SEQ ID NO: 1609 herein), QRGNNEAATADVNT (SEQ ID NO: 63 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1610 herein), QRGNNPAATADVNT (SEQ ID NO: 64 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1611 herein), QRGNNHAATADVNT (SEQ ID NO: 65 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1612 herein), QEENNIAATPGVNT (SEQ ID NO: 66 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1613 herein), QPPNNMAATHEVNT (SEQ ID NO: 67 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1614 herein), QHHNNSAATTIVNT (SEQ ID NO: 68 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1615 herein), QTTNNRAAFNMVET (SEQ ID NO: 69 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1616 herein) ), QKKNNNAASKKVAT (SEQ ID NO: 70 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1617 herein), QGGNNKAADDAVKT (SEQ ID NO: 71 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1618 herein), QAAKGGAADDAVKT (SEQ ID NO: 72 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1619 herein), QDDRAAAANESVDT (SEQ ID NO: 73 in U.S. Patent Application Publication No. 20160369298;SEQ ID NO: 1620 herein), QQQHDDAAYQRVHT (SEQ ID NO: 74 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1621 herein), QSSSSLAAVSTVQT (SEQ ID NO: 75 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1622 herein), QNNQTTAAIRNVTT (SEQ ID NO: 76 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1623 herein), NYNKKSDNVDFT (SEQ ID NO: 77 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1624 herein), NYNKKSENVDFT (SEQ ID NO: 77 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1625 herein), 8; SEQ ID NO: 1625 herein), NYNKKSLNVDFT (SEQ ID NO: 79 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1626 herein), NYNKKSPNVDFT (SEQ ID NO: 80 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1627 herein), NYSKKSHCVDFT (SEQ ID NO: 81 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1628 herein), NYRKTIYVDFT (SEQ ID NO: 1629 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1629 herein), SEQ ID NO: 82; SEQ ID NO: 1629 herein), NYKEKKDVHFT (SEQ ID NO: 83 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1630 herein), NYGHRAIVQFT (SEQ ID NO: 84 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1631 herein), NYANHQFVVCT (SEQ ID NO: 85 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1632 herein), NYDDDPTGVLLT (SEQ ID NO: 86 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1633 herein), NYDDDPTGVLLT (SEQ ID NO: 87 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1634 herein), NYDDDPTGVLLT (SEQ ID NO: 88 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1635 herein), NYDDDPTGVLLT (SEQ ID NO: 89 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1636 herein), NYDDDPTGVLLT (SEQ ID NO: 89 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1637 herein), NYDDDPTGVLLT (SEQ ID NO: 89 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1638 herein), NYDDDPTGVLLT (SEQ ID NO: 89 in U.S. Patent SEQ ID NO: 86 in the specification; SEQ ID NO: 1633 herein), NYDDPTGVLLT (SEQ ID NO: 87 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1634 herein), NFEQQNSVEWT (SEQ ID NO: 88 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1635 herein), SQSGASN (SEQ ID NO: 89 and SEQ ID NO: 241 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1636 herein), NNGSQA (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1636 herein), No. 8, SEQ ID NO: 90; SEQ ID NO: 1637 herein), YYLSRTNTPSGTTTWSRLQFSQAGA (SEQ ID NO: 91 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1638 herein), SKTSADNNNSEYSWTG (SEQ ID NO: 92 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1639 herein), HKDDEEKF (SEQ ID NOs: 93, 209, 214, 219, 224, 234, 239, and 244 in U.S. Patent Application Publication No. 20160369298;SEQ ID NO: 1640 herein), KQGSEKTNVDIEEV (SEQ ID NO: 94 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1641 herein), QRGNNQAATADVNT (SEQ ID NO: 95 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1642 herein), NYNKKSVNVDFT (SEQ ID NO: 96 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1643 herein), SQSGASNYNTPSGTTTQSRLQFSTS ADNNNSEYSWTGATKYH (SEQ ID NO: 106 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1644 herein), SASGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 107 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1645 herein), SQSGASNYNTPSGTTTQSRLQFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 1 in U.S. Patent Application Publication No. 20160369298 08; SEQ ID NO: 1646 herein), SASGASNYNTPSGTTTQSRLQFSTSADNNNSEFSWPGATTYH (SEQ ID NO: 109 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1647 herein), SQSGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 110 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1648 herein), SASGASNYNTPSGSLTQSSLGFSTDGE NNNSDFSWTGATKYH (SEQ ID NO: 111 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1649 herein), SQSGASNYNTPSGTTTQSRLQFSTSADNNNSDFSWTGATKYH (SEQ ID NO: 112 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1650 herein), SGAGASNFNSEGGSLTQSSLGFSTDGENNNSDFSWTGATKYH (SEQ ID NO: 113 in US Patent Application Publication No. 20160369298;SEQ ID NO: 1651 herein), SGAGASN (SEQ ID NO: 176 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1652 herein), NSEGGSLTQSSLGFS (SEQ ID NOs: 177, 185, 193, and 202 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1653 herein), TDGENNNSDFS (SEQ ID NO: 178 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1654 herein), SEFSWPGATT (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1655 herein), SEQ ID NO: 179 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1655 herein), TSADNNNSDFSWT (SEQ ID NO: 180 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1656 herein), SQSGASNY (SEQ ID NOs: 181, 187, and 198 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1657 herein), NTPSGTTTQSRLQFS (SEQ ID NOs: 182, 188, 191, and 199 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1657 herein), SEQ ID NO: 1658), TSADNNNSEYSWTGATKYH (SEQ ID NO: 183 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1659 herein), SASGASNF (SEQ ID NO: 184 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1660 herein), TDGENNNSDFSWTGATKYH (SEQ ID NOs: 186, 189, 194, 197, and 203 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1661 herein), SASGASNY (U.S. Patent Application SEQ ID NO:190 and SEQ ID NO:195 in US Patent Application Publication No. 20160369298; SEQ ID NO:1662 herein), TSADNNNSEFSWPGATTYH (SEQ ID NO:192 in US Patent Application Publication No. 20160369298; SEQ ID NO:1663 herein), NTPSGSLTQSSLGFS (SEQ ID NO:196 in US Patent Application Publication No. 20160369298; SEQ ID NO:1664 herein), TSADNNNSDFSWTGATKYH (SEQ ID NO:200 in US Patent Application Publication No. 20160369298;SEQ ID NO: 1665 herein), SGAGASNF (SEQ ID NO: 201 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1666 herein), CTCCAGVVSVVSMRSRVCVNSGCAGCTDHCVVSRNSGTCVMSACACAA (SEQ ID NO: 204 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1667 herein), CTCCAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAA (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1668 herein), GAGAGAGGCAACAGACAAGCAGCTACCGCAGATGTCAACACACAA (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1669 ... 298; SEQ ID NO: 1668 herein), SAAGASN (SEQ ID NO: 206 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1669 herein), YFLSRTNTESGSTTQSTLRFSQAG (SEQ ID NO: 207 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1670 herein), SKTSADNNNSDFS (SEQ ID NOs: 208, 228, and 253 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1671 herein), KQGSEKTDVDIDKV (SEQ ID NO: 210 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1672 herein), STAGASN (SEQ ID NO: 211 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1673 herein), YFLSRTNTTSGIETQSTLRFSQAG (SEQ ID NO: 212 and SEQ ID NO: 247 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1674 herein), SKTDGENNNSDFS (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1675 herein), No. 8, SEQ ID NO: 213 and SEQ ID NO: 248; SEQ ID NO: 1675 herein), KQGAAADDVEIDGV (SEQ ID NO: 215 and SEQ ID NO: 250 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1676 herein), SEAGASN (SEQ ID NO: 216 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1677 herein), YYLSRTNTPSGTTTQSRLQFSQAG (SEQ ID NOs: 217, 232, and 242 in U.S. Patent Application Publication No. 20160369298;SEQ ID NO: 1678 herein), SKTSADNNNSEYS (SEQ ID NOs: 218, 233, 238, and 243 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1679 herein), KQGSEKTNVDIEKV (SEQ ID NOs: 220, 225, and 245 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1680 herein), YFLSRTNDASGSDTKSTLLFSQAG (SEQ ID NO: 222 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1681 herein), ), STTPSENNNSEYS (SEQ ID NO: 223 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1682 herein), SAAGATN (SEQ ID NO: 226 and SEQ ID NO: 251 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1683 herein), YFLSRTNGEAGSATLSELRFSQAG (SEQ ID NO: 227 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1684 herein), HGDDADRF (SEQ ID NO: 1685 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1686 herein), No. 229 and SEQ ID NO: 254; SEQ ID NO: 1685 herein), KQGAEKSDVEVDRV (SEQ ID NO: 230 and SEQ ID NO: 255 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1686 herein), KQDSGGDNIDIDQV (SEQ ID NO: 235 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1687 herein), SDAGASN (SEQ ID NO: 236 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1688 herein), YFLSRTNTEGGHDT QSTLRFSQAG (SEQ ID NO: 237 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1689 herein), KEDGGGSDVAIDEV (SEQ ID NO: 240 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1690 herein), SNAGASN (SEQ ID NO: 246 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1691 herein), and YFLSRTNGEAGSATLSELRFSQPG (SEQ ID NO: 252 in US Patent Application Publication No. 20160369298;Non-limiting examples of nucleotide sequences that may encode the amino acid mutation site include the following: AGCVVMDCAGGARSCASCAAC (SEQ ID NO: 97 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1693 herein), AACRACRRSMRSMAGGCA (SEQ ID NO: 98 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1694 herein), CACRRGGACRRC; RMSRRSARSTTT (SEQ ID NO: 99 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1695 herein), TATTTCTTGAGCAGAACAAACRVCVVSRSCGGAMNCVHSACGMHSTCAVVSCTTVDSTTTTCTCAGSBCRGSGCG (SEQ ID NO: 100 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1696 herein), TCAAMAMMAVNSRVCSRSAACAACAACAGTRASTTCTCGTGGMMAGGA (SEQ ID NO: 101 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1697 herein), AAGSAARCRSCRVSRVARVCRATRYCGMSNHCRVMVRSGTC (SEQ ID NO: 102 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1698 herein), CAGVVSVVSMRSRVCVNSGCAGCTDHCVVSRNSGTCVMSACA (SEQ ID NO: 103 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1699 herein) , AACTWCRVSVASMVSVHSDDTGTGSWSTKSACT (SEQ ID NO: 104 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1700 herein), TTGTTGAACATCACCACGTGACGCACGTTC (SEQ ID NO: 256 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1701 herein), TCCCCGTGGTTCTACTACATAATGTGGCCG (SEQ ID NO: 257 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1701 herein), SEQ ID NO: 1702 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1703 herein), TTCCACACTCCGTTTTGGATAATGTTGAAC (SEQ ID NO: 258 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1703 herein), AGGGACATCCCCAGCTCCATGCTGTGGTCG (SEQ ID NO: 259 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1704 herein), AGGGACAACCCCTCCGACTCGCCCTAATCC (SEQ ID NO: 260 in US Patent Application Publication No. 20160369298;SEQ ID NO: 1705 herein), TCCTAGTAGAAGACACCCTCTCACTGCCCG (SEQ ID NO: 261 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1706 herein), AGTACCATGTACACCCACTCTCCCAGTGCC (SEQ ID NO: 262 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1707 herein), ATATGGACGTTCATGCTGATCACCATACCG (U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1708 herein), 9298; SEQ ID NO: 1708 herein), AGCAGGAGCTCCTTGGCCTCAGCGTGCGAG (SEQ ID NO: 264 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1709 herein), ACAAGCAGCTTCACTATGACAACCACTGAC (SEQ ID NO: 265 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1710 herein), CAGCCTAGGAACTGGCTTCCTGGACCCTGTTACCGCCAGCAGAGAGTCTCAAMAMMAVNSRVCSRSAACAACAACAGTRASTTCTCCTGGMMAGGAGCTACCAAGTACCACCTCAATGGCAGAGACTCTCTGGTGAATCCCGGACCAGCTATGGCAAGCCACRRGGACRRCRMSRRSARSTTTTTTCCTCAGAGCGGGGTTCTCATCTTTGGGAAGSAARRCRSCRVSRVARVCRATRYCGMSNHCRVMVRSGTCATGATT ACAGACGAAGAGGAGATCTGGAC (SEQ ID NO: 266 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1711 herein), TGGGACAATGGCGGTCGTCTCTCAGAGTTKTKKT (SEQ ID NO: 267 in US Patent Application Publication No. 20160369298; SEQ ID NO: 1712 herein), AGAGGACCKKTCCTCGATGGTTCATGGTGGAGTTA (SEQ ID NO: 268 in US Patent Application Publication No. 20160369298;CCACTTAGGGCCTGGTCGATACCGTTCGGTG (SEQ ID NO: 269 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1714 herein), or TCTCGCCCCAAGAGTAGAAACCCTTCSTTYYG (SEQ ID NO: 270 in U.S. Patent Application Publication No. 20160369298; SEQ ID NO: 1715 herein);

[0121] In some embodiments, the AAV serotype may comprise an ocular cell targeting peptide as described in WO2016134375 (the contents of which are incorporated by reference in their entirety), such as, but not limited to, SEQ ID NO: 9 or SEQ ID NO: 10 of WO2016134375. Additionally, any of the ocular cell targeting peptides or amino acids described in WO2016134375 may be inserted into any parent AAV serotype, such as, but not limited to, AAV2 (SEQ ID NO: 8 of WO2016134375; SEQ ID NO: 1716 herein), or AAV9 (SEQ ID NO: 11 of WO2016134375; SEQ ID NO: 1717 herein). In some embodiments, modifications such as insertions are made at positions P34 to A35, T138 to A139, A139 to P140, G453 to T454, N587 to R588, and / or R588 to Q589 of the AAV2 protein. In certain embodiments, insertions are made at positions D384, G385, 1560, T561, N562, E563, E564, E565, N704, and / or Y705 of the AAV9 protein. The ocular cell targeting peptide may be, but is not limited to, any of the following amino acid sequences: GSTPPPM (SEQ ID NO: 1 in WO2016134375; SEQ ID NO: 1718 herein) or GETRAPL (SEQ ID NO: 4 in WO2016134375; SEQ ID NO: 1719 herein).

[0122] In some embodiments, the AAV serotype may be modified as described in U.S. Patent Application Publication No. 20170145405, the contents of which are incorporated by reference in their entirety. AAV serotypes may include modified AAV2 (e.g., Y444F, Y500F, Y730F, and / or S662V modifications), modified AAV3 (e.g., Y705F, Y731F, and / or T492V modifications), and modified AAV6 (e.g., S663V and / or T492V modifications).

[0123] In some embodiments, the AAV serotype may be modified as described in International Publication No. WO2017083722, the contents of which are incorporated by reference in their entirety. AAV serotypes may include AAV1 (Y705+731F+T492V), AAV2 (Y444+500+730F+T491V), AAV3 (Y705+731F), AAV5, AAV5 (Y436+693+719F), AAV6 (VP3 variant Y705F / Y731F / T492V), AAV8 (Y733F), AAV9, AAV9 (VP3 variant Y731F), and AAV10 (Y733F).

[0124] In some embodiments, the AAV serotype may comprise an engineered epitope comprising amino acids SPAKFA (SEQ ID NO: 24 in WO2017015102; SEQ ID NO: 1720 herein) or NKDKLN (SEQ ID NO: 2 in WO2017015102; SEQ ID NO: 1721 herein), as described in WO2017015102 (the contents of which are incorporated by reference in their entirety). The epitope may be inserted in the region of amino acids 665-670 based on the numbering of the VP1 capsid of AAV8 (SEQ ID NO: 3 in WO2017015102) and / or in the region of residues 664-668 of AAV3B (SEQ ID NO: 3).

[0125] In some embodiments, the AAV serotype is selected from the group consisting of, but not limited to, one or more (e.g., 2, 3, 4, 5, 6, or 7) amino acid residues 262-268, 370-379, 451-459, 472-473, 493-500, 528-534, 547-552, 588-597, 709-710, or 716-722 of AAV1, or AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAV14, AAV15, AAV16, AAV17, AAV18, AAV19, AAV20, AAV21, AAV22, AAV23, AAV24, AAV25, AAV26, AAV27, AAV28, AAV29, AAV30, AAV31, AAV32, AAV33, AAV34, AAV35, AAV36, AAV37, AAV38, AAV39, AAV40, AAV41, AAV42, AAV43, AAV44, AAV45, AAV46, AAV47, AAV48, AAV49, AAV50, AAV51, AAV52, AAV53, AAV54, AAV55, AAV56, AAV57, AAV58, AAV59, AAV60, AAV61, AAV62, AAV63, AAV64, AAV65, AAV66, AAV67, AAV68, AAV69, AAV70, AAV71, AAV72, AAV73, AAV74, AAV The AAV variants may be or have the sequences as described in WO2017058892 (the contents of which are incorporated herein by reference in their entirety), such as AAV variants having capsid proteins that may contain substitutions at the equivalent amino acid residues of AAV8, AAV9, AAV10, AAV11, AAV12, AAVrh8, AAVrh10, AAVrh32.33, bovine AAV, or avian AAV. The amino acid substitutions may be, but are not limited to, any of the amino acid sequences described in WO2017058892.In some embodiments, the AAV comprises the following residues: 256L, 258K, 259Q, 261S, 263A, 264S, 265T, 266G, 272H, 385S, 386Q, S472R, V473D, N500E, 547S, 709A, 710N, 716D, 717N, 718N, 720L, A456T, Q457T, N458Q, K459S, T492S, K493A, S586R, S587G, S588N, T589R and / or 722 of AAV1 (SEQ ID NO: 1 of WO2017058892). any combination of 244N, 246Q, 248R, 249E, 250I, 251K, 252S, 253G, 254S, 255V, 256D, 263Y, 377E, 378N, 453L, 456R, 532Q, 533P, 535N, 536P, 537G, 538T, 539T, 540A, 541T, 542Y, 543L, 546N, 653V, 654P, 656S, 697Q, 698F, 704D, 705S, 706T, 707G, 708E of AAV5 (SEQ ID NO: 5 in WO2017058892); any combination of 709Y and / or 710R, any combination of 248R, 316V, 317Q, 318D, 319S, 443N, 530N, 531S, 532Q, 533P, 534A, 535N, 540A, 541T, 542Y, 543L, 545G, 546N, 697Q, 704D, 706T, 708E, 709Y and / or 710R of AAV5 (SEQ ID NO: 5 in WO2017058892), 264S, 2 The AAV may contain amino acid substitutions in any combination of: 66G, 269N, 272H, 457Q, 588S, and / or 589I; in AAV8 (SEQ ID NO: 8 in WO2017058892), 457T, 459N, 496G, 499N, 500N, 589Q, 590N, and / or 592A; and in AAV9 (SEQ ID NO: 9 in WO2017058892), 451I, 452N, 453G, 454S, 455G, 456Q, 457N, and / or 458Q.

[0126] In some embodiments, the AAV may comprise a sequence of amino acids at VP1 positions 155, 156, and 157 or VP2 positions 17, 18, 19, and 20, as described in WO2017066764 (the contents of which are incorporated by reference in their entirety). The amino acid sequence may be, but is not limited to, NSS, SXS, SSY, NXS, NSY, SXY, or NXY, where N, X, and Y are independently, but are not limited to, non-serine or non-threonine amino acids, and the AAV may be, but is not limited to, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or AAV12. In some embodiments, the AAV may contain a deletion of at least one amino acid at position 156, 157, or 158 of VP1 or position 19, 20, or 21 of VP2, and the AAV may be, but is not limited to, AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, or AAV12.

[0127] In some embodiments, the AAV can be a serotype generated by Cre-recombination-based AAV targeted evolution (CREATE) as described by Deverman et al. (Nature Biotechnology, Vol. 34, No. 2, pp. 204-209, 2016), Chan et al. (Nature Neuroscience, Vol. 20, No. 8, pp. 1172-1179, 2017), and WO 2015038958 and WO 2017100671, the contents of each of which are incorporated by reference in their entirety. In some embodiments, AAV serotypes generated in this manner have improved CNS transduction and / or neuronal and astrocyte tropism compared to AAV serotypes not generated in this manner. As a non-limiting example, an AAV serotype may comprise a targeting peptide, such as, but not limited to, PHP.B, PHP.B2, PHP.B3, PHP.A, PHP.S, PHP.N, G2A12, G2A15, G2A3, G2B4, or G2B5. In some embodiments, these AAV serotypes may be derivatives of AAV9 (SEQ ID NO: 136) or AAV9 K449R (SEQ ID NO: 9) with an amino acid insert between amino acids 588 and 589. Non-limiting examples of these amino acid inserts include TLAVPFK (PHP.B; SEQ ID NO: 1260), SVSKPFL (PHP.B2; SEQ ID NO: 1268), FTLTTPK (PHP.B3; SEQ ID NO: 1269), YTLSQGW (PHP.A; SEQ ID NO: 1275), QAVRTSL (PHP.S; SEQ ID NO: 1319), LAKERLS (G2A3; SEQ ID NO: 1320), MNSTKNV (G2B4; SEQ ID NO: 1321), VSGGHHS (G2B5; SEQ ID NO: 1322) and / or DGTLAVPFKAQ (PHP.N; SEQ ID NO: 1289).

[0128] In some embodiments, the AAV serotype may be as described in Jackson et al. (Frontiers in Molecular Neuroscience, Vol. 9, No. 154, 2016), the contents of which are incorporated by reference herein in their entirety.

[0129] In some embodiments, the AAV serotype is AAV9 (SEQ ID NO: 135 or 136). In some embodiments, the AAV serotype is AAV9 with a peptide insert.

[0130] In some embodiments, the AAV serotype is the K449R AAV9 variant (SEQ ID NO: 9). AAV9 K449R has the same function as wild-type AAV9. In some embodiments, the AAV serotype is AAV9 K449R with a peptide insert.

[0131] In some embodiments, the AAV serotype is PHP.B (e.g., as described in WO2015038958). In some embodiments, the AAV serotype is paired with a synapsin promoter to enhance neurotransmission compared to the use of a more ubiquitous promoter (i.e., CBA or CMV).

[0132] In some embodiments, the AAV serotype is PHP.N (e.g., as described in WO2017100671). In some embodiments, the AAV serotype is a serotype that includes an AAVPHP.N (PHP.N) peptide or a variant thereof.

[0133] In some embodiments, the AAV serotype is a serotype that includes an AAVPHP.B (PHP.B) peptide or a variant thereof. In some embodiments, the AAV serotype is a serotype that includes an AAVPHP.A (PHP.A) peptide or a variant thereof.

[0134] In some embodiments, the AAV serotype is a serotype that includes the PHP.S peptide or a variant thereof. In some embodiments, the AAV serotype is a serotype that includes a PHP.B2 peptide or a variant thereof.

[0135] In some embodiments, the AAV serotype is a serotype that includes the PHP.B3 peptide or a variant thereof. In some embodiments, the AAV serotype is a serotype that includes a G2B4 peptide or a variant thereof.

[0136] In some embodiments, the AAV serotype is a serotype that includes a G2B5 peptide or a variant thereof. In some embodiments, the AAV serotype is VOY101 or a variant thereof. In some embodiments, VOY101 comprises the amino acid sequence of SEQ ID NO: 1. In some embodiments, the capsid sequence comprises the nucleic acid sequence of SEQ ID NO: 1722.

[0137] In some embodiments, the AAV serotype is VOY201 or a variant thereof. In some embodiments, VOY201 comprises the amino acid sequence of SEQ ID NO: 1724. In some embodiments, the capsid sequence comprises the nucleic acid sequence of SEQ ID NO: 1723.

[0138] In some embodiments, the AAV capsid is capable of penetrating the blood-brain barrier after intravenous administration. Non-limiting examples of such AAV capsids include AAV9, AAV9 K449R, VOY101, VOY201, or AAV capsids containing a peptide insert, such as, but not limited to, AAVPHP.N (PHP.N), AAVPHP.B (PHP.B), PHP.S, G2A3, G2B4, G2B5, G2A12, G2A15, PHP.B2, PHP.B3, or AAVPHP.A (PHP.A).

[0139] In some embodiments, an AAV serotype may comprise a capsid amino acid sequence having 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of those described above. In some embodiments, an AAV serotype comprises a capsid amino acid sequence at least 80% identical to SEQ ID NO: 1, 2, 3, 9, 136 or 1724. In some embodiments, an AAV serotype comprises a capsid amino acid sequence at least 85% identical to SEQ ID NO: 1, 2, 3, 9, 136 or 1724. In some embodiments, an AAV serotype comprises a capsid amino acid sequence at least 90% identical to SEQ ID NO: 1, 2, 3, 9, 136 or 1724. In some embodiments, an AAV serotype comprises a capsid amino acid sequence at least 95% identical to SEQ ID NO: 1, 2, 3, 9, 136 or 1724. In some embodiments, an AAV serotype comprises a capsid amino acid sequence at least 99% identical to SEQ ID NO: 1, 2, 3, 9, 136 or 1724. In some embodiments, an AAV serotype comprises the capsid amino acid sequence of SEQ ID NO: 1, 2, 3, 9, 136 or 1724.

[0140] In some embodiments, the AAV serotype may be encoded by a capsid nucleic acid sequence having 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of those described above. In some embodiments, an AAV serotype comprises a capsid nucleic acid sequence that is at least 80% identical to SEQ ID NO: 4, 135, 1722 or 1723. In some embodiments, an AAV serotype comprises a capsid nucleic acid sequence that is at least 85% identical to SEQ ID NO: 4, 135, 1722 or 1723. In some embodiments, an AAV serotype comprises a capsid nucleic acid sequence that is at least 90% identical to SEQ ID NO: 4, 135, 1722 or 1723. In some embodiments, an AAV serotype comprises a capsid nucleic acid sequence that is at least 95% identical to SEQ ID NO: 4, 135, 1722 or 1723. In some embodiments, an AAV serotype comprises a capsid nucleic acid sequence that is at least 99% identical to SEQ ID NO: 4, 135, 1722 or 1723. In some embodiments, an AAV serotype comprises a capsid nucleic acid sequence of SEQ ID NO: 4, 135, 1722 or 1723.

[0141] In some embodiments, the initiation codon for translation of the AAV VP1 capsid protein can be CTG, TTG, or GTG as described in U.S. Pat. No. 8,163,543, the contents of which are incorporated by reference herein in their entirety.

[0142] This disclosure refers to structural capsid proteins (including VP1, VP2, and VP3) encoded by capsid (Cap) genes. These capsid proteins form the outer proteinaceous structural shell (i.e., capsid) of viral vectors such as AAV. VP capsid proteins synthesized from Cap polynucleotides generally contain a methionine (Met1) as the first amino acid in the peptide sequence, relative to the initiation codon (AUG or ATG) in the corresponding Cap nucleotide sequence. However, the first methionine (Met1) residue, or generally any first amino acid (AA1), is typically cleaved by a protein processing enzyme, such as Met-aminopeptidase, after or during polypeptide synthesis. This "Met / AA clipping" process is often associated with the corresponding acetylation of a second amino acid (e.g., alanine, valine, serine, threonine, etc.) in the polypeptide sequence. Met clipping typically occurs in VP1 and VP3 capsid proteins, but may also occur in VP2 capsid proteins.

[0143] Incomplete Met / AA clipping may result in a mixture of one or more (one, two, or three) VP capsid proteins comprising the viral capsid, some of which may contain the Met1 / AA1 amino acid (Met+ / AA+) and some of which may lack the Met1 / AA1 amino acid as a result of Met / AA clipping (Met- / AA-). Further discussion of Met / AA clipping in capsid proteins is provided by Jin et al., "Direct Liquid Chromatography / Mass Spectrometry Analysis for Complete Characterization of Recombinant Adeno-Associated Virus Capsid Proteins," in ... See, for example, Hwang et al., "N-Terminal Acetylation of Cellular Proteins Creates Specific Degradation Signals," Hum Gene Ther Methods, October 2017, Vol. 28, No. 5, pp. 255-267; Hwang et al., "N-Terminal Acetylation of Cellular Proteins Creates Specific Degradation Signals," Science, February 19, 2010, Vol. 327, No. 5968, pp. 973-977 (the contents of each of which are incorporated by reference herein in their entirety).

[0144] According to the present disclosure, reference to a capsid protein is not limited to either a clipped (Met- / AA-) or an unclipped (Met+ / AA+) sequence, and may, within context, refer to an individual capsid protein, a viral capsid composed of a mixture of capsid proteins, and / or a polynucleotide sequence (or fragment thereof) that encodes, describes, produces, or results in a capsid protein of the present disclosure. Also, direct reference to a "capsid protein" or "capsid polypeptide" (such as VP1, VP2, or VP3) can include VP capsid proteins that contain the Met1 / AA1 amino acids (Met+ / AA+), as well as the corresponding VP capsid protein lacking the Met1 / AA1 amino acids as a result of Met / AA clipping (Met- / AA-).

[0145] Furthermore, according to the present disclosure, reference to specific sequence numbers (whether protein or nucleic acid) each comprising or encoding one or more capsid proteins comprising the Met1 / AA1 amino acids (Met+ / AA+) should be understood to teach VP capsid proteins lacking the Met1 / AA1 amino acids, as upon review of the sequence, it is readily apparent that any sequence simply lacks the first recited amino acid (whether methionine or not).

[0146] As a non-limiting example, reference to a VP1 polypeptide sequence that is 736 amino acids in length and that includes a "Met1" amino acid (Met+) encoded by an AUG / ATG start codon can also be understood to teach a VP1 polypeptide sequence that is 735 amino acids in length and that does not include the "Met1" amino acid (Met-) of the 736 amino acid Met+ sequence. As a second non-limiting example, reference to a VP1 polypeptide sequence that is 736 amino acids in length and that includes an "AA1" amino acid (AA1+) encoded by an optional NNN start codon can also be understood to teach a VP1 polypeptide sequence that is 735 amino acids in length and that does not include the "AA1" amino acid (AA1-) of the 736 amino acid AA1+ sequence.

[0147] Reference to a viral capsid formed from VP capsid proteins (such as a reference to a particular AAV capsid serotype) can incorporate VP capsid proteins that contain Met1 / AA1 amino acids (Met+ / AA1+), the corresponding VP capsid protein lacking the Met1 / AA1 amino acids as a result of Met / AA1 clipping (Met- / AA1-), or combinations thereof (Met+ / AA1+ and Met- / AA1-).

[0148] As non-limiting examples, AAV capsid serotypes can include VP1(Met+ / AA1+), VP1(Met- / AA1-), or a combination of VP1(Met+ / AA1+) and VP1(Met- / AA1-). AAV capsid serotypes can also include VP3(Met+ / AA1+), VP3(Met- / AA1-), or a combination of VP3(Met+ / AA1+) and VP3(Met- / AA1-), as well as the similar optional combination of VP2(Met+ / AA1) and VP2(Met- / AA1-).

[0149] Expression vector In some embodiments, the AAV particles of the present disclosure serve as expression vectors encoding FXN. The expression vector is not limited to AAV, but may be an adenovirus, retrovirus, lentivirus, plasmid, vector, or any variant thereof.

[0150] In some embodiments, the AAV particle expression vector may include, from ITR to ITR, listed from 5' to 3', an ITR, a promoter, an intron, a nucleic acid sequence encoding FXN, a polyA sequence, and an ITR.

[0151] Inverted Terminal Repeat (ITR) The AAV particle of the present disclosure comprises a viral genome having at least one ITR region and a payload region encoding FXN. As used herein, a "viral genome" or "vector genome" is a polynucleotide comprising at least one terminal inverted ITR and at least one coding payload. In one embodiment, the viral genome has two ITRs. These two ITRs flank the payload region at the 5' and 3' ends. The ITRs serve as origins of replication containing recognition sites for replication. The ITRs contain sequence regions that can be complementary and symmetrically arranged. The ITRs incorporated into the viral genome of the present disclosure can comprise naturally occurring or recombinantly derived polynucleotide sequences.

[0152] The ITRs may be from the same serotype as the capsid, selected from any of the serotypes listed in Table 1 or derivatives thereof. The ITRs may be of a different serotype from the capsid. In some embodiments, the AAV particles have two or more ITRs. In some embodiments, the AAV particles have a viral genome comprising two ITRs. In some embodiments, the ITRs are of the same serotype as each other. In some embodiments, the ITRs are of different serotypes. Non-limiting examples include zero, one, or both of the ITRs having the same serotype as the capsid. In some embodiments, both ITRs in the viral genome of the AAV particle are AAV2 ITRs.

[0153] Independently, each ITR may be about 100 to about 150 nucleotides in length. The ITRs may be about 100-105, 106-110, 111-115, 116-120, 121-125, 126-130, 131-135, 136-140, 141-145, or 146-150 nucleotides in length. In some embodiments, the ITRs are 140-142 nucleotides in length. Non-limiting examples of ITR lengths are 102, 105, 119, 130, 140, 141, 142, or 145 nucleotides in length, and those having at least 95% identity thereto.

[0154] In some embodiments, one or more ITRs are AAV2 ITRs or fragments or variants thereof. In some embodiments, both the 5' ITR and the 3' ITR are AAV2 ITRs or fragments or variants thereof. In some embodiments, one or more ITRs are 141 nucleotides in length. In some embodiments, both the 5' ITR and the 3' ITR are 141 nucleotides in length. In some embodiments, the 5' ITR comprises a sequence at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1811. In some embodiments, the 3' ITR comprises a sequence at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1812. In some embodiments, the 5' ITR comprises a sequence at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1811, and the 3' ITR comprises a sequence at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1812. In some embodiments, the viral genome comprises the 5' and 3' ITRs described above and a payload region encoding frataxin, e.g., encoding SEQ ID NO: 1725 or a variant thereof having at least 90% sequence identity. In some embodiments, the viral genome comprises the 5' and 3' ITRs described above and a payload region encoding frataxin, e.g., SEQ ID NO: 1824 or a variant thereof having at least 90% sequence identity, e.g., a variant that retains one or more functional properties of wild-type frataxin.

[0155] Promoter Those skilled in the art will recognize that target cells may require specific promoters, including, but not limited to, species-specific, inducible, tissue-specific, or cell cycle-specific promoters (Parr et al., Nat. Med. 3:1145-9 (1997); the contents of which are incorporated herein by reference in their entirety).

[0156] In some embodiments, delivery of AAV particles to cells of the central nervous system (e.g., parenchyma) comprises a composition in which the AAV genome further comprises a cell-specific promoter region, hi some embodiments, delivery comprises a composition in which the AAV genome further comprises a ubiquitous promoter region.

[0157] In some embodiments, the promoter is effective to drive expression of a payload or transgene, hi some embodiments, the promoter is effective to drive expression of FXN.

[0158] In some embodiments, the FXN promoter is used in the viral genome of an AAV particle encoding FXN or a variant thereof. Certain embodiments provide that the FXN promoter is engineered for optimal FXN expression.

[0159] In some embodiments, the promoter is a weak promoter that provides expression of a payload, e.g., FXN, over a period of time in a target tissue, for example, but not limited to, nervous system tissue (e.g., CNS tissue). Expression can be at 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20 hours, 21 hours, 22 hours, 23 hours, 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 1 week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 15 days, 16 days, 17 days, 18 days, 19 days, 20 days, 3 weeks, 2 ... The period may be 2 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days, 31 days, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 1 year, 13 months, 14 months, 15 months, 16 months, 17 months, 18 months, 19 months, 20 months, 21 months, 22 months, 23 months, 2 years, 3 years, 4 years, 5 years, 6 years, 7 years, 8 years, 9 years, 10 years or more than 10 years. Expression may be for 1-5 hours, 1-12 hours, 1-2 days, 1-5 days, 1-2 weeks, 1-3 weeks, 1-4 weeks, 1-2 months, 1-4 months, 1-6 months, 2-6 months, 3-6 months, 3-9 months, 4-8 months, 6-12 months, 1-2 years, 1-5 years, 2-5 years, 3-6 years, 3-8 years, 4-8 years, or 5-10 years. In some embodiments, the promoter is a weak promoter that directs sustained expression of the payload in neural tissue.

[0160] In some embodiments, the promoter may be a promoter that is less than 1 kb in size. , 505, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, or more than 800 nucleotides in length. The promoter may have a length of 50 to 100, 100 to 150, 150 to 200, 200 to 300, 200 to 400, 200 to 500, 200 to 600, 200 to 700, 200 to 800, 300 to 400, 300 to 500, 300 to 600, 300 to 700, 300 to 800, 400 to 500, 400 to 600, 400 to 700, 400 to 800, 500 to 600, 500 to 700, 500 to 800, 600 to 700, 600 to 800, or 700 to 800 nucleotides.

[0161] In some embodiments, the promoter may be a combination of two or more components, such as, but not limited to, CMV and CBA. Each component may be 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 381, 382, ​​383, 384, 385, 386, 387, 388, 389, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, 810, 820, 830, 840, 850, 860, 870, 880, 890, 900, 910, 920, 930, 940, 950, 960, 970, 980, 990, 1000, 1010, 1020, 1030, 1040, 1 It may have a length of 80, 490, 500, 510, 520, 530, 540, 550, 560, 570, 580, 590, 600, 610, 620, 630, 640, 650, 660, 670, 680, 690, 700, 710, 720, 730, 740, 750, 760, 770, 780, 790, 800, or more than 800 nucleotides. Each component may have a length of 200-300, 200-400, 200-500, 200-600, 200-700, 200-800, 300-400, 300-500, 300-600, 300-700, 300-800, 400-500, 400-600, 400-700, 400-800, 500-600, 500-700, 500-800, 600-700, 600-800, or 700-800 nucleotides. In some embodiments, the promoter is a combination of a 382 nucleotide CMV enhancer sequence and a 260 nucleotide CBA promoter sequence. In some embodiments, the promoter is a combination of a 380 nucleotide CMV enhancer sequence and a 260 nucleotide CBA promoter sequence.

[0162] In some embodiments, the vector genome comprises at least one element that enhances target specificity and expression of FXN (see, e.g., Powell et al., Viral Expression Cassette Elements to Enhance Transgene Target Specificity and Expression in Gene Therapy, 2015; the contents of which are incorporated herein by reference in their entirety). Non-limiting examples of expression-enhancing elements include promoters, endogenous miRNAs, posttranscriptional regulatory elements (PREs), polyadenylation (polyA) signal sequences, upstream enhancers (USEs), CMV enhancers, and / or introns. In certain embodiments, elements used to enhance target specificity and / or expression of FXN are referred to as "enhancers" or "enhancer sequences." In some embodiments, a promoter may include an enhancer sequence. In some embodiments, an enhancer may be a component of the viral genome separate from the promoter. In some embodiments, the enhancer may be 5' to the promoter sequence in the viral genome. In some embodiments, the enhancer may be 3' to the promoter sequence in the viral genome. In some embodiments, the enhancer comprises or consists of SEQ ID NO: 1777.

[0163] As used herein, "intron" or "intron sequence" includes a full-length intron or a fragment thereof. As used herein, "exon" or "exon sequence" includes a full-length exon or a fragment thereof. In some embodiments, an enhancer may include at least one intron or exon sequence. In some embodiments, an enhancer may include at least one intron sequence. In some embodiments, an enhancer may include at least one exon sequence. In some embodiments, an enhancer includes one intron sequence and one exon sequence. In some embodiments, an enhancer sequence includes two intron sequences. In some embodiments, an enhancer sequence includes two exon sequences. In some embodiments, an enhancer sequence includes two intron sequences and two exon sequences. In some embodiments, an enhancer includes SEQ ID NO: 1818. In some embodiments, an enhancer may include two intron sequences and two exon sequences. In some embodiments, the enhancer may include an iE1 exon (e.g., exon 1), an iE1 intron (e.g., intron 1), a human beta-globin intron (e.g., intron 2), and a human beta-globin exon (e.g., exon 3). In some embodiments, the enhancer may include, from 5' to 3', SEQ ID NOs: 1817, 1819, 1820, 1821. In some embodiments, the enhancer may include SEQ ID NO: 1816.

[0164] Promoters that drive expression in most tissues include, but are not limited to, human elongation factor 1 α-subunit (EF1α), immediate early cytomegalovirus (CMV), chicken β-actin (CBA) and its derivatives CAG, β-glucuronidase (GUSB), or ubiquitin C (UBC). Tissue-specific expression elements can be used to restrict expression to specific cell types, such as, but not limited to, neural promoters that can be used to restrict expression to neurons, astrocytes, or oligodendrocytes. Non-limiting examples of neuronal tissue-specific expression elements include neuron-specific enolase (NSE), platelet-derived growth factor (PDGF), platelet-derived growth factor B chain (PDGF-β), synapsin (Syn), methyl-CpG-binding protein 2 (MeCP2), CaMKII, mGluR2, NFL, NFH, nβ2, PPE, Enk, and EAAT2 promoters. Non-limiting examples of tissue-specific expression elements for astrocytes include the glial fibrillary acidic protein (GFAP) and EAAT2 promoters. Non-limiting examples of tissue-specific expression elements for oligodendrocytes include the myelin basic protein (MBP) promoter.

[0165] In some embodiments, the vector genome contains a ubiquitous promoter. Non-limiting examples of ubiquitous promoters include H1, U6, CMV, CBA (including derivatives such as CAG and CBh), EF-1α, PGK, UBC, GUSB (hGBp), and UCOE (promoter of HNRPA2B1-CBX3). Yu et al. (Molecular Pain, 2011, Vol. 7, p. 63; the contents of which are incorporated herein by reference in their entirety) used lentiviral vectors to evaluate the expression of eGFP under the CAG, EF-1α, PGK, and UBC promoters in rat DRG cells and primary DRG cells. They found that UBC showed weaker expression than the other three promoters, with only 10-12% glial cell expression observed for all promoters. Söderblom et al. (E. Neuro, 2015; the contents of which are incorporated herein by reference in their entireties) studied eGFP expression in AAV8 containing the CMV and UBC promoters and AAV2 containing the CMV promoter after injection in the motor cortex. Intranasal administration of plasmids containing the UBC or EFIα promoters demonstrated higher sustained airway expression than expression from the CMV promoter (see, e.g., Gill et al., Gene Therapy, 2001, Vol. 8, pp. 1539-1546, the contents of which are incorporated herein by reference in their entireties). Husain et al. (Gene Therapy, 2009; the contents of which are incorporated herein by reference in their entireties) evaluated an HβH construct containing the hGUSB promoter, HSV-1 LAT promoter, and NSE promoter and found that this HβH construct showed weaker expression than NSE in the mouse brain. Passini and Wolfe (J. Virol., 2001, pp. 12382-12392, the contents of which are incorporated herein by reference in their entireties) evaluated the long-term effects of HβH vectors after intracerebroventricular injection in neonatal mice and found that there was sustained expression for at least one year.Xu et al. (Gene Therapy, 2001, Vol. 8, pp. 1323-1332; the contents of which are incorporated herein by reference in their entirety) found that when the NF-L and NF-H promoters were used, lower expression was found in all brain regions compared to CMV-lacZ, CMV-luc, EF, GFAP, hENK, nAChR, PPE, PPE+wpre, NSE(0.3kb), NSE(1.8kb), and NSE(1.8kb+wpre). Xu et al. found that the promoter activity, in descending order, was NSE(1.8kb), EF, NSE(0.3kb), GFAP, CMV, hENK, PPE, NFL, and NFH. NFL is a 650-nucleotide promoter, and NFH is a 920-nucleotide promoter; both are absent from the liver, but NFH is abundant in proprioceptive neurons, the brain, and the spinal cord, and NFH is present in the heart. Scn8a is a 470-nucleotide promoter expressed throughout the DRG, spinal cord, and brain, with particularly high expression in hippocampal neurons and cerebellar Purkinje cells, the cortex, the thalamus, and the hypothalamus (see, e.g., Drews et al., 2007, and Raymond et al., 2004; the contents of each of which are incorporated by reference in their entirety).

[0166] In some embodiments, the vector genome comprises a UBC promoter. The UBC promoter may have a size of 300 to 350 nucleotides. In some embodiments, the UBC promoter is 332 nucleotides in length.

[0167] In some embodiments, the vector genome comprises a GUSB promoter. The GUSB promoter may have a size of 350-400 nucleotides. In some embodiments, the GUSB promoter is 378 nucleotides in length. In some embodiments, the construct may be AAV-promoter-CMV / globin intron-FXN-RBG, where the AAV may be self-complementary and may be an AAV6, AAVrhlO, or AAVDJ serotype.

[0168] In some embodiments, the vector genome comprises an NFL promoter. The NFL promoter may have a size of 600 to 700 nucleotides. In some embodiments, the NFL promoter is 650 nucleotides in length.

[0169] In some embodiments, the vector genome comprises an NFH promoter. The NFH promoter may have a size of 900 to 950 nucleotides. In some embodiments, the NFH promoter is 920 nucleotides in length.

[0170] In some embodiments, the vector genome comprises a scn8a promoter. The scn8a promoter may have a size of 450 to 500 nucleotides. In some embodiments, the scn8a promoter is 470 nucleotides in length.

[0171] In some embodiments, the vector genome comprises the FXN promoter. In some embodiments, the vector genome comprises a PGK promoter. In some embodiments, the vector genome comprises a CBA promoter.

[0172] In some embodiments, the vector genome comprises a CMV promoter. In some embodiments, the vector genome comprises an H1 promoter. In some embodiments, the vector genome comprises a U6 promoter.

[0173] In some embodiments, the vector genome comprises a liver or skeletal muscle promoter. Non-limiting examples of liver promoters include hAAT and TBG. Non-limiting examples of skeletal muscle promoters include desmin, MCK, and C5-12.

[0174] In some embodiments, the AAV vector comprises an enhancer element, a promoter, and / or a 5'UTR intron. The enhancer may be, but is not limited to, a CMV enhancer; the promoter may be, but is not limited to, a CMV, CBA, FXN, UBC, GUSB, NSE, synapsin, MeCP2, or GFAP promoter; and the 5'UTR / intron may be, but is not limited to, an SV40 or CBA-MVM promoter. In some embodiments, the enhancers, promoters and / or introns used in combination may be: (1) a CMV enhancer, a CMV promoter, and an SV40 5'UTR intron; (2) a CMV enhancer, a CBA promoter, and an SV40 5'UTR intron; (3) a CMV enhancer, a CBA promoter, and a CBA-MVM 5'UTR intron; (4) a UBC promoter; (5) a GUSB promoter; (6) an NSE promoter; (7) a synapsin promoter; (8) an MeCP2 promoter; (9) a GFAP promoter; (10) an H1 promoter; and / or (11) a U6 promoter.

[0175] In some embodiments, the AAV vector has an engineered promoter. In some embodiments, the AAV vector comprises a promoter comprising a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1734-1777. In some embodiments, the promoter is or is derived from a CMV promoter and comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1743-1751, 1767, 1772-1774, and 1777. In some embodiments, the promoter is or is derived from a CBA promoter and comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1734-1742, 1760-1766, 1768, and 1775-1776. In some embodiments, the promoter is or is derived from the FXN promoter and comprises a sequence having at least 90%, at least 95%, at least 99% or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1752-1759 and 1769-1770.

[0176] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1738. In some embodiments, the promoter is SEQ ID NO: 1738. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0177] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1738. In some embodiments, the promoter is SEQ ID NO: 1740. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0178] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1742. In some embodiments, the promoter is SEQ ID NO: 1742. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0179] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1750. In some embodiments, the promoter is SEQ ID NO: 1750. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0180] In some embodiments, the viral genome comprises an enhancer, such as an immediate-early "ie" enhancer or a CMV / globin enhancer. In some embodiments, the enhancer comprises ie1 exon 1 and ie1 intron 1 or a fragment thereof. In some embodiments, the enhancer comprises ie1 exon 1, ie1 intron 1 or a fragment thereof, human beta-globin intron 2, and human beta-globin exon 3. In some embodiments, the enhancer comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to a sequence provided by any of SEQ ID NOs: 1815-1821. In some embodiments, the enhancer comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to a sequence provided by SEQ ID NO: 1816. In some embodiments, the viral genome comprises an enhancer as described above and a payload region encoding frataxin, e.g., encoding SEQ ID NO: 1725 or a variant thereof having at least 90% sequence identity thereto, or comprising the nucleic acid sequence SEQ ID NO: 1824 or a variant thereof having at least 90% sequence identity thereto.

[0181] Introns In some embodiments, the vector genome comprises at least one intron or a fragment or derivative thereof, in which the at least one intron can enhance expression of FXN (see, e.g., Powell et al., Viral Expression Cassette Elements to Enhance Transgene Target Specificity and Expression in Gene Therapy, 2015, the contents of which are incorporated herein by reference in their entirety). Non-limiting examples of introns include MVM (67-97 bp), F.IX truncated intron 1 (300 bp), β-globin SD / immunoglobulin heavy chain splice acceptor (250 bp), adenovirus splice donor / immunoglobin splice acceptor (500 bp), SV40 late splice donor / splice acceptor (19S / 16S) (180 bp), and hybrid adenovirus splice donor / IgG splice acceptor (230 bp).

[0182] In some embodiments, the intron may be 100 to 500 nucleotides in length. The intron may have a length of 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, or 500 nucleotides. The intron may have a length of 80 to 100, 80 to 120, 80 to 140, 80 to 160, 80 to 180, 80 to 200, 80 to 250, 80 to 300, 80 to 350, 80 to 400, 80 to 450, 80 to 500, 200 to 300, 200 to 400, 200 to 500, 300 to 400, 300 to 500, or 400 to 500 nucleotides.

[0183] In some embodiments, the AAV vector may include an SV40 intron or a fragment or variant thereof. In some embodiments, the promoter may be CMV. In some embodiments, the promoter may be CBA. In some embodiments, the promoter may be H1.

[0184] In some embodiments, the AAV vector may comprise one or more beta-globin introns or fragments or variants thereof. In some embodiments, the intron comprises one or more human beta-globin sequences (e.g., including fragments / variants thereof).

[0185] In some embodiments, the intron comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to the sequence provided by any of SEQ ID NOS: 1815-1821. In some embodiments, the viral genome comprises the intron described above and a payload region encoding frataxin, e.g., encoding SEQ ID NO: 1725 or a variant thereof having at least 90% sequence identity thereto, or comprising the nucleic acid sequence SEQ ID NO: 1824 or a variant thereof having at least 90% sequence identity thereto. In some embodiments, the promoter may be CMV. In some embodiments, the promoter may be CBA. In some embodiments, the promoter may be H1.

[0186] In some embodiments, the encoded FXN may be located downstream of an intron in an expression vector, such as, but not limited to, an SV40 intron or a beta globin intron, or others known in the art. Furthermore, the encoded FXN may be located upstream of a polyadenylation sequence in an expression vector. In some embodiments, the encoded FXN may be located within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more than 30 nucleotides downstream of a promoter having an intron in an expression vector and / or upstream of a polyadenylation sequence. In some embodiments, the encoded FXN may be located within 1-5, 1-10, 1-15, 1-20, 1-25, 1-30, 5-10, 5-15, 5-20, 5-25, 5-30, 10-15, 10-20, 10-25, 10-30, 15-20, 15-25, 15-30, 20-25, 20-30, or 25-30 nucleotides downstream from an intron and / or upstream of a polyadenylation sequence in an expression vector. In some embodiments, the encoded FXN may be located within the first 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, or more than 25% of nucleotides downstream from an intron and / or upstream of a polyadenylation sequence in an expression vector. In some embodiments, the encoded FXN may be located within the first 1-5%, 1-10%, 1-15%, 1-20%, 1-25%, 5-10%, 5-15%, 5-20%, 5-25%, 10-15%, 10-20%, 10-25%, 15-20%, 15-25%, or 20-25% of the sequence downstream from the intron and / or upstream of the polyadenylation sequence in the expression vector.

[0187] In certain embodiments, the intron sequence is not an enhancer sequence. In certain embodiments, the intron sequence is not a subcomponent of the promoter sequence. Viral genome components: untranslated regions (UTRs) By definition, the wild-type untranslated regions (UTRs) of a gene are transcribed but not translated. Generally, the 5' UTR begins at the transcription start site and ends at the start codon, and the 3' UTR begins immediately after the stop codon and continues until the transcription termination signal.

[0188] Features typically found in abundantly expressed genes of specific target organs can be engineered into UTRs to enhance stability and protein production. As a non-limiting example, 5'UTRs derived from mRNAs normally expressed in the liver (e.g., albumin, serum amyloid A, apolipoprotein A / B / E, transferrin, alpha-fetoprotein, erythropoietin, or factor VIII) can be used in the viral genome of the AAV particles of the present disclosure to enhance expression in hepatocyte cell lines or the liver.

[0189] Without wishing to be bound by theory, wild-type 5' untranslated regions (UTRs) contain features that play a role in translation initiation. The Kozak sequence is widely known to be involved in the process by which the ribosome initiates translation of many genes and is typically contained in 5' UTRs. Kozak sequences have the consensus CCR(A / G)CCAUGG, where R is a purine (adenine or guanine) three bases upstream of the start codon (ATG), followed by another "G."

[0190] In some embodiments, the 5'UTR of the viral genome comprises a Kozak sequence. In some embodiments, the 5'UTR of the viral genome does not contain a Kozak sequence. Without wishing to be bound by theory, wild-type 3'UTRs are known to have adenosine and uridine stretches embedded therein. These AU-rich signatures are particularly prevalent in genes with high turnover rates. Based on their sequence characteristics and functional properties, AU-rich elements (AREs) can be separated into three classes (Chen et al., 1995, the contents of which are incorporated herein by reference in their entirety): Class I AREs, such as, but not limited to, c-Myc and MyoD, contain several dispersed copies of the AUUUA motif within the U-rich region; Class II AREs, such as, but not limited to, GM-CSF and TNF-α, have two or more overlapping UUAUUUA(U / A)(U / A) nonamers; Class III AREs, such as, but not limited to, c-Jun and myogenin, contain several dispersed copies of the AUUUA motif within the U-rich region. ARES are less well defined. These U-rich regions do not contain the AUUUA motif. Most proteins that bind to AREs are known to destabilize messengers, but members of the ELAV family, most notably HuR, have been demonstrated to increase mRNA stability. HuR binds to all three classes of AREs. Engineering a HuR-specific binding site into the 3'UTR of a nucleic acid molecule will result in HuR binding and thus message stabilization in vivo.

[0191] The introduction, removal or modification of 3'UTR AU-rich element (ARE) can be used to adjust the stability of polynucleotide.When manipulating specific polynucleotide, such as the payload region of a viral genome, one or more copies of ARE can be introduced to make the polynucleotide more unstable, thereby reducing translation and reducing the production of the resulting protein.Similarly, ARE can be identified and removed or mutated to increase intracellular stability, thus increasing the translation and production of the resulting protein.

[0192] In some embodiments, the 3'UTR of the viral genome may contain an oligo(dT) sequence for templated addition of a polyA tail. Any UTR from any gene known in the art can be incorporated into the viral genome of AAV particles.These UTRs or their parts can be placed in the same orientation as the gene from which they are selected, or their orientation or position can be changed.In some embodiments, the UTR used in the viral genome of AAV particles can be inverted, shortened, extended, or made with one or more other 5'UTR or 3'UTR known in the art.As used herein, the term "modified" when referring to UTR means that the UTR is changed in some way relative to the reference sequence.For example, the 3' or 5'UTR can be changed relative to the wild-type or natural UTR by changing the orientation or position as taught above, or by inserting additional nucleotides, deleting nucleotides, exchanging or rearranging nucleotides.

[0193] In some embodiments, the viral genome of the AAV particle comprises at least one artificial UTR that is not a variant of a wild-type UTR. In some embodiments, the viral genome of the AAV particle comprises UTRs whose proteins are selected from a family of transcripts that share a common function, structure, feature, or characteristic.

[0194] miRNA target site In some embodiments, the viral genome may contain at least one miRNA binding site. MicroRNAs (or miRNAs or miRs) are 19-25 nucleotide non-coding RNAs that bind to nucleic acid target sites and downregulate gene expression by either reducing nucleic acid molecule stability or inhibiting translation. In some embodiments, the 3'UTR of the viral genome may be engineered to contain at least one miRNA binding site.

[0195] In some embodiments, the viral genome comprises at least one sequence encoding miRNA target site that reduces the expression of transgene in specific tissue.MiRNA and its target tissue are well known in the art.In some embodiments, miR-122 miRNA target site (miR-122TS) or its tandem copy can be encoded in the viral genome to reduce the expression of viral genome in the liver where miR-122 is abundantly expressed.

[0196] In some embodiments, the viral genome comprises at least one miR122 binding site. In some embodiments, the miR122 binding site comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1827. In some embodiments, the AAV vector genome comprises three copies of the miR122 binding site, e.g., three copies of SEQ ID NO: 1827 or a variant thereof having at least 90% sequence identity. In some embodiments, the miR122 binding site series comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1826. In some embodiments, the viral genome comprises one, two, or three miR122 binding sites as described above and a payload region encoding frataxin, e.g., encoding SEQ ID NO: 1725 or a variant thereof having at least 90% sequence identity thereto, or comprising the nucleic acid sequence SEQ ID NO: 1824 or a variant thereof having at least 90% sequence identity thereto. In some embodiments, the viral genome comprises the three miR122 binding sites described above and a payload region encoding frataxin, e.g., encoding SEQ ID NO: 1725 or a variant thereof having at least 90% sequence identity thereto, or comprising the nucleic acid sequence SEQ ID NO: 1824 or a variant thereof having at least 90% sequence identity thereto.

[0197] skeleton In certain embodiments, cis-elements, such as vector backbones, are incorporated into viral particles that encode FXN. Backbone sequences can regulate transcription during virus production. Backbone sequences can contribute to the stability of FXN expression. Backbone sequences can contribute to the level of FXN expression, which can be cloned into pAAVsc or pcDNA3.1 vector backbones.

[0198] Polyadenylation sequence In some embodiments, the viral genome of an AAV particle of the present disclosure comprises at least one polyadenylation sequence. The viral genome of an AAV particle may comprise a polyadenylation sequence between the 3' end of the payload coding sequence and the 5' end of the 3' UTR.

[0199] In some embodiments, polyadenylation sequences or "poly A sequences" may range from absent to about 500 nucleotides in length. Polyadenylation sequences include, but are not limited to, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75 , 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 18 0, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183, 184, 185, 186, 187, 188, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 1 99, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220, 221, 222, 223, 224, 225, 226, 227, 228, 229, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 242, 243, 244, 245, 246, 247, 248, 249, 250, 251, 252, 253, 254, 255, 256, 257,258, 259, 260, 261, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287, 288 , 289, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308, 309, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319 , 320, 321, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, 344, 345, 346, 347, 348, 349, 35 0, 351, 352, 353, 354, 355, 356, 357, 358, 359, 360, 361, 362, 363, 364, 365, 366, 367, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380, 381 1, 382, ​​383, 384, 385, 386, 387, 388, 389, 390, 391, 392, 393, 394, 395, 396, 397, 398, 399, 400, 401, 402, 403, 404, 405, 406, 407, 408, 409, 410, 411, 412, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 4 12, 413, 414, 415, 416, 417, 418, 419, 420, 421, 422, 423, 424, 425, 426, 427, 428, 429, 430, 431, 432, 433, 434, 435, 436, 437, 438, 439, 440, 441, 442, 443 43, 444, 445, 446, 447, 448, 449, 450, 451, 452, 453, 454, 455, 456, 457, 458, 459, 460, 461, 462, 463, 464, 465, 466, 467, 468, 469, 470, 471, 472, 473, 474, 475, 476, 477, 478, 479, 480, 481, 482, 483, 484, 485, 486, 487, 488, 489, 490, 491, 492, 493, 494, 495, 496, 497, 498, 499, or 500 nucleotides in length.

[0200] In some embodiments, the polyadenylation sequence is 50-100 nucleotides in length. In some embodiments, the polyadenylation sequence is 50 to 150 nucleotides in length. In some embodiments, the polyadenylation sequence is 50 to 160 nucleotides in length.

[0201] In some embodiments, the polyadenylation sequence is 50 to 200 nucleotides in length. In some embodiments, the polyadenylation sequence is 60-100 nucleotides in length. In some embodiments, the polyadenylation sequence is 60 to 150 nucleotides in length.

[0202] In some embodiments, the polyadenylation sequence is 60 to 160 nucleotides in length. In some embodiments, the polyadenylation sequence is between 60 and 200 nucleotides in length. In some embodiments, the polyadenylation sequence is 70-100 nucleotides in length.

[0203] In some embodiments, the polyadenylation sequence is 70 to 150 nucleotides in length. In some embodiments, the polyadenylation sequence is 70 to 160 nucleotides in length. In some embodiments, the polyadenylation sequence is 70 to 200 nucleotides in length.

[0204] In some embodiments, the polyadenylation sequence is 80-100 nucleotides in length. In some embodiments, the polyadenylation sequence is 80 to 150 nucleotides in length. In some embodiments, the polyadenylation sequence is 80 to 160 nucleotides in length.

[0205] In some embodiments, the polyadenylation sequence is 80 to 200 nucleotides in length. In some embodiments, the polyadenylation sequence is 90-100 nucleotides in length. In some embodiments, the polyadenylation sequence is 90 to 150 nucleotides in length.

[0206] In some embodiments, the polyadenylation sequence is 90 to 160 nucleotides in length. In some embodiments, the polyadenylation sequence is 90 to 200 nucleotides in length. In some embodiments, the encoded FXN may be located upstream of a polyadenylation sequence in an expression vector. Furthermore, the encoded FXN may be located downstream of a promoter having an SV40 intron in an expression vector, such as, but not limited to, a CMV, U6, CBA, or CBA promoter, or a fragment thereof (e.g., one disclosed herein). In some embodiments, the encoded FXN may be located within 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, or more than 30 nucleotides downstream of the promoter and / or upstream of the polyadenylation sequence in an expression vector. In some embodiments, the encoded FXN may be located within 1-5, 1-10, 1-15, 1-20, 1-25, 1-30, 5-10, 5-15, 5-20, 5-25, 5-30, 10-15, 10-20, 10-25, 10-30, 15-20, 15-25, 15-30, 20-25, 20-30, or 25-30 nucleotides downstream from the promoter and / or upstream of the polyadenylation sequence in an expression vector. In some embodiments, the encoded FXN may be located within the first 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, or more than 25% of nucleotides downstream from the promoter and / or upstream of the polyadenylation sequence in an expression vector. In some embodiments, the encoded FXN may be located within the first 1-5%, 1-10%, 1-15%, 1-20%, 1-25%, 5-10%, 5-15%, 5-20%, 5-25%, 10-15%, 10-20%, 10-25%, 15-20%, 15-25%, or 20-25% of the sequence downstream from the promoter and / or upstream from the polyadenylation sequence in the expression vector.

[0207] In some embodiments, the viral genome comprises a human growth hormone (hGH) polyA sequence. In some embodiments, the viral genome comprises a polyA sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1828. In some embodiments, the viral genome comprises an hGH polyA as described above and a payload region encoding frataxin, e.g., encoding SEQ ID NO: 1725 or a variant thereof having at least 90% sequence identity thereto, or comprising the nucleic acid sequence SEQ ID NO: 1824 or a variant thereof having at least 90% sequence identity thereto.

[0208] Filler Arrangement In some embodiments, the viral genome comprises one or more filler sequences. The filler sequences may be wild-type sequences or engineered sequences. The filler sequences may be variants of wild-type sequences. In one embodiment, the filler sequence is a derivative of human albumin.

[0209] In some embodiments, the viral genome comprises one or more filler sequences to optimize the length of the viral genome for packaging. In some embodiments, the viral genome comprises at least one filler sequence to optimize the length of the viral genome to about 2.3 kb. In some embodiments, the viral genome comprises at least one filler sequence to optimize the length of the viral genome to about 4.6 kb.

[0210] In some embodiments, the viral genome is a single-stranded (ss) viral genome, including but not limited to, 0.1 kb, 0.2 kb, 0.3 kb, 0.4 kb, 0.5 kb, 0.6 kb, 0.7 kb, 0.8 kb, 0.9 kb, 1 kb, 1.1 kb, 1.2 kb, 1.3 kb, 1.4 kb, 1.5 kb, 1.6 kb, 1.7 kb, 1.8 kb, 1.9 kb The vector genome may comprise one or more filler sequences, independently or together, having a length of about 0.1 kb to 3.8 kb, such as 2 kb, 2.1 kb, 2.2 kb, 2.3 kb, 2.4 kb, 2.5 kb, 2.6 kb, 2.7 kb, 2.8 kb, 2.9 kb, 3 kb, 3.1 kb, 3.2 kb, 3.3 kb, 3.4 kb, 3.5 kb, 3.6 kb, 3.7 kb, or 3.8 kb. In some embodiments, the total length of the filler sequences in the vector genome is 3.1 kb. In some embodiments, the total length of the filler sequences in the vector genome is 2.7 kb. In some embodiments, the total length of the filler sequences in the vector genome is 0.8 kb. In some embodiments, the total length of the filler sequences in the vector genome is 0.4 kb. In some embodiments, the length of each filler sequence in the vector genome is 0.8 kb. In some embodiments, the length of each filler sequence in the vector genome is 0.4 kb.

[0211] In some embodiments, the viral genome is a self-complementary (sc) viral genome and includes one or more filler sequences, independently or together, having a length of about 0.1 kb to 1.5 kb, such as, but not limited to, 0.1 kb, 0.2 kb, 0.3 kb, 0.4 kb, 0.5 kb, 0.6 kb, 0.7 kb, 0.8 kb, 0.9 kb, 1 kb, 1.1 kb, 1.2 kb, 1.3 kb, 1.4 kb, or 1.5 kb. In some embodiments, the total length of the filler sequences in the vector genome is 0.8 kb. In some embodiments, the total length of the filler sequences in the vector genome is 0.4 kb. In some embodiments, the length of each filler sequence in the vector genome is 0.8 kb. In some embodiments, the length of each filler sequence in the vector genome is 0.4 kb.

[0212] In some embodiments, the viral genome comprises any portion of the filler sequence. The viral genome may comprise 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the filler sequence.

[0213] In some embodiments, the viral genome is a single-stranded (ss) viral genome and includes one or more filler sequences to bring the length of the viral genome to about 4.6 kb. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 3' of the 5' ITR sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 5' of the promoter sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 3' of the polyadenylation signal sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 5' of the 3' ITR sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located between two intron sequences. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located within an intron sequence. In some embodiments, the viral genome comprises two filler sequences, a first filler sequence located 3' of the 5' ITR sequence and a second filler sequence located 3' of the polyadenylation signal sequence. In some embodiments, the viral genome comprises two filler sequences, a first filler sequence located 5' of the promoter sequence and a second filler sequence located 3' of the polyadenylation signal sequence. In some embodiments, the viral genome comprises two filler sequences, a first filler sequence located 3' of the 5' ITR sequence and a second filler sequence located 5' of the 5' ITR sequence.

[0214] In some embodiments, the viral genome is a self-complementary (sc) viral genome and includes one or more filler sequences to bring the length of the viral genome to approximately 2.3 kb. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 3' of the 5' ITR sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 5' of the promoter sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 3' of the polyadenylation signal sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located 5' of the 3' ITR sequence. In some embodiments, the viral genome includes at least one filler sequence, the filler sequence located between two intron sequences. As a non-limiting example, the viral genome includes at least one filler sequence, the filler sequence located within an intron sequence. In some embodiments, the viral genome comprises two filler sequences, a first filler sequence located 3' of the 5' ITR sequence and a second filler sequence located 3' of the polyadenylation signal sequence. In some embodiments, the viral genome comprises two filler sequences, a first filler sequence located 5' of the promoter sequence and a second filler sequence located 3' of the polyadenylation signal sequence. In some embodiments, the viral genome comprises two filler sequences, a first filler sequence located 3' of the 5' ITR sequence and a second filler sequence located 5' of the 5' ITR sequence.

[0215] In some embodiments, the viral genome may include one or more filler sequences between one of more regions of the viral genome. In some embodiments, the filler region may be located before a region such as, but not limited to, a payload region, an inverted terminal repeat (ITR), a promoter region, an intron region, an enhancer region, a polyadenylation signal sequence region, and / or an exon region. In some embodiments, the filler region may be located after a region such as, but not limited to, a payload region, an inverted terminal repeat (ITR), a promoter region, an intron region, an enhancer region, a polyadenylation signal sequence region, and / or an exon region. In some embodiments, the filler region may be located before and after a region such as, but not limited to, a payload region, an inverted terminal repeat (ITR), a promoter region, an intron region, an enhancer region, a polyadenylation signal sequence region, and / or an exon region.

[0216] In some embodiments, the viral genome may include one or more filler sequences that bisect at least one region of the viral genome. The bisected region of the viral genome may comprise 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% of the region 5' of the filler sequence region. In some embodiments, the filler sequence may bisect at least one region such that 10% of the region is located 5' of the filler sequence and 90% of the region is located 3' of the filler sequence. In some embodiments, the filler sequence may bisect at least one region such that 20% of the region is located 5' of the filler sequence and 80% of the region is located 3' of the filler sequence. In some embodiments, the filler sequence may bisect at least one region such that 30% of the region is located 5' of the filler sequence and 70% of the region is located 3' of the filler sequence. In some embodiments, the filler sequence may bisect at least one region such that 40% of the region is located 5' of the filler sequence and 60% of the region is located 3' of the filler sequence. In some embodiments, the filler sequence may bisect at least one region such that 50% of the region is located 5' of the filler sequence and 50% of the region is located 3' of the filler sequence. In some embodiments, the filler sequence may bisect at least one region such that 60% of the region is located 5' of the filler sequence and 40% of the region is located 3' of the filler sequence. In some embodiments, the filler sequence may bisect at least one region such that 70% of the region is located 5' of the filler sequence and 30% of the region is located 3' of the filler sequence. In some embodiments, the filler sequence may bisect at least one region such that 80% of the region is located 5' of the filler sequence and 20% of the region is located 3' of the filler sequence, hi some embodiments, the filler sequence may bisect at least one region such that 90% of the region is located 5' of the filler sequence and 10% of the region is located 3' of the filler sequence.

[0217] In some embodiments, the viral genome comprises a filler sequence after the 5' ITR. In some embodiments, the viral genome comprises a filler sequence after the promoter region. In some embodiments, the viral genome comprises a filler sequence after the payload region. In some embodiments, the viral genome comprises a filler sequence after the intron region. In some embodiments, the viral genome comprises a filler sequence after the enhancer region. In some embodiments, the viral genome comprises a filler sequence after the polyadenylation signal sequence region. In some embodiments, the viral genome comprises a filler sequence after the exon region.

[0218] In some embodiments, the viral genome comprises a filler sequence before the promoter region. In some embodiments, the viral genome comprises a filler sequence before the payload region. In some embodiments, the viral genome comprises a filler sequence before the intron region. In some embodiments, the viral genome comprises a filler sequence before the enhancer region. In some embodiments, the viral genome comprises a filler sequence before the polyadenylation signal sequence region. In some embodiments, the viral genome comprises a filler sequence before the exon region.

[0219] In some embodiments, the viral genome comprises a filler sequence before the 3' ITR. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a 5'ITR and a promoter region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a 5'ITR and a payload region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a 5'ITR and an intron region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a 5'ITR and an enhancer region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a 5'ITR and a polyadenylation signal sequence region.

[0220] In some embodiments, the filler sequence may be located between two regions, such as, but not limited to, the 5' ITR and an exon region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a promoter region and a payload region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a promoter region and an intron region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a promoter region and an enhancer region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a promoter region and a polyadenylation signal sequence region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a promoter region and an exon region. In some embodiments, a filler sequence may be located between two regions, such as, but not limited to, a promoter region and a 3' ITR.

[0221] In some embodiments, the filler sequence may be located between two regions, such as, but not limited to, a payload region and an intron region. In some embodiments, the filler sequence may be located between two regions, such as, but not limited to, a payload region and an enhancer region. In some embodiments, the filler sequence may be located between two regions, such as, but not limited to, a payload region and a polyadenylation signal sequence region. In some embodiments, the filler sequence may be located between two regions, such as, but not limited to, a payload region and an exon region.

[0222] In some embodiments, the filler sequence may be located between two regions, for example, but not limited to, between the payload region and the 3' ITR. Self-complementary and single-stranded vectors In some embodiments, the AAV vectors used in this disclosure are single-stranded vectors (ssAAV).

[0223] In some embodiments, AAV vectors can be self-complementary AAV vectors (scAAV).See, for example, U.S. Patent No. 7,465,583.scAAV vectors contain DNA strands that anneal together to form double-stranded DNA.By skipping the synthesis of the second strand, scAAV can be rapidly expressed in cells.

[0224] In some embodiments, the AAV vector used in this disclosure is a scAAV. Methods for producing and / or modifying AAV vectors have been disclosed in the art, such as pseudotyped AAV vectors (WO200028004; WO200123001; WO2004112727; WO2005005610 and WO2005072364, the contents of each of which are incorporated herein by reference in their entirety).

[0225] Genome size In some embodiments, the viral genome of the AAV particles of the present disclosure may be single-stranded or double-stranded. The size of the vector genome may be small, medium, large, or maximum size.

[0226] In some embodiments, a vector genome comprising a nucleic acid sequence encoding FXN described herein may be a small, single-stranded vector genome. The small, single-stranded vector genome may be about 2.7 kb to about 3.5 kb in size, such as about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, or about 3.5 kb in size. In some embodiments, the small, single-stranded vector genome may be 3.2 kb in size.

[0227] In some embodiments, a vector genome comprising a nucleic acid sequence encoding FXN described herein may be a small, double-stranded vector genome. The small, double-stranded vector genome may be about 1.3 to about 1.7 kb in size, such as about 1.3, about 1.4, about 1.5, about 1.6, or about 1.7 kb in size. In some embodiments, the small, double-stranded vector genome may be 1.6 kb in size.

[0228] In some embodiments, a vector genome comprising a nucleic acid sequence encoding FXN described herein may be a medium-sized single-stranded vector genome. The medium-sized single-stranded vector genome may be about 3.6 to about 4.3 kb in size, such as about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, or about 4.3 kb in size. In some embodiments, the medium-sized single-stranded vector genome may be 4.0 kb in size.

[0229] In some embodiments, the vector genome comprising the nucleic acid sequence encoding FXN described herein may be a medium-sized, double-stranded vector genome. The medium-sized, double-stranded vector genome may be about 1.8 to about 2.1 kb in size, such as about 1.8, about 1.9, about 2.0, or about 2.1 kb in size. In some embodiments, the medium-sized, double-stranded vector genome may be 2.0 kb in size. In addition, the vector genome may include a promoter and a poly(A) tail.

[0230] In one embodiment, a vector genome comprising a nucleic acid sequence encoding FXN described herein may be a large single-stranded vector genome. The large single-stranded vector genome may be 4.4 to 6.0 kb in size, such as approximately 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, and 6.0 kb in size. As a non-limiting example, the large single-stranded vector genome may be 4.7 kb in size. As another non-limiting example, the large single-stranded vector genome may be 4.8 kb in size. As yet another non-limiting example, the large single-stranded vector genome may be 6.0 kb in size.

[0231] In one embodiment, the vector genome containing the nucleic acid sequence encoding FXN described herein may be a large double-stranded vector genome. The large double-stranded vector genome may be 2.2 to 3.0 kb in size, such as approximately 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, and 3.0 kb in size. As a non-limiting example, the large double-stranded vector genome may be 2.4 kb in size.

[0232] payload In some embodiments, the present disclosure provides constructs that improve the expression of FXN delivered by a gene therapy vector.

[0233] In some aspects, the present disclosure relates to compositions containing or comprising nucleic acid sequence(s) encoding frataxin (FXN) or functional fragment(s) thereof, and methods of administering these compositions in vitro or in vivo in human and / or animal models of disease.

[0234] The AAV particles of the present disclosure may contain a nucleic acid molecule encoding at least one "payload." As used herein, "payload" or "payload region" refers to one or more polynucleotides or polynucleotide regions encoded by or within the viral genome, or the expression product of such a polynucleotide or polynucleotide region, e.g., a transgene, a polypeptide, or a multipolypeptide, e.g., a polynucleotide encoding FXN or a variant thereof. The payload may include any nucleic acid known in the art to be useful for expression of FXN (by supplementation of the protein product or gene replacement using a regulatory nucleic acid) in target cells transduced or contacted with an AAV particle carrying the payload.

[0235] The payload construct may comprise a combination of coding and non-coding nucleic acid sequences. Any section, fragment, or entire viral genome, and payload constructs therein, may be codon optimized.

[0236] In some embodiments, the nucleic acid sequence of the AAV particle may be a payload construct that includes at least one portion encoding FXN. In some embodiments, the payload construct encodes more than one payload. By way of a non-limiting example, a payload construct encoding more than one payload can be replicated and packaged into a viral particle. A target cell transduced with a viral particle containing more than one payload can express each of the payloads in a single cell.

[0237] In some embodiments, the payload construct may encode a coding or non-coding RNA. In certain embodiments, the adeno-associated viral vector particle further comprises at least one cis-element selected from the group consisting of a Kozak sequence, a backbone sequence, and an intron sequence.

[0238] In some embodiments, the payload is a polypeptide, which may be a peptide or protein. The protein encoded by the payload construct may include a secreted protein, an intracellular protein, an extracellular protein, and / or a membrane protein. The encoded protein may be structural or functional. The protein encoded by the payload construct includes, but is not limited to, a mammalian protein. In certain embodiments, the AAV particle contains a viral genome encoding FXN or a variant thereof. AAV particles encoding a payload may be useful in the fields of human disease, veterinary applications, and various in vivo and in vitro settings.

[0239] In some embodiments, the payload may include a polypeptide that serves as a marker protein for assessing cell transformation and expression, a fusion protein, a polypeptide with a desired biological activity, a gene product that can complement a genetic defect, an RNA molecule, a transcription factor, and other gene products of interest for regulation and / or expression. In some embodiments, the payload may include a nucleotide sequence (e.g., a transposon, a transcription factor) that provides a desired effect or regulatory function.

[0240] The encoded payload may include a gene therapy product. Gene therapy products may include, but are not limited to, polypeptides, RNA molecules, or other gene products that provide a desired therapeutic effect when expressed in target cells. In some embodiments, gene therapy products may include substitutes for non-functional genes, or genes that are absent, expressed in insufficient amounts, or mutated. In some embodiments, gene therapy products may include substitutes for non-functional proteins or polypeptides, or proteins or polypeptides that are absent, expressed in insufficient amounts, misfolded, degraded too quickly, or mutated. For example, gene therapy products may include FXN polypeptides or polynucleotides encoding FXN polypeptides for treating FXN deficiency or FA.

[0241] In some embodiments, the payload encodes messenger RNA (mRNA). As used herein, the term "messenger RNA" (mRNA) refers to any polynucleotide that encodes a polypeptide of interest and can be translated to produce the encoded polypeptide of interest in vitro, in vivo, in situ, or ex vivo. Certain embodiments provide mRNA that encodes FXN or its variants.

[0242] Components of mRNA include, but are not limited to, the coding region, 5'-UTR (untranslated region), 3'-UTR, 5'-cap, and polyA tail. In some embodiments, the encoded mRNA or any portion of the AAV genome may be codon-optimized.

[0243] In some embodiments, the protein or polypeptide encoded by a payload construct encoding FXN or a variant thereof is about 50 to about 4500 amino acid residues in length (in this context, hereinafter "X amino acids in length" refers to X amino acid residues). In some embodiments, the encoded protein or polypeptide is 50 to 2000 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 1000 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 1500 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 1000 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 800 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 600 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 400 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 200 amino acids in length. In some embodiments, the encoded protein or polypeptide is 50 to 100 amino acids in length.

[0244] The payload construct encoding the payload may contain or encode a selectable marker. A selectable marker may comprise a gene sequence in the host cell, or a protein or polypeptide encoded by a gene sequence expressed in the host cell, that allows for identification, selection, and / or purification of the host cell from a population of cells that may or may not express the selectable marker. In some embodiments, the selectable marker provides resistance to survive a selection process that would otherwise kill the host cell, such as treatment with an antibiotic. In some embodiments, the antibiotic selectable marker may comprise one or more antibiotic resistance factors, including, but not limited to, neomycin resistance (e.g., neo), hygromycin resistance, kanamycin resistance, and / or puromycin resistance.

[0245] In some embodiments, any nucleic acid sequence encoding a protein or polypeptide can be used as a selectable marker, including recognition by a specific antibody. In some embodiments, the payload construct encoding the payload may include a selectable marker, including, but not limited to, β-lactamase, luciferase, β-galactosidase, or a cell surface marker, e.g., CD4 or any other receptor gene, including truncated nerve growth factor (NGFR), as that term is understood in the art (for GFP, see WO 96 / 23810; Heim et al., Current Biology 2:178-182 (1996); Heim et al., Proc. Natl. Acad. Sci. USA (1995); or Heim et al., Science 373:663-664 (1995); for β-lactamase, see WO 96 / 30540); the contents of each of which are incorporated herein by reference in their entireties.

[0246] In some embodiments, the payload construct encoding a selectable marker may comprise a fluorescent protein. The fluorescent proteins described herein may include any fluorescent marker, including but not limited to green, yellow, and / or red fluorescent proteins (GFP, YFP, and / or RFP). In some embodiments, the payload construct encoding a selectable marker may comprise a human influenza hemagglutinin (HA) tag.

[0247] In certain embodiments, the nucleic acid for expression of the payload in the target cell will be incorporated into the viral genome and located between the two ITR sequences. Payload: Frataxin In some embodiments, the payload is a frataxin protein. As used herein, the terms "frataxin protein" or "FXN protein" are used interchangeably with "frataxin polypeptide" or "FXN polypeptide" and encompass wild-type FXN as well as its functional variants. A functional variant is a variant that retains some or all of the activity of its wild-type counterpart to achieve a desired therapeutic effect. For example, in some embodiments, a functional variant is effectively used in gene therapy to treat disorders or conditions, such as FXN deficiency or FA. Unless otherwise indicated, the FXN variants described herein (e.g., in connection with the constructs, vectors, genomes, methods, kits, compositions, etc. of the present disclosure) are functional variants.

[0248] Friedreich's ataxia (FA) is an autosomal recessive disorder that occurs when the frataxin (FXN) gene contains an amplified intronic GAA repeat (an example of a trinucleotide repeat expansion). See Parkinson et al., Journal of Neurochemistry, 2013, 126(Suppl. 1), 103-117 (the contents of which are incorporated herein by reference in their entirety). The GAA repeat expansion within the gene causes a decrease in FXN protein levels. FXN is an iron-binding protein responsible for iron-sulfur cluster formation. One consequence of FXN protein deficiency is mitochondrial iron overload, which can cause damage to many proteins. See Nageshwaran and Festenstein, Frontiers in Neurology, Vol. 6, Art. 262 (2015), the contents of which are incorporated herein by reference in their entirety. The FXN gene is located on chromosome 9. See Sandi et al., Frontiers in Genetics, Vol. 5, Art. 165 (June 2014), the contents of which are incorporated herein by reference in their entirety.

[0249] The mutant gene contains an expanded GAA triplet repeat in the first intron, and in a few cases, point mutations have been detected. Because the deletion is located in an intron (which is removed from the mRNA transcript between transcription and translation), the mutation does not result in the production of abnormal FXN protein. See Nageshwaran and Festenstein, Frontiers in Neurology, Vol. 6, Art. 262 (2015). Instead, the mutation causes gene silencing (i.e., the mutation reduces gene transcription) through the induction of heterochromatin structure in a manner similar to the variegated position effect. In addition to reducing FXN protein expression, long tracts of GAA repeats induce chromosome breakage in in vivo yeast studies.

[0250] Low levels of FXN protein result in insufficient biosynthesis of iron-sulfur clusters, necessary for mitochondrial electron transport and the assembly of functional aconitase, as well as dysregulated iron metabolism throughout the cell. See Nageshwaran and Festenstein, Frontiers in Neurology, Vol. 6, Art. 262 (2015). In normal individuals, the FXN gene encodes the mitochondrial matrix FXN protein. This globular protein, consisting of two alpha helices and seven beta strands, is highly conserved and present in all eukaryotes and some prokaryotes. The FXN protein has various known functions; most notably, it supports iron-sulfur protein synthesis in the electron transport chain, ultimately generating adenosine triphosphate (ATP), the energy currency required to perform metabolic functions in the cell. The FXN protein also regulates iron translocation in mitochondria to provide adequate amounts of reactive oxygen species (ROS) to maintain normal processes. Without the FXN protein, there is a lack of energy in the mitochondria and excess iron causes the production of excess ROS, resulting in further cell damage.

[0251] Other disorders of the central nervous system may eventually be found to be associated with abnormal expression of FXN protein or defects in its quantity or function. Such disorders may include, but are not limited to, neurological or neuromuscular disorders, such as Alzheimer's disease, Huntington's disease, autism, Parkinson's disease and spinal muscular atrophy, or other neurological or neuromuscular diseases, disorders or conditions described herein.

[0252] As used herein, "associated with decreased frataxin protein levels" or "associated with decreased expression" means that one or more symptoms of a disease are caused by lower-than-normal levels of frataxin protein in a target tissue or biofluid, such as blood. A disease or condition associated with decreased frataxin protein levels or expression may be a disorder of the central nervous system. Such a disease or condition may be a neuromuscular or neurological disorder or condition. For example, a disease associated with decreased frataxin protein levels may be FA or another neurological or neuromuscular disorder described herein.

[0253] The present disclosure addresses a need in the art by providing FXN-related therapies deliverable by AAV-based compositions and complexes for the treatment of FA. Although delivery is exemplified in relation to AAV, other viral vectors, non-viral vectors, nanoparticles, or liposomes can similarly be used to deliver therapeutic FXN, including, but not limited to, vector genomes of any AAV serotype, or other viral delivery vehicles or lentiviruses. The observations and teachings extend to any macromolecular structure, including modified cells, that are introduced into the CNS in the manner described herein.

[0254] Sequence identifiers for representative polynucleotide and polypeptide sequences of frataxin that can be used in the viral genomes disclosed herein and that can constitute the frataxin payload are provided in Table 2. Functional variants, e.g., those that retain at least about 90% or at least 95% sequence identity to the sequences shown in Table 2, can also be used. Codon-optimized and other variants (e.g., those with at least about 90% amino acid sequence identity) that encode the same or essentially the same FXN amino acid sequence can also be used.

[0255] [Table 2]

[0256] In some embodiments, the viral genome comprises a payload region encoding a frataxin protein. The encoded frataxin may be from any species, including, but not limited to, human, non-human primate, or rodent.

[0257] In some embodiments, the viral genome comprises a payload region encoding human (Homo sapiens) frataxin or a variant thereof. Various embodiments of the present disclosure provide adeno-associated virus (AAV) particles comprising a viral genome, the viral genome comprising at least one inverted terminal repeat region and a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to the human frataxin (hFXN) sequence of SEQ ID NO: 1725, 1726 and / or 1727, or a variant thereof.

[0258] In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to SEQ ID NO: 1725. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to SEQ ID NO: 1725. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence encoding a polypeptide having at least 98% sequence identity to SEQ ID NO: 1725. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence encoding a polypeptide having at least 99% sequence identity to SEQ ID NO: 1725. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence encoding SEQ ID NO: 1725.

[0259] In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 1728, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1728, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 98% sequence identity to SEQ ID NO: 1728, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 99% sequence identity to SEQ ID NO: 1728, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence of SEQ ID NO: 1728, or a fragment thereof. In some embodiments, the fragment of SEQ ID NO: 1728 comprises nucleotides 221-853 of SEQ ID NO: 1728.

[0260] In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 1823, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1823, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 98% sequence identity to SEQ ID NO: 1823, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 99% sequence identity to SEQ ID NO: 1823, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and the nucleic acid sequence of SEQ ID NO: 1823, or a fragment thereof. In some embodiments, the nucleic acid sequence further comprises a stop codon.

[0261] In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 1824, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1824, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 98% sequence identity to SEQ ID NO: 1824, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 99% sequence identity to SEQ ID NO: 1824, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence of SEQ ID NO: 1824, or a fragment thereof.

[0262] In some embodiments, the FXN polypeptide is derived from a non-human primate, such as the cynomolgus monkey Macaca fascicularis (cynoFXN) FXN sequence. Particular embodiments provide FXN polypeptides such as the humanized Macaca fascicularis (HcynoFXN) sequence. In some embodiments, the FXN polypeptide sequence has at least about 90% sequence identity to the art-accepted canonical human FXN amino acid sequence of SEQ ID NO: 1725, which may be encoded by the nucleic acid sequence of SEQ ID NO: 1728. In some embodiments, the FXN polypeptide sequence has at least about 90% sequence identity to the art-accepted canonical human FXN amino acid sequence of SEQ ID NO: 1726, which is encoded by the nucleic acid sequence of SEQ ID NO: 1729. In some embodiments, the FXN polypeptide sequence has at least about 90% sequence identity to the art-accepted canonical human FXN amino acid sequence of SEQ ID NO: 1727, which is encoded by the nucleic acid sequence of SEQ ID NO: 1730.

[0263] In some embodiments, the viral genome comprises a payload region encoding cynomolgus or crab-eating (long-tailed) monkey (Macaca fascicularis) frataxin or a variant thereof.

[0264] In some embodiments, the viral genome comprises a payload region encoding rhesus monkey (Macaca mulatta) frataxin or a variant thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 1822, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 1822, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 98% sequence identity to SEQ ID NO: 1822, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence having at least 99% sequence identity to SEQ ID NO: 1822, or a fragment thereof. In some embodiments, the AAV viral genome comprises at least one inverted terminal repeat region and a nucleic acid sequence of SEQ ID NO: 1822, or a fragment thereof.

[0265] In some embodiments, a frataxin polypeptide may comprise an amino acid sequence having 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of those described above.

[0266] In some embodiments, the frataxin polypeptide may be encoded by a nucleic acid sequence having 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to any of the foregoing.

[0267] Viral genome: promoter In some embodiments, the payload region of the viral genome comprises elements that enhance or regulate payload expression, such as, but not limited to, a promoter. The promoter may be a wild-type or engineered promoter, or a combination thereof. In some embodiments, the viral genome comprises at least one promoter. In some embodiments, the viral genome comprises more than one promoter.

[0268] In some embodiments, the promoter is a wild-type frataxin promoter or a derivative thereof (e.g., a truncation or variant). A suitable derivative of the wild-type frataxin promoter is one that is functional, e.g., effective to express a payload at at least a minimally detectable level.

[0269] In some embodiments, the promoter is an engineered frataxin promoter. Shorter variants of the frataxin promoter are included herein. The frataxin promoter may be 200 to 1400 nt in length, or any length therebetween. In some embodiments, the frataxin promoter variant may be 223, 363, 534, 747, 906, 1060, 1226, or 1353 nucleotides in length. The frataxin promoter variant may be shorter than the wild-type frataxin promoter sequence due to deletions in any region of the promoter sequence, for example, but not limited to, the 5' end of the promoter sequence, the 3' end of the promoter sequence, or within the promoter sequence.

[0270] In some embodiments, the promoter is a combination of one or more of any of the promoters described herein. In some embodiments, the promoter is used in conjunction with an enhancer sequence. In some embodiments, the enhancer sequence may be derived from a cytomegalovirus immediate early gene (CMVie). In some embodiments, the enhancer may be located upstream (5') of the promoter. In some embodiments, the enhancer comprises SEQ ID NO: 1777.

[0271] In some embodiments, the promoter is a CBA promoter or a derivative thereof (e.g., a truncation or variant). It is understood that a suitable derivative of a CBA promoter is functional, e.g., effective to express a payload.

[0272] In some embodiments, the CBA promoter comprises, listed from 5' to 3', a CMVie enhancer, a backbone sequence, and a CB promoter sequence. Each of the three components (CMVie enhancer, backbone, and CB sequences) may be of different lengths between variants.

[0273] In some embodiments, the CBA promoter comprises, when listed from 5' to 3', a backbone sequence and a CB promoter sequence. In some embodiments, the CBA promoter comprises a CB promoter sequence.

[0274] In some embodiments, the CBA promoter may be 100 to 700 nt in length, or any length therebetween. In some embodiments, the CBA promoter variant may be 100, 180, 260, 270, 332, 412, 492, or 572 nucleotides in length. The CBA promoter variant may be shorter than the wild-type CBA promoter sequence due to deletions in the enhancer, backbone, or any region of the promoter sequence, including, but not limited to, the 5' end of the promoter sequence, the 3' end of the promoter sequence, or within the promoter sequence.

[0275] In some embodiments, the promoter is a CMV promoter or a derivative thereof (e.g., a truncation or variant). It is understood that a suitable derivative of a CMV promoter is functional, e.g., effective for expressing a payload. The CMV promoter may include a CMV enhancer and a CMV promoter sequence, or only a CMV promoter sequence. The CMV enhancer and the CMV promoter sequence may be different lengths between promoter variants.

[0276] In some embodiments, the CMV promoter may be 50 to 700 nt in length, or any length therebetween. In some embodiments, the CMV promoter variant may be 55, 109, 163, 217, 289, 361, 433, or 505 nucleotides in length. The CMV promoter variant may be shorter than the wild-type CMV promoter sequence due to deletions in any region of the enhancer or promoter sequence, including, but not limited to, the 5' end of the promoter sequence, the 3' end of the promoter sequence, or within the promoter sequence.

[0277] In some embodiments, the promoter is a deletion variant of the parent promoter sequence, in which one or more nucleotides have been removed from the parent sequence. In some embodiments, the promoter is an insertional variant of the parent promoter sequence, in which one or more nucleotides have been added to the parent sequence.

[0278] In some embodiments, the promoter comprises one or more mutations compared to the parent promoter sequence. In some embodiments, the promoter is modified in one or more ways (eg, deletion, mutation, and / or insertion) to create a promoter variant.

[0279] In some embodiments, the promoter may comprise a sequence, fragment, or variant thereof of any of the sequences in Table 3. For example, the promoter may comprise a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1734-1777, e.g., a sequence having the specified percent identity and providing some or all of the same function as a sequence selected from the group consisting of SEQ ID NOs: 1734-1777. In some embodiments, the promoter is or is derived from a CMV promoter and comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1743-1751, 1767, 1772-1772, and 1777. In some embodiments, the promoter is or is derived from a CBA promoter and comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1734-1742, 1760-1766, 1768, and 1775-1776. In some embodiments, the promoter is or is derived from an FXN promoter and comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to a sequence selected from the group consisting of SEQ ID NOs: 1752-1759 and 1769-1770.

[0280] In some embodiments, the promoter may comprise a combination of more than one of any of those listed in Table 3. In some embodiments, the promoter sequence may further comprise at least one of the intron / exon sequences given in Table 6.

[0281] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1738. In some embodiments, the promoter is SEQ ID NO: 1738. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824). In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0282] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1740. In some embodiments, the promoter is SEQ ID NO: 1740. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824). In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0283] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1742. In some embodiments, the promoter is SEQ ID NO: 1742. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824). In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0284] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1750. In some embodiments, the promoter is SEQ ID NO: 1750. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824). In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0285] In some embodiments, the promoter used in the viral genomes disclosed herein comprises any one of the promoter sequences in Table 3. In Table 3, CMV represents "cytomegalovirus"; CBA represents "chicken beta-actin" which may have a CMV IE ("immediate early") enhancer region and a promoter region; CAG represents a CMV enhancer, a CBA promoter, and a rabbit beta-globin splice acceptor site; FXN represents "frataxin"; and mCBA represents a variant of the CBA promoter generated using PCR.

[0286] [Table 3]

[0287] In some embodiments, a promoter is used to regulate frataxin expression in a target cell. In certain embodiments, a promoter can be used to increase frataxin expression in a target cell to a level higher than the level of normal endogenous frataxin expression. In certain embodiments, a promoter can be used to induce frataxin expression in a target cell to a level close to or equal to the level of normal endogenous frataxin expression.

[0288] In some embodiments, junction sequences can be used in combination with promoters described herein, including, but not limited to, those listed in Table 3. In certain embodiments, the junction sequence can be located 5' to the promoter in the viral genome. In certain embodiments, the junction sequence can be located 3' to the promoter in the viral genome. In certain embodiments, the viral genome can include more than one junction sequence. As a non-limiting example, the viral genome can include a junction sequence at the 5' end of the promoter and at the 3' end of the promoter. The junction sequences can be the same sequence, two different sequences, or sequences split on either side of the promoter sequence. In certain embodiments, the junction sequence comprises SEQ ID NO: 1813. In certain embodiments, the junction sequence comprises SEQ ID NO: 1814.

[0289] In some embodiments, promoters are used to enhance frataxin expression in target cells (e.g., nervous system or cardiac tissue). Frataxin expression can be increased 0.01 to 100 (0.01 to 100x) times the endogenous frataxin expression for that target cell. In some embodiments, promoters are used to maintain frataxin expression in target cells at 0.5 to 3x (e.g., 0.5 to 1x, 1 to 1.5x, 1.5 to 2x, 2 to 2.5x, 2.5 to 3x) the endogenous frataxin expression (i.e., normal human levels or approximately 5.5 to 32.8 ng / mg protein).

[0290] In some embodiments, a promoter, e.g., a promoter in Table 3, is used in an AAV vector genome further comprising a sequence encoding a frataxin polypeptide sequence, e.g., a human frataxin polypeptide sequence. In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1742. In some embodiments, the promoter is SEQ ID NO: 1742. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1742 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0291] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1750. In some embodiments, the promoter is SEQ ID NO: 1750. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1750 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0292] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1738. In some embodiments, the promoter is SEQ ID NO: 1738. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1738 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0293] In some embodiments, the promoter comprises a sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1740. In some embodiments, the promoter is SEQ ID NO: 1740. In some embodiments, the AAV vector genome comprises a promoter sequence having at least 90% sequence identity to SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 90% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 90% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence having at least 95% sequence identity to SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having an amino acid sequence at least 95% identical to SEQ ID NO: 1725 (e.g., a payload region comprising a nucleic acid sequence at least 95% identical to SEQ ID NO: 1824). In some embodiments, the AAV vector genome comprises a promoter sequence of SEQ ID NO: 1740 and a payload region encoding a frataxin polypeptide having the amino acid sequence of SEQ ID NO: 1725 (e.g., a payload region comprising SEQ ID NO: 1824), and / or further comprising one or more of the sequences provided in Tables 5-11 or a 95% identical variant thereof. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1728 or a fragment thereof, optionally comprising nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824.

[0294] In various embodiments, any of the promoters disclosed herein, e.g., a promoter from Table 3 or a promoter having 90% or greater homology thereto, may be paired in an AAV viral vector genome, alone or in combination with additional sequences, e.g., filler sequence(s), with one or more of the components disclosed in Tables 5-11 or a component(s) having 90% or greater homology thereto. In some embodiments, the AAV vector genome may contain multiple copies (e.g., two, three, or more copies) of one or more viral genome components described herein. In some embodiments, the viral genome contains two miR binding sites (e.g., two miR122 binding sites). In some embodiments, the viral genome contains three miR binding sites (e.g., three miR122 binding sites). In some embodiments, the viral genome comprises any of the promoters disclosed herein, e.g., a promoter from Table 3, or a promoter having 90% or more homology thereto, along with one or more components provided in any of Tables 5-11 or otherwise described herein, in the 5' to 3' order shown in any of Tables 4, 12, 13, 14, 15, 16, or 17. In some embodiments, the viral genome comprises a promoter provided in Table 3, along with one or more components provided in any of Tables 5-11 or otherwise described herein, in the 5' to 3' order shown in any of Tables 4, 12, 13, 14, 15, 16, or 17. In some embodiments, the viral genome comprises all of the components in the 5' to 3' order shown in any of Tables 4, 12, 13, 14, 15, 16, or 17.

[0295] For example, a promoter comprising or having 90% or greater homology thereto, SEQ ID NO: 1742, may be paired in an AAV vector genome with any of the components in Tables 5-11 (or component(s) having 90% or greater homology thereto), e.g., a promoter located between the 5' ITR sequence and ie1 exon 1 (e.g., directly contacting these two other components or separated by one or more non-coding sequences). In some embodiments, the viral genome comprises three miR122 binding sites. In some embodiments, the viral genome further comprises a payload region, e.g., one encoding a frataxin protein.

[0296] In another example, a promoter comprising or having 90% or greater homology thereto, SEQ ID NO: 1750, may be paired in an AAV vector genome with a promoter located between any of the components in Tables 5-11 (or component(s) having 90% or greater homology thereto), e.g., the 5' ITR sequence and ie1 exon 1 (e.g., directly contacting these two other components or separated by one or more non-coding sequences). In some embodiments, the viral genome comprises three miR122 binding sites. In some embodiments, the viral genome further comprises a payload region, e.g., one encoding a frataxin protein.

[0297] For example, a promoter comprising or having 90% or greater homology thereto, SEQ ID NO: 1738, may be paired in an AAV vector genome with any of the components in Tables 5-11 (or component(s) having 90% or greater homology thereto), e.g., a promoter located between the 5' ITR sequence and ie1 exon 1 (e.g., directly contacting the two other components or separated by one or more non-coding sequences). In some embodiments, the viral genome comprises three miR122 binding sites. In some embodiments, the viral genome further comprises a payload region, e.g., one encoding a frataxin protein.

[0298] In another example, a promoter comprising or having 90% or greater homology thereto may be paired in an AAV vector genome with any of the components in Tables 5-11 (or component(s) having 90% or greater homology thereto), e.g., a promoter located between the 5' ITR sequence and ie1 exon 1 (e.g., directly contacting these two other components or separated by one or more non-coding sequences). In some embodiments, the viral genome comprises three miR122 binding sites. In some embodiments, the viral genome further comprises a payload region, e.g., one encoding a frataxin protein.

[0299] In some embodiments, the promoter is or is derived from a CBA promoter. The CBA promoter can drive expression of a payload in various tissues of a subject. As a non-limiting example, expression of FXN using the CBA promoter (the promoter is set forth as SEQ ID NO: 1776, and ITR-to-ITR is provided as SEQ ID NO: 1778) is shown in Example 4, including Tables 16-28, of co-owned International Patent Application No. PCT / US2019 / 032387, the contents of which are incorporated herein by reference in their entirety. Expression of FXN in mice after IV injection is shown in Table 16 of co-owned International Patent Application No. PCT / US2019 / 032387, where expression is seen in the cortex, lumbar spinal cord, lumbar dorsal root ganglion, trigeminal ganglion, heart, and liver with VOY101 particles having a CBA promoter. Expression of FXN in NHPs after IV injection is shown in Table 18 of co-owned International Patent Application No. PCT / US2019 / 032387, where expression is seen in the brainstem, cervical spinal cord, thoracic spinal cord, lumbar spinal cord, cervical DRG, thoracic DRG, lumbar / sacral DRG, ventricle, atrium, liver, soleus muscle, and jejunum with VOY101 particles carrying the CBA promoter. 11 VG / kg, 2 x 10 12 VG / kg or 2 x 10 13Expression of FXN in NHPs after IV injection at different doses (6.7 x 10 VG / kg) is shown in Table 19 of co-owned International Patent Application No. PCT / US2019 / 032387, where expression is seen in the brainstem, cerebellum, cervical spinal cord, thoracic spinal cord, lumbar spinal cord, cervical DRG, thoracic DRG, lumbar / sacral DRG, ventricle, atrium, liver, kidney, lung, soleus muscle, and / or spleen with VOY201 particles bearing the CBA promoter. 12 VG / kg or 4.89 x 10 13 Expression of FXN in NHPs after IV injection at 1000kJ / kg (VG / kg) is shown in Table 20 of co-owned International Patent Application No. PCT / US2019 / 032387, where expression is seen in the brainstem, cerebellum, cortex, cervical spinal cord, thoracic spinal cord, lumbar spinal cord, cervical DRG, thoracic DRG, lumbar / sacral DRG, ventricle, atrium, liver, kidney, soleus muscle, thoracic sympathetic chain ganglia, and / or adrenal glands with VOY101 particles bearing the CBA promoter. Distribution of vector genomes after IV injection in mice is shown in Table 17 of co-owned International Patent Application No. PCT / US2019 / 032387, where distribution is seen in the cortex, lumbar spinal cord, thoracic dorsal root ganglia, trigeminal ganglia, heart, and liver with VOY101 and AAV9 particles bearing the CBA promoter. The distribution of vector genomes after IV injection in NHPs is shown in Table 18 of co-owned International Patent Application No. PCT / US2019 / 032387, where distribution is seen in the frontal cortex, striatum, brainstem, cerebellum, cervical spinal cord, thoracic spinal cord, cervical dorsal root ganglia, thoracic dorsal root ganglia, lumbar / sacral dorsal root ganglia, ventricle, atrium, liver, kidney, lung, soleus muscle, jejunum, and spleen for VOY101 particles with a CBA promoter. 11 VG / kg, 2 x 10 12 VG / kg or 2 x 10 13The distribution of vector genomes in NHPs after IV injection at different doses (6.7 x 10 VG / kg) is shown in Table 19 of co-owned International Patent Application No. PCT / US2019 / 032387, where distribution is seen in the frontal cortex, striatum, brainstem, cerebellum, cervical spinal cord, thoracic spinal cord, lumbar spinal cord, cervical DRG, thoracic DRG, lumbar / sacral DRG, ventricle, atrium, liver, kidney, lung, soleus muscle, jejunum, and / or spleen for VOY201 particles with a CBA promoter. 12 VG / kg or 4.89 x 10 13 The distribution of vector genomes in NHPs after IV injection at 1000kJ / kg (VG / kg) is shown in Table 20 of co-owned International Patent Application No. PCT / US2019 / 032387, where distribution is seen in the motor cortex, sensorimotor cortex, striatum, brainstem, cerebellar cortex, cervical spinal cord, thoracic spinal cord, lumbar spinal cord, thoracic spinal cord, cervical DRG, thoracic DRG, lumbar / sacral DRG, ventricle, atrium, liver, kidney, soleus muscle, jejunum, spleen, thoracic sympathetic trunk ganglion and / or adrenal gland for VOY101 particles with a CBA promoter.

[0300] In some embodiments, the promoter is or is derived from a promoter, including the CMVie enhancer, CBA, CMV, frataxin promoter, truncated CBA, and / or truncated CMV promoter. The promoter can drive expression of the payload in various tissues of a subject. As a non-limiting example, a mouse model of Friedreich's ataxia can be used to evaluate the in vivo distribution, expression, and efficacy of IV administration of VOY101 particles carrying FXN, as shown in Example 5 of co-owned International Patent Application No. PCT / US2019 / 032387 (the contents of which are incorporated herein by reference in their entirety). In certain embodiments, promoters, such as, but not limited to, those in Table 3, can be evaluated for driving expression of FXN in mice, as outlined in Example 5 of co-owned International Patent Application No. PCT / US2019 / 032387. As another non-limiting example, a Friedreich's ataxia NHP model for evaluating the in vivo distribution and expression of IV dosing of VOY101 particles carrying FXN is shown in Example 5 of co-owned International Patent Application No. PCT / US2019 / 032387, the contents of which are incorporated herein by reference in their entirety. In certain embodiments, promoters, such as, but not limited to, those in Table 3, can be evaluated for driving expression of FXN in NHPs, as outlined in Example 5 of co-owned International Patent Application No. PCT / US2019 / 032387.

[0301] In some embodiments, the promoter is or is derived from a CBA promoter, which can drive expression of the payload in various tissues of a subject. As a non-limiting example, expression of FXN using the CBA promoter (the promoter is set forth as SEQ ID NO: 1776, and ITR-to-ITR is provided as SEQ ID NO: 1778) is shown in Example 14, including Tables 33-34, of co-owned International Patent Application No. PCT / US2019 / 032387, the contents of which are incorporated herein by reference in their entirety. Expression of FXN in mice after IV injection is shown in Table 33 of co-owned International Patent Application No. PCT / US2019 / 032387, where expression is seen in the cortex, striatum, hippocampus, brainstem, thoracic spinal cord, thoracic DRG, heart, and / or liver with VOY101, VOY801, and / or VOY1101 particles bearing the CBA promoter. The distribution of vector genomes after IV injection in mice is shown in Table 34 of co-owned International Patent Application No. PCT / US2019 / 032387, where distribution is seen in the cortex, striatum, hippocampus, brainstem, thoracic spinal cord, heart, and liver for VOY101, VOY801, and / or VOY1101 particles with the CBA promoter. As another non-limiting example, expression of FXN using the CBA promoter (the promoter is set forth as SEQ ID NO: 1776, and the ITR-to-ITR sequence is provided as SEQ ID NO: 1778) in VOY701 and VOY101 capsids is shown in Example 14, including Tables 35-36, of co-owned U.S. Provisional Patent Application No. 62 / 839,889, the contents of which are incorporated herein by reference in their entirety. Expression of FXN in mice after IV injection is shown in Table 35 of co-owned U.S. Provisional Patent Application No. 62 / 839,889, where expression is seen in the cortex, striatum, hippocampus, brainstem, thoracic spinal cord and / or liver for VOY701 and / or VOY101 particles with a CBA promoter. Distribution of vector genomes after IV injection in mice is shown in Table 36 of co-owned U.S. Provisional Patent Application No. 62 / 839,889, where distribution is seen in the cortex, striatum, hippocampus, brainstem, thoracic spinal cord and / or liver for VOY701 and / or VOY101 particles with a CBA promoter.

[0302] In some embodiments, the AAV particles described herein comprise a viral genome having a payload region encoding a frataxin protein, which may be engineered to optimize frataxin expression in target cells.

[0303] Viral genome: ITR-to-ITR sequence containing the frataxin payload Any of the components described herein can be used to design and optimize the ITR-to-ITR sequence of a viral genome for desired frataxin expression. The viral genome can include any number of components, including, but not limited to, one or more of ITRs, enhancers, promoters, introns, UTRs, payload regions, tags or selectable markers, miR binding or target sites, framework regions, polyA sequences, and / or filler sequences. Each of these components can be present 0, 1, 2, or more than 2 times in a given viral genome.

[0304] Each of the ITRs, promoters, enhancers, introns, exons, payloads, tags, miR binding sites, polyA and / or filler components may be selected independently or in any combination from the sequences provided in Tables 3 and 5-11.

[0305] In some embodiments, the AAV viral genome comprises a 5' ITR, an enhancer, an intron, a payload region, an optional tag, up to three miR binding sites, a polyA sequence, an optional filler sequence, and a 3' ITR. In some embodiments, the 5' ITR is an AAV2 ITR. In some embodiments, the 5' ITR comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1811. In some embodiments, the enhancer comprises ie1 exon 1 and ie1 intron 1 or a fragment thereof. In some embodiments, the enhancer comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1817 and / or 1819. In some embodiments, the enhancer comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to one or more human beta-globin sequences, e.g., SEQ ID NO: 1816, 1820, and / or 1821. In some embodiments, the enhancer comprises SEQ ID NOs: 1817, 1819, 1820 and 1821. In some embodiments, the enhancer comprises SEQ ID NO: 1816.

[0306] In some embodiments, the payload region comprises a nucleic acid sequence encoding a polypeptide having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1725, 1726, 1727, 1731, 1732, or 1733, e.g., at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1725. In some embodiments, the payload region comprises a nucleic acid sequence having at least 90%, at least 95%, at least 99%, or 100% sequence identity to SEQ ID NO: 1728, 1729, 1730, or a fragment thereof. In some embodiments, a fragment of SEQ ID NO: 1728 comprises nucleotides 221-853 of SEQ ID NO: 1728. In some embodiments, the frataxin polypeptide is encoded by a nucleic acid sequence comprising SEQ ID NO: 1822, 1823, or 1824. In some embodiments, no tag is present. In some embodiments, a tag is present and is a human influenza hemagglutinin HA tag. In some embodiments, the HA tag comprises SEQ ID NO: 1825. In some embodiments, no miR binding site is present. In some embodiments, at least one miR binding site is present and comprises a miR122 binding site. In some embodiments, the miR122 binding site comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1827. In some embodiments, the AAV vector genome comprises three copies of a miR122 binding site, e.g., three copies of SEQ ID NO: 1827 or a variant thereof having at least 90% sequence identity. In some embodiments, the miR binding site series comprising three copies of a miR122 binding site comprises SEQ ID NO: 1826. In some embodiments, the viral genome comprises a human growth hormone polyA sequence. In some embodiments, the viral genome comprises a polyA sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1828. In some embodiments, the AAV viral genome further comprises a filler sequence, e.g., an albumin filler sequence.In some embodiments, the filler sequence comprises any of the sequences given by SEQ ID NOs: 1829-1842. In some embodiments, the 3' ITR is an AAV2 ITR. In some embodiments, the 3' ITR comprises a sequence at least 90%, at least 95%, at least 99%, or 100% identical to SEQ ID NO: 1812.

[0307] In certain embodiments, the AAV particle comprises at least one cis-element, including, but not limited to, a Kozak sequence, a scaffold sequence, and / or an intron sequence. Certain embodiments provide that the AAV particle further comprises a promoter region. For example, the promoter may comprise one from a CBA, CMV, FXN, and / or SV40 gene, or any of their variants. Non-limiting examples of ITR-to-ITR sequences of AAV particles comprising a viral genome with a payload region encoding a frataxin protein are listed in Table 4.

[0308] In Table 4, cFXN indicates cynomolgus monkey (Macaca fascicularis) frataxin, hFXN indicates human (Homo sapiens) frataxin, hβglobin indicates human beta-globin, HA indicates human influenza hemagglutinin HA tag, and hGH indicates human growth hormone. Alb indicates albumin. The number after alb indicates the length of the albumin filler. miR-122 BS is the miR-122 binding site. A "-" sign indicates that the construct does not have that component or sequence. A "+" sign indicates that the construct has that component or sequence.

[0309] [Table 4]

[0310] In some embodiments, the AAV particle comprises a viral genome comprising a sequence having a certain percent identity to any of SEQ ID NOs: 1778-1810. The viral genome may have 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 99% or 100% identity to any of SEQ ID NOs: 1778-1810. The viral genome may have 1 to 10%, 10 to 20%, 30 to 40%, 50 to 60%, 50 to 70%, 50 to 80%, 50 to 90%, 50 to 99%, 50 to 100%, 60 to 70%, 60 to 80%, 60 to 90%, 60 to 99%, 60 to 100%, 70 to 80%, 70 to 90%, 70 to 99%, 70 to 100%, 80 to 85%, 80 to 90%, 80 to 95%, 80 to 99%, 80 to 100%, 90 to 95%, 90 to 99%, or 90 to 100% identity to any of SEQ ID NOs: 1778 to 1810. In some embodiments, the viral genome comprises a sequence having at least 80% identity to any of SEQ ID NOs: 1778 to 1810. In some embodiments, the viral genome comprises a sequence having at least 85% identity to any of SEQ ID NOs: 1778-1810. In some embodiments, the viral genome comprises a sequence having at least 90% identity to any of SEQ ID NOs: 1778-1810. In some embodiments, the viral genome comprises a sequence having at least 95% identity to any of SEQ ID NOs: 1778-1810. In some embodiments, the viral genome comprises a sequence having at least 99% identity to any of SEQ ID NOs: 1778-1810.

[0311] In some embodiments, the viral genome comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1797. In some embodiments, the viral genome comprises SEQ ID NO: 1797. In some embodiments, the viral genome comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1801. In some embodiments, the viral genome comprises SEQ ID NO: 1801. In some embodiments, the viral genome comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1808. In some embodiments, the viral genome comprises a sequence having at least 95% sequence identity to SEQ ID NO: 1809. In some embodiments, the viral genome comprises SEQ ID NO: 1809. In some embodiments, the viral genome of an AAV particle of the present disclosure may comprise any combination of sequence regions described in Tables 2-11 or otherwise described herein, encapsulated in any of the capsids listed in Table 1 or described herein.

[0312] In some embodiments, the AAV particle viral genome may comprise at least one sequence region set forth in Tables 2-11. These regions may be located before or after any of the other sequence regions described herein. The viral genome may further comprise more than one copy of one or more sequence regions set forth in Tables 2-11.

[0313] Viral genome: Inverted terminal repeats (ITRs) In some embodiments, the AAV particle viral genome may comprise at least one inverted terminal repeat (ITR) region. The ITR region(s) may independently be, for example, but not limited to, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 300, 310, 311, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321, 322, 323, 324, 325, 326, 327, 330, 331, 332, 333, 334, 335, 336, 337, 338, 339, 340, 341, 342, 343, and 175 nucleotides in length. The lengths of the ITR regions for viral genomes are 75-80, 75-85, 75-100, 80-85, 80-90, 80-105, 85-90, 85-95, 85-110, 90-95, 90-100, 90-115, 95-100, 95-105, 95-120, 100-105, 100-110, 100-125, 105-110, 105-115, 105-130, 110-115, 110-120, 110-135, 115-120, 115-125, 115-140, 120-125, 120-135, 120-140, 120-14 ... The 5' ITR may be 130, 120-145, 125-130, 125-135, 125-150, 130-135, 130-140, 130-155, 135-140, 135-145, 135-160, 140-145, 140-150, 140-165, 145-150, 145-155, 145-170, 150-155, 150-160, 150-175, 155-160, 155-165, 160-165, 160-170, 165-170, 165-175, and 170-175 nucleotides. As a non-limiting example, the viral genome includes a 5' ITR that is...

Claims

1. A composition for use in delivering a nucleotide sequence encoding a frataxin protein to a cell, comprising: the composition comprises an adeno-associated virus (AAV) genome; the AAV genome comprises an engineered promoter and a nucleotide sequence encoding the frataxin protein; The composition, wherein the engineered promoter consists of a nucleotide sequence at least 90% identical to SEQ ID NO:1742.

2. The composition described in claim 1, wherein the manipulated promoter consists of a nucleotide sequence that is at least 95% identical to sequence number 1742.

3. The composition described in claim 1 or 2, wherein the manipulated promoter consists of a nucleotide sequence at least 99% identical to sequence number 1742.

4. A composition described in any one of claims 1 to 3, wherein the manipulated promoter consists of the nucleotide sequence of SEQ ID NO: 1742.

5. A composition described in any one of claims 1 to 4, wherein the frataxin protein comprises an amino acid sequence that is at least 90%, at least 95%, or at least 99% identical to SEQ ID NO: 1725, 1726, or 1727.

6. A composition described in any one of claims 1 to 5, wherein the frataxin protein comprises the amino acid sequence of SEQ ID NO: 1725.

7. A composition described in any one of claims 1 to 6, wherein the nucleotide sequence encoding the frataxin protein comprises a nucleotide sequence that is at least 90%, at least 95%, or at least 99% identical to SEQ ID NO: 1823 or 1824.

8. A composition described in any one of claims 1 to 7, wherein the nucleotide sequence encoding the frataxin protein comprises the nucleotide sequence of SEQ ID NO: 1823 or 1824.

9. A composition described in any one of claims 1 to 8, wherein the AAV genome further comprises a 5' inverted repeat repeat (ITR).

10. The composition of claim 9, wherein the 5' ITR is an AAV2 ITR.

11. The composition of claim 9 or 10, wherein the AAV genome further comprises a 3' ITR.

12. The composition of claim 11, wherein the 3' ITR is an AAV2 ITR.

13. A composition described in any one of claims 1 to 12, wherein the AAV genome further comprises the nucleotide sequence of SEQ ID NO: 1827, or a nucleotide sequence that is at least 95% identical to said sequence.

14. A composition described in any one of claims 1 to 12, wherein the AAV genome further comprises the nucleotide sequence of SEQ ID NO: 1826, or a nucleotide sequence that is at least 95% identical to said sequence.

15. A composition described in any one of claims 1 to 14, wherein the AAV genome further comprises one or more of the following components: intron / exon regions, polyadenylation (polyA) sequences or filler sequences.

16. A composition described in any one of claims 1 to 15, wherein the AAV genome further comprises an intron / exon region and a polyA sequence.

17. The composition described in claim 15 or 16, wherein the AAV genome comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1815 to 1821, or a nucleotide sequence that is at least 95% identical to said sequence.

18. The composition described in claim 17, wherein the AAV genome comprises the nucleotide sequence of SEQ ID NO: 1816, or a nucleotide sequence that is at least 95% identical to said sequence.

19. A composition described in any one of claims 15 to 18, wherein the AAV genome comprises the nucleotide sequence of SEQ ID NO: 1828, or a nucleotide sequence that is at least 95% identical to said sequence.

20. A composition for use in delivering a nucleotide sequence encoding a frataxin protein to a cell, comprising: the composition comprises an adeno-associated virus (AAV) genome; The AAV genome (i) the nucleotide sequence of SEQ ID NO: 1811, or a nucleotide sequence at least 95% identical thereto; (ii) a nucleotide sequence at least 95% identical to SEQ ID NO: 1742; (iii) the nucleotide sequence of SEQ ID NO: 1816, or a nucleotide sequence at least 95% identical thereto; (iv) a nucleotide sequence comprising the nucleotide sequence of SEQ ID NO: 1823, or a nucleotide sequence at least 95% identical thereto; (v) the nucleotide sequence of SEQ ID NO: 1826, or a nucleotide sequence at least 95% identical thereto; (vi) the nucleotide sequence of SEQ ID NO: 1828, or a nucleotide sequence at least 95% identical thereto; (vii) the nucleotide sequence of SEQ ID NO: 1841, or a nucleotide sequence at least 95% identical thereto; and (viii) the nucleotide sequence of SEQ ID NO: 1812, or a nucleotide sequence at least 95% identical thereto; A composition comprising:

21. A composition described in any one of claims 1 to 20, wherein the AAV genome comprises a nucleotide sequence that is at least 90% identical to sequence number 1797.

22. A composition described in any one of claims 1 to 21, wherein the AAV genome comprises a nucleotide sequence that is at least 95% identical to sequence number 1797.

23. A composition described in any one of claims 1 to 22, wherein the AAV genome comprises a nucleotide sequence that is at least 99% identical to sequence number 1797.

24. A composition for use in delivering a nucleotide sequence encoding a frataxin protein to a cell, comprising: The composition comprises an adeno-associated virus (AAV) genome comprising the nucleotide sequence of SEQ ID NO: 1797.

25. A composition described in any one of claims 1 to 24, wherein the composition further comprises an AAV particle comprising the AAV genome and AAV capsid proteins.

26. The composition described in claim 25, wherein the AAV capsid protein is an AAV5 capsid protein or a variant thereof or an AAV9 capsid protein or a variant thereof.

27. ​​A composition described in any one of claims 1 to 26, wherein the cell is a mammalian cell or an insect cell.

28. A composition described in any one of claims 1 to 27, wherein the cells are human cells.

29. A composition described in any one of claims 1 to 27, wherein the cells are HEK293 cells or Sf9 cells.

30. A composition described in any one of claims 1 to 28, wherein the cells are cells of the dorsal root ganglion, dorsal column, Clarke's column, nucleus gracilis and nucleus cuneate, dentate nucleus of the cerebellum, corticospinal tract, and / or heart.

31. A composition described in any one of claims 1 to 28 and 30, wherein the cell is a neuron.

32. A composition described in any one of claims 1 to 28 and 30 to 31, wherein the cell is a proprioceptive sensory neuron or a Betz cell.

33. A composition described in any one of claims 1 to 32, wherein the cells are within a subject.

34. The composition described in claim 33, wherein the subject has a neurological or neuromuscular disorder.

35. The composition described in claim 34, wherein the neurological or neuromuscular disorder is associated with reduced frataxin protein levels.

36. A composition described in any one of claims 33 to 35, wherein the subject has Friedreich's ataxia (FA).

37. A composition described in any one of claims 1 to 36, wherein the composition is a pharmaceutical composition.

38. The composition of any one of claims 1 to 37, wherein the composition is formulated for intravenous administration.

39. Use of an adeno-associated virus (AAV) genome in the manufacture of a medicament for delivery of a nucleotide sequence encoding a frataxin protein to a cell, comprising: the AAV genome comprises an engineered promoter and a nucleotide sequence encoding the frataxin protein; The use wherein the engineered promoter consists of a nucleotide sequence at least 90% identical to SEQ ID NO: 1742.

40. The use of claim 39, wherein the AAV genome comprises the nucleotide sequence of SEQ ID NO: 1797, or a nucleotide sequence that is at least 90% identical, at least 95% identical, or at least 99% identical to SEQ ID NO: 1797.

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