Recombinant adeno associated virus (RAAV) encoding gjb2 and uses thereof

Abstract

The present disclosure, at least in part, relates to compositions (e.g., isolated nucleic acid and rAAVs) and methods for treating Non-syndromic hearing loss and deafness (DFNB1) by delivering gap junction beta 2 (GJB2) protein to inner ear cells that normally express GJB2 (e.g., fibrocytes and supporting cells of the organ of Corti and nearby regions). The isolated nucleic acid of the present disclosure comprises an expression cassette, wherein the expression cassette comprises a gap junction beta 2 (GJB2) gene regulatory element (GRE) (e.g., GJB2 enhancers, GJB2 promoters, GJB2 5' UTR, and / or GJB2 3' UTR), and a nucleotide sequence encoding a GJB2 protein.

Description

RECOMBINANT ADENO ASSOCIATED VIRUS (rAAV) ENCODING GJB2 AND USES THEREOFREEATED APPEICATIONS

[0001] This application claims priority under 35 U.S.C. § 119(e) to U.S. Provisional Application, U.S.S.N. 63 / 161,619, filed March 16, 2021, and to U.S. Provisional Application, U.S.S.N. 63 / 078,233, filed September 14, 2020, each of which is incorporated herein by reference.FEDERAEEY SPONSORED RESEARCH

[0002] This invention was made with Government support under DA048787 awarded by the National Institutes of Health. The Government has certain rights in the invention.BACKGROUND

[0003] Loss of gap junction beta 2 (GJB2) expression in the inner ear underlies a disorder termed nonsyndromic Hearing Loss and Deafness, (DFNB1), characterized by recessive, mild-to-profound sensorineural hearing impairment. Many of these patients are born with profound hearing loss, which is probably irreversible even at birth. Two-thirds have some residual hearing at birth, and the majority of those lose hearing over the next few years. Therefore, these patients are potential candidates for treatment of DFNB1. Previous gene replacement therapy of GJB2 failed to rescue hearing even though gene addition of the GJB2 gene rescued cell survival and the gap junction network. Effective GJB2 gene replacement therapy for hearing rescuing has not been developed.SUMMARY

[0004] The present disclosure, at least in part, relates to an isolated nucleic acid comprising an expression cassette, wherein the expression cassette comprises a gap junction beta 2 (GJB2) gene regulatory element (GRE), and a nucleotide sequence encoding a GJB2 protein. In some embodiments, the expression cassette further comprises a promoter (e.g., GJB2 promoter). In some embodiments, the expression cassette is flanked by two adeno-associated virus (AAV) inverted terminal repeats (ITRs). The presence of native GJB2 regulatory elements (GREs) in the isolated nucleic acid prevents promiscuous GJB2 gene expression in the inner ear, which is toxic and damages hearing. Accordingly, in some embodiments, the isolated nucleic acid described herein is capable of expressing the GJB2 protein in inner earcells that normally express the GJB2 gene (e.g., connective tissue cells of the cochlea and supporting cells of the organ of Corti and nearby regions), but not in the cell that do not normally express GJB2 (e.g., hair cells and spiral ganglion neurons).

[0005] In some aspects, the present disclosure provides an isolated nucleic acid comprising an expression cassette, wherein the expression cassette comprises a gap junction beta 2 (GJB2) gene regulatory element (GRE), and a nucleotide sequence encoding a GJB2 protein.

[0006] In some embodiments, the GJB2 protein is a human GJB2 protein. In some embodiments, the GJB2 protein comprises an amino acid sequence at least 80% identical to SEQ ID NO: 1. In some embodiments, the nucleotide sequence encoding a human GJB2 protein comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 2.

[0007] In some embodiments, the expression cassette further comprises a promoter operably linked to the nucleotide sequence encoding a GJB2 protein. In some embodiments, the promoter is a human GJB2 promoter. In some embodiments, the promoter comprises 500 nucleotides of a human GJB2 promoter. In some embodiments, the promoter comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 5. In some embodiments, the promoter comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 102. In some embodiments, the promoter comprises a nucleic acid sequence 100% identical to SEQ ID NO: 102.

[0008] In some embodiments, the promoter is a human GJB2 basal promoter. In some embodiments, the human GJB2 basal promoter comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 47.

[0009] In some embodiments, the expression cassette comprises a nucleotide sequence encoding a 5' UTR. In some embodiments, the 5' UTR is positioned between the promoter and the nucleotide sequence encoding the GJB2 protein. In some embodiments, the 5' UTR comprises about 300 nucleotides of a human GJB2 gene 5' UTR. In some embodiments, the promoter and the 5' UTR comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 30.

[0010] In some embodiments, the GJB2 gene regulatory element comprises an enhancer. In some embodiments, the enhancer is positioned 5' to the promoter. In some embodiments, the enhancer is normally present within approximately 200 kb upstream or downstream of a GJB2 gene. In some embodiments, the enhancer is normally present within approximately 95 kb of a GJB2 gene. In some embodiments, the GJB2 GRE comprises one or more enhancers. In some embodiments, the one or more enhancers are the same enhancers or different enhancers. In some embodiments, the enhancer comprises a nucleotide sequence at least 80%identical to nucleotide sequence or a fragment thereof as set forth in any one of SEQ ID NOs: 6 to 29. In some embodiments, the enhancer comprises a nucleotide sequence at least 80% identical to a GJB2 enhancer as set forth in any of SEQ ID NOs: 37-46 and 55-60. In some embodiments, the enhancer comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 42.

[0011] In some aspects, the present disclosure also provides an isolated nucleic acid comprising an expression cassette, wherein the expression cassette comprises a Gap Junction beta 2 (GJB2 ) promoter, and a nucleotide sequence encoding a GJB2 protein.

[0012] In some embodiments, the GJB2 promoter comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 102. In some embodiments, the GJB2 promoter comprises a nucleic acid sequence 100% identical to SEQ ID NO: 102.

[0013] In some embodiments, the expression cassette further comprises a 5 ' UTR. In some embodiments, the 5' UTR comprises: a first nucleic acid sequence at least 80% identical to SEQ ID NO: 103; and / or a second nucleic acid sequence at least 80% identical to SEQ ID NO: 104. In some embodiments, the expression cassette further comprises a 5 ' UTR. In some embodiments, the 5' UTR comprises: a first nucleic acid sequence 100% identical to SEQ ID NO: 103; and / or a second nucleic acid sequence 100% identical to SEQ ID NO: 104.

[0014] In some embodiments, the isolated nucleic acid comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 105. In some embodiments, the isolated nucleic acid comprises a nucleic acid sequence 100% identical to SEQ ID NO: 105.

[0015] In some embodiments, the isolated nucleic acid is capable of expressing GJB2 in cells that normally express the GJB2 gene. In some embodiments, the isolated nucleic acid is capable of expressing GJB2 in cochlear connective tissue cells and supporting cells of the organ of Corti. In some embodiments, the supporting cell of the organ of Corti are pillar cells, Deiter cells, Hensen’s cells, Claudius cells, inner phalangeal cells, and border cells. In some embodiments, the cochlear connective tissue cells are strial intermediate cells, fibrocytes of the lateral wall and suprastrial zone, basal cells of the stria vascularis, fibrocytes in the spiral ligament, fibrocytes in the spiral limbus, mesenchymal cells lining the bony otic capsule facing the scala vestibuli, and supralimbal dark cells.

[0016] In some embodiments, the expression cassette is flanked by two adeno-associated virus inverted terminal repeats (ITRs). In some embodiments, the AAV ITR is from a serotype selected from the group consisting of AAV1 ITR, AAV2 ITR, AAV3 ITR, AAV4 ITR, AAV5 ITR, and AAV6 ITR. In some embodiments, the AAV ITR is AAV2 ITR.

[0017] In some embodiments, the expression cassette comprises: a 5' ITR having a nucleotide sequence at least 80% identical to SEQ ID NO: 106; and / or a 3 ' ITR having a nucleotide sequence at least 80% identical to SEQ ID NO: 107. In some embodiments, the expression cassette comprises: a 5 ' ITR having a nucleotide sequence 100% identical to SEQ ID NO: 106; and / or a 3 ' ITR having a nucleotide sequence 100% identical to SEQ ID NO: 107.

[0018] In some embodiments, the expression cassette further comprises a Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) 3' to the nucleotide sequence encoding the GJB2 protein.

[0019] In some embodiments, the WPRE comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 108. In some embodiments, the WPRE comprises a nucleotide sequence 100% identical to SEQ ID NO: 108.

[0020] In some embodiments, the expression cassette further comprises a nucleotide sequence encoding a 3' UTR located 5' of the WPRE. In some embodiments, the 3' UTR is a GJB2 exon 2 3' UTR. In some embodiments, the GJB2 exon 2 3' UTR comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 32.

[0021] In some embodiments, the expression cassette further comprises one or more miRNA binding site positioned in the 3' UTR. In some embodiments, the miRNA binding site is a neuron-associated miRNA binding site. In some embodiments, the neuron- associated miRNA is selected from: miR-124, miR-127, miR-129, miR-129*, miR-136, miR-136*, miR-137, miR-154, miR-3OO-3p, miR-323, miR-329, miR-341, miR-369-5p, miR-376a, miR-376b-3p, miR-376c, miR-379, miR-382, miR-382*, miR-410, miR-411, miR-433, miR-434, miR-495, miR-541, miR-543*, miR-551b, miR-143, miR-449a, miR-219-2-3p, miR-126, miR-126*, miR-141, miR-142-3p, miR-142-5p, miR-146a, miR-150, miR-200c, and miR-223.In some embodiments, the neuron-associated miRNA is miR-124. In some embodiments, the miRNA binding site is a cochlear hair cell- associated miRNA binding site. In some embodiments, the cochlear hair cell-associated miRNA binding site is selected from: miR-124, miR-96, miR- 182, and miR-183.

[0022] In some embodiments, the expression cassette further comprises a poly A signal. In some embodiments, the poly A signal is a bovine growth hormone poly A signal.

[0023] In some embodiments, the poly A signal comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 109. In some embodiments, the poly A signal comprises a nucleotide sequence 100% identical to SEQ ID NO: 109.

[0024] In some aspects, the present disclosure also provides an isolated nucleic acid comprising a nucleotide sequence 100% identical to SEQ ID NO: 110 or 111. In someaspects, the present disclosure also provides an isolated nucleic acid comprising a nucleotide sequence at least 80% identical to SEQ ID NO: 110 or 111.

[0025] In some aspects, the present disclosure also provides a vector comprising the isolated nucleic acid as described herein. In some embodiments, the vector is a plasmid or a viral vector. In some embodiments, the viral vector is an AAV vector.

[0026] In some aspects, the present disclosure also provides a vector comprising from 5' to 3': (a) an AAV 5' ITR; (b) a GJB2 promoter, or a basal GJB2 promoter sequence thereof; (c) a GJB2 5' UTR (e.g., a GJB2 exon 1 5' UTR); (d) a nucleotide sequence encoding a GJB2 protein; (e) a GJB2 3' UTR (e.g., a GJB2 exon 2 3' UTR), optionally the GJB2 3' UTR comprises one or more miR-124 binding site; (f) a bovine growth hormone poly A signal; and (g) an AAV 3’ ITR.

[0027] In some aspects, the present disclosure also provides a vector comprising from 5' to 3': (a) an AAV 5' ITR; (b) a GJB2 enhancer; (c) a GJB2 promoter, or a basal GJB2 promoter sequence thereof; (d) a GJB2 5' UTR (e.g., a GJB2 exon 1 5' UTR); (e) a nucleotide sequence encoding a GJB2 protein; (f) a GJB2 3' UTR (e.g., a GJB2 exon 2 3' UTR), optionally the GJB2 3' UTR comprises one or more miR-124 binding site; (g) a bovine growth hormone poly A signal; and (h) an AAV 3' ITR.

[0028] In some embodiments, the vector comprises a nucleotide sequence at least 80% identical to any one of SEQ ID NOs: 36, 48-62 and 61-83. In some embodiments, the vector is an AAV vector. In some embodiments, the vector is capable of expressing a GJB2 gene in cells that normally express GJB2.

[0029] In some aspects, the present disclosure also provides a recombinant adeno-associated virus (rAAV) comprising: (i) a capsid protein; and (ii) the isolated nucleic acid described herein.

[0030] In some aspects, the present disclosure also provides a recombinant adeno-associated virus (rAAV) comprising: (i) a capsid protein; and (ii) an isolated nucleic acid comprising: (a) an AAV 5' ITR (e.g., a GJB2 exon 1 5' UTR); (b) a GJB2 promoter, or a basal GJB2 promoter sequence thereof; (c) a GJB2 5' UTR (e.g., a GJB2 exon 2 3' UTR), optionally the GJB2 exon 2 3' UTR comprises one or more miR-124 binding site; (d) a nucleotide sequence encoding a GJB2 protein; (e) a GJB2 3' UTR; (f) a bovine growth hormone poly A signal; and (g) an AAV 3' ITR.

[0031] In some aspects, the present disclosure also provides a recombinant adeno-associated virus (rAAV) comprising: (i) a capsid protein; and (ii) an isolated nucleic acid comprising: (a) an AAV 5' ITR; (b) a GJB2 enhancer; (c) a GJB2 promoter, or a basal GJB2 promotersequence thereof; (d) a GJB2 5' UTR (e.g., a GJB2 exon 1 5' UTR); (e) a nucleotide sequence encoding a GJB2 protein; (f) a GJB2 3' UTR (e.g., a GJB2 exon 2 3' UTR), optionally the GJB2 exon 2 3' UTR comprises one or more miR-124 binding site; (g) a bovine growth hormone poly A signal; and (h) an AAV 3' ITR.

[0032] In some embodiments, the rAAV has tropism for a subset of cochlea cells that normally express the GJB2 gene. In some embodiments, the rAAV has tropism for cells of the inner ear.

[0033] In some embodiments, the capsid protein is an AAV1 capsid protein, an AAV2 capsid protein, an AAV5 capsid protein, an AAV7 capsid protein, an AAV8 capsid protein, an AAV9 capsid protein, an AAV-S capsid protein, or a variant thereof. In some embodiments, the AAV capsid is AAV9.PHP.B, AAV9.PHP.eB, or AAV-S. In some embodiments, the AAV capsid protein is AAV-S.

[0034] In some aspects, the present disclosure provides a host cell comprising the isolated nucleic acid, the vector, or the rAAV as described herein.

[0035] In some aspects, the present disclosure provides a pharmaceutical composition comprising the isolated nucleic acid, the vector, the rAAV, or the host cell as described herein. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0036] In some aspects, the present disclosure provides a method for specifically expressing GJB2 in cells that normally expresses the GJB2 gene in a subject, the method comprising administering to the subject an effective amount of the isolated nucleic acid, the vector, the rAAV, the host cell, or the pharmaceutical composition as described herein.

[0037] In some aspects, the present disclosure provides a method for treating Non-syndromic Hearing Loss and Deafness (DFNB1) in a subject, the method comprising administering to the subject an effective amount of the isolated nucleic acid, the vector, the rAAV, the host cell, or the pharmaceutical composition as described herein.

[0038] A method for treating a GJB2-associated disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the isolated nucleic acid, the vector, the rAAV, the host cell, or the pharmaceutical composition as described herein.

[0039] In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human mammal. In some embodiments, the non-human mammal is mouse, rat, or non-human primate.

[0040] In some embodiments, the hearing loss is associated with a mutation in the GJB2gene. In some embodiments, the mutation in the GJB2 gene is a point mutation, a missense mutation, a nonsense mutation, a splice-altering mutation, a synonymous mutation, a deletion, an insertion, or a combination thereof. In some embodiments, the subject is human; and the mutation is a mutation listed in Table 2 (below) or a combination thereof. In some embodiments, the mutation is NM_004004.6 C.101T>C (GRCh37 / hgl9 Chrl3:20763620A>G) or c.35delG (GRCh37 / hgl9 chrl3:20763685AC>A).

[0041] In some embodiments, the administration results in expression of GJB2 protein in the cochlea connective tissue cells and supporting cells of the organ of Corti and nearby regions. In some embodiments, the supporting cell of the organ of Corti are pillar cells, Deiters’ cells, Hensen’s cells, Claudius cells, inner phalangeal cells, and border cells. In some embodiments, the connective tissue cells are strial intermediate cells, fibrocytes of the lateral wall and suprastrial zone, basal cells of the stria vascularis, fibrocytes in the spiral ligament, fibrocytes in the spiral limbus, mesenchymal cells lining the bony otic capsule facing the scala vestibuli, and supralimbal dark cells.

[0042] In some embodiments, the administration is via injection. In some embodiments, the injection is through round window membrane of the cochlea, into the scala media of the cochlea, into the scala tympani of the cochlea, into the scala vestibuli of the cochlea, into a semicircular canal of the inner ear, or into the saccule or the utricle of the inner ear.

[0043] The details of one or more embodiments of the invention are set forth in the description below. Other features or advantages of the present invention will be apparent from the following drawing and detailed description of certain embodiments and also from the appended claims.BRIEF DESCRIPTION OF THE DRAWINGS

[0044] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate certain embodiments, and together with the written description, serve to provide non-limiting examples of certain aspects of the compositions and methods disclosed herein.

[0045] FIGs. 1A-1C show the structure and expression distribution of GJB2, and how loss of GJB2 expression affects the patients. FIG. 1A shows the structure of the GJB2 hemichannel. Six subunits of GJB2 protein, each with four trans-membrane helices, assemble in the plane of the membrane to form a large central pore. GJB2 hemichannels from adjacent cells join to create a channel from the cytoplasm of one cell to the cytoplasm of the other. Gap junctions are formed by hundreds or thousands of channels packed in a junctional plaque. FIGs. 1B-1Cshow the network of fibrocytes and epithelial cells in which GJB2 is expressed (FIG. IB), and the inner and outer hair cells, in which GJB2 is not expressed (FIG. 1C). FIG. ID shows that many patients carrying GJB2 mutation(s) who have some residual hearing at birth show further hearing loss over the next 3-6 years. A window for treatment is present for 1-5 years after birth, with -10,000 affected children in the United State aged 0-5 possibly treatable.

[0046] FIGs. 2A-2B show the delivery of viral vector to the cochlea by direct injection through the round window membrane (RWM) and the deleterious effect of promiscuous expression of Gjb2 to the hearing of injected mice. FIG. 2A is a cartoon illustrating the round window membrane (RWM) injection. FIG. 2B shows that promiscuous expression of Gjb2 in the inner ear damaged hearing in wild-type mice.

[0047] FIGs. 3A-3N show the identification of cis-regulatory elements (e.g., enhancers) that are critical for GJB2 expression in the subset of cochlea cells that naturally express the GJB2 gene. FIGs. 3A-3B show that certain patients with GJB2-associated deafness have upstream deletions occurring in trans with GJB2 coding sequence mutations, which suggests that some patients carry mutation(s) in the cis-regulatory element, and the region next to the CRYL1 gene is of particular importance for identification of such cis-regulatory elements. FIG. 3C (top) shows the identification of gene regulatory elements (GREs), in UCSC Genome Browser views of ATAC-Seq from mouse cochlea at developmental stages P2, P5 and P8, over -300 kb in the region of the mouse Gjb2 gene. Shaded regions mark regions containing putative GREs. X-axis is the genomic region on chrl4 in the mouse genome. Y-axis is the number of reads from the AT AC- Seq that align to a specific region in the genome. Light shading denotes regions of open chromatin, which are the hallmarks of transcriptionally active regions that are enriched for read pile up, suggesting higher activity in these regions. Regions A and B mark the transcriptionally active sequences within mouse Gjb2 itself. Regions C-M are regions that are transcriptionally active around Gjb2 that might be part of a cis-regulatory network. FIG. 3C (bottom) shows transcriptionally active regions in and around the light-shaded regions that have been tested as specific GREs (dark highlight). Note that the GREs were initially identified in mouse. Human GJB2 GREs were identified in silico by modeling the mouse GREs. Human GJB2 GREs were tested in subsequent experiments. FIGs. 3D-3E show various vector designs with or without incorporation of GJB2 promoter and / or enhancers. These vectors were tested in mouse inner ear. The C15 vector, which is the GJB2 enhancer vector, concatenates 500 bp of the human GJB2 promoter, the human GJB2 5’ UTR followed by a coding sequence for GFP and human GJB2 3’ UTR, and three human GJB2 enhancers that match mouse sequences identified by ATAC-seq. Vectors c20-23 wereconstructed to test the toxicity of promiscuous expression of Gjb2 in mouse. Vector c20 was lethal at doses over 2xl09genomic copies. FIG. 3F shows a segment of the mouse cochlea, from the lateral wall (top) to the interdental cells (bottom). Cells transduced with the AAV9- PHP.B-C15 vector and expressing the GFP marker gene under Gjb2 enhancers are shown in the left panel. Cells normally expressing GJB2 are shown in the middle panel. In the right panel, IHCs and OHCs (indicated) are also identified by labeling actin with fluorescent phalloidin. The expression pattern of GFP, which was driven by the cl5 construct, is consistent with native Gjb2 expression reported in Kikuchi el al., 1995 using the same antibody against GJB2. Notably, cl5 does not drive GFP expression in hair cells. FIG. 3G shows the expression of Gjb2 in inner hair cells driven by construct c20. 3D reconstruction of the organ of Corti in an uninjected mouse cochlea, with outer hair cells and inner hair cells is shown in the top panel. GJB2-containing gap junctions in supporting cells were labeled with an antibody to GJB2 protein. Hair cells do not make gap junctions. Vector c20, with a promiscuous promoter, drives GJB2 expression in inner hair cells and other cell types (see bottom panel). FIG. 3H shows that promiscuous Gjb2 expression damages hearing in wildtype mice, but targeted expression rescues hearing in Gjb2 knockout mice. However, a C70 construct, which includes GJB2 promoter / enhancer based on preliminary results from the ATAC-Seq, was capable of rescuing hearing by 15-20 dB, and did not damage hearing in the wild-type. FIGs. 3I-3L shows the map of the c70 vector plasmid encoding mouse GJB2 or human GJB2 with or without an HA tag. FIG. 3M shows schematics of vector c.70 encoding mouse GJB2 or human GJB2 with or without the HA tag. FIG. 3N shows additional vectors that were created and tested.

[0048] FIG. 4 shows that AAV-S encoding eGFP with a CBA promoter efficiently transduces hair cells, supporting cells, and cells of the lateral wall, in both neonatal mouse and juvenile NHP cochlea.

[0049] FIGs. 5A-5V show vector maps of the AAV vectors including the identified GJB2 GREs 1, 2, 3, 4, 5, 7, 8, and 9, respectively. The vectors include, from 5' to 3 ', a 5' ITR, a human GJB2 GRE, a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, a nucleotide sequence encoding an eGFR, and GJB2 exon 2 3 ' UTR. FIG. 5A shows vector c.81.1, which includes human GJB2 GRE1, and encodes human GJB2; FIG. 5B shows vector c.81.1, which includes human GJB2 GRE1, and encodes mouse GJB2; FIG. 5C shows vector c.81.2, which includes human GJB2 GRE2, and encodes eGFP; FIG. 5D shows vector c.81.2, which includes human GJB2 GRE2, and encodes human GJB2; FIG. 5E shows vector c.81.2, which includes human GJB2 GRE2, and encodes mouse GJB2; FIG. 5F shows vector c.81.3, whichincludes human GJB2 GRE3, and encodes eGFP; FIG. 5G shows vector c.81.3, which includes human GJB2 GRE3, and encodes human GJB2; FIG. 5H shows vector c.81.3, which includes human GJB2 GRE3, and encodes mouse GJB2; FIG. 51 shows vector c.81.4, which includes human GJB2 GRE4, and encodes human GJB2; FIG. 5J shows vector c.81.4, which includes human GJB2 GRE4, and encodes mouse GJB2; FIG. 5K shows vector c.81.5, which includes human GJB2 GRE5, and encodes eGFP; FIG. 5L shows vector c.81.5, which includes human GJB2 GRE5, and encodes human GJB2; FIG. 5M shows vector c.81.5, which includes human GJB2 GRE5, and encodes mouse GJB2; FIG. 5N shows vector c.81.7, which includes human GJB2 GRE7, and encodes eGFP; FIG. 50 shows vector c.81.7, which includes human GJB2 GRE7, and encodes human GJB2; FIG. 5P shows vector c.81.7, which includes human GJB2 GRE7, and encodes mouse GJB2; FIG. 5Q shows vector c.81.8, which includes human GJB2 GRE8, and encodes human GJB2; FIG. 5R shows vector c.81.8, which includes human GJB2 GRE8, and encodes mouse GJB2; FIG. 5S shows vector c.81.9, which includes human GJB2 GRE9, and encodes eGFP; FIG. 5T shows vector c.81.9, which includes human GJB2 GRE9, and encodes human GJB2; FIG. 5U shows vector c.81.9, which includes human GJB2 GRE9, and encodes mouse GJB2. FIG. 5V shows schematics of c81.2, c81.3, c81.5, c81.7 and c81.9 encoding eGFP, mouse GJB2 and human GJB2 as described above.

[0050] FIGs. 6A-6D show GFP expression by vector c81.5 in the cells of the organ of Corti FIG. 6A shows a fluorescent image of GFP expressing cells, including a variety of supporting cells in, and medial to, the organ of Corti. FIG. 6B shows antibody label of endogenous GJB2 in the region of the organ of Corti. Gjb2 expression largely overlapped that of exogenous GFP. FIG. 6C is an overlay of FIGs. 6A and 6B, with a third staining of actin, which revealed stereocilia of hair cells. No GFP was expressed in the hair cells. FIG. 6D shows a frozen section immunofluorescence image of GFP and a protein marker for hair cells, MY07A. GFP was expressed in a variety of supporting cells in the organ of Corti, but did not overlap with MY07A expression, which was expressed in hair cells.

[0051] FIGs. 7A-7E show GFP expression pattern by vector 81.5 in the lateral wall of the cochlea. FIG. 7A shows GFP expression in cells including fibrocytes of the lateral wall. FIG. 7B shows an antibody labeling of endogenous Gjb2 in the region of the lateral wall. GJB2 expression largely overlaps that of exogenous GFP. FIG. 7C is an overlay image of FIGs. 7A and 7B. Note that GFP was expressed in the cells expressing Gjb2. FIGs. 7D-7E show frozen section immunofluorescences of GFP (FIG. 7D) and GJB2 in supporting cells of the organ of Corti and fibrocytes of the lateral wall (FIG. 7E).

[0052] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate certain embodiments, and together with the written description, serve to provide non-limiting examples of certain aspects of the compositions and methods disclosed herein.DETAILED DESCRIPTION

[0053] The present disclosure, at least in part, relates to an isolated nucleic acid comprising an expression cassette, wherein the expression cassette comprises a gap junction beta 2 (GJB2) gene regulatory element (GRE), and a nucleotide sequence encoding a GJB2 protein. In some embodiments, the expression cassette further comprises a promoter (e.g., GJB2 promoter). In some embodiments, the expression cassette is flanked by two adeno-associated virus (AAV) inverted terminal repeats (ITRs). The presence of native GJB2 regulatory elements (GREs) in the isolated nucleic acid prevents promiscuous GJB2 gene expression in the inner ear, which is toxic and damages hearing. Accordingly, in some embodiments, the isolated nucleic acid described herein is capable of expressing the GJB2 protein in inner ear cells that normally express the GJB2 gene (e.g., connective tissue cells of the cochlea and supporting cells of the organ of Corti and nearby regions), but not in the cell that do not normally express GJB2 gene (e.g., hair cells and spiral ganglion neurons).I. Isolated Nucleic Acid

[0054] In some aspects, the present disclosure relates to compositions and methods for treating certain autosomal recessive genetic diseases, for example, non-syndromic hearing loss (DFNB1). DFNB1 is caused by mutations in the GJB2 gene. The GJB2 gene encodes the GJB2 protein, also known as connexin 26. Connexin 26 is a member of the connexin protein family. GJB2 protein forms channels in clusters called gap junctions, which allow communication between neighboring cells, including cells in the inner ear. Mutations in the GJB2 gene eliminate or change the structure of gap junctions and affect the function or survival of cells that are needed for hearing. Gene replacement therapy (e.g., gene therapy by recombinant adeno-associated virus (rAAVs)) is attractive due to the small size of the GJB2 gene coding sequence (less than 700 bp). However, restoration of GJB2 expression in the inner ear using the currently available gene therapy does not lead to the restoration of hearing.

[0055] Accordingly, the present disclosure is based, in part, on the surprising discovery that successful GJB2 gene therapy requires GJB2 expression in cells that normally express theGJB2 protein (e.g., connective tissue cells of the cochlea and supporting cells of the organ of Corti and nearby regions) and not in other cells (e.g., hair cells and spiral ganglion neurons). Excluding sensory cells, most cells in the cochlea are connected via gap junctions, and these gap junctions appear to play a critical role in cochlear function. GJB2 protein occurs in gap junctions connecting most cell classes in the cochlea. There are two independent systems of cells, which are defined by interconnecting gap junctions. The first system, the epithelial cell gap junction system, is mainly composed of all organs of Corti supporting cells e.g., epithelial cells of the inner and outer sulcus, and interdental cells), and also includes interdental cells in the spiral limbus and root cells within the spiral ligament. In the inner ear, the sensory region of the cochlea, termed the organ of Corti, includes one row of inner hair cells (IHC) and three to four rows of outer hair cells (OHC) that are surrounded by various supporting cells. The supporting cells play crucial roles in the development, function, and maintenance of inner ear sensory epithelia. Unlike hair cells, which contact only the lumenal surface of the epithelium, supporting cells span the entire depth of the epithelium, from the basal lamina to the lumen. Supporting cells are linked to each other and to hair cells by tight and adherens junctions; they communicate directly with other supporting cells by gap junctions (e.g., Wan et al., Inner ear supporting cells: Rethinking the silent majority, Semin Cell Dev Biol. 2013 May; 24(5): 448-459). Non-limiting examples of supporting cells for the organ of Corti include pillar cells, Deiters’ cells, Hensen’s cells, Claudius cells, inner phalangeal cells, and border cells. The second system, the connective tissue cell gap junction system, includes strial intermediate cells, fibrocytes of the lateral wall and suprastrial zone, basal cells of the stria vascularis, fibrocytes in the spiral ligament, fibrocytes in the spiral limbus, mesenchymal cells lining the bony otic capsule facing the scala vestibuli, and supralimbal dark cells. In some embodiments, in the cochlea, GJB2 is normally expressed in supporting cells of the organ of Corti and nearby regions (e.g., pillar cells, Deiters’ cells, Hensen’s cells, Claudius cells, inner phalangeal cells; and border cells), and the connective tissue system comprising strial intermediate cells, fibrocytes of the lateral wall and suprastrial zone, basal cells of the stria vascularis, fibrocytes in the spiral ligament, fibrocytes in the spiral limbus, mesenchymal cells lining the bony otic capsule facing the scala vestibuli, and supralimbal dark cells (See, e.g., Kikuchi et al. (1995) Gap junctions in the rat cochlea: immunohistochemical and ultrastructural analysis. Anat Embry ol (Berl) 191:101-118; and Kikuchi et al., Gap junction systems in the mammalian cochlea, Brain Res Brain Res Rev. 2000 Apr;32(l): 163-6. doi: 10.1016 / s0165-0173(99)00076-4.).

[0056] GJB2 expression is critical for cochlear function. For example, the K+that enters haircells through transduction channels and leaves through basal K+channels is shuttled away from the organ of Corti by the epithelial system and conveyed by the cytoplasmic system to the stria, where it is pumped back into endolymph. Further, GJB2 plays a role in the development of the cochlea, as mice lacking GJB2 protein in the inner ear have reduced endocochlear potential and profound apoptotic loss of hair cells and supporting cells by postnatal day 30 (P30), even though hair cells do not express Gjb2 (Cohen-Salmon et al., 2002; Wang et al., 2009; Sun et al., 2009; Crispino et al., 2011; Johnson et al., 2017). If Gjb2 is deleted after P6, the phenotype is much milder (Chang et al., 2015). However there remains a long-term requirement for GJB2 protein: hair cell loss occurs after months even with deletion as late as P14 (Ma et al., 2020). Not wishing to be bound by any particular theory, GJB2’s function in shuttling K+may be related to its role in the development of the cochlea: If K+is not carried away from hair cells by a gap junction network, K+accumulation could depolarize hair cells, leading to Ca2+influx and eventual cell death. The gap junction network may also be required to transport glucose and nutrients from blood vessels to the sensory epithelium, and its absence could lead to cell death.

[0057] In some embodiments, the present disclosure provides an isolated nucleic acid comprising two adeno-associated virus (AAV) inverted terminal repeats (ITRs) flanking an expression cassette, wherein the expression cassette comprises a promoter (e.g., a human GJB2 promoter) operably linked to a nucleotide sequence encoding a GJB2 gene regulatory element (GRE), and a nucleotide sequence encoding a gap junction beta 2 (GJB2) protein. Incorporation of the native GJB2 gene regulatory element and / or tissue / cell-specific promoter in the isolated nucleic acid facilitates the expression of the GJB2 gene in cells that normally express it (e.g., connective tissue cells of the cochlea including fibrocytes and supporting cells of the organ of Corti and nearby regions). An expression cassette, as used herein, refers to component of vector DNA comprising a protein coding sequence to be expressed by a cell having the vector and its regulatory sequences. Once delivered to the target cell, the expression cassette directs the cell’s machinery to make RNA and / or protein(s) (e.g., GJB2 protein).

[0058] A “nucleic acid” sequence refers to a DNA or RNA sequence. In some embodiments, proteins and nucleic acids of the disclosure are isolated. As used herein, the term “isolated” means artificially produced. As used herein with respect to nucleic acids, the term “isolated” means: (i) amplified in vitro by, for example, the polymerase chain reaction (PCR); (ii) recombinantly produced by cloning; (iii) purified, for example, by cleavage and gel separation; or (iv) synthesized by, for example, chemical synthesis. An isolated nucleic acidis one which is readily manipulable by recombinant DNA techniques well known in the art. Thus, a nucleotide sequence contained in a vector in which 5 ' and 3 ' restriction sites are known or for which polymerase chain reaction (PCR) primer sequences have been disclosed is considered isolated but a nucleic acid sequence existing in its native state in its natural host is not. An isolated nucleic acid may be substantially purified, but need not be. For example, a nucleic acid that is isolated within a cloning or expression vector is not pure in that it may comprise only a tiny percentage of the material in the cell in which it resides. Such a nucleic acid is isolated, however, as the term is used herein because it is readily manipulable by standard techniques known to those of ordinary skill in the art. As used herein with respect to proteins or peptides, the term “isolated” refers to a protein or peptide that has been isolated from its natural environment or artificially produced (e.g., by chemical synthesis, by recombinant DNA technology, etc.).

[0059] In some embodiments, the GJB2 protein is a human GJB2 protein. In some embodiments, the human GJB2 protein comprises an amino acid sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 1.

[0060] An exemplary human GJB2 protein sequence is set forth in SEQ ID NO: 1:MDWGTLQTILGGVNKHSTS IGKIWLTVLFIFRIMILVVAAKEVWGDEQADFVCNTLQPGCKN VCYDHYFP I SHIRLWALQLIFVSTPALLVAMHVAYRRHEKRKFIKGEIKSEFKDIEEIKTQK VRIEGSLWWTYTSS IFFRVIFEAAFMYVFYVMYDGFSMQRLVKCNAWPCPNTVDCFVSRPTE KTVFTVFMIAVSGICILLNVTELCYLLIRYCSGKSKKPV

[0061] In some embodiments, the expression cassette of the isolated nucleic acid encodes a human GJB2 protein having the amino acid sequence as set forth in SEQ ID NO: 1. In some embodiments, the nucleotide sequence encoding a human GJB2 protein comprises a nucleotide sequence at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 2.

[0062] An exemplary nucleotide sequence encoding a human GJB2 protein is set forth in SEQ ID NO: 2: ATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCCACCAGCATTGG AAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGTGGCTGCAAAGG AGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAGGCTGCAAGAAC GTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTGCAGCTGATCTT CGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACATGAGAAGAAGA GGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGATCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTCTTCCGGGTCAT CTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTCCATGCAGCGGC TGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGTCCCGGCCCACG GAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATCCTGCTGAATGT CACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAAGCCAGTT

[0063] In some embodiments, the GJB2 protein is a mouse GJB2 protein. In some embodiments, the mouse GJB2 protein comprises an amino acid sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 3.

[0064] An exemplary mouse GJB2 protein sequence is set forth in SEQ ID NO: 3:MDWGTLQS ILGGVNKHSTS IGKIWLTVLFIFRIMILVVAAKEVWGDEQADFVCNTLQPGCKN VCYDHHFP I SHIRLWALQLIMVSTPALLVAMHVAYRRHEKKRKFMKGEIKNEFKDIEEIKTQ KVRIEGSLWWTYTTS IFFRVIFEAVFMYVFYIMYNGFFMQRLVKCNAWPCPNTVDCFI SRPT EKTVFTVFMI SVSGICILLNITELCYLFVRYCSGKSKRPV

[0065] In some embodiments, the isolated nucleic acid comprises a nucleotide sequence encoding a mouse GJB2 protein having an amino acid sequence as set forth in SEQ ID NO: 3. In some embodiments, the nucleotide sequence encoding a mouse GJB2 protein comprises a nucleotide sequence at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 4.

[0066] An exemplary nucleotide sequence encoding a mouse GJB2 protein is set forth in SEQ ID NO: 4:ATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCCACCAGCATTGG AAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGTGGCTGCAAAGG AGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGGCTGCAAGAAT GTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTGCAGCTGATCAT GGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACATGAAAAGAAAC GGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGATCAAAACCCAG AAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCGGGTCAT CTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGCAACGTC TGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTTCCAGGCCCACA GAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATTCTGCTAAATAT CACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAGTC

[0067] In some embodiments, the nucleotide sequence encoding the GJB2 protein is codon optimized for expression in a host (e.g., a human). “Codon optimization” as described herein, refers to the design process of altering codons to codons known to increase maximum protein expression efficiency in a desired cell. In some alternatives, codon optimization is described,wherein codon optimization can be performed by using algorithms that are known to those skilled in the art to create synthetic genetic transcripts optimized for high protein yield. Programs containing algorithms for codon optimization are known to those skilled in the art. Programs can include, for example, OptimumGene™, GeneGPS® algorithms, etc. Additionally, synthetic codon optimized sequences can be obtained commercially, for example from Integrated DNA Technologies and other commercially available DNA sequencing services.

[0068] As used herein, the term “sequence identity” refers to the percentage of amino acid (or nucleic acid) residues of a candidate sequence that are identical to the amino acid (or nucleic acid) residues of a reference sequence, e.g., GJB2 protein disclosed herein and its coding sequences, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent identity e.g., gaps can be introduced in one or both of the candidate and reference sequences for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). Alteration of the amino acid sequence or nucleic acid coding sequences can be obtained by deletion, addition, or substitution of residues of the reference sequence. Alignment for purposes of determining percent identity can be achieved in various ways that are within the skill of one in the art, for instance, using publicly available computer software, such as BLAST, BLAST-2, BLAST-P, BLAST-N, BLAST-X, WU- B LAST-2, ALIGN, ALIGN-2, CLUSTAL, or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. For instance, the percent amino acid (or nucleic acid) sequence identity of a given candidate sequence to, with, or against a given reference sequence (which can alternatively be phrased as a given candidate sequence that has or includes a certain percent amino acid (or nucleic acid) sequence identity to, with, or against a given reference sequence) is calculated as follows:100 x (fraction of A / B) where A is the number of amino acid (or nucleic acid) residues scored as identical in the alignment of the candidate sequence and the reference sequence, and where B is the total number of amino acid (or nucleic acid) residues in the reference sequence. In particular, a reference sequence aligned for comparison with a candidate sequence can show that the candidate sequence exhibits from, e.g., 50% to 100% identity across the full length of the candidate sequence or a selected portion of contiguous amino acid (or nucleic acid) residues of the candidate sequence. The length of the candidate sequence aligned for comparisonpurpose is at least 30%, e.g., at least 40%, e.g., at least 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or 100% of the length of the reference sequence. When a position in the candidate sequence is occupied by the same amino acid (or nucleic acid) residue as the corresponding position in the reference sequence (e.g., GJB2 amino acid sequences, coding sequences, nucleotide sequences for GJB2 gene regulatory elements (GREs), or any other sequences described herein), then the molecules are identical at that position.

[0069] An expression cassette of an isolated nucleic acid sequence described herein e.g., the expression cassette of the isolated nucleic acid comprising a nucleotide sequence encoding a GJB2 protein) may further comprise a promoter operably linked to the coding sequence (e.g., GJB2 protein coding sequence). A “promoter” refers to a DNA sequence recognized by the synthetic machinery of the cell, or introduced synthetic machinery, required to initiate the transcription of a gene. The phrases “operatively linked,” “under control,” or “under transcriptional control” means that the promoter is in the correct location and orientation in relation to the nucleic acid to control RNA polymerase initiation and expression of the gene. A promoter may be a constitutive promoter, inducible promoter, or a tissue- specific promoter.

[0070] In some embodiments, the promoter is a tissue / cell-specific promoter. A tissue / cell specific promoter, as used herein, refers to a promoter that has activity in only certain cell types. In some embodiments, the promoter used in the isolated nucleic acid described herein has activity in cochlear cells that normally express the GJB2 gene. Use of a tissue / cell- specific promoter in the isolated nucleic acid described herein can restrict unwanted transgene (e.g., GJB2 gene) expression as well as facilitate persistent transgene expression. In some embodiments, the expression cassette of the isolated nucleic acid comprises a tissue / cell specific promoter. In some embodiments, the expression cassette of the isolated nucleic acid comprises a GJB2 promoter (e.g., a GJB2 promoter for any species where cell specific GJB2 expression is desired). In some embodiments, the expression cassette of the isolated nucleic acid comprises a human GJB2 promoter. In some embodiments, the expression cassette of the isolated nucleic acid comprises at least 300 bp (e.g., 300 bp, 400 bp, 500 bp, 600 bp, 700 bp, or more) of any consecutive nucleotides of a human GJB2 promoter. In some embodiments, the expression cassette of the isolated nucleic acid comprises a promoter having 500 bp consecutive nucleotides of a human GJB2 promoter. In some embodiments, the expression cassette of the isolated nucleic acid comprises a promoter having a nucleotide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or100% identical to SEQ ID NO: 5. An exemplary nucleotide sequence of 500 bp of a human GJB2 promoter is set forth in SEQ ID NO: 5: ACCTGTCTCCCGCCGTGGCGCCTTTTAACCGCACCCCACACCCCGCCTCTTCCCTCGGAGAC TGGGAAAGTTACGGAGGGGGCGGCGCCGCGGGCGGAGCGCGCCCGGCCTCTGGGTCCTCAGA GCTTCCCGGGTCCGCGAACCCCCGACCGCCCCCGAAAGCCCCGAACCCCCCAAGTCCCCTTC GAGGTCCCGATCTCCTAGTTCCTTTGAGCCCCCATGAGTTCCCCAAGTGCCCCCAGCGCCCT GAGTCTCCCCCGGTTACCCCGAGCGCCGCCTCCCCCAGCCCCTTGGCGGCCCGGGTGAAGCG GGGGCGGCTGAGAGTCGGGACCCCCCAGGAAGCGGCGCCCCAGACCCCGGCTCCGGCGCTGT GCCGTGGGCGGGGTTCAGGGATGGCTGTGGTCGTTGTCCTCTGTACTCCGCATAGTGCGAGA GGACTTGGCATTTATGAGCGCTTCTTTAATTTTTTATTGTTAGAGAAACAGGCATTCCTCCA AGGA

[0071] In some embodiments, the expression cassette of the isolated nucleic acid comprises a GJB2 basal promoter (e.g., a human GJB2 basal promoter). A GJB2 basal promoter is a promoter region of a GJB2 gene highly conserved across different species (e.g., human and mouse). The GJB2 basal promoter has been previous described, for example, in Tu, Z. J., and Kiang, D. T. (1998). Mapping and characterization of the basal promoter of the human connexin26 gene. Biochim. Biophys. Acta 1443, 169-181; Kiang, D. T., Jin, N., Tu, Z. J., and Lin, H. H. (1997). Upstream genomic sequence of the human connexin26 gene. Gene 199, 165-171; and Castillo et al., DFNB1 Non-syndromic Hearing Impairment: Diversity of Mutations and Associated Phenotypes, Front. Mol. Neurosci., 22 December 2017, each of which is incorporated herein by reference. In some embodiments, the expression cassette of the isolated nucleic acid comprises a GJB2 basal promoter having a nucleotide sequence at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 47. An exemplary nucleotide sequence of a human GJB2 basal promoter is set forth in SEQ ID NO: 47: GGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGA GGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAG AGGCGGGGTGT

[0072] Examples of constitutive promoters include, without limitation, the retroviral Rous sarcoma virus (RSV) long terminal repeat (LTR) promoter (optionally with the RSV enhancer), the cytomegalovirus (CMV) promoter (optionally with the CMV enhancer) (See, e.g., Boshart et al., Cell, 41:521-530 (1985)) the simian vacuolating virus 40 (SV40) promoter, the dihydrofolate reductase promoter, the P-actin promoter, the phosphoglycerol kinase (PGK) promoter, and the elongation factor 1 -alpha 1 (EFla) promoter. In some embodiments, the promoter is a chicken beta-actin (CBA) promoter. In some embodiments,the promoter is an enhanced chicken P-actin promoter. In some embodiments, the promoter is a U6 promoter. Since the CBA promoter is constitutively active in all cell types, using a CBA promoter in the isolated nucleic acid described herein leads to promiscuous expression of GJB2 protein in all cell types, including cells that do notnormally express GJB2 protein (e.g., hair cells of the cochlea). Accordingly, in some embodiments, a CBA promoter is not used in the isolated nucleic acid described herein.

[0073] Inducible promoters allow regulation of gene expression and can be regulated by exogenously supplied compounds, environmental factors such as temperature, or the presence of a specific physiological state, e.g., acute phase, a particular differentiation state of the cell, or in replicating cells only. Inducible promoters and inducible systems are available from a variety of commercial sources, including, without limitation, Invitrogen, Clontech, and Ariad. Many other promoters have been described and can be readily selected by one of skill in the art. Examples of inducible promoters regulated by exogenously supplied promoters include the zinc-inducible sheep metallothionine (MT) promoter, the dexamethasone (Dex) -inducible mouse mammary tumor virus (MMTV) promoter, the T7 polymerase promoter system (WO 98 / 10088); the ecdysone insect promoter (No et al., Proc. Natl. Acad. Sci. USA, 93:3346- 3351 (1996)), the tetracycline-repressible system (Gossen et al., Proc. Natl. Acad. Sci. USA, 89:5547-5551 (1992)), the tetracycline-inducible system (Gossen et al., Science, 268:1766- 1769 (1995), see also Harvey et al., Curr. Opin. Chem. Biol., 2:512-518 (1998)), the RU486- inducible system (Wang et al., Nat. Biotech., 15:239-243 (1997) and Wang et al., Gene Ther., 4:432-441 (1997)) and the rapamycin-inducible system (Magari et al., J. Clin. Invest., 100:2865-2872 (1997)).

[0074] In some embodiments, the isolated nucleic acid comprises a gene regulatory element (GRE) (e.g., GJB2 GRE). Gene regulatory elements, as used herein, refer to a variety of DNA sequences that are involved in the regulation of gene expression. For example, a GRE may rely on the interactions involving DNA, cellular proteins (e.g., histones), and transcription factors to regulate gene expression.

[0075] In some embodiments, the isolated nucleic acid comprises gene regulatory elements which are cis-regulatory elements (e.g., cis-regulatory elements for the GJB2 gene). Cis- regulatory elements are regions of non-coding DNA which regulate the transcription of neighboring genes. Cis-regulatory elements are found in the vicinity of the genes that they regulate. Cis-regulatory elements typically regulate gene transcription by binding to transcription factors. In some embodiments, the gene regulatory elements impart cell- specific gene expression capabilities (e.g., cell specific GJB2 gene expression). In someembodiments, the gene regulatory elements are cis-regulatory elements associated with the GJB2 gene.

[0076] In some embodiments, the cis-regulatory elements of the GJB2 gene are enhancers. An enhancer, as used herein, refers to DNA sequences, which are located more distal to the transcription start site as compared to a promoter, capable of interacting with site- specific transcription factors to regulate gene expression in a cell-type specific manner. Enhancers confer cell-specific gene expression regulation by binding to the collection of transcription factors in a cell, which leads to transcriptional activation or inhibition through various mechanisms, e.g., recruitment of epigenetic enzymes that catalyze post-translational histone modifications, and recruitment of cofactors that promote DNA looping. Enhancers can be identified in the vicinity of the gene they regulate, or at a distance of hundreds of kilobases from their target genes. Multiple enhancers can act additively and redundantly to regulate gene expression e.g., Doane el al, Regulatory elements in molecular networks, Wiley Interdiscip Rev Syst Biol Med. 2017 May; 9(3)). In some embodiments, the enhancers described herein are enhancers capable of regulating genomic GJB2 gene expression. In some embodiments, the GJB2 enhancers are identified in the transcriptionally active sequences of the GJB2 gene. A transcriptionally active sequence, as used herein, refers to a region of DNA in a chromosome in which the DNA is in open chromatin confirmation such that the sequence is exposed, thereby allowing binding of transcription factors and transcription to take place. In some embodiments, the GJB2 enhancers are identified within approximately 1000 kb of a genomic GJB2 gene (e.g., within 1000 kb, within 900 kb, within 800 kb, within 700 kb, within 600 kb, within 500 kb, within 450 kb, within 400 kb, within 350 kb, within 300 kb, within 250 kb, within 200 kb, within 150 kb, within 100 kb, within 95 kb, within 90 kb, within 85 kb, within 85 kb, within 80 kb, within 75 kb, within 70 kb, within 65 kb, within 60 kb, within 55 kb, within 50 kb, within 45 kb, within 40 kb, within 35 kb, within 30 kb, within 25 kb, within 20 kb, within 15 kb, within 10 kb, or less upstream or downstream of the GJB2 gene). In some embodiments, the GJB2 enhancers are identified within approximately 200 kb of the GJB2 gene. In some embodiments, the GJB2 enhancers are identified within approximately 95 kb of the GJB2 gene (e.g., regions C-M listed in FIG. 3C) In some embodiments, the GJB2 enhancers are within the regions of DNA sequences near the GJB2 gene (FIG. 3C) listed in Table 1.Table 1. Human and mouse DNA regions that include GJB2 enhancers.

[0077] In some embodiments, a GJB2 GRE (e.g., a GJB2 enhancer) sequences can be identified from the regional sequence listed in Table 2. In some embodiments, a GJB2 GRE (e.g., a GJB2 enhancer) comprises at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900 at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400, at least 2500, at least 2600, at least 2700, at least 2800, at least 2800, at least 2900, at least 3000, at least 3100, at least 3200, at least 3300, at least 3400, at least 3500, at least 3600, at least 3700, at least 3800, at least 3900, at least 4000, at least 4100, at least 4200, at least 4200, at least 4400, at least 4500, at least 4600, at least 4700, at least 4800, at least 4900, at least 5000, or more consecutive nucleotides in any of the regional sequences described in Table 2 (e.g., human GJB2 regions A-M or mouse Gjb2 regions A-M). In some embodiments, a GJB2 GRE (e.g., a GJB2 enhancer) is identified with the transcriptionally active regions of the GJB2 gene (e.g., regions A and / or B). In some embodiments, the GJB2 GRE (e.g., a GJB2 enhancer) comprises at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900 at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400, at least 2500, at least 2600, at least 2700, at least 2800, at least 2800, at least 2900, at least 3000, at least 3100, at least 3200, at least 3300, at least 3400, at least 3500, at least 3600, at least 3700, at least 3800, at least 3900, at least 4000, at least 4100, at least 4200, at least 4200, at least 4400, at least 4500, at least 4600, at least 4700, at least 4800, at least 4900, at least 5000, or more consecutive nucleotides in within regions A and / or B. In some embodiments, the GJB2 GRE (e.g., a GJB2 enhancer) comprises at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900 at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400, at least 2500, at least 2600, at least 2700, at least 2800, at least 2800, at least 2900, at least 3000, at least 3100, at least 3200, at least 3300, at least 3400, at least 3500, at least 3600, at least 3700, at least 3800, at least 3900, at least 4000, at least 4100, at least 4200, at least 4200, at least 4400, at least 4500, at least 4600, at least 4700, at least 4800, at least 4900, at least 5000, or more consecutive nucleotides in within regions C-M. In some embodiments, the GJB2 GRE (e.g., a GJB2 enhancer) comprises nucleotide sequences out of the regions listed in Table 3.

[0078] In some embodiments, a GJB2 GRE (e.g., a GJB2 enhancer) is located on the sense strand of the GJB2 coding sequence in the genome. In some embodiments, GJB2 GRE (e.g., a GJB2 enhancer) is located on the reverse complement strand of the GJB2 coding sequence in the genome. It is within the skill of one in the art to select the appropriate sequence (e.g., GRE sequence on the sense strand, or GRE sequences on the reverse complement strand) when designing a vector using the enhancer sequences as described herein.

[0079] In some embodiments, a GJB2 GRE (e.g., a GJB2 enhancer) comprises at least 200, at least 250, at least 300, at least 350, at least 400, at least 450, at least 500, at least 550, at least 600, at least 650, at least 700, at least 750, at least 800, at least 850, at least 900 at least 1000, at least 1100, at least 1200, at least 1300, at least 1400, at least 1500, at least 1600, at least 1700, at least 1800, at least 1900, at least 2000, at least 2100, at least 2200, at least 2300, at least 2400, at least 2500, at least 2600, at least 2700, at least 2800, at least 2800, at least 2900, at least 3000, at least 3100, at least 3200, at least 3300, at least 3400, at least 3500, at least 3600, at least 3700, at least 3800, at least 3900, at least 4000, at least 4100, at least 4200, at least 4200, at least 4400, at least 4500, at least 4600, at least 4700, at least 4800, at least 4900, at least 5000, or more nucleotides. In some embodiments, a GJB2 GRE (e.g., a GJB2 enhancer) comprises 200-500 nucleotides or any number of nucleotides in between, 300-600 nucleotides or any number of nucleotides in between, 400-700 nucleotides or any number of nucleotides in between, 500-800 nucleotides or any number of nucleotides in between, 600-900 nucleotides or any number of nucleotides in between, 700-1000 nucleotides or any number of nucleotides in between, 1000-1500 nucleotides or any number of nucleotides in between, 1500-2000 nucleotides or any number of nucleotides in between. In some embodiments, a GJB2 GRE (e.g., a GJB2 enhancer) comprises 700 nucleotides.

[0080] In some embodiments, the GJB2 GRE is a human GJB2 enhancer. In some embodiments, the GJB2 GRE (e.g., a human GJB2 enhancer) comprises nucleotide sequence at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%,at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to any one of the GRE sequences as listed in Table 3.Table 3: Human GJB2 Enhancer Sequences

[0081] In some embodiments, the GJB2 GRE is a non-human primate (e.g., Cynomolgus macaque) GJB2 enhancer. In some embodiments, the GJB2 GRE (e.g., a Cynomolgus macaque GJB2 enhancer) comprises nucleotide sequence at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to any one of the GRE sequences as listed in Table 4.Table 4: Cynomolgus macaque GJB2 (mfGJB2) Enhancer Sequences

[0082] In some embodiments, the human GJB2 GREs share homology with the mfGJB2GREs. In some embodiments, the human GJB2 GREs correspond to mfGJB2 GREs as set forth in Table 5:Table 5: Homology between Human GJB2 GREs and mfGJB2 GREs

[0083] In some embodiments, the isolated nucleic acid comprises one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 9, or more) enhancers e.g., GJB2 enhancers). In some embodiments, the isolated nucleic acid comprises more than one enhancer, and the more than one enhancer are the same enhancers or different enhancers. . In some embodiments, the GJB2 GRE is positioned 5’ to the promoter. In other embodiments, the GJB2 GRE is positioned 3’ to the promoter. In some embodiments, the presence of the GJB2 enhancer(s) in the isolated nucleic acid facilitates cell-type specific expression of the GJB2 protein encoded by the isolated nucleic acid. In some embodiments, cells that normally express the GJB2 gene e.g., fibrocytes and supporting cells of the organ of Corti and nearby regions) have the transcriptional network to activate GJB2 expression regulated by the GJB2 enhancers, but not in cells that do not normally express GJB2 (e.g., hair cells and spiral ganglion neurons).

[0084] In some embodiments, the expression cassette of the isolated nucleic acid further comprises a 5 ' UTR. In some embodiments, the 5' UTR is a native 5' UTR of the genomic GJB2 gene. The 5' untranslated region (5' UTR) (also known as a leader sequence or leader RNA) is the region of an mRNA that is directly upstream of the initiation codon. The 5' UTR plays important roles in both transcriptional and translational regulation of the downstream gene (e.g., the GJB2 gene). In some embodiments, the isolated nucleic acid comprising a nucleotide sequence encoding a GJB2 5 ' UTR is also capable of expression GJB2 in a cell-specific manner (e.g., expressing GJB2 in cells that normally express it). In some embodiments, the nucleotide sequence encoding the GJB2 5 ' UTR comprises a portion of a nucleotide sequence encoding a full-length human GJB2 gene 5 ' UTR. In some embodiments, the 5 ' UTR is a human GJB2 gene exon 1 5 ' UTR. In some embodiments, the nucleotide sequence encoding a 5' UTR comprises at least 100 consecutive nucleotides, at least 200 consecutive nucleotides, at least 300 consecutive nucleotides, at least 400 consecutive nucleotides, at least 500 consecutive nucleotides, at least 600 consecutive nucleotides, at least 700 consecutive nucleotides, at least 800 consecutive nucleotides, at least 900 consecutive nucleotides, at least 1000 consecutive nucleotides, or more of a native full-length 5' UTR e.g., the human GJB2 gene exon 1 5' UTR). In some embodiments, the expression cassette comprises a nucleotide sequence encoding the 5' UTR having at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleotide sequence encoding a human GJB2 gene 5' UTR (e.g., human GJB2 exon 1 5' UTR). In some embodiments, the expression cassette comprises a nucleotide sequence encoding the 5' UTR having at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleotide sequence encoding a consecutive 300 bp of a human GJB2 gene 5' UTR (e.g., the human GJB2 gene exon 1 5' UTR) as set forth in SEQ ID NO: 53. In some embodiments, an exemplary nucleotide sequence encoding the 300 bp of the human GJB2 gene exon 1 5' UTR has a nucleotide sequence as set forth in SEQ ID NO: 53: GGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCG CTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAG GAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCC GACGCAGAGCAAACCGCCCAGAGTAG

[0085] In some embodiments, the cell specific GJB2 expression is achieved by incorporation of a nucleotide sequence encoding a basal promoter and a GJB2 5 ' UTR or a portion thereof (basal promoter / 5 ' UTR). In some embodiments, an expression cassette (e.g., GJB2 expression cassette) comprises a nucleotide sequence encoding a 5 ' UTR. In some embodiments, the isolated nucleic acid can further comprise additional nucleotide sequence encoding one or more GJB2 GREs (e.g., GJB2 enhancers). The nucleotide sequence encoding the GJB2 GREs and the nucleotide sequence encoding the basal promoter / 5 ' UTR can be placed in any order. In some embodiments, the nucleotide sequence encoding the GJB2GREs is placed 5 ' to the nucleotide sequence encoding the basal promoter / 5 ' UTR. In some embodiments, the isolated nucleic acid comprising a nucleotide sequence encoding a GJB2 basal promoter / 5 ' UTR is also capable of expressing GJB2 in a cell-specific manner (e.g., expressing GJB2 in cells that normally express it). In some embodiments, the nucleotide sequence encoding the basal promoter / 5 ' UTR comprises a portion of a nucleotide sequence encoding a full-length human GJB2 gene 5 ' UTR. In some embodiments, the 5' UTR comprises at least 100 consecutive nucleotides, at least 200 consecutive nucleotides, at least 300 consecutive nucleotides, at least 400 consecutive nucleotides, at least 500 consecutive nucleotides, at least 600 consecutive nucleotides, at least 700 consecutive nucleotides, at least 800 consecutive nucleotides, at least 900 consecutive nucleotides, at least 1000 consecutive nucleotides, or more of a native full-length 5' UTR e.g., the GJB2 5' UTR). In some embodiments, the 5 ' UTR is a human GJB2 gene exon 1 5 ' UTR. In some embodiments, the expression cassette comprises a nucleotide sequence encoding a basal promoter / 5' UTR having at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to a nucleotide sequence encoding the basal promoter and about 300 bp of a human GJB2 gene 5' UTR (e.g., the human GJB2 gene exon 1 5' UTR) (SEQ ID NO: 30). In some embodiments, an exemplary nucleotide sequence encoding the 300 bp of the human GJB2 gene basal promoter / exon 1 5' UTR has a nucleotide sequence as set forth in SEQ ID NO: 30:GGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGA GGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAG AGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCA GCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCC TAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCG CGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAG

[0086] In some embodiments, a nucleotide sequence encoding a basal promoter / 5 ' UTR (e.g., a human GJB2 basal promoter / exon 1 5 ' UTR) within the expression cassette (e.g., GJB2 expression cassette) further comprises an intron or a portion thereof. In some embodiments, the expression cassette of the isolated nucleic acid (e.g., GJB2 expression cassette) further comprises a conserved sequence of intron 1 of the GJB2 gene. In some embodiments, the nucleotide sequence encoding an intron (e.g., human GJB2 intron 1) has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 54.An exemplary nucleotide sequence encoding the conserved sequence of GJB2 intron 1 is set forth in SEQ ID NO: 54:AAGCAGGTGAGTTTGTGGTGTCGCCGATGTCCCTTCGGGGTACTCTAGCGCAGCCGCCTGGC TACTTGACCCACTGCCACCAAACGTTTTAAATTCACCGAAAGCTTAGCTTCGAAGCAAAGCT CCGTTTCGCCGGTGAAGCAGGAAGCCTTCGCTGCAGGAACTGACCTTTACCTCTTGGAGCGG CTTCTGCAGAAAAATCCCCGGGCAGAGATTTGGGCGGAGTTTGCCTAGAACTAACGCGGAGC CAGCCGATCCCGGCCTACCCCGGGGCCAAGATTTTAAGGGGTGAAGAGTCCCTTTTGCCTTT TCTGGATCCTGGTGATTCACCTAGTGTCTTCCCTAAGGAACTGAACCAACTCCTCCGCTGGC CTCTGGCAGCCCTCCAGGCGGTGCAGGATGGCGTGGGCCCGGTAGGAAGCTGCATGTAACCG CCCAGGGTCGGGAGGCCAGGAGGGCAGCTCCTCCTCTGACTTGAATATTGAAAACAAGAGGA TGCTTTTAAGAAAAAGAAGAAGGAGGATTCACTACCAGCTCTGAAGGGTGGAAAAGAGATGA TTCATCCGGATTGTGGAGAGGGTGGAATCTTGTTTAGGAGAGCGTTGGTTGTGGCAGGCAGG GTGTAACTATGAATCAGTGAAGACAATTCACATCCTGGGATGAAAAGAAGGCCATGGGCTCA CAGGAGATTATCCACTGGCCTCTCCACATCCGCTTGCAGTAAGGAGTGTGGGACTCTCCCAA GCTTCAGCGCTGAACTGCAATGCAGTGACGTCGCTTAAGA

[0087] In some embodiments, the nucleotide sequence encoding a basal promoter / 5 'UTR / intron has at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 31. An exemplary nucleotide sequence encoding human GJB2 basal promoter / 5 'UTR / conserved sequence of intron 1 is set forth in SEQ ID NO: 31:GGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGA GGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAG AGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCA GCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCC TAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCG CGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAA GCAGGTGAGTTTGTGGTGTCGCC GATGTCCCTTCGGGGTACTCTAGCGCAGCCGCCTGGCTACTTGACCCACTGCCACCAAACGT TTTAAATTCACCGAAAGCTTAGCTTCGAAGCAAAGCTCCGTTTCGCCGGTGAAGCAGGAAGC CTTCGCTGCAGGAACTGACCTTTACCTCTTGGAGCGGCTTCTGCAGAAAAATCCCCGGGCAG AGATTTGGGCGGAGTTTGCCTAGAACTAACGCGGAGCCAGCCGATCCCGGCCTACCCCGGGG CCAAGATTTTAAGGGGTGAAGAGTCCCTTTTGCCTTTTCTGGATCCTGGTGATTCACCTAGT GTCTTCCCTAAGGAACTGAACCAACTCCTCCGCTGGCCTCTGGCAGCCCTCCAGGCGGTGCA GGATGGCGTGGGCCCGGTAGGAAGCTGCATGTAACCGCCCAGGGTCGGGAGGCCAGGAGGGC AGCTCCTCCTCTGACTTGAATATTGAAAACAAGAGGATGCTTTTAAGAAAAAGAAGAAGGAG GATTCACTACCAGCTCTGAAGGGTGGAAAAGAGATGATTCATCCGGATTGTGGAGAGGGTGG AATCTTGTTTAGGAGAGCGTTGGTTGTGGCAGGCAGGGTGTAACTATGAATCAGTGAAGACA ATTCACATCCTGGGATGAAAAGAAGGCCATGGGCTCACAGGAGATTATCCACTGGCCTCTCC ACATCCGCTTGCAGTAAGGAGTGTGGGACTCTCCCAAGCTTCAGCGCTGAACTGCAATGCAG TGACGTCGCTTAAGA

[0088] In some embodiments, the expression cassette (e.g., GJB2 expression cassette) comprises a nucleotide sequence encoding a proximal promoter of the human GJB2 gene. Insome embodiments, the proximal promoter of the human GJB2 gene has at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 102. In some embodiments, an exemplary nucleotide sequence encoding the human GJB2 gene proximal promoter has a nucleotide sequence as set forth in SEQ ID NO: 102. In some embodiments, the expression cassette (e.g., GJB2 expression cassette) comprises SEQ ID NO: 102: GACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCG GGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGT

[0089] In some embodiments, the expression cassette (e.g., GJB2 expression cassette) comprises a nucleotide sequence encoding a 5' UTR of the human GJB2 gene. In some embodiments, the 5' UTR of the human GJB2 gene has at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 103 or CC. In some embodiments, an exemplary nucleotide sequence encoding the human GJB2 gene 5' UTR has a nucleotide sequence as set forth in SEQ ID NO: 103 or CC. In some embodiments, an exemplary nucleotide sequence encoding the human GJB2 gene 5' UTR has a nucleotide sequence comprising SEQ ID NO: 103 and SEQ ID NO: 104. In some embodiments, the expression cassette (e.g., GJB2 expression cassette) comprises SEQ ID NO: 103:AACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCG GGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCG GCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCC GAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAG

[0090] In some embodiments, the expression cassette (e.g., GJB2 expression cassette) comprises SEQ ID NO: 104:AGCAAACCGCCCAGAGTAGAAG

[0091] In some embodiments, the expression cassette (e.g., GJB2 expression cassette) comprises a nucleotide sequence encoding a proximal promoter and a 5' UTR of the human GJB2 gene. In some embodiments, the proximal promoter and the 5' UTR of the human GJB2 gene has at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to the nucleotide sequence set forth in SEQ ID NO: 105. In some embodiments, an exemplary nucleotide sequence encoding the humanGJB2 gene proximal promoter and 5' UTR has a nucleotide sequence as set forth in SEQ ID NO: 105. In some embodiments, the expression cassette (e.g., GJB2 expression cassette) comprises SEQ ID NO: 105: GACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCG GGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACT TTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAG ACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGG CGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGA CCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAG

[0092] An isolated nucleic acid described herein may also contain an artificial intron, desirably located between the promoter / enhancer sequence and the protein coding sequence (e.g., nucleotide sequence encoding GJB2 protein). In some embodiments, an intron is a synthetic or artificial (e.g., heterologous) intron. Examples of synthetic introns include an intron sequence derived from SV-40 (referred to as the SV-40 T intron sequence) and intron sequences derived from chicken beta-actin gene. In some embodiments, a transgene described by the disclosure comprises one or more (1, 2, 3, 4, 5, or more) artificial introns. In some embodiments, the one or more artificial introns are positioned between a promoter and a nucleotide sequence encoding the GJB2 protein.

[0093] In some embodiments, the expression cassette (e.g., the GJB2) further comprises a nucleotide sequence encoding a 3 ' UTR located 3 ' of the nucleotide sequence encoding the GJB2 protein. In some embodiments, the 3' UTR is a GJB2 gene 3 ' UTR. In some embodiments, the 3 ’UTR is a GJB2 gene exon 2 3 ' UTR. In some embodiments, the nucleotide sequence encoding the 3 ' UTR has at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 32. An exemplary nucleotide sequence encoding GJB2 gene exon 2 3' UTR is set forth in SEQ ID NO: 32:CGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCT CAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACC ATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCT AAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTT CCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAG GATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACA CAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGA ACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTG CCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATG TAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGT AAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTG TAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCC TCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTA CTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCAT CGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATG GGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGA CTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTA CCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAG CTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTAT GCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTG TTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCT TGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTAT AATAA

[0094] In some embodiments, the expression cassette of the isolated nucleic acid comprises a de-targeting agent that restricts or reduces the transgene expression (e.g., GJB2 expression) in a cell type (e.g., hair cell or spiral ganglion neurons). In some embodiments, incorporation of one or more miRNA binding sites into an expression allows for de-targeting of transgene expression in a cell-type specific manner (e.g., in hair cell or spiral ganglion neurons). In some embodiments, one or more miRNA binding sites are positioned in the 3 ' UTR (e.g., GJB2 exon 2 3 ' UTR of the expression cassette of the isolated nucleic acid).

[0095] In some embodiments, an expression cassette comprises one or more (e.g., 1, 2, 3, 4, 5, or more) miRNA binding sites that de-target expression of GJB2 from cells that do not normally express GJB2 (e.g., hair cell or spiral ganglion neurons). In some embodiments, the expression cassette of the isolated nucleic acid comprises one or more miR binding sites for detargeting neuron cells (e.g., spiral ganglion neurons), e.g., binding sites for neuron enriched miRs as described in Jovicic et al., Comprehensive Expression Analyses of Neural Cell- Type-Specific miRNAs Identify New Determinants of the Specification and Maintenance of Neuronal Phenotypes, J Neurosci. 2013 Mar 20; 33(12): 5127-5137, which is incorporated herein by reference. Non-limiting examples of neuron enriched miRs include miR- 124, miR- 127, miR-129, miR-129*, miR-136, miR-136*, miR-137, miR-154, miR-3OO-3p, miR-323, miR-329, miR-341, miR-369-5p, miR-376a, miR-376b-3p, miR-376c, miR-379, miR-382, miR-382*, miR-410, miR-411, miR-433, miR-434, miR-495, miR-541, miR-543*, miR- 551b, miR-143, miR-449a, miR-219-2-3p, miR-126, miR-126*, miR-141, miR-142-3p, miR-142-5p, miR-146a, miR-150, miR-200c, or miR-223. In some embodiments, the expression cassette of the isolated nucleic acid comprises one or more miR binding sites for detargeting hair cells (e.g., inner or outer hair cell), e.g., binding sites for hair cell enriched miRs as described in Li et al., MicroRNAs in hair cell development and deafness, Curr OpinOtolaryngol Head Neck Surg. 2010 Oct; 18(5): 459-465, which is incorporated herein by reference. Non-limiting examples of neuron enriched miRs include miR-96, miR-182, miR- 183, miR-18a, or miR-99a. In some embodiments, the GJB2 exon 2 3 ' UTR of the expression cassette comprises one or more miR binding sites for detargeting neuron cells and hair cells. In some embodiments, the GJB2 exon 2 3 ' UTR of the expression cassette comprises one or more miR binding sites for miR- 124.

[0096] Aspects of the disclosure relate to gene therapy vectors comprising an isolated nucleic acid as described herein. A gene therapy vector may be a viral vector (e.g., a lentiviral vector, an adeno-associated virus vector, an adenoviral (Ad) vector, etc.), a plasmid, a closed- ended DNA (e.g., ceDNA), a lipid / DNA nanoparticle, etc. In some embodiments, a gene therapy vector is a viral vector. In some embodiments, an expression cassette encoding a protein (e.g., GJB2 protein) is flanked by one or more viral replication sequences, for example, lentiviral long terminal repeats (LTRs) or adeno-associated virus (AAV) inverted terminal repeats (ITRs).

[0097] The isolated nucleic acids of the disclosure may be recombinant adeno-associated virus (AAV) vectors (rAAV vectors). In some embodiments, an isolated nucleic acid as described by the disclosure comprises two adeno-associated virus (AAV) inverted terminal repeat (ITR) sequences, or variants thereof. The isolated nucleic acid (e.g., the recombinant AAV vector) may be packaged into a capsid protein and administered to a subject and / or delivered to a selected target cell. “Recombinant AAV (rAAV) vectors” are typically composed of, at a minimum, an expression cassette (e.g., expression cassette for GJB2), and 5 ' and 3 ' AAV inverted terminal repeats (ITRs). The isolated nucleic acids may also comprise a region encoding, for example, 5 ' and 3 ' untranslated regions (UTRs), and / or an expression control sequence (e.g., a poly- A tail).

[0098] Generally, ITR sequences are about 145 bp in length. Preferably, substantially the entire sequence encoding the ITR is used in the isolated nucleic acid, although some degree of minor modification of these sequences is permissible. The ability to modify these ITR sequences is within the skill of one in the art. (See, e.g., texts such as Sambrook et al., Molecular Cloning. A Laboratory Manual, 2d ed., Cold Spring Harbor Laboratory, New York (1989); and K. Fisher et al., J Virol., 70:520 532 (1996)). An example of such a molecule employed in the present invention is an isolated nucleic acid comprising an expression cassette encoding a GJB2 protein, in which the expression cassette comprising the nucleotide sequences GJB2 protein and GJB2 gene regulatory elements (GREs) are flanked by the 5 ' and 3 ' AAV ITR sequences. The AAV ITR sequences may be obtained from anyknown AAV, including presently identified mammalian AAV types. In some embodiments, the isolated nucleic acid (e.g., the rAAV vector) comprises at least one ITR having a serotype selected from AAV1, AAV2, AAV5, AAV6, AAV6.2, AAV7, AAV8, AAV9, AAV10, AAV11, and variants thereof. In some embodiments, the isolated nucleic acid comprises a region e.g., a first region) encoding an AAV2 ITR.

[0099] In some embodiments, the isolated nucleic acid further comprises a region (e.g., a second region, a third region, a fourth region, etc.) comprising a second AAV ITR. In some embodiments, the second AAV ITR has a serotype selected from AAV1, AAV2, AAV5, AAV6, AAV6.2, AAV7, AAV8, AAV9, AAV10, AAV11, and variants thereof. In some embodiments, the second AAV ITR is an AAV2 ITR. In some embodiments, the second ITR is a mutant ITR that lacks a functional terminal resolution site (TRS). The term “lacking a terminal resolution site” can refer to an AAV ITR that comprises a mutation (e.g., a sense mutation such as a non-synonymous mutation, or missense mutation) that abrogates the function of the terminal resolution site (TRS) of the ITR, or to a truncated AAV ITR that lacks a nucleic acid sequence encoding a functional TRS (e.g., a ATRS ITR, or AfTR). Without wishing to be bound by any particular theory, an rAAV vector comprising an ITR lacking a functional TRS produces a self-complementary rAAV vector, for example, as described by McCarthy (2008) Molecular Therapy 16( 10): 1648- 1656. In some embodiments, the isolated nucleic acid comprises a 5 ' AAV2 ITR and a 3 ' AAV2 ITR.

[0100] An exemplary 5 ' AAV2 ITR nucleotide sequence is set forth in SEQ ID NO: 34:TTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCG ACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCA ACTCCATCACTAGGGGTTCCTAGAT

[0101] An exemplary 5 ' ITR nucleotide sequence is set forth in SEQ ID NO: 106:CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGG TCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGG GTTCCT

[0102] exemplary 3 ' AAV2 ITR nucleotide sequence is set forth in SEQ ID NO: 35:CCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCG GGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCA GAGAGGGAGTGGCCA

[0103] An exemplary 3 ' ITR nucleotide sequence is set forth in SEQ ID NO: 107:AGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCC GGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGC GCGCAG

[0104] In some embodiments, the isolated nucleic acid (e.g., rAAV vector) described herein comprises a 5' ITR sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 34 or 106.

[0105] In some embodiments, the isolated nucleic acid e.g., rAAV vector) described herein comprises a 3' ITR sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 35 or 107.

[0106] In some embodiments, the isolated nucleic acid (e.g., rAAV vector) described herein comprises a posttranscriptional response element. As used herein, the term “posttranscriptional response element” refers to a nucleic acid sequence that, when transcribed, adopts a tertiary structure that enhances expression of a gene. Examples of posttranscriptional regulatory elements include, but are not limited to, woodchuck hepatitis virus posttranscriptional regulatory element (WPRE), mouse RNA transport element (RTE), constitutive transport element (CTE) of the simian retrovirus type 1 (SRV-1), the CTE from the Mason-Pfizer monkey virus (MPMV), and the 5' untranslated region of the human heat shock protein 70 (Hsp70 5' UTR). In some embodiments, the isolated nucleic acid (e.g., rAAV vector) comprises a woodchuck hepatitis virus posttranscriptional regulatory element (WPRE).

[0107] In some embodiments, the isolated nucleic acid (e.g., rAAV vector) described herein comprises a posttranscriptional response element having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 108. An exemplary posttranscriptional response element is set forth in SEQ ID NO: 108: GATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGC TCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTA TGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGG CCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTG GGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCA CGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACT GACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGC CACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACC TTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAG ACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCATCGGACTAG

[0108] In some embodiments, the vector further comprises conventional control elements which are operably linked with elements of the GJB2 coding sequence in a manner that permits its transcription, translation, and / or expression in a cell transfected with the vector or infected with the virus produced by the disclosure. Expression control sequences include appropriate transcription initiation, termination; efficient RNA processing signals, such as splicing and polyadenylation (polyA) signals; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (e.g., Kozak consensus sequence); sequences that enhance protein stability. A polyadenylation sequence generally is inserted following the coding sequences and optionally before a 3 ' AAV ITR sequence. A rAAV construct useful in the disclosure may also contain an intron, desirably located between the promoter / enhancer sequence and the transgene.

[0109] In some embodiments, the isolated nucleic acid e.g., rAAV vector) described herein comprises a polyadenylation signal sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 109. An exemplary polyadenylation signal sequence is set forth in SEQ ID NO: 109: GTCGACTAGAGCTCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTT GCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAA AATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGG GCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGA

[0110] In some embodiments, an AAV vector described herein comprises a GJB2 proximal promoter (e.g., SEQ ID NO: 102), a GJB2 5' UTR (e.g., SEQ ID NO: 103 and CC), a nucleotide sequence encoding a GJB2 gene product (e.g., SEQ ID NO: 2), a GJB2 3 ' UTR (e.g., SEQ ID NO: 32), a WPRE (e.g., SEQ ID NO: 108), and a bovine growth hormone poly A signal (e.g., SEQ ID NO: 109). In some embodiments, an AAV vector described herein comprises a nucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 110. An exemplary AAV vector sequence is set forth in SEQ ID NO: 110: GACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCG GGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACT TTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAG ACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGG CGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGA CCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCG GATCCGCCACCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCC ACCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAGGCTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTGCAGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACATGAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGATCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTCTTCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTCCATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGTCCCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATCCTGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAAGCCAGTTTACCCATACGATGTTCCAGATTACGCTTAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCATCGGACTAGGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGGTCGACTAGAGCTCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGA

[0111] In some embodiments, an AAV vector described herein comprises a 5 ' ITR (e.g.,SEQ ID NO: 106), a GJB2 proximal promoter e.g., SEQ ID NO: 102), a GJB2 5' UTR (e.g.,SEQ ID NO: 103 and CC), a nucleotide sequence encoding a GJB2 gene product (e.g., SEQ ID NO: 2), a GJB2 3 ' UTR (e.g., SEQ ID NO: 32), a WPRE (e.g., SEQ ID NO: 108), a bovine growth hormong poly A signal (e.g., SEQ ID NO: 109), and a 3 ' ITR (e.g., SEQ ID NO: 107). In some embodiments, an AAV vector described herein comprises a nucleotide sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 111. An exemplary AAV vector sequence is set forth in SEQ ID NO: 111:CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGG TCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGG GTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACCAGGGTAATGGGGATCCTC TAGAACGCGTTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGT CGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGC GCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAA AAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGA CTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCA GAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAA CCGCCCAGAGTAGAAGCGGATCCGCCACCATGGATTGGGGCACGCTGCAGACGATCCTGGGG GGTGTGAACAAACACTCCACCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCG CATTATGATCCTCGTTGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCT GCAACACCCTGCAGCCAGGCTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCAC ATCCGGCTATGGGCCCTGCAGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCA CGTGGCCTACCGGAGACATGAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAAT TTAAGGACATCGAGGAGATCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACC TACACAAGCAGCATCTTCTTCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGT CATGTACGACGGCTTCTCCATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACA CTGTGGACTGCTTTGTGTCCCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCA GTGTCTGGAATTTGCATCCTGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTG TTCTGGGAAGTCAAAAAAGCCAGTTTACCCATACGATGTTCCAGATTACGCTTAAGGCGCGC CACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGG GATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATT CTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACA ATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTT CTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACT TTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGC CAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTT TCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAG TGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTA TGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAG GCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGT CTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCA TAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGC TTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGAC TGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTG ACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTA AAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTT CAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAAC ATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAA CCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTG AGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAA TAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACA TTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCA AGCTTATCGATGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTT AACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTAT TGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATG AGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACC CCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCT CCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGC TGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTC GCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAA TCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCC TTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCATCGGACTAGGAATTCA TCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGGTCGACTAGAGC TCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCG TGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATT GCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAA GGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCG GTACCAAACCTAGGTAATACCCATTACCCTGGTAGATAAGTAGCATGGCGGGTTAATCATTA ACTACAAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACT GAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGA GCGAGCGCGCAG

[0112] In some embodiments, an AAV vector described herein comprises 5 ' ITR, a GJB2 basal promoter, a 5 ' UTR (e.g., GJB2 exon 1 5 ' UTR), Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), an optional HA tag, a 3 ' UTR (e.g., GJB2 exon 2 3 ' UTR), a WPRE, a bovine growth hormone poly A signal, and a 3 ' ITR (e.g., vector c70). In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 36. An exemplary nucleotide sequence for vector c70 encoding a mouse GJB2 protein with an HA tag is set forth in SEQ ID NO: 36 (mouse GJB2 coding sequence in boldface; HA tag underlined):TTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCG CCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCG CGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTTTAATTAA GACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCG GGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACT TTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCGGATCCGCCACCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCCACCAGCATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGGCTGCAAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTGCAGCTGATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACATGAAAAGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGATCAAAACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCGGGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGCAACGTCTGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTTCCAGGCCCACAGAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATTCTGCTAAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAGTCTACCCATACGATGTTCCAGATTACGCTTAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGC AAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCG CAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAG CGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATT TCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTT CGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGG GGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGA GTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGG GCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCAC TCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCG CTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGG CCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAG GTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTC CTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGA AGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTA TTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGC TGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGA AGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGC AACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAA TAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACT GGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTA TGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAG GATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGT TCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGA TCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATA CTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTAC CGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTT CGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAG GGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTC CTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTT

[0113] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% sequence identity to SEQ ID NO: 61. An exemplary nucleotide sequence for vector c70 encoding a human GJB2 protein with an HA tag is set forth in SEQ ID NO: 61 (human GJB2 coding sequence in boldface; HA tag underlined):TTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCG CCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCG CGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTTTAATTAA GACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCG GGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACT TTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAG ACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGG CGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGA CCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCG GATCCGCCACCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCC ACCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGT GGCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAG GCTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTG CAGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACA TGAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGA TCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTC TTCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTC CATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGT CCCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATC CTGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAA GCCAGTTTACCCATACGATGTTCCAGATTACGCTTAAGGCGCGCCACCCCTGCAGGGAATTC CGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCT CAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACC ATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCT AAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTT CCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAG GATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACA CAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGA ACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTG CCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATG TAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGT AAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACT TTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTG TAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCC TCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTA CTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCAT CGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATG GGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGA CTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTA CCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAG CTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTAT GCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTG TTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCT TGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTAT AATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAAC CTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTT CGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAG CTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAG GGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTC CTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGG AGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTT

[0114] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 62. An exemplary nucleotide sequence for vector c70 encoding a mouse GJB2 protein with an HA tag is set forth in SEQ ID NO: 62 (mouse GJB2 coding sequence in boldface; no HA tag):CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAG GCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTG GTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAAT TTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAG TTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTT TTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTT TGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATA CCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGT GTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACA GCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAA GCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTT TATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGG GGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCT GGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGG GCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAG AGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTTTAATTAAGACCT CGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAG CTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGG TGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCC GGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCC GCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCGGATCCGCCACCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCCACCAG CATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGTGGCTG CAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGGCTGC AAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTGCAGCT GATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACATGAAA AGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGATCAAA ACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCG GGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGC AACGTCTGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTTCCAGG CCCACAGAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATTCTGCTAAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAGTCTAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCG GCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACC GTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATG TCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACC CCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTG ATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCC TTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAA GTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAG CGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAG TTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGC ATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGA TGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCA ACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGG GATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGA GCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAAC TACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGA CCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGA GCGTGGGTCTCGCGGTATCATTG

[0115] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 63. An exemplary nucleotide sequence for vector c70 encoding a mouse GJB2 protein with a HA tag is set forth in SEQ ID NO: 63 (human GJB2 coding sequence in boldface; no HA tag): CAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAG GCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTG GTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAAT TTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAG TTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTT TTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTT TGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATA CCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGT GTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACG GGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACA GCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAA GCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTT TATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGG GGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCT GGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGG GCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAG AGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTTTAATTAAGACCT CGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAG CTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCC AGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGG TGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCC GCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCGGATCC GCCACCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCCACCAG CATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGTGGCTG CAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAGGCTGC AAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTGCAGCT GATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACATGAGA AGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGATCAAA ACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTCTTCCG GGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTCCATGC AGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGTCCCGG CCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATCCTGCT GAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAAGCCAG TTTAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGA CAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCC AACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACT CCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCC TGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTG GTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACA AGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCT TTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTT AATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAA AACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCC CCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAAT TTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTAT TCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTT CCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTA AGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATC TCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTG GGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAG TTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAG CTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAAT ATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTAT AGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCA CATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGT AATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAAT ACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGA ACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACT GGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTA TCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGT CTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCT GACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGC TTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAG GGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCT TGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTC GGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGC GTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCAT CGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAG TTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTC CCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCT ATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCC CTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCT TTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGG TATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTAC GCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTAC ACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCG CCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTA CGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTG ATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCC AAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCG ATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAA AATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGT TAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCG GCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACC GTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATG TCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACC CCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTG ATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCC TTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAA GTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAG CGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAG TTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGC ATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGA TGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCA ACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGG GATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGA GCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAAC TACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGA CCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGA GCGTGGGTCTCGCGGTATCATTG

[0116] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE1 ), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 2 3 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c81.1).

[0117] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 64. An exemplary nucleotide sequence for vector c81.1 encoding eGFP is set forth in SEQ ID NO: 64 (hGJB2 GRE1 underlined; eGFP coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTA AGAGAGC AC T TGGGAAGAGCCCCCGAGGGCAGCCGGGGCTTGCCGCCTCACCCTTTTGGTTT CACATCCCAGAAATCAGTAAGGCAGGAATTGGAGGCTGCTTCTTGCCTTAGCAACTCGGTGA CCTTAGGCAGAACAGTTCAGCCTTCTGAGTGTCCTTCCTCTTCTGTAAGGGGAGCGTAAACC GTCCTCCATGCAGAACGTGTACTGTGCCTGGCACAGCACTGGGGCATTAGGATCTCCAAATT AAAGGCTCACTCTGCGGGATGGAGGCAGCCACAGCTGGAAGAAGGAACATTTGGGGCCAGAA GTCCCCCTACCTCCGTCCTAAGAGAGAAGATGGGAATAACGACCCTCGCTGAAATGATTGCT CTCTGGCCAGCTCGCCTCGCATCCACATCCAAATCTGGGAGGCACAGAGCGCATCAGGACAT CGGGTTCTGTCAGTGTAATGGGCGTGGCTCCTGACCTTCTGTCTGTATCAGAGAAGATAAGG GAGAACATTTGAAAGAAAGGAGAAAGAAGATAGCCACTGGAGAACAGAGCAAAGGAGCCAGC AGAAAAAGACGAGACGGCTGTAGCCCCACAGGAAGCAGAAACCGATAGGCTAAGTAGGATAC ACACAAAGAAAAGTAGATCCCGAGAGGCATTTCCCCGAGGGCTTTCATGTGGTTTCTCGTGA GGAGAAGCTGACTGCAGGGTGTTTGAAAGAACGACTTATGCAGCCATAAAAAATGATGAGTT CATGTCCTTTGTAGGGACATGGATGATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGG GGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGC GCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGG TGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGC GCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCG CAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCT CCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTG TCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCAC CGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCT TCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGC TACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGT GAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGG ACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATG GCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGG CAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGC TGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGC GATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCT GTACAAGTAATAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTA AGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTA GCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACA ATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCC GTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACG CTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGA TCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCA CTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTC GGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCA TCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACA CTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCAC AACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACC AAACGACGAGCGTGACACCA

[0118] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 65. An exemplary nucleotide sequence for vector c81.1 encoding human GJB2 is set forth in SEQ ID NO: 65 hGJB2 GRE1 underlined; human GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTA AGAGAGC AC T TGGGAAGAGCCCCCGAGGGCAGCCGGGGCTTGCCGCCTCACCCTTTTGGTTT CACATCCCAGAAATCAGTAAGGCAGGAATTGGAGGCTGCTTCTTGCCTTAGCAACTCGGTGA CCTTAGGCAGAACAGTTCAGCCTTCTGAGTGTCCTTCCTCTTCTGTAAGGGGAGCGTAAACC GTCCTCCATGCAGAACGTGTACTGTGCCTGGCACAGCACTGGGGCATTAGGATCTCCAAATT AAAGGCTCACTCTGCGGGATGGAGGCAGCCACAGCTGGAAGAAGGAACATTTGGGGCCAGAA GTCCCCCTACCTCCGTCCTAAGAGAGAAGATGGGAATAACGACCCTCGCTGAAATGATTGCT CTCTGGCCAGCTCGCCTCGCATCCACATCCAAATCTGGGAGGCACAGAGCGCATCAGGACAT CGGGTTCTGTCAGTGTAATGGGCGTGGCTCCTGACCTTCTGTCTGTATCAGAGAAGATAAGG GAGAACATTTGAAAGAAAGGAGAAAGAAGATAGCCACTGGAGAACAGAGCAAAGGAGCCAGCAGAAAAAGACGAGACGGCTGTAGCCCCACAGGAAGCAGAAACCGATAGGCTAAGTAGGATAC ACACAAAGAAAAGTAGATCCCGAGAGGCATTTCCCCGAGGGCTTTCATGTGGTTTCTCGTGA GGAGAAGCTGACTGCAGGGTGTTTGAAAGAACGACTTATGCAGCCATAAAAAATGATGAGTT CATGTCCTTTGTAGGGACATGGATGATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGG GGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGC GCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGG TGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGC GCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCG CAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCT CCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGATTGGGGCACGCTGCAGACGATCC TGGGGGGTGTGAACAAACACTCCACCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATT TTTCGCATTATGATCCTCGTTGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTT TGTCTGCAACACCCTGCAGCCAGGCTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCT CCCACATCCGGCTATGGGCCCTGCAGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCC ATGCACGTGGCCTACCGGAGACATGAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAG TGAATTTAAGGACATCGAGGAGATCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGT GGACCTACACAAGCAGCATCTTCTTCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTC TATGTCATGTACGACGGCTTCTCCATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCC CAACACTGTGGACTGCTTTGTGTCCCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGA TTGCAGTGTCTGGAATTTGCATCCTGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGA TATTGTTCTGGGAAGTCAAAAAAGCCAGTTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGC ATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAG CTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATT TGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAA GCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCA CTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGAT ATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAG AGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACA TTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCT TAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAA AGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAA TGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTG CAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAG CCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCA TGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTA CCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGG AAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGG AGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTG GACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCC CTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTA AGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCT TGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTG TCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGT AACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAAT AATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTC TGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTA TGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTT CTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGC AACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACC ACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCAT CGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTG CGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGG CCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCT CCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATA GCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGT GCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTG CATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAG GGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAG GAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGG GCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGC GCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCA CACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGT GTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGC TTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGC TCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGT GATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTC CACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCT ATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATT TAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCT CAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTG ACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCC GGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCT CGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTG GCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAAT ATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAG TATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTG TTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGA GTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGA ACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTG ACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTAC TCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGC CATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGG AGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCG GAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0119] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 66. An exemplary nucleotide sequence for vector c81.1 encoding mouse GJB2 is set forth in SEQ ID NO: 66 hGJB2 GRE1 underlined; mouse GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTAAGAGAGCACTTGGGAAGAGCCCCCGAGGGCAGCCGGGGCTTGCCGCCTCACCCTTTTGGTTTCACATCCCAGAAATCAGTAAGGCAGGAATTGGAGGCTGCTTCTTGCCTTAGCAACTCGGTGACCTTAGGCAGAACAGTTCAGCCTTCTGAGTGTCCTTCCTCTTCTGTAAGGGGAGCGTAAACCGTCCTCCATGCAGAACGTGTACTGTGCCTGGCACAGCACTGGGGCATTAGGATCTCCAAATTAAAGGCTCACTCTGCGGGATGGAGGCAGCCACAGCTGGAAGAAGGAACATTTGGGGCCAGAAGTCCCCCTACCTCCGTCCTAAGAGAGAAGATGGGAATAACGACCCTCGCTGAAATGATTGCTCTCTGGCCAGCTCGCCTCGCATCCACATCCAAATCTGGGAGGCACAGAGCGCATCAGGACATCGGGTTCTGTCAGTGTAATGGGCGTGGCTCCTGACCTTCTGTCTGTATCAGAGAAGATAAGGGAGAACATTTGAAAGAAAGGAGAAAGAAGATAGCCACTGGAGAACAGAGCAAAGGAGCCAGCAGAAAAAGACGAGACGGCTGTAGCCCCACAGGAAGCAGAAACCGATAGGCTAAGTAGGATACACACAAAGAAAAGTAGATCCCGAGAGGCATTTCCCCGAGGGCTTTCATGTGGTTTCTCGTGAGGAGAAGCTGACTGCAGGGTGTTTGAAAGAACGACTTATGCAGCCATAAAAAATGATGAGTTCATGTCCTTTGTAGGGACATGGATGATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCCACCAGCATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGGCTGCAAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTGCAGCTGATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACATGAAAAGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGATCAAAACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCGGGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGCAACGTCTGGTGAAATGCAACGCTTGGCCCTGCCC CAATACAGTGGACTGCTTCATTTCCAGGCCCACAGAAAAGACTGTCTTCACCGTGTTTATGA TTTCTGTGTCTGGAATTTGCATTCTGCTAAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAGTCTACCCATACGATGTTCCAGATTACGCTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTT AATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAA GGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAA GATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGC TATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACAC TATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCAT GACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTAC TTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCAT GTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGA CACCA

[0120] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE2), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 2 3 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c81.2).

[0121] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 48. An exemplary nucleotide sequence for vector c81.2 encoding eGFP is set forth in SEQ ID NO: 48 hGJB2 GRE2 underlined; eGFP coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC AGTGATGCCTGAAACCTCAGATGGTACTGAACCCTCTATATAATCTGTTTTTTCCTATACAT ACAAACCTACCATAAGGCTTAATGGTAAGAGATTAACAATAAAGAATAATAAAACAACACTT ATAACAATGTATAACAATATATTGTAATATAAGTTTTTGGATGCAGTCTCTCTCTCAAAATG CTATCATATTTTCCAACTGTGGTTGACTACAGGTAACTGGAACCACAAAAATGAAACAGTGGATAAGAGGGCGACTCCTGTACCAAAGAAAAAAATAGAGTGTTGCAGCTGTAACATAGTTGAA TGACTGAGTTAGACTGCATAACTGACACACAAAACCACATAAATATAAATGAAGGAATCTCT GGGTGTAATCTGGTGCAAAGGTGACTGTGTTAATCATTAATCCACAAGTTGCTATCCTGAAG TGTGCCAAATGCTTTATGTTTATTTCATCACATAGCTCTATAAAGAAAGGATTTGTAATTCC TTTCTACAGAAGTGGAAAGTAAGTCTTAAGACTCAAAAAACTTTAAAAACTACAATGAAGTA ACAACTTTTATTAATTTATTTTGTGTCTTTCCAGAATTTCTATATATATAGGAATGTGATAT GAATCTATATGTGAATTGAATCTACATGAATATTGATGACTTTTATTTCCCCTTTTGCACAT AAGATAGAATATTTTACCTACTATTCCACACTTTGCTTTTCTTAACATATCATGGGATC T T T TTATATAAGTGAACAAAGAGTTTCTTCATTCTTTCACACAGTTTAATTAAGACCTCGAAGGG GACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAG GACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCC GAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCG GCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAG GACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGG CCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGTGAGCAAG GGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGG CCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGA AGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACC TACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTC CGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACA AGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGC ATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCA CAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCC ACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGC GACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGA CCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTC TCGGCATGGACGAGCTGTACAAGTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCC AGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAA GGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACC CCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAA AACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGAC CCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCA TTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGT TTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTC TTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTC TGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTT CTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATG TCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACA GCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAAT CGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAA TATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAA TGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAAC GCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAA AATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTA AAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGC CTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTA GATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATG GTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAA TAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAA ATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAA AATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGAT ACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCC TTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGG CGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTC AGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCC TGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTC GGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGA CGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTG CCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTG GGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCA TCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCC TTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCA TTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGG ATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCC TAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCA AAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTG CCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCA TACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGG TTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTC CCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTT AGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTT CACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTC TTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTT TGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAA AATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACA ATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCC CTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCT GCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATA CGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTT TCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATC CGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGT ATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGC TCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTT ACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTT CCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGG GCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAG TCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACC ATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAAC CGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGA ATGAAGCCATACCAAACGACGAGCGTGACACCA

[0122] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 67. An exemplary nucleotide sequence for vector c81.2 encoding human GJB2 is set forth in SEQ ID NO: 67 hGJB2 GRE2 underlined; human GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC AGTGATGCCTGAAACCTCAGATGGTACTGAACCCTCTATATAATCTGTTTTTTCCTATACAT ACAAACCTACCATAAGGCTTAATGGTAAGAGATTAACAATAAAGAATAATAAAACAACACTT ATAACAATGTATAACAATATATTGTAATATAAGTTTTTGGATGCAGTCTCTCTCTCAAAATG CTATCATATTTTCCAACTGTGGTTGACTACAGGTAACTGGAACCACAAAAATGAAACAGTGG ATAAGAGGGCGACTCCTGTACCAAAGAAAAAAATAGAGTGTTGCAGCTGTAACATAGTTGAA TGACTGAGTTAGACTGCATAACTGACACACAAAACCACATAAATATAAATGAAGGAATCTCT GGGTGTAATCTGGTGCAAAGGTGACTGTGTTAATCATTAATCCACAAGTTGCTATCCTGAAG TGTGCCAAATGCTTTATGTTTATTTCATCACATAGCTCTATAAAGAAAGGATTTGTAATTCC TTTCTACAGAAGTGGAAAGTAAGTCTTAAGACTCAAAAAACTTTAAAAACTACAATGAAGTA ACAACTTTTATTAATTTATTTTGTGTCTTTCCAGAATTTCTATATATATAGGAATGTGATAT GAATCTATATGTGAATTGAATCTACATGAATATTGATGACTTTTATTTCCCCTTTTGCACAT AAGATAGAATATTTTACCTACTATTCCACACTTTGCTTTTCTTAACATATCATGGGATC T T T TTATATAAGTGAACAAAGAGTTTCTTCATTCTTTCACACAGTTTAATTAAGACCTCGAAGGG GACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAG GACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCC GAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCG GCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAG GACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGG CCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGATTGGGGC ACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCCACCAGCATTGGAAAGATCTGGCT CACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGTGGCTGCAAAGGAGGTGTGGGGAG ATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAGGCTGCAAGAACGTGTGCTACGAT CACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTGCAGCTGATCTTCGTGTCCACGCC AGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACATGAGAAGAAGAGGAAGTTCATCA AGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGATCAAAACCCAGAAGGTCCGCATC GAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTCTTCCGGGTCATCTTCGAAGCCGC CTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTCCATGCAGCGGCTGGTGAAGTGCA ACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGTCCCGGCCCACGGAGAAGACTGTC TTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATCCTGCTGAATGTCACTGAATTGTG TTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAAGCCAGTTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGA GGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGAC CTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGA GCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTC ACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCAT ATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGT TCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCC ACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGAC AAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATA GGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCA GATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCA CCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATG ATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGAT GTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTG TGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACA GTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAA GTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTG AAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGA ATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGT AAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCT TTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTT ATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGT TGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCC GTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTG TGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGG TTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGC ACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGT TGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGG ACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCT CAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCT CGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAA TAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGT GGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCG GACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCT CGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCA GTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGC ATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAG CGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAA GCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAA AAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCC CTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTT AAAAAAT GAGC T GATT T AAC AAAAAT T T AACGCGAAT T T T AAC AAAAT AT T AACGT T T AC AA TTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACAC CCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCG CGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTT TCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTT CTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAAT ATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCG GCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGA TCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGA GTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCG GTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAA TGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAG AATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACG ATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCT TGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0123] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 68. An exemplary nucleotide sequence for vector c81.2 encoding mouse GJB2 is set forth in SEQ ID NO: 68 (hGJB2 GRE2 underlined; mouse GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC AGTGATGCCTGAAACCTCAGATGGTACTGAACCCTCTATATAATCTGTTTTTTCCTATACAT ACAAACCTACCATAAGGCTTAATGGTAAGAGATTAACAATAAAGAATAATAAAACAACACTT ATAACAATGTATAACAATATATTGTAATATAAGTTTTTGGATGCAGTCTCTCTCTCAAAATG CTATCATATTTTCCAACTGTGGTTGACTACAGGTAACTGGAACCACAAAAATGAAACAGTGG ATAAGAGGGCGACTCCTGTACCAAAGAAAAAAATAGAGTGTTGCAGCTGTAACATAGTTGAA TGACTGAGTTAGACTGCATAACTGACACACAAAACCACATAAATATAAATGAAGGAATCTCT GGGTGTAATCTGGTGCAAAGGTGACTGTGTTAATCATTAATCCACAAGTTGCTATCCTGAAGTGTGCCAAATGCTTTATGTTTATTTCATCACATAGCTCTATAAAGAAAGGATTTGTAATTCC TTTCTACAGAAGTGGAAAGTAAGTCTTAAGACTCAAAAAACTTTAAAAACTACAATGAAGTA ACAACTTTTATTAATTTATTTTGTGTCTTTCCAGAATTTCTATATATATAGGAATGTGATAT GAATCTATATGTGAATTGAATCTACATGAATATTGATGACTTTTATTTCCCCTTTTGCACAT AAGATAGAATATTTTACCTACTATTCCACACTTTGCTTTTCTTAACATATCATGGGATC T T T TTATATAAGTGAACAAAGAGTTTCTTCATTCTTTCACACAGTTTAATTAAGACCTCGAAGGG GACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAG GACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCC GAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCG GCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAG GACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGG CCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGATTGGGGC ACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCCACCAGCATTGGAAAGATCTGGCT CACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGTGGCTGCAAAGGAGGTGTGGGGAG ATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGGCTGCAAGAATGTATGCTACGAC CACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTGCAGCTGATCATGGTGTCCACGCC AGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACATGAAAAGAAACGGAAGTTCATGA AGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGATCAAAACCCAGAAGGTCCGTATC GAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCGGGTCATCTTTGAAGCCGT CTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGCAACGTCTGGTGAAATGCA ACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTTCCAGGCCCACAGAAAAGACTGTC TTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATTCTGCTAAATATCACAGAGCTGTG CTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAGTCTACCCATACGATGTTC CAGATTACGCTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATT AAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCT AGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTC AGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTA ATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTA GGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTC TCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTC CTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTT TGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTT TGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATT ATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACT ATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTG TCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAG ACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTA ATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGT CCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAG AGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGG GAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGAT TAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTT AAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGA GCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAA TGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAA AGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAA TGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGG TTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGA AAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAA TGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCC TGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCG GGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGC TGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATC GTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCT ACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGG CCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCC GCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTG TGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAA GGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAG GTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACA ATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGT TGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGA CGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCG CCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCA ACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCG TGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTC GCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATT TAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGC CATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGA CTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGG GATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGA ATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGAT GCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTG TCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGA GGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTA TAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGT GCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGAC AATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTC CGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAAC GCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGG ATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGC ACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACT CGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGC ATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAAC ACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCA CAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATAC CAAACGACGAGCGTGACACCA

[0124] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE3), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 2 3 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c.81.3).

[0125] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 49. An exemplary nucleotide sequencefor vector c.81.3 is set forth in SEQ ID NO: 49 (hGJB2 GRE3 underlined; eGFP coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTG CTATCTATCATCTTGAAGGGCTTCTGGAACAAGTTAGAATAGAGTCAACACTCATGAACTGC TGTAGCAAAAAAAACTATAGATGTAGGATTGACAAGGGCAATAGAGCGATGACTCCCTGGCT GTGTTGTATTTGATGGACGGCAGTAGCTTTTCACAAAATGCTCATTTGGATGTTTCAAATTA AAACGTTTCACTTTCTAGAACCAATTACGTGGTCAGTTTAGCTCCTGAGGTCCCAGTCAGAG GGGTATTCTGTAGCTTGCAAAGCCTCTCTTTGGGGACTGGACATGGAGTCTGTGGTCTTAGA ATTCAGAACCGGGAGAATGTGTTAGCCACTCATCTAAGCTATTCCTTAAACGCTTTCAGAGC CATCTCCACTGTGGGGAAAGAAGTTCTTTGTGTTCTCTGACTTAGTCTCATTCTAAAAAAAA AAAAAAAAAAAAAAAAAAAGCAATTGCAATACCCAGAGCGCACAGTAGATGGCACTGAGACT TGTCGGAAAGCTGGACGCACTCAAGAGGTGGCAGAAAAATCTATAGGTAAGCTTTTCTTCTA GTCTGGTGTTGCTGCTCCTGACCTTATTAATGGGCTGAGAAATAGATTTCTTTCCTTTCCTT TTCTTTTTTATATGAAATTAAATGAAGTATAAAAGAATATGAGAATGTGTTGCTATTAGCAA GGATAAGTAATGCTTTAGGAAACGTTTGGTTCATGTGTGTGTTTTCAGACTGATGTGTGTCC TGGATCCAGTGTAAAATGTACTTCTGTCTGTAGGTCTCTGCCACAGAAAAGTTGGAAAGCCA TTGTTGTATTCCATTTCCAGGGCAACAAAAGATACCACTGTCACTTCATGTGAAATGGTGTT GTTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTT CGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCC CTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCC ACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGC CCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCC AACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAG AGTAGAAGCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCG AGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCC ACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCC CACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGA AGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTC TTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGT GAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATC AAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTA CCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCA CCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTC GTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACC TTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAG CTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATA TTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTT CCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTG GTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCAC CTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGT TGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGC CACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGT TTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAAT AAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCA TTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGC GCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAA AAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCA ACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTA AAAAAT GAGC T GATT T AAC AAAAAT T T AACGCGAAT T T T AAC AAAAT AT T AACGT T T AC AAT TTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACC CGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAA GCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGC GAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTT CTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTC TAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATA TTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGG CATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGAT CAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAG TTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGG TATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAAT GACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGA ATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGA TCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTT GATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0126] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 70. An exemplary nucleotide sequence for vector c.81.3 is set forth in SEQ ID NO: 70 hGJB2 GRE3 underlined; human GJB2 coding sequence in bold face): CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTG CTATCTATCATCTTGAAGGGCTTCTGGAACAAGTTAGAATAGAGTCAACACTCATGAACTGC TGTAGCAAAAAAAACTATAGATGTAGGATTGACAAGGGCAATAGAGCGATGACTCCCTGGCTGTGTTGTATTTGATGGACGGCAGTAGCTTTTCACAAAATGCTCATTTGGATGTTTCAAATTA AAACGTTTCACTTTCTAGAACCAATTACGTGGTCAGTTTAGCTCCTGAGGTCCCAGTCAGAG GGGTATTCTGTAGCTTGCAAAGCCTCTCTTTGGGGACTGGACATGGAGTCTGTGGTCTTAGA ATTCAGAACCGGGAGAATGTGTTAGCCACTCATCTAAGCTATTCCTTAAACGCTTTCAGAGC CATCTCCACTGTGGGGAAAGAAGTTCTTTGTGTTCTCTGACTTAGTCTCATTCTAAAAAAAA AAAAAAAAAAAAAAAAAAAGCAATTGCAATACCCAGAGCGCACAGTAGATGGCACTGAGACT TGTCGGAAAGCTGGACGCACTCAAGAGGTGGCAGAAAAATCTATAGGTAAGCTTTTCTTCTA GTCTGGTGTTGCTGCTCCTGACCTTATTAATGGGCTGAGAAATAGATTTCTTTCCTTTCCTT TTCTTTTTTATATGAAATTAAATGAAGTATAAAAGAATATGAGAATGTGTTGCTATTAGCAA GGATAAGTAATGCTTTAGGAAACGTTTGGTTCATGTGTGTGTTTTCAGACTGATGTGTGTCC TGGATCCAGTGTAAAATGTACTTCTGTCTGTAGGTCTCTGCCACAGAAAAGTTGGAAAGCCA TTGTTGTATTCCATTTCCAGGGCAACAAAAGATACCACTGTCACTTCATGTGAAATGGTGTT GTTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTT CGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCC CTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCC ACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGC CCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCC AACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAG AGTAGAAGCCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCCA CCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGTG GCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAGG CTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTGC AGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACAT GAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGAT CAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTCT TCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTCC ATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGTC CCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATCC TGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAAG CCAGTTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAA ATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCAT TTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTG AAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCT ATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGT TATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGA GGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGG GTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAA GTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAA GTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATG TTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGAT TTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGT TGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAG AAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTT GTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAAC ACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTC GCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGG AGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAG ATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTG AAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGA TCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTA CTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTAT CAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTA ATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGAT TGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTT GCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGT TTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACT TTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCT TTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTC CCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCT TCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCT TCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGC CACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTC ATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCC ACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCC GGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGA TGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACC GCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCAC GTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTG CTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTT GTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTT TGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTT AACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGC TCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTT TCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGT TAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAA CCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGT CGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGG TGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTT TAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTC GCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTT ACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACA TGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAAC GACGAGCGTGACACCA

[0127] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 71. An exemplary nucleotide sequencefor vector c.81.3 is set forth in SEQ ID NO: 71 (hGJB2 GRE3 underlined; mouse GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTG CTATCTATCATCTTGAAGGGCTTCTGGAACAAGTTAGAATAGAGTCAACACTCATGAACTGC TGTAGCAAAAAAAACTATAGATGTAGGATTGACAAGGGCAATAGAGCGATGACTCCCTGGCT GTGTTGTATTTGATGGACGGCAGTAGCTTTTCACAAAATGCTCATTTGGATGTTTCAAATTA AAACGTTTCACTTTCTAGAACCAATTACGTGGTCAGTTTAGCTCCTGAGGTCCCAGTCAGAG GGGTATTCTGTAGCTTGCAAAGCCTCTCTTTGGGGACTGGACATGGAGTCTGTGGTCTTAGA ATTCAGAACCGGGAGAATGTGTTAGCCACTCATCTAAGCTATTCCTTAAACGCTTTCAGAGC CATCTCCACTGTGGGGAAAGAAGTTCTTTGTGTTCTCTGACTTAGTCTCATTCTAAAAAAAA AAAAAAAAAAAAAAAAAAAGCAATTGCAATACCCAGAGCGCACAGTAGATGGCACTGAGACT TGTCGGAAAGCTGGACGCACTCAAGAGGTGGCAGAAAAATCTATAGGTAAGCTTTTCTTCTA GTCTGGTGTTGCTGCTCCTGACCTTATTAATGGGCTGAGAAATAGATTTCTTTCCTTTCCTT TTCTTTTTTATATGAAATTAAATGAAGTATAAAAGAATATGAGAATGTGTTGCTATTAGCAA GGATAAGTAATGCTTTAGGAAACGTTTGGTTCATGTGTGTGTTTTCAGACTGATGTGTGTCC TGGATCCAGTGTAAAATGTACTTCTGTCTGTAGGTCTCTGCCACAGAAAAGTTGGAAAGCCA TTGTTGTATTCCATTTCCAGGGCAACAAAAGATACCACTGTCACTTCATGTGAAATGGTGTT GTTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTT CGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCC CTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCC ACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGC CCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCC AACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAG AGTAGAAGCCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCCA CCAGCATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGTG GCTGCAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGG CTGCAAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTGC AGCTGATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACAT GAAAAGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGAT CAAAACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCGGGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGCAACGTCTGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTTCCAGGCCCACAGAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATTCTGCTAAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAGTCTACCCATACGATGTTCCAGATTACGCTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGG GCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTG ATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCAC TCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCG CTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTC TCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGG CCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAG GTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCA AATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAA GAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTC CTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCA CGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGA AGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTA TTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAG TACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGC TGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGA AGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAA CCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0128] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE4), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 2 3 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c.81.4).

[0129] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 72. An exemplary nucleotide sequence for vector c.81.4 is set forth in SEQ ID NO: 72 hGJB2 GRE4 underlined; eGFP coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTAACTGGGCAATGCGTTAAACTGGCTTTTTTGACTTCCCAGAACAATATCTAATTAGCAAATAACACAATTCAGTGACATTCAGCAGGATGCAAATTCCAGACACTGCAATCATGAACACTGTGAAGACAGTCTTCTCCGTGGGCCGGGACACAAAGCAGTCCACAGTGTTGGGACAAGGCCAGGCGTTGCACTTCACCAGCCGCTGCATGGAGAAGCCGTCGTACATGACATAGAAGACGTACATGAAGGCGGCTTCGAAGATGACCCGGAAGAAGATGCTGCTTGTGTAGGTCCACCACAGGGAGCCTTCGATGCGGACCTTCTGGGTTTTGATCTCCTCGATGTCCTTAAATTCACTCTTTATCTCCCCCTTGATGAACTTCCTCTTCTTCTCATGTCTCCGGTAGGCCACGTGCATGGCCACTAGGAGCGCTGGCGTGGACACGAAGATCAGCTGCAGGGCCCATAGCCGGATGTGGGAGATGGGGAAGTAGTGATCGTAGCACACGTTCTTGCAGCCTGGCTGCAGGGTGTTGCAGACAAAGTCGGCCTGCTCATCTCCCCACACCTCCTTTGCAGCCACAACGAGGATCATAATGCGAAAAATGAAGAGGACGGTGAGCCAGATCTTTCCAATGCTGGTGGAGTGTTTGTTCACACCCCCCAGGATCGTCTGCAGCGTGCCCCAATCCATCTTCTACTCTGGGCGGTTTGCTCTGGAAAAGACGAATGCACACAACACAG GAATCACTAGCTAGGACAGAACAGGGAGACTTCTCTGAGTCTGGGTAAGCAAGCATGCT T AA ATCTCTTCCTGAGCAAACACCAACTCTTACACAACCTCACCAAAACAGGTGAAGACAGAACC AACTTAGTTTGTCATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACC ACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTC CTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAATA AAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTG AAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCA TTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0130] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 73. An exemplary nucleotide sequence for vector c.81.4 is set forth in SEQ ID NO: 73 hGJB2 GRE4 underlined; human GJB2 coding sequence in bold face): CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTA ACTGGGCAATGCGTTAAACTGGCTTTTTTGACTTCCCAGAACAATATCTAATTAGCAAATAA CACAATTCAGTGACATTCAGCAGGATGCAAATTCCAGACACTGCAATCATGAACACTGTGAA GACAGTCTTCTCCGTGGGCCGGGACACAAAGCAGTCCACAGTGTTGGGACAAGGCCAGGCGT TGCACTTCACCAGCCGCTGCATGGAGAAGCCGTCGTACATGACATAGAAGACGTACATGAAG GCGGCTTCGAAGATGACCCGGAAGAAGATGCTGCTTGTGTAGGTCCACCACAGGGAGCCTTC GATGCGGACCTTCTGGGTTTTGATCTCCTCGATGTCCTTAAATTCACTCTTTATCTCCCCCT TGATGAACTTCCTCTTCTTCTCATGTCTCCGGTAGGCCACGTGCATGGCCACTAGGAGCGCT GGCGTGGACACGAAGATCAGCTGCAGGGCCCATAGCCGGATGTGGGAGATGGGGAAGTAGTG ATCGTAGCACACGTTCTTGCAGCCTGGCTGCAGGGTGTTGCAGACAAAGTCGGCCTGCTCAT CTCCCCACACCTCCTTTGCAGCCACAACGAGGATCATAATGCGAAAAATGAAGAGGACGGTG AGCCAGATCTTTCCAATGCTGGTGGAGTGTTTGTTCACACCCCCCAGGATCGTCTGCAGCGT GCCCCAATCCATCTTCTACTCTGGGCGGTTTGCTCTGGAAAAGACGAATGCACACAACACAG GAATCACTAGCTAGGACAGAACAGGGAGACTTCTCTGAGTCTGGGTAAGCAAGCATGCT T AA ATCTCTTCCTGAGCAAACACCAACTCTTACACAACCTCACCAAAACAGGTGAAGACAGAACC AACTTAGTTTGTCATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGG CGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCG CGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCC CCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAG CGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAG CAAACCGCCCAGAGTAGAAGCCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGA ACAAACACTCCACCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATG ATCCTCGTTGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACAC CCTGCAGCCAGGCTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGC TATGGGCCCTGCAGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCC TACCGGAGACATGAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGA CATCGAGGAGATCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAA GCAGCATCTTCTTCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTAC GACGGCTTCTCCATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGA CTGCTTTGTGTCCCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTG GAATTTGCATCCTGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGG AAGTCAAAAAAGCCAGTTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTG TTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTC AGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGT AGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAA AGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAG GCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGT TTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGG TGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCA TTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTA CACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGA TACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATT CGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGA GAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGT GAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTT AGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACA GGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGA TTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGG GGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTC TGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAAT T T T T TAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGT T TAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTC CAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCA TTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAAT TTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCT AATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAA TTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCT GCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTA TAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGG TGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTC CTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCT TGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGA AATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCC TTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGC TCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCG CCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAG CTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGAC CCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGG GAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTG ATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGT CGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGC AGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGT CAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACG CGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTC CTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGT TCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGT AGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAA TAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATT TATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTT AACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTG CTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGAC GGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATG TGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCT ATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGG GAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTC ATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCA ACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACC CAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATC GAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAAT GATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAG AGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACA GAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAG TGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTT TTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAA GCCATACCAAACGACGAGCGTGACACCA

[0131] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 74. An exemplary nucleotide sequence for vector c.81.4 is set forth in SEQ ID NO: 74 hGJB2 GRE4 underlined; mouse GJB2 coding sequence in bold face): CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTA ACTGGGCAATGCGTTAAACTGGCTTTTTTGACTTCCCAGAACAATATCTAATTAGCAAATAA CACAATTCAGTGACATTCAGCAGGATGCAAATTCCAGACACTGCAATCATGAACACTGTGAA GACAGTCTTCTCCGTGGGCCGGGACACAAAGCAGTCCACAGTGTTGGGACAAGGCCAGGCGT TGCACTTCACCAGCCGCTGCATGGAGAAGCCGTCGTACATGACATAGAAGACGTACATGAAG GCGGCTTCGAAGATGACCCGGAAGAAGATGCTGCTTGTGTAGGTCCACCACAGGGAGCCTTC GATGCGGACCTTCTGGGTTTTGATCTCCTCGATGTCCTTAAATTCACTCTTTATCTCCCCCT TGATGAACTTCCTCTTCTTCTCATGTCTCCGGTAGGCCACGTGCATGGCCACTAGGAGCGCT GGCGTGGACACGAAGATCAGCTGCAGGGCCCATAGCCGGATGTGGGAGATGGGGAAGTAGTG ATCGTAGCACACGTTCTTGCAGCCTGGCTGCAGGGTGTTGCAGACAAAGTCGGCCTGCTCAT CTCCCCACACCTCCTTTGCAGCCACAACGAGGATCATAATGCGAAAAATGAAGAGGACGGTG AGCCAGATCTTTCCAATGCTGGTGGAGTGTTTGTTCACACCCCCCAGGATCGTCTGCAGCGT GCCCCAATCCATCTTCTACTCTGGGCGGTTTGCTCTGGAAAAGACGAATGCACACAACACAG GAATCACTAGCTAGGACAGAACAGGGAGACTTCTCTGAGTCTGGGTAAGCAAGCATGCT T AA ATCTCTTCCTGAGCAAACACCAACTCTTACACAACCTCACCAAAACAGGTGAAGACAGAACC AACTTAGTTTGTCATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGG CGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCG CGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGG TTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCC CCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAG CGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAG CAAACCGCCCAGAGTAGAAGCCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCA ACAAACACTCCACCAGCATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATG ATCCTCGTGGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGGCTGCAAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGC TCTGGGCTCTGCAGCTGATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCC TACCGGAGACATGAAAAGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGA CATCGAAGAGATCAAAACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCA CCAGCATCTTCTTCCGGGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTAC AATGGCTTCTTCATGCAACGTCTGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGA CTGCTTCATTTCCAGGCCCACAGAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTG GAATTTGCATTCTGCTAAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGA AAGTCCAAAAGACCAGTCTACCCATACGATGTTCCAGATTACGCTTAAAGGCGCGCCACCCC TGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAG GCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACC TTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAG CTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCA CTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATA TTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTT CCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCA CGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACA AAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAG GTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTG GTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCAC CTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGA TAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATG TACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGT GGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAG TACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACA GATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAG TTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGA AAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAA TATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTA AGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTT TAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAA TATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTA TCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTT GCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCG TATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGT GGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGT TGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGC CACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCA CTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTT GCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGA CCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTC AGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTC GAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGT TTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAAT AAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTG GGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGG ACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTC GCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAG TGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCAT CTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCA TTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGC GCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAG CTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAA AAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCC TTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCA ACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTA AAAAAT GAGC T GATT T AAC AAAAAT T T AACGCGAAT T T T AAC AAAAT AT T AACGT T T AC AAT TTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACC CGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAA GCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGC GAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTT CTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTC TAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATA TTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGG CATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGAT CAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAG TTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGG TATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAAT GACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0132] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE5), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 2 3 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c.81.5).

[0133] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 50. An exemplary nucleotide sequence for vector c.81.5 is set forth in SEQ ID NO: 50 hGJB2 GRE5 underlined; eGFP coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTA AGCTACTAACTACAACCACGAGATTATAGATGTTTGCTGATATTGTTCTCAGTTTGGTTATT GTGTTGTTTATGAAT GAAAGTAGTGTATGTTTGTGTGAATTTTTGTTTTTAATTTTTTATGA GTGCCCTAACAAAGATTACAAATTGGGAATACAAACTCCAGAGCAATGGAGACAGTGACACT TTTGTGGAGGGGTACATGTGGCTGTTCGGGTGGTTATTAACACAGGCTGCTGCCCCTGCCCT GCAATGGGAATCCCCAGGGCATTGGAGGATTCAACCTCTTGCAGTTACCTCTTGTAAGACAG CAGATGGCAGCAGAGAGAGGCTTTGCACATCCCTGCAGGTTCTAGTTTGCACAAAGGGCTTC TGAGAGACCTATCAACCAATTATAACATCAAGTGGCAAAAAGAGTCCTTGATAAGTTATTTC GCTTCTCAAAGAAACCGAAAACGCCAAACTAATCACTAGTCTTGTTTTTTTTTTTCCTGGCA AAAGCCTGCTATCTTTCATGATTTAGCTTTCATGAAATTGTTCCTGAAGACCCCCAAAAGAAACAATTTCATGCCCCGAACTCTGTTCAGAGACTTTGCTGTGCCTGTCATGTCCAGCTTGCCA TATCCTGTTTTGTAAAGTAGCCACCTTATATACACACCTGCTGTCTGCACTGTGACCTCCTT TCAAAATCATCTTTGGTTCTTCAGAGGCCTGGAATAATGCTCTGCCCAGATGAAGATCTCCG TAAATGTGTTTTTGAAATGGCTAATCAAATAATGGATACCCTTAGGTAT T T T TGCAGAAACA CTTGGCAGCCTTCCATAATATCCCTACTATGAAATGGAAACTTGTGAATGAGATGTGGCTTT AATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCG GACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCC GTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGG CGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCT CGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACG CCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTA GAAGCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCT GGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCT ACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACC CTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCA GCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCA AGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAAC CGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGA GTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGG TGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAG CAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCA GTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGA CCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAAGGCGCGCCACCCCTGCA GGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAA CCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAA ATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCT GCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTA AGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTT AAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCAC AGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTT AAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGT TACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTA TTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTG TAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTA TGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAA CAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGC AAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACC ACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTA GCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACC ATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATT TGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTG TTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAG AATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATT GCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTA TTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAAT GATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATA ATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGA TAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTC CTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATG GCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCC CGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACG GCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGA CAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCA CCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTT CCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGAC GAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGA GATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGC CCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAA TGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGC AGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCG AGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTC ACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAG CGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGT GCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAA GCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCC GCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCT AAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAAC TTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTG ACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCC TATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAA AT GAGC T GATT T AAC AAAAAT T T AACGCGAAT T T T AAC AAAAT AT T AACGT T T AC AAT T T T A TGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCC AACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTG TGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGA CGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTA GACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAA TACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGA AAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATT TTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGT TGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTT CGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATT ATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACT TGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTA TGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGG AGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATC GTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0134] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 75. An exemplary nucleotide sequence for vector c.81.5 is set forth in SEQ ID NO: 75 hGJB2 GRE5 underlined; human GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTA AGCTACTAACTACAACCACGAGATTATAGATGTTTGCTGATATTGTTCTCAGTTTGGTTATT GTGTTGTTTATGAAT GAAAGTAGTGTATGTTTGTGTGAATTTTTGTTTTTAATTTTTTATGA GTGCCCTAACAAAGATTACAAATTGGGAATACAAACTCCAGAGCAATGGAGACAGTGACACT TTTGTGGAGGGGTACATGTGGCTGTTCGGGTGGTTATTAACACAGGCTGCTGCCCCTGCCCT GCAATGGGAATCCCCAGGGCATTGGAGGATTCAACCTCTTGCAGTTACCTCTTGTAAGACAG CAGATGGCAGCAGAGAGAGGCTTTGCACATCCCTGCAGGTTCTAGTTTGCACAAAGGGCTTC TGAGAGACCTATCAACCAATTATAACATCAAGTGGCAAAAAGAGTCCTTGATAAGTTATTTC GCTTCTCAAAGAAACCGAAAACGCCAAACTAATCACTAGTCTTGTTTTTTTTTTTCCTGGCA AAAGCCTGCTATCTTTCATGATTTAGCTTTCATGAAATTGTTCCTGAAGACCCCCAAAAGAA ACAATTTCATGCCCCGAACTCTGTTCAGAGACTTTGCTGTGCCTGTCATGTCCAGCTTGCCA TATCCTGTTTTGTAAAGTAGCCACCTTATATACACACCTGCTGTCTGCACTGTGACCTCCTT TCAAAATCATCTTTGGTTCTTCAGAGGCCTGGAATAATGCTCTGCCCAGATGAAGATCTCCG TAAATGTGTTTTTGAAATGGCTAATCAAATAATGGATACCCTTAGGTAT T T T TGCAGAAACA CTTGGCAGCCTTCCATAATATCCCTACTATGAAATGGAAACTTGTGAATGAGATGTGGCTTT AATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCG GACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCC GTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGG CGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCT CGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACG CCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCCACCAG CATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGTGGCTG CAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAGGCTGC AAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTGCAGCT GATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACATGAGA AGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGATCAAA ACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTCTTCCG GGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTCCATGC AGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGTCCCGG CCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATCCTGCT GAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAAGCCAG TTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAG ACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCC CAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAAC TCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCC CTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAA AGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACA GCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAA GTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCG CATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGG ATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCC AACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGG GGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACG AGCGTGACACCA

[0135] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 76. An exemplary nucleotide sequence for vector c.81.5 is set forth in SEQ ID NO: 76 hGJB2 GRE5 underlined; mouse GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTA AGCTACTAACTACAACCACGAGATTATAGATGTTTGCTGATATTGTTCTCAGTTTGGTTATT GTGTTGTTTATGAAT GAAAGTAGTGTATGTTTGTGTGAATTTTTGTTTTTAATTTTTTATGA GTGCCCTAACAAAGATTACAAATTGGGAATACAAACTCCAGAGCAATGGAGACAGTGACACT TTTGTGGAGGGGTACATGTGGCTGTTCGGGTGGTTATTAACACAGGCTGCTGCCCCTGCCCT GCAATGGGAATCCCCAGGGCATTGGAGGATTCAACCTCTTGCAGTTACCTCTTGTAAGACAG CAGATGGCAGCAGAGAGAGGCTTTGCACATCCCTGCAGGTTCTAGTTTGCACAAAGGGCTTC TGAGAGACCTATCAACCAATTATAACATCAAGTGGCAAAAAGAGTCCTTGATAAGTTATTTC GCTTCTCAAAGAAACCGAAAACGCCAAACTAATCACTAGTCTTGTTTTTTTTTTTCCTGGCA AAAGCCTGCTATCTTTCATGATTTAGCTTTCATGAAATTGTTCCTGAAGACCCCCAAAAGAA ACAATTTCATGCCCCGAACTCTGTTCAGAGACTTTGCTGTGCCTGTCATGTCCAGCTTGCCATATCCTGTTTTGTAAAGTAGCCACCTTATATACACACCTGCTGTCTGCACTGTGACCTCCTT TCAAAATCATCTTTGGTTCTTCAGAGGCCTGGAATAATGCTCTGCCCAGATGAAGATCTCCG TAAATGTGTTTTTGAAATGGCTAATCAAATAATGGATACCCTTAGGTAT T T T TGCAGAAACA CTTGGCAGCCTTCCATAATATCCCTACTATGAAATGGAAACTTGTGAATGAGATGTGGCTTT AATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCG GACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCC GTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGG CGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCT CGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACG CCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTA GAAGCCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCCACCAG CATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGTGGCTG CAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTGGCTGC AAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTGCAGCT GATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACATGAAA AGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGATCAAA ACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCG GGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGC AACGTCTGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTTCCAGG CCCACAGAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATTCTGCT AAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAG TCTACCCATACGATGTTCCAGATTACGCTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCA TTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGC TGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTT GAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAG CCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCAC TGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATA TCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGA GAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACAT TGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTT AAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAA GATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAAT GGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGC AGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGC CTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCAT GTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTAC CTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGA AAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGA GGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGG ACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCC TTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAA GGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTT GACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGT CAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTA ACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATA ATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCT GGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTAT GTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTC TCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCA ACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCA CCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGT GTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGC GCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGC CTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTC CCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAG CGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTG CCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGC ATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGG GGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGG AACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGG CGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCG CAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCAC ACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTG TGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCT TTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCT CCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTG ATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCC ACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTA TTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTT AACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTC AGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGA CGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCG GGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTC GTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGG CACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATA TGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGT TTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAG TGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAA CGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGA CGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACT CACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCC ATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGA GCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGG AGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0136] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE7), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 2 3 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c.81.7).

[0137] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 51. An exemplary nucleotide sequencefor vector c.81.7 is set forth in SEQ ID NO: 51 (hGJB2 GRE7 underlined; eGFP coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC TCAGCTGGAGTGACGCACCTCATCCATGCGGGCCTGGCGTCTGGAAGGTGGCTGGGTCTCTC GGGC T T GAGCACCATCATCTTAGCTCCAACATGTCATTATTCCTTCCTCACTGAGGACTTTT CTGCTTCCTAATTGGTTGTTGAAGATGAGGCCCCCATGCTCTTTTAAGAAAACCTGTTGTGC CCCAGGCTTGGCTGTGATGGGCACTGACTCATACAGAAGTAGAAAGGCCTGCTGAGTCATCA ACACTCGTGCGACGCCCTCGCATTTTCATTAATGATGGCCTCCCTGCCACACGTGAATCACT CCAGCCCGAGATCTGAAACCAGGACACACCCCAGGGGCGAGGTGACGCTGAGTGAGCCCAGC TGTGTCCCTTTCATGAGAACTCAGAGCACAGGGCTCTGTGTGCATGGCCGTCCCCTCCAGAG AGGAGGAAGTAAATGCCGGGATTAGTGGAAGATCATTTCCTTCTATTTGCCTTGGCTTACGT CTTTCAGAATTCAAACACGTGCACTGTTGACCCTGCAATGGTGGAGTTTTTGGATTTTCCTT CAGTCCGATTGCTAAAATACTTCCCTCTCATGTGAGCTGTTGTGAAAGTCATCAGCCAGATA CCATTCTAAAAACAAAGAATGTGCTTCTCGTATGTTGCATGCTGGTTACTGAAATATTAGGG AATTACATAAAGGTTTTCTGGGGCACATATTCAAGCTGAATGATAAAATTGAAGGTCACACA AAGCTAAGGTCTTTCAAATCCTGACCCAATTAGCT CTCTGTTAGCTCTCT GAG T T T GGAC AA GCTGTCTGGTCCTCTGAAGCATACTTTGTTCGCCCTGGGTAGGGGCCCTCTGTTTTAACAGC GTTTGGCATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCG GGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGC CGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAA GGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACT CGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGA GACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACC GCCCAGAGTAGAAGCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCC TGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGC GATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCC CTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACC ACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACC ATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACAC CCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGA CCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACC TGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTT CAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAAC ATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCC GTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGAC CCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAA CACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTAT TGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTT TACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCC GACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTAC AGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAA ACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAA TGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTA TTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCA ATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTT TTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCT GAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCT TGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTG GCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCT CAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGT AAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGA CAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACT CGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0138] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 77. An exemplary nucleotide sequence for vector c.81.7 is set forth in SEQ ID NO: 77 hGJB2 GRE7 underlined; human GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC TCAGCTGGAGTGACGCACCTCATCCATGCGGGCCTGGCGTCTGGAAGGTGGCTGGGTCTCTC GGGCTTGAGCACCATCATCTTAGCTCCAACATGTCATTATTCCTTCCTCACTGAGGACTTTTCTGCTTCCTAATTGGTTGTTGAAGATGAGGCCCCCATGCTCTTTTAAGAAAACCTGTTGTGC CCCAGGCTTGGCTGTGATGGGCACTGACTCATACAGAAGTAGAAAGGCCTGCTGAGTCATCA ACACTCGTGCGACGCCCTCGCATTTTCATTAATGATGGCCTCCCTGCCACACGTGAATCACT CCAGCCCGAGATCTGAAACCAGGACACACCCCAGGGGCGAGGTGACGCTGAGTGAGCCCAGC TGTGTCCCTTTCATGAGAACTCAGAGCACAGGGCTCTGTGTGCATGGCCGTCCCCTCCAGAG AGGAGGAAGTAAATGCCGGGATTAGTGGAAGATCATTTCCTTCTATTTGCCTTGGCTTACGT CTTTCAGAATTCAAACACGTGCACTGTTGACCCTGCAATGGTGGAGTTTTTGGATTTTCCTT CAGTCCGATTGCTAAAATACTTCCCTCTCATGTGAGCTGTTGTGAAAGTCATCAGCCAGATA CCATTCTAAAAACAAAGAATGTGCTTCTCGTATGTTGCATGCTGGTTACTGAAATATTAGGG AATTACATAAAGGTTTTCTGGGGCACATATTCAAGCTGAATGATAAAATTGAAGGTCACACA AAGCTAAGGTCTTTCAAATCCTGACCCAATTAGCT CTCTGTTAGCTCTCT GAG T T T GGAC AA GCTGTCTGGTCCTCTGAAGCATACTTTGTTCGCCCTGGGTAGGGGCCCTCTGTTTTAACAGC GTTTGGCATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCG GGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGC CGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAA GGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACT CGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGA GACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACC GCCCAGAGTAGAAGCCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAAC ACTCCACCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTC GTTGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCA GCCAGGCTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGG CCCTGCAGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGG AGACATGAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGA GGAGATCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCA TCTTCTTCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGC TTCTCCATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTT TGTGTCCCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTT GCATCCTGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCA AAAAAGCCAGTTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGAT TAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGC TAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCT CAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCT AATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTT AGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTT CTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCT CCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCT TTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTT TTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCAT TATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTAC TATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCT GTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACA GACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGT AATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTG TCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAA GAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGG GGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGA TTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTT TAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTG AGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAA ATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTA ATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATG GTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTG AAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATC CTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCA CTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCC GGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCAT CGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGC TACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCG GCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCT GTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGA AGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTA GGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGAC AATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAG TTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCG ACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGC GCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGC AACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCT CGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGAT TTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGG CCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAG GGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCG AATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGA TGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAG AGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTT ATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATG TGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTT CCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAA CGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTG GATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAAC TCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAG CATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAA CACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATA CCAAACGACGAGCGTGACACCA

[0139] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 78. An exemplary nucleotide sequencefor vector c.81.7 is set forth in SEQ ID NO: 78 (hGJB2 GRE7 underlined; mouse GJB2 coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC TCAGCTGGAGTGACGCACCTCATCCATGCGGGCCTGGCGTCTGGAAGGTGGCTGGGTCTCTC GGGC T T GAGCACCATCATCTTAGCTCCAACATGTCATTATTCCTTCCTCACTGAGGACTTTT CTGCTTCCTAATTGGTTGTTGAAGATGAGGCCCCCATGCTCTTTTAAGAAAACCTGTTGTGC CCCAGGCTTGGCTGTGATGGGCACTGACTCATACAGAAGTAGAAAGGCCTGCTGAGTCATCA ACACTCGTGCGACGCCCTCGCATTTTCATTAATGATGGCCTCCCTGCCACACGTGAATCACT CCAGCCCGAGATCTGAAACCAGGACACACCCCAGGGGCGAGGTGACGCTGAGTGAGCCCAGC TGTGTCCCTTTCATGAGAACTCAGAGCACAGGGCTCTGTGTGCATGGCCGTCCCCTCCAGAG AGGAGGAAGTAAATGCCGGGATTAGTGGAAGATCATTTCCTTCTATTTGCCTTGGCTTACGT CTTTCAGAATTCAAACACGTGCACTGTTGACCCTGCAATGGTGGAGTTTTTGGATTTTCCTT CAGTCCGATTGCTAAAATACTTCCCTCTCATGTGAGCTGTTGTGAAAGTCATCAGCCAGATA CCATTCTAAAAACAAAGAATGTGCTTCTCGTATGTTGCATGCTGGTTACTGAAATATTAGGG AATTACATAAAGGTTTTCTGGGGCACATATTCAAGCTGAATGATAAAATTGAAGGTCACACA AAGCTAAGGTCTTTCAAATCCTGACCCAATTAGCT CTCTGTTAGCTCTCT GAG T T T GGAC AA GCTGTCTGGTCCTCTGAAGCATACTTTGTTCGCCCTGGGTAGGGGCCCTCTGTTTTAACAGC GTTTGGCATTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCG GGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGC CGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAA GGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACT CGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGA GACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACC GCCCAGAGTAGAAGCCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAAC ACTCCACCAGCATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTC GTGGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCA GCCTGGCTGCAAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGG CTCTGCAGCTGATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGG AGACATGAAAAGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGA AGAGATCAAAACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTCTTCCGGGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTTCATGCAACGTCTGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTTCCAGGCCCACAGAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATTCTGCTAAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAGACCAGTCTACCCATACGATGTTCCAGATTACGCTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTA TCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAAT GAGC T GATT T AAC AAAAAT T T AACGCGAAT T T T AAC AAAAT AT T AACGT T T AC AAT T T T AT G GTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAA CACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTG ACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACG AAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGA CGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATA CATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAA AAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTT GCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTG GGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCG CCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTAT CCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTG GTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATG CAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAG GACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGT TGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0140] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE8), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 23 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c.81.8).

[0141] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 79. An exemplary nucleotide sequence for vector c.81.8 is set forth in SEQ ID NO: 79 (hGJB2 GRE8 underlined; eGFP coding sequence in bold face):

[0142] CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTT ACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACT TCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTG GGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATC TACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGC CTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATT TAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACC AAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGG ATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGC TACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGC TTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTT CAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTG CCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCG CAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACAC CGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTCGACCTGAACGATTAAGGCAAAACTTCGAAATGTGCCCCAGCAGAGATTTATTTTTCAGGGGGTGTTTTGCATTCCAGCCCCTCTGCCTTCCTGGCGTTTAGTGCGATTTGTTTAGCCATGTGCTCCCTGGTGTGTGTTTTTGAATGTGTGTGAGATGGGTTGTCTCTCGGGACCTGGCAGGTGCGGCCACCAGGTCAGGGCTGCCCCCCAACCCTGTGCCTCCTTCCTCCTAGACTCTGGCCCCCTCAGTGCTGAGGGTGATACAGAGCACTTTTCAAGCTGGATTTGGAATGTGGCCTCTCCCCTCCAAACTCCTGGAGATCATGCAAAGGCCTTTGGAGCCAGCCAGTCACCTGGAAGGTGACATTCCCACCAGCTGAGGCCTCACCTTCAGCGGGGGCTGGGCAGCTTTGGAGCCTGGGGCCAGCCAAGCTCACTCTGCCCATATCCCTGCCACGTGTGGCCCAGCGGATGATCACCTGTCTTCATCTGCGTACTGGGCCACATCCCTCCTGCCGTCCCCCACTTCCCTGATGACACCTACAGCAAGCCCCTACCCAAGTGTTCTGTGATCCCCTGTAAATGTGGCCTCCCTAGCTACTTGCTTTTATGAAACCAACAATCCTGGGGACACAGTTTTCGGCTGTCTCAAGACGGGGCAACCACTCTTTTCCCCAGGCCTGTGGGTCCCAGGCCTGGAGCTAGGGTTGGCATTCTTGCCTGAATTCTCCACTCTATCCCAACCCCTGAGGCCGCCTGAGGAGGCTCAGACTGTGTCAGGCTAGGAGGACAGTCAAACCACAAAAACATGCCTTTTAAGAAGTATAAGCACAAATCCCTCTTTGATGTTATATAAAAGCTCAGTGTCACTTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAATAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0143] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 80. An exemplary nucleotide sequence for vector c.81.8 is set forth in SEQ ID NO: 80 (hGJB2 GRE8 underlined; human GJB2 coding sequence in bold face): CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC GACCTGAACGATTAAGGCAAAACTTCGAAATGTGCCCCAGCAGAGATTTATTTTTCAGGGGG TGTTTTGCATTCCAGCCCC TCTGCCTTCCTGGCGTTTAGTGCGATTTGTTTAGCCATGTGCT CCCTGGTGTGTGTTTTTGAATGTGTGTGAGATGGGTTGTCTCTCGGGACCTGGCAGGTGCGG CCACCAGGTCAGGGCTGCCCCCCAACCCTGTGCCTCCTTCCTCCTAGACTCTGGCCCCCTCA GTGCTGAGGGTGATACAGAGCACTTTTCAAGCTGGATTTGGAATGTGGCCTCTCCCCTCCAA ACTCCTGGAGATCATGCAAAGGCCTTTGGAGCCAGCCAGTCACCTGGAAGGTGACATTCCCA CCAGCTGAGGCCTCACCTTCAGCGGGGGCTGGGCAGCTTTGGAGCCTGGGGCCAGCCAAGCT CACTCTGCCCATATCCCTGCCACGTGTGGCCCAGCGGATGATCACCTGTCTTCATCTGCGTA CTGGGCCACATCCCTCCTGCCGTCCCCCACTTCCCTGATGACACCTACAGCAAGCCCCTACC CAAGTGTTCTGTGATCCCCTGTAAATGTGGCCTCCCTAGCTACTTGCTTTTATGAAACCAAC AATCCTGGGGACACAGTTTTCGGCTGTCTCAAGACGGGGCAACCACTCTTTTCCCCAGGCCT GTGGGTCCCAGGCCTGGAGCTAGGGTTGGCATTCTTGCCTGAATTCTCCACTCTATCCCAAC CCCTGAGGCCGCCTGAGGAGGCTCAGACTGTGTCAGGCTAGGAGGACAGTCAAACCACAAAA ACATGCCTTTTAAGAAGTATAAGCACAAATCCCTCTTTGATGTTATATAAAAGCTCAGTGTC ACTTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGT TCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCC CCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGATTGGGGCACGCTGCAGACGATCCTGGGGGGTGTGAACAAACACTCCACCAGCATTGGAAAGATCTGGCTCACCGTCCTCTTCATTTTTCGCATTATGATCCTCGTTGTGGCTGCAAAGGAGGTGTGGGGAGATGAGCAGGCCGACTTTGTCTGCAACACCCTGCAGCCAGGCTGCAAGAACGTGTGCTACGATCACTACTTCCCCATCTCCCACATCCGGCTATGGGCCCTGCAGCTGATCTTCGTGTCCACGCCAGCGCTCCTAGTGGCCATGCACGTGGCCTACCGGAGACATGAGAAGAAGAGGAAGTTCATCAAGGGGGAGATAAAGAGTGAATTTAAGGACATCGAGGAGATCAAAACCCAGAAGGTCCGCATCGAAGGCTCCCTGTGGTGGACCTACACAAGCAGCATCTTCTTCCGGGTCATCTTCGAAGCCGCCTTCATGTACGTCTTCTATGTCATGTACGACGGCTTCTCCATGCAGCGGCTGGTGAAGTGCAACGCCTGGCCTTGTCCCAACACTGTGGACTGCTTTGTGTCCCGGCCCACGGAGAAGACTGTCTTCACAGTGTTCATGATTGCAGTGTCTGGAATTTGCATCCTGCTGAATGTCACTGAATTGTGTTATTTGCTAATTAGATATTGTTCTGGGAAGTCAAAAAAGCCAGTTTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAG CAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGC CACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCC CGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTG ATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCA TAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGAC CGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCA CGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGT GCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATC GCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCT TGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATT TTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTT TAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCG CATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTG CTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTT TTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGG TTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGC GGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATA ACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTG TCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTG GTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCT CAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTT TTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGT CGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCT TACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTG CGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAAC ATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAA CGACGAGCGTGACACCA

[0144] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 81. An exemplary nucleotide sequence for vector c.81.8 is set forth in SEQ ID NO: 81 hGJB2 GRE8 underlined; mouse GJB2 coding sequence in bold face): CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTC GACCTGAACGATTAAGGCAAAACTTCGAAATGTGCCCCAGCAGAGATTTATTTTTCAGGGGG TGTTTTGCATTCCAGCCCC TCTGCCTTCCTGGCGTTTAGTGCGATTTGTTTAGCCATGTGCT CCCTGGTGTGTGTTTTTGAATGTGTGTGAGATGGGTTGTCTCTCGGGACCTGGCAGGTGCGG CCACCAGGTCAGGGCTGCCCCCCAACCCTGTGCCTCCTTCCTCCTAGACTCTGGCCCCCTCA GTGCTGAGGGTGATACAGAGCACTTTTCAAGCTGGATTTGGAATGTGGCCTCTCCCCTCCAA ACTCCTGGAGATCATGCAAAGGCCTTTGGAGCCAGCCAGTCACCTGGAAGGTGACATTCCCA CCAGCTGAGGCCTCACCTTCAGCGGGGGCTGGGCAGCTTTGGAGCCTGGGGCCAGCCAAGCT CACTCTGCCCATATCCCTGCCACGTGTGGCCCAGCGGATGATCACCTGTCTTCATCTGCGTA CTGGGCCACATCCCTCCTGCCGTCCCCCACTTCCCTGATGACACCTACAGCAAGCCCCTACC CAAGTGTTCTGTGATCCCCTGTAAATGTGGCCTCCCTAGCTACTTGCTTTTATGAAACCAAC AATCCTGGGGACACAGTTTTCGGCTGTCTCAAGACGGGGCAACCACTCTTTTCCCCAGGCCT GTGGGTCCCAGGCCTGGAGCTAGGGTTGGCATTCTTGCCTGAATTCTCCACTCTATCCCAAC CCCTGAGGCCGCCTGAGGAGGCTCAGACTGTGTCAGGCTAGGAGGACAGTCAAACCACAAAA ACATGCCTTTTAAGAAGTATAAGCACAAATCCCTCTTTGATGTTATATAAAAGCTCAGTGTC ACTTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGT TCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCC CCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGC CACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAG CCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCC CAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCA GAGTAGAAGCCATGGATTGGGGCACACTCCAGAGCATCCTCGGGGGTGTCAACAAACACTCC ACCAGCATTGGAAAGATCTGGCTCACGGTCCTCTTCATCTTCCGCATCATGATCCTCGTGGT GGCTGCAAAGGAGGTGTGGGGAGATGAGCAAGCCGATTTTGTCTGCAACACGCTCCAGCCTG GCTGCAAGAATGTATGCTACGACCACCACTTCCCCATCTCTCACATCCGGCTCTGGGCTCTG CAGCTGATCATGGTGTCCACGCCAGCCCTCCTGGTAGCTATGCATGTGGCCTACCGGAGACA TGAAAAGAAACGGAAGTTCATGAAGGGAGAGATAAAGAACGAGTTTAAGGACATCGAAGAGA TCAAAACCCAGAAGGTCCGTATCGAAGGGTCCCTGTGGTGGACCTACACCACCAGCATCTTC TTCCGGGTCATCTTTGAAGCCGTCTTCATGTACGTCTTTTACATCATGTACAATGGCTTCTT CATGCAACGTCTGGTGAAATGCAACGCTTGGCCCTGCCCCAATACAGTGGACTGCTTCATTT CCAGGCCCACAGAAAAGACTGTCTTCACCGTGTTTATGATTTCTGTGTCTGGAATTTGCATT CTGCTAAATATCACAGAGCTGTGCTATTTGTTCGTTAGGTATTGCTCAGGAAAGTCCAAAAG ACCAGTCTACCCATACGATGTTCCAGATTACGCTTAAAGGCGCGCCACCCCTGCAGGGAATT CCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGC TCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAAC CATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCC TAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGT TCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGA GGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGAC ACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTG AACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGT GCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGAT GTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATG TAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGAGATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGCCCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCGAGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTTATGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCCAACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTGTGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGACGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCA

[0145] In some embodiments, an AAV vector described herein comprises an AAV 5 ' ITR, a GJB2 GRE enhancer (hGJB2 GRE9), a GJB2 basal promoter, a GJB2 exon 1 5 ' UTR, Kozak sequence, nucleotide sequence encoding a gene product (e.g., GJB2 or GFP), a GJB2 exon 2 3 ' UTR, a WPRE, a bovine growth hormone poly A signal, and an AAV 3 ' ITR (e.g., vector c.81.9).

[0146] In some embodiments, an AVV vector described herein comprises a nucleotide sequence at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 52. An exemplary nucleotide sequence for vector c.81.9 is set forth in SEQ ID NO: 52 hGJB2 GRE9 underlined; eGFP coding sequence in bold face):CGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTA GCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCG CTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTC GCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACG ACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACT GATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAAC TTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATC CCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTC TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAG CGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGC AGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAA CTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTG GCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGG TCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACA GGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAAC GCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTG ATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCC TGGCCTTTTGCTGGCCTTTTGCTCACATGTCCTGCAGGCAGCTGCGCGCTCGCTCGCTCACT GAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGA GCGAGCGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTGCGGCCGCACGCGTT CTAGGTAGACAACTAAGATGTTCATCTTATGGTTTAATGTTTAGTTGTAAAGGTTGTTTGCT TCTCATTTGGTTCCAAGAAAGAGTATTTAGGCCAATTTCAGGGAGAAATATGTGTATAGATA TATTCATATGTCAAACTGATTAGTGCTGAATGTCACATTTCCATATTCTAATAACATTTCTA GCAAAGAAGAGGACACAGTGAAGAGAGAATTGCCCGCATTGTCATTGTCTCTTTCTGAGCCT AGAACGCCTAACACTTGGGTGTGGAGAGACTCAGCCTCAATTCACTTTCTAGCAGCCACTGA GATGTGCTTGCCTGGGGTGCCCCCTGGCAGGCAGGGCTGGAACTGCTTTCCAGTACCCACAC GGACTGTGAACGAATCTTTCTTTGTGCTTTGTGTACAGAATGGAAGTTCAACAAATATTTGT TGAATGTGTATGTCCTTCCAATACGCAGCAGCCCAGAGCAAACGTGGTAATCTTGTGTGTGT TCATGTGAAAGCAGAATTTAATGGTGCTTTTAAGCACCAAAGTTTAAGATGCACGAGAAAAC TGTATCTCCATTTTTTCCTTTTCGTTTACAATTACTTGTATAAGCCAGGCACGGTGGTGGCTCACGCCTGTAATCCCAGCACTTTGGGAGGCCGAGGCGGGCGGATCACATGAGGTCGGGAGTTAATTAAGACCTCGAAGGGGACTTGGGGGGTTCGGGGCTTTCGGGGGCGGTCGGGGGTTCGCGGACCCGGGAAGCTCTGAGGACCCAGAGGCCGGGCGCGCTCCGCCCGCGGCGCCGCCCCCTCCGTAACTTTCCCAGTCTCCGAGGGAAGAGGCGGGGTGTGGGGTGCGGTTAAAAGGCGCCACGGCGGGAGACAGGTGTTGCGGCCCCGCAGCGCCCGCGCGCTCCTCTCCCCGACTCGGAGCCCCTCGGCGGCGCCCGGCCCAGGACCCGCCTAGGAGCGCAGGAGCCCCAGCGCAGAGACCCCAACGCCGAGACCCCCGCCCCGGCCCCGCCGCGCTTCCTCCCGACGCAGAGCAAACCGCCCAGAGTAGAAGCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCGAGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAAAGGCGCGCCACCCCTGCAGGGAATTCCGCATTGCCCAGTTGTTAGATTAAGAAATAGACAGCATGAGAGGGATGAGGCAACCCGTGCTCAGCTGTCAAGGCTCAGTCGCTAGCATTTCCCAACACAAAGATTCTGACCTTAAATGCAACCATTTGAAACCCCTGTAGGCCTCAGGTGAAACTCCAGATGCCACAATGGAGCTCTGCTCCCCTAAAGCCTCAAAACAAAGGCCTAATTCTATGCCTGTCTTAATTTTCTTTCACTTAAGTTAGTTCCACTGAGACCCCAGGCTGTTAGGGGTTATTGGTGTAAGGTACTTTCATATTTTAAACAGAGGATATCGGCATTTGTTTCTTTCTCTGAGGACAAGAGAAAAAAGCCAGGTTCCACAGAGGACACAGAGAAGGTTTGGGTGTCCTCCTGGGGTTCTTTTTGCCAACTTTCCCCACGTTAAAGGTGAACATTGGTTCTTTCATTTGCTTTGGAAGTTTTAATCTCTAACAGTGGACAAAGTTACCAGTGCCTTAAACTCTGTTACACTTTTTGGAAGTGAAAACTTTGTAGTATGATAGGTTATTTTGATGTAAAGATGTTCTGGATACCATTATATGTTCCCCCTGTTTCAGAGGCTCAGATTGTAATATGTAAATGGTATGTCATTCGCTACTATGATTTAATTTGAAATATGGTCTTTTGGTTATGAATACTTTGCAGCACAGCTGAGAGGCTGTCTGTTGTATTCATTGTGGTCATAGCACCTAACAACATTGTAGCCTCAATCGAGTGAGACAGACTAGAAGTTCCTAGTGATGGCTTATGATAGCAAATGGCCTCATGTCAAATATTTAGATGTAATTTTGTGTAAGAAATACAGACTGGATGTACCACCAACTACTACCTGTAATGACAGGCCTGTCCAACACATCTCCCTTTTCCATGACTGTGGTAGCCAGCATCGGAAAGAACGCTGATTTAAAGAGGTCGCTTGGGAATTTTATTGACACAGTACCATTTAATGGGGAGGACAAAATGGGGCAGGGGAGGGAGAAGTTTCTGTCGTTAAAAACAGATTTGGAAAGACTGGACTCTAAAGTCTGTTGATTAAAGATGAGCTTTGTCTACTTCAAAAGTTTGTTTGCTTACCCCTTCAGCCTCCAATTTTTTAAGTGAAAATATAGCTAATAACATGTGAAAAGAATAGAAGCTAAGGTTTAGATAAATATTGAGCAGATCTATAGGAAGATTGAACCTGAATATTGCCATTATGCTTGACATGGTTTCCAAAAAATGGTACTCCACATATTTCAGTGAGGGTAAGTATTTTCCTGTTGTCAAGAATAGCATTGTAAAAGCATTTTGTAATAATAAAGAATAGCTTTAATGATATGCTTGTAACTAAAATAATTTTGTAATGTATCAAATACATTTAAAACATTAAAATATAATCTCTATAATAATTTAAAATCTAATATGGTTTTAATAGAACAGCGATATCAAGCTTATCGATAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAATCATCGTCCTTTCCTTGGCTGCTCGCCTATGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGAC GAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCGAATTCATCGATACCGAGCGCTGCTCGAGA GATCTGTGATAGCGGCCATCAAGCTGGCTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGC CCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAA TGAGGAAATTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGC AGGACAGCAAGGGGGAGGATTGGGAAGACAATAGCAGGCATGCTGGGGACACGTGCGGACCG AGCGGCCGCAGGAACCCCTAGTGATGGAGTTGGCCACTCCCTCTCTGCGCGCTCGCTCGCTC ACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAG CGAGCGAGCGCGCAGCTGCCTGCAGGGGCGCCTGATGCGGTATTTTCTCCTTACGCATCTGT GCGGTATTTCACACCGCATACGTCAAAGCAACCATAGTACGCGCCCTGTAGCGGCGCATTAA GCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCC GCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCT AAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAAC TTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTG ACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCC TATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAA AT GAGC T GATT T AAC AAAAAT T T AACGCGAAT T T T AAC AAAAT AT T AACGT T T AC AAT T T T A TGGTGCACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGCCCCGACACCCGCC AACACCCGCTGACGCGCCCTGACGGGCTTGTCTGCTCCCGGCATCCGCTTACAGACAAGCTG TGACCGTCTCCGGGAGCTGCATGTGTCAGAGGTTTTCACCGTCATCACCGAAACGCGCGAGA CGAAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTA GACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTT...

Claims

CLAIMSWhat is claimed is:

1. An isolated nucleic acid comprising an expression cassette, wherein the expression cassette comprises a Gap Junction beta 2 (GJB2) gene regulatory element (GRE), and a nucleotide sequence encoding a GJB2 protein.

2. The isolated nucleic acid of claim 1, wherein the GJB2 protein is a human GJB2 protein.

3. The isolated nucleic acid of claim 2, wherein the GJB2 protein comprises an amino acid sequence at least 80% identical to SEQ ID NO: 1.

4. The isolated nucleic acid of any one of claims 1-3, wherein the nucleotide sequence encoding a GJB2 protein comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 2.

5. The isolated nucleic acid of any one of claims 1-4, wherein the expression cassette further comprises a promoter operably linked to the nucleotide sequence encoding a GJB2 protein.

6. The isolated nucleic acid of claim 5, wherein the promoter is a human GJB2 promoter.

7. The isolated nucleic acid of claim 6, wherein the promoter comprises 500 nucleotides of a human GJB2 promoter.

8. The isolated nucleic acid of claim 7, wherein the promoter comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 5.

9. The isolated nucleic acid of claim 6, wherein the promoter comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 102, optionally 100% identical to SEQ ID NO: 102.

10. The isolated nucleic acid of any one of claims 1-9, wherein the Gap Junction beta 2 (GJB2) gene regulatory element (GRE) comprises a nucleotide sequence encoding a 5' UTR.

11. The isolated nulcleic acid of claim 9, wherein the 5' UTR is positioned between the promoter and the nucleotide sequence encoding a GJB2 protein.

12. The isolated nucleic acid of claims 10 or 11, wherein the 5' UTR comprises about 300 nucleotides of a human GJB2 gene 5' UTR.

13. The isolated nucleic acid of claim 12, wherein the promoter and the 5' UTR comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 30.

14. The isolated nucleic acid of any one of claims 1-13, wherein the GJB2 gene regulatory element further comprises an enhancer.

15. The isolated nucleic acid of claim 14, wherein the enhancer is positioned 5' to the promoter.

16. The isolated nucleic acid of claims 14 or 15, wherein the enhancer is normally present within approximately 200 kb upstream or downstream of a GJB2 gene.

17. The isolated nucleic acid of any one of claims 14-16, wherein the enhancer is normally present within approximately 95 kb of a GJB2 gene.

18. The isolated nucleic acid of any one of claims 14-17, wherein the GJB2 GRE comprises one or more enhancers.

19. The isolated nucleic acid of claim 18, wherein the one or more enhancers are the same enhancers or different enhancers.

20. The isolated nucleic acid of any one of claims 14-19, wherein the enhancer comprises a nucleotide sequence at least 80% identical to a nucleotide sequence or a fragment thereof as set forth in any one of SEQ ID NOs: 6 to 29.

21. The isolated nucleic acid of any one of claims 14-20, wherein the enhancer comprises a nucleotide sequence at least 80% identical to a GJB2 enhancer as set forth in any one of SEQ ID NOs: 37-46.

22. The isolated nucleic acid of claim 21, wherein the enhancer comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 42.

23. An isolated nucleic acid comprising an expression cassette, wherein the expression cassette comprises a Gap Junction beta 2 (GJB2) promoter, and a nucleotide sequence encoding a GJB2 protein.

24. The isolated nucleic acid of claim 23, wherein the GJB2 promoter comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 102, optionally 100% identical to SEQ ID NO: 102.

25. The isolated nucleic acid of claim 23 or 24, wherein the expression cassette further comprises a 5 ' UTR.

26. The isolated nucleic acid of claim 25, wherein the 5 ' UTR comprises: a first nucleic acid sequence at least 80% identical to SEQ ID NO: 103, optionally 100% identical to SEQ ID NO: 103; and / or a second nucleic acid sequence at least 80% identical to SEQ ID NO: 104, optionally 100% identical to SEQ ID NO: 104.

27. The isolated nucleic acid of any one of claims 23-27, wherein the isolated nucleic acid comprises a nucleic acid sequence at least 80% identical to SEQ ID NO: 105, optionally 100% identical to SEQ ID NO: 105.

28. The isolated nucleic acid of any one of claim 1-27, wherein the isolated nucleic acid is capable of expressing GJB2 in cells that normally express the GJB2 gene.

29. The isolated nucleic acid of claim 28, wherein the isolated nucleic acid is capable of expressing GJB2 in cochlear connective tissue cells and supporting cells of the organ of Corti.

30. The isolated nucleic acid of claim 29, wherein the supporting cell of the organ of Corti are pillar cells, Deiters’ cells, Hensen’s cells, Claudius cells, inner phalangeal cells, and border cells.

31. The isolated nucleic acid of claim 29, wherein the cochlear connective tissue cells are strial intermediate cells, fibrocytes of the lateral wall and suprastrial zone, basal cells of the stria vascularis, fibrocytes in the spiral ligament, fibrocytes in the spiral limbus, mesenchymal cells lining the bony otic capsule facing the scala vestibuli, and supralimbal dark cells.32 The isolated nucleic acid of any one of claims 1-31, wherein the expression cassette is flanked by two adeno-associated virus inverted terminal repeats (ITRs).

33. The isolated nucleic acid of claim 32, wherein the AAV ITR is from a serotype selected from the group consisting of AAV1 ITR, AAV2 ITR, AAV3 ITR, AAV4 ITR, AAV5 ITR, and AAV6 ITR.

34. The isolated nucleic acid of claims 32 or 33, wherein the AAV ITR is AAV2 ITR.

35. The isolated nucleic acid of claims 32 or 33, wherein the expression cassette comprises: a 5 ' ITR having a nucleotide sequence at least 80% identical to SEQ ID NO: 106, optionally 100% identical to SEQ ID NO: 106; and / or a 3 ' ITR having a nucleotide sequence at least 80% identical to SEQ ID NO: 107, optionally 100% identical to SEQ ID NO: 107.

36. The isolated nucleic acid of any one of claims 1-35, wherein the expression cassette further comprises a Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) 3' to the nucleotide sequence encoding the GJB2 protein.

37. The isolated nucleic acid of claim 36, wherein the WPRE comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 108, optionally 100% identical to SEQ IDNO: 108.

38. The isolated nucleic acid of any one of claims 1-37, wherein the expression cassette further comprises a nucleotide sequence encoding a 3' UTR located 3' of the WPRE.

39. The isolated nucleic acid of claim 38, wherein the 3' UTR is a GJB2 3' UTR.

40. The isolated nucleic acid of claim 39, wherein the GJB2 3' UTR comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 32.

41. The isolated nucleic acid of any one of claims 1-40, wherein the expression cassette further comprises a poly A signal.

42. The isolated nucleic acid of claim 41, wherein the poly A signal is a bovine growth hormone poly A signal.

43. The isolated nucleic acid of claim 41, wherein the poly A signal comprises a nucleotide sequence at least 80% identical to SEQ ID NO: 109, optionally 100% identical to SEQ ID NO: 109.

44. An isolated nucleic acid comprising a nucleotide sequence at least 80% identical to SEQ ID NO: 110 or 111, optionally 100% identical to SEQ ID NO: 110 or 111.

45. A vector comprising the isolated nucleic acid of any one of claims 1-44.

46. The vector of claim 45, wherein the vector is a plasmid or a viral vector.

47. The vector of claim 46, wherein the viral vector is an AAV vector.

48. A vector comprising from 5' to 3':(a) a 5' ITR;(b) a GJB2 promoter, or a basal GJB2 promoter sequence thereof;(c) aGJB25'UTR;(d) a nucleotide sequence encoding a GJB2 protein;(e) aGJB23'UTR;(f) a bovine growth hormone poly A signal; and(g)a3'ITR. r comprising from 5' to 3':(a) a5'ITR;(b) a GJB2 enhancer;(c) a GJB2 promoter, or a basal GJB2 promoter sequence thereof;(d) a GJB25' UTR;(e) a nucleotide sequence encoding a GJB2 protein;(f) a GJB23' UTR;(g) a bovine growth hormone poly A signal; and(h) a3'ITR. binant adeno-associated virus (rAAV) comprising:(i) a capsid protein; and(ii) the isolated nucleic acid of any one of claims 1-44. binant adeno-associated virus (rAAV) comprising:(i) a capsid protein; and(ii) an isolated nucleic acid comprising:(a) a5'ITR;(b) a GJB2 promoter, or a basal GJB2 promoter sequence thereof;(c) aGJB25'UTR;(d) a nucleotide sequence encoding a GJB2 protein;(e) aGJB23'UTR;(f) a bovine growth hormone poly A signal; and(g)a3'ITR. binant adeno-associated virus (rAAV) comprising:(i) a capsid protein; and(ii) an isolated nucleic acid comprising:(a) a 5' ITR;(b) a GJB2 enhancer;(c) a GJB2 promoter, or a basal GJB2 promoter sequence thereof;(d) a GJB2 5' UTR;(e) a nucleotide sequence encoding a GJB2 protein;(f) a GJB2 3' UTR;(g) a bovine growth hormone poly A signal; and(h) a 3 ' ITR.

53. The rAAV of any one of claims claim 50-52, wherein the rAAV has tropism for a subset of cochlear cells that normally express the GJB2 gene.

54. The rAAV of any one of claims 50-53, wherein the rAAV has tropism for cells of the inner ear.

55. The rAAV of any one of claims 50-54, wherein the capsid protein is an AAV1 capsid protein, an AAV2 capsid protein, an AAV5 capsid protein, an AAV7 capsid protein, an AAV8 capsid protein, an AAV9 capsid protein, an AAV-S capsid protein, or a variant thereof.

56. The rAAV of any one of claims 50-55, wherein the AAV capsid is AAV9.PHP.B, AAV9.PHP.eB, or AAV-S.

57. The rAAV of claim 56, wherein the AAV capsid protein is AAV-S.

58. A cell comprising the isolated nucleic acid of any one of claims 1-44, the vector of any one of claims 45-49, or the rAAV of any one of claims 50-57.

59. A pharmaceutical composition comprising the isolated nucleic acid of any one of claims 1-44, the vector of any one of claims 45-49, the rAAV of any one of claims 50-57, or the cell of claim 58.

60. The pharmaceutical composition of claim 59 further comprising a pharmaceutically acceptable carrier.

61. A method for specifically expressing GJB2 in cells that normally expresses the GJB2 gene in a subject, the method comprising administering to the subject an effective amount of the isolated nucleic acid of any one of claims 1-44, the vector of any one of claims 45-49, the rAAV of any one of claims 50-57, the cell of claim 58, or the pharmaceutical composition of claim 59 or 60.

62. A method for treating Non-syndromic Hearing Loss and Deafness (DFNB1) in a subject in need thereof, the method comprising administering to the subject an effective amount of the isolated nucleic acid of any one of claims 1-44, the vector of any one of claims 45-49, the rAAV of any one of claims 50-57, the cell of claim 58, or the pharmaceutical composition of claims 59 or 60.

63. A method for treating a GJB2-associated disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the isolated nucleic acid of any one of claims 1-44, the vector of any one of claims 45-49, the rAAV of any one of claims 50-57, the cell of claim 58, or the pharmaceutical composition of claims 59 or 60.

64. The method of any one of claims 61-63, wherein the subject is a mammal.

65. The method of claim 64, wherein the mammal is a human.

66. The method of claim 64, wherein the mammal is a non-human mammal.

67. The method of claim 66, wherein the non-human mammal is mouse, rat, or non- human primate.

68. The method of any one of claims 61-67, wherein the hearing loss is associated with a mutation in the GJB2 gene.

69. The method of claim 68, wherein the mutation in the GJB2 gene is a point mutation, a missense mutation, a nonsense mutation, a deletion, an insertion, or a combination thereof.

70. The method of claim 69, wherein the subject is human; and the mutation is a mutation listed in Table 2 or a combination thereof.

71. The method of claims 69 or 70, wherein the mutation is c 101. T>C or Del35G.

72. The method of any one of claims 61-71, wherein the step of administrating results in expression of GJB2 protein in cochlear connective tissue cells and supporting cells of the organ of Corti.

73. The method of claim 72, wherein the supporting cells of the organ of Corti are pillar cells, Deiters’ cells, Hensen’s cells, Claudius cells, inner phalangeal cells, and border cells.

74. The method of claim 72, wherein the cochlear connective tissue cells are strial intermediate cells, fibrocytes of the lateral wall and suprastrial zone, basal cells of the stria vascularis, fibrocytes in the spiral ligament, fibrocytes in the spiral limbus, mesenchymal cells lining the bony otic capsule facing the scala vestibuli, and supralimbal dark cells.

75. The method of any one of claims 61-74, wherein the administration is via injection.

76. The method of claim 75, wherein the injection is through the round window membrane of the cochlea, into the scala media of the cochlea, into the scala vestibuli of the cochlea, into a semicircular canal of the inner ear, or into the saccule or the utricle of the inner ear.

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