Patent Publication Number: US-2023149566-A1

Title: Compositions and methods for treating macular dystrophy

Description:
RELATED APPLICATIONS 
     This application claims the benefit of provisional application U.S. Ser. No. 62/653,131, filed Apr. 5, 2018, the contents of which are herein incorporated by reference in their entirety. 
    
    
     INCORPORATION OF SEQUENCE LISTING 
     The contents of the text file named “NIGH-011 002US SeqList.txt,” which was created on Apr. 4, 2019 and is 72 KB in size, are hereby incorporated by reference in their entirety. 
     FIELD OF THE DISCLOSURE 
     The invention relates to the fields of molecular biology, neurobiology and gene therapy treatments for degenerative eye diseases. 
     BACKGROUND 
     Macular degeneration is a medical condition, which may result in blurred or no vision in the center of the visual field. In macular degeneration, the photoreceptors in the part of the retina called the macula, which is responsible for central vision, degenerate or die. In some cases, macular degeneration is caused by mutations in the Bestrophin-1 gene (BEST1, also called VMD2). There is currently no treatment for this devastating disease. There is thus a long felt need in the art for additional therapeutic approaches to treat macular degeneration. The disclosure provides compositions and methods of treatment for macular degeneration. 
     SUMMARY 
     The disclosure provides a composition comprising a nucleic acid sequence comprising: (a) a sequence encoding a vitelliform macular dystrophy-2 (VMD2) promoter, and (b) a sequence encoding a Bestrophin-1 (BEST1) protein. In some embodiments, the sequence encoding the VMD2 promoter encodes a human VMD2 promoter. In some embodiments, the sequence encoding the BEST1 protein encodes a human BEST1 protein. In some embodiments, the sequence encoding the BEST1 protein comprises a coding sequence. In some embodiments, the sequence encoding the BEST1 protein comprises a cDNA sequence. 
     The disclosure provides a composition comprising a nucleic acid sequence comprising: (a) a sequence encoding a ubiquitous promoter, and (b) a sequence encoding a Bestrophin-1 (BEST1) protein. In some embodiments, the sequence encoding the BEST1 protein encodes a human BEST1 protein. In some embodiments, the sequence encoding the BEST1 protein comprises a coding sequence. In some embodiments, the sequence encoding the BEST1 protein comprises a cDNA sequence. In some embodiments, the sequence encoding a ubiquitous promoter comprises a sequence encoding a CAG promoter. 
     In some embodiments of the compositions of the disclosure, the nucleic acid sequence further comprises: (c) a sequence encoding a posttranscriptional regulatory element (PRE). In some embodiments, the sequence encoding the PRE comprises a sequence isolated or derived from a naturally occurring sequence. In some embodiments, the sequence encoding the PRE comprises a sequence isolated or derived from a non-naturally-occurring sequence. In some embodiments, the sequence encoding the PRE comprises a sequence isolated or derived from a viral sequence. In some embodiments, the sequence encoding the PRE comprises a sequence isolated or derived from a woodchuck hepatitis virus (WPRE). 
     In some embodiments of the compositions of the disclosure, the nucleic acid sequence further comprises: (d) a sequence encoding a polyadenylation (polyA) signal. In some embodiments, the sequence encoding the polyA signal comprises a sequence isolated or derived from a naturally occurring sequence. In some embodiments, the sequence encoding the polyA signal comprises a sequence isolated or derived from a non-naturally-occurring sequence. In some embodiments, the sequence encoding the polyA signal comprises a sequence isolated or derived from a mammalian sequence. In some embodiments, the sequence encoding the polyA signal comprises a sequence isolated or derived from a human sequence. In some embodiments, the sequence encoding the polyA signal comprises a sequence isolated or derived from a mammalian Bovine Growth Hormone (BGH) gene. 
     In some embodiments of the compositions of the disclosure, the nucleic acid sequence further comprises: (e) a sequence encoding a 5′ untranslated region (UTR). In some embodiments, the sequence encoding the 5′ UTR comprises a sequence isolated or derived from a naturally occurring sequence. In some embodiments, the sequence encoding the 5′ UTR comprises a sequence isolated or derived from a non-naturally-occurring sequence. In some embodiments, the sequence encoding the 5′ UTR comprises a sequence isolated or derived from a mammalian sequence. In some embodiments, the sequence encoding the 5′ UTR comprises a sequence isolated or derived from a human sequence. In some embodiments, the sequence encoding the 5′ UTR comprises a sequence isolated or derived from a viral sequence. In some embodiments of the compositions of the disclosure, the nucleic acid sequence further comprises: (f) a sequence encoding an intron, and (g) a sequence encoding an exon, wherein the sequence encoding the intron and the sequence encoding the exon are operably linked. In some embodiments, the intron is located between the sequence encoding the VMD2 promoter and the sequence encoding the exon, wherein the sequence encoding the exon is located between the sequence encoding the intron and the sequence encoding the 5′ UTR, and wherein the sequence encoding the intron is spliced by a mammalian cell. In some embodiments, the sequence encoding the exon comprises a sequence isolated or derived from a mammalian gene. In some embodiments, the sequence encoding the exon comprises a sequence isolated or derived from a rabbit (Oryctolagus  cuniculus ) beta globin gene. In some embodiments, the sequence encoding the intron comprises a non-naturally occurring sequence. In some embodiments, the sequence encoding the intron comprises a fusion sequence. In some embodiments, the sequence encoding the intron comprises a sequence encoding a splice donor site, and a sequence encoding a splice branch point and acceptor site. In some embodiments, the sequence encoding the splice donor site comprises a sequence isolated or derived from a vertebrate gene. In some embodiments, the sequence encoding the splice donor site comprises a sequence isolated or derived from a chicken ( Gallus gallus ) beta actin gene (CBA). In some embodiments, the sequence encoding the splice branch point and acceptor site comprises a sequence isolated or derived from a vertebrate gene. In some embodiments, the sequence encoding the splice branch point and acceptor site comprises a sequence isolated or derived from a rabbit (Oryctolagus  cuniculus ) beta globin gene. 
     In some embodiments of the compositions of the disclosure, the sequence encoding the 5′ UTR comprises a sequence encoding a Kozak sequence or a portion thereof. In some embodiments, the sequence encoding a Kozak sequence has at least 50% identity to the nucleic acid sequence of GCCRCCATGG. In some embodiments, the sequence encoding a Kozak sequence comprises or consists of the nucleic acid sequence of GGCACCATGA. 
     In some embodiments of the compositions of the disclosure, the sequence encoding the human VMD2 promoter comprises or consists of 
     
       
         
           
               
            
               
                 (SEQ ID NO: 1) 
               
            
           
           
               
               
            
               
                 1 
                 AATTCTGTCA TTTTACTAGG GTGATGAAAT 
               
               
                   
               
               
                   
                 TCCCAAGCAA CACCATCCTT TTCAGATAAG 
               
               
                   
               
               
                 61 
                 GGCACTGAGG CTGAGAGAGG AGCTGAAACC 
               
               
                   
               
               
                   
                 TACCCGGGGT CACCACACAC AGGTGGCAAG 
               
               
                   
               
               
                 121 
                 GCTGGGACCA GAAACCAGGA CTGTTGACTG 
               
               
                   
               
               
                   
                 CAGCCCGGTA TTCATTCTTT CCATAGCCCA 
               
               
                   
               
               
                 181 
                 CAGGGCTGTC AAAGACCCCA GGGCCTAGTC 
               
               
                   
               
               
                   
                 AGAGGCTCCT CCTTCCTGGA GAGTTCCTGG 
               
               
                   
               
               
                 241 
                 CACAGAAGTT GAAGCTCAGC ACAGCCCCCT 
               
               
                   
               
               
                   
                 AACCCCCAAC TCTCTCTGCA AGGCCTCAGG 
               
               
                   
               
               
                 301 
                 GGTCAGAACA CTGGTGGAGC AGATCCTTTA 
               
               
                   
               
               
                   
                 GCCTCTGGAT TTTAGGGCCA TGGTAGAGGG 
               
               
                   
               
               
                 361 
                 GGTGTTGCCC TAAATTCCAG CCCTGGTCTC 
               
               
                   
               
               
                   
                 AGCCCAACAC CCTCCAAGAA GAAATTAGAG 
               
               
                   
               
               
                 421 
                 GGGCCATGGC CAGGCTGTGC TAGCCGTTGC 
               
               
                   
               
               
                   
                 TTCTGAGCAG ATTACAAGAA GGGACTAAGA 
               
               
                   
               
               
                 481 
                 CAAGGACTCC TTTGTGGAGG TCCTGGCTTA 
               
               
                   
               
               
                   
                 GGGAGTCAAG TGACGGCGGC TCAGCACTCA 
               
               
                   
               
               
                 541 
                 CGTGGGCAGT GCCAGCCTCT AAGAGTGGGC 
               
               
                   
               
               
                   
                 AGGGGCACTG GCCACAGAGT CCCAGGGAGT 
               
               
                   
               
               
                 601 
                 CCCACCAGCC TAGTCGCCAG ACC. 
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, the sequence encoding the CAG promoter comprises or consists of 
     
       
         
           
               
            
               
                 (SEQ ID NO: 2) 
               
            
           
           
               
               
            
               
                 1 
                 CCATTGACGT CAATAATGAC GTATGTTCCC 
               
               
                   
               
               
                   
                 ATAGTAACGC CAATAGGGAC TTTCCATTGA 
               
               
                   
               
               
                 61 
                 CGTCAATGGG TGGAGTATTT ACGGTAAACT 
               
               
                   
               
               
                   
                 GCCCACTTGG CAGTACATGA AGTGTATCAT 
               
               
                   
               
               
                 121 
                 ATGCCAAGTA CGCCCCCTAT TGACGTCAAT 
               
               
                   
               
               
                   
                 GACGGTAAAT GGCCCGCCTG GCATTATGCC 
               
               
                   
               
               
                 181 
                 CAGTACATGA CCTTATGGGA CTTTCCTACT 
               
               
                   
               
               
                   
                 TGGCAGTACA TCTACGTATT AGTCATCGCT 
               
               
                   
               
               
                 241 
                 ATTACCATGG TCGAGGTGAG CCCCACGTTC 
               
               
                   
               
               
                   
                 TGCTTCACTC TCCCCATCTC CCCCCCCTCC 
               
               
                   
               
               
                 301 
                 CCACCCCCAA TTTTGTATTT ATTTATTTTT 
               
               
                   
               
               
                   
                 TAATTATTTT GTGCAGCGAT GGGGGGGGGG 
               
               
                   
               
               
                 361 
                 GGGGGGGGGG GGCGCGCGCC AGGCGGGGCG 
               
               
                   
               
               
                   
                 GGGGGGGGGG AGGGGCGGGG CGGGGCGAGG 
               
               
                   
               
               
                 421 
                 CGGAGAGGTG CGGCGGCAGC CAATCAGAGC 
               
               
                   
               
               
                   
                 GGCGCGCTCC GAAAGTTTCC TTTTATGGCG 
               
               
                   
               
               
                 481 
                 AGGCGGCGGC GGCGGCGGCC CTATAAAAAG 
               
               
                   
               
               
                   
                 CGAAGCGCGC GGGGGGGGGG AGTCGCTGCG 
               
               
                   
               
               
                 541 
                 CGCTGCCTTC GCCCCGTGCC CCGCTCCGCC 
               
               
                   
               
               
                   
                 GCCGCCTCGC GCCGCCCGCC CCGGCTCTGA 
               
               
                   
               
               
                 601 
                 CTGACCGCGT TACTCCCACA GGTGAGCGGG 
               
               
                   
               
               
                   
                 CGGGACGGCC CTTCTCCTCC GGGCTGTAAT 
               
               
                   
               
               
                 661 
                 TAGCGCTTGG TTTAATGACG GCTTGTTTCT 
               
               
                   
               
               
                   
                 TTTCTGTGGC TGCGTGAAAG CCTTGAGGGG 
               
               
                   
               
               
                 721 
                 CTCCGGGAGG GCCCTTTGTG CGGGGGGAGC 
               
               
                   
               
               
                   
                 GGCTCGGGGC TGTCCGCGGG GGGACGGCTG 
               
               
                   
               
               
                 781 
                 CCTTCGGGGG GGACGGGGCA GGGCGGGGTT 
               
               
                   
               
               
                   
                 CGGCTTCTGG CGTGTGACCG GCGGCTCTAG 
               
               
                   
               
               
                 841 
                 AGCCTCTGCT AACCATGTTC ATGCCTTCTT 
               
               
                   
               
               
                   
                 CTTTTTCCTA CAGCTCCTGG GCAACGTGCT 
               
               
                   
               
               
                 901 
                 GGTTATTGTG CTGTCTCATC ATTTTGGCAA 
               
               
                   
               
               
                   
                 AGAATTGGAT C. 
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, the sequence encoding the human BEST1 protein comprises or consists of 
     
       
         
           
               
            
               
                 (SEQ ID NO: 3) 
               
            
           
           
               
               
            
               
                 1 
                 ATGACCATCA CTTACACAAG CCAAGTGGCT 
               
               
                   
               
               
                   
                 AATGCCCGCT TAGGCTCCTT CTCCCGCCTG 
               
               
                   
               
               
                 61 
                 CTGCTGTGCT GGCGGGGCAG CATCTACAAG 
               
               
                   
               
               
                   
                 CTGCTATATG GCGAGTTCTT AATCTTCCTG 
               
               
                   
               
               
                 121 
                 CTCTGCTACT ACATCATCCG CTTTATTTAT 
               
               
                   
               
               
                   
                 AGGCTGGCCC TCACGGAAGA ACAACAGCTG 
               
               
                   
               
               
                 181 
                 ATGTTTGAGA AACTGACTCT GTATTGCGAC 
               
               
                   
               
               
                   
                 AGCTACATCC AGCTCATCCC CATTTCCTTC 
               
               
                   
               
               
                 241 
                 GTGCTGGGCT TCTACGTGAC GCTGGTCGTG 
               
               
                   
               
               
                   
                 ACCCGCTGGT GGAACCAGTA CGAGAACCTG 
               
               
                   
               
               
                 301 
                 CCGTGGCCCG ACCGCCTCAT GAGCCTGGTG 
               
               
                   
               
               
                   
                 TCGGGCTTCG TCGAAGGCAA GGACGAGCAA 
               
               
                   
               
               
                 361 
                 GGCCGGCTGC TGCGGCGCAC GCTCATCCGC 
               
               
                   
               
               
                   
                 TACGCCAACC TGGGCAACGT GCTCATCCTG 
               
               
                   
               
               
                 421 
                 CGCAGCGTCA GCACCGCAGT CTACAAGCGC 
               
               
                   
               
               
                   
                 TTCCCCAGCG CCCAGCACCT GGTGCAAGCA 
               
               
                   
               
               
                 481 
                 GGCTTTATGA CTCCGGCAGA ACACAAGCAG 
               
               
                   
               
               
                   
                 TTGGAGAAAC TGAGCCTACC ACACAACATG 
               
               
                   
               
               
                 541 
                 TTCTGGGTGC CCTGGGTGTG GTTTGCCAAC 
               
               
                   
               
               
                   
                 CTGTCAATGA AGGCGTGGCT TGGAGGTCGA 
               
               
                   
               
               
                 601 
                 ATCCGGGACC CTATCCTGCT CCAGAGCCTG 
               
               
                   
               
               
                   
                 CTGAACGAGA TGAACACCTT GCGTACTCAG 
               
               
                   
               
               
                 661 
                 TGTGGACACC TGTATGCCTA CGACTGGATT 
               
               
                   
               
               
                   
                 AGTATCCCAC TGGTGTATAC ACAGGTGGTG 
               
               
                   
               
               
                 721 
                 ACTGTGGCGG TGTACAGCTT CTTCCTGACT 
               
               
                   
               
               
                   
                 TGTCTAGTTG GGCGGCAGTT TCTGAACCCA 
               
               
                   
               
               
                 781 
                 GCCAAGGCCT ACCCTGGCCA TGAGCTGGAC 
               
               
                   
               
               
                   
                 CTCGTTGTGC CCGTCTTCAC GTTCCTGCAG 
               
               
                   
               
               
                 841 
                 TTCTTCTTCT ATGTTGGCTG GCTGAAGGTG 
               
               
                   
               
               
                   
                 GCAGAGCAGC TCATCAACCC CTTTGGAGAG 
               
               
                   
               
               
                 901 
                 GATGATGATG ATTTTGAGAC CAACTGGATT 
               
               
                   
               
               
                   
                 GTCGACAGGA ATTTGCAGGT GTCCCTGTTG 
               
               
                   
               
               
                 961 
                 GCTGTGGATG AGATGCACCA GGACCTGCCT 
               
               
                   
               
               
                   
                 CGGATGGAGC CGGACATGTA CTGGAATAAG 
               
               
                   
               
               
                 1021 
                 CCCGAGCCAC AGCCCCCCTA CACAGCTGCT 
               
               
                   
               
               
                   
                 TCCGCCCAGT TCCGTCGAGC CTCCTTTATG 
               
               
                   
               
               
                 1081 
                 GGCTCCACCT TCAACATCAG CCTGAACAAA 
               
               
                   
               
               
                   
                 GAGGAGATGG AGTTCCAGCC CAATCAGGAG 
               
               
                   
               
               
                 1141 
                 GACGAGGAGG ATGCTCACGC TGGCATCATT 
               
               
                   
               
               
                   
                 GGCCGCTTCC TAGGCCTGCA GTCCCATGAT 
               
               
                   
               
               
                 1201 
                 CACCATCCTC CCAGGGCAAA CTCAAGGACC 
               
               
                   
               
               
                   
                 AAACTACTGT GGCCCAAGAG GGAATCCCTT 
               
               
                   
               
               
                 1261 
                 CTCCACGAGG GCCTGCCCAA AAACCACAAG 
               
               
                   
               
               
                   
                 GCAGCCAAAC AGAACGTTAG GGGCCAGGAA 
               
               
                   
               
               
                 1321 
                 GACAACAAGG CCTGGAAGCT TAAGGCTGTG 
               
               
                   
               
               
                   
                 GACGCCTTCA AGTCTGCCCC ACTGTATCAG 
               
               
                   
               
               
                 1381 
                 AGGCCAGGCT ACTACAGTGC CCCACAGACG 
               
               
                   
               
               
                   
                 CCCCTCAGCC CCACTCCCAT GTTCTTCCCC 
               
               
                   
               
               
                 1441 
                 CTAGAACCAT CAGCGCCGTC AAAGCTTCAC 
               
               
                   
               
               
                   
                 AGTGTCACAG GCATAGACAC CAAAGACAAA 
               
               
                   
               
               
                 1501 
                 AGCTTAAAGA CTGTGAGTTC TGGGGCCAAG 
               
               
                   
               
               
                   
                 AAAAGTTTTG AATTGCTCTC AGAGAGGGAT 
               
               
                   
               
               
                 1561 
                 GGGGCCTTGA TGGAGCACCC AGAAGTATCT 
               
               
                   
               
               
                   
                 CAAGTGAGGA GGAAAACTGT GGAGTTTAAC 
               
               
                   
               
               
                 1621 
                 CTGACGGATA TGCCAGAGAT CCCCGAAAAT 
               
               
                   
               
               
                   
                 CACCTCAAAG AACCTTTGGA ACAATCACCA 
               
               
                   
               
               
                 1681 
                 ACCAACATAC ACACTACACT CAAAGATCAC 
               
               
                   
               
               
                   
                 ATGGATCCTT ATTGGGCCTT GGAAAACAGG 
               
               
                   
               
               
                 1741 
                 GATGAAGCAC ATTCCTAA. 
               
            
           
         
       
     
     The disclosure provides a vector comprising a composition of the disclosure. In some embodiments, the vector is a plasmid. 
     The disclosure provides a delivery vector comprising the vector of the disclosure. In some embodiments, the delivery vector is a viral delivery vector. In some embodiments, the delivery vector comprises a single stranded viral genome. In some embodiments, the delivery vector comprises a double stranded viral genome. In some embodiments, the delivery vector comprises an RNA molecule. 
     The disclosure provides a delivery vector comprising the vector of the disclosure. In some embodiments, the delivery vector comprises a sequence isolated or derived from an adeno-associated virus (AAV) vector. In some embodiments, the delivery vector comprises a sequence isolated or derived from an AAV vector of serotype AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11 or any combination thereof. In some embodiments, the delivery vector comprises a sequence isolated or derived from an AAV vector of serotype AAV2. In some embodiments, the delivery vector comprises a sequence isolated or derived from an AAV vector of serotype AAV8. In some embodiments, the delivery vector comprises a sequence encoding a first inverted terminal repeat (ITR) and a second ITR isolated or derived from an AAV vector of serotype AAV2 and a sequence encoding a viral gene isolated or derived from an AAV vector of serotype AAV2. In some embodiments, the delivery vector comprises a sequence encoding a first inverted terminal repeat (ITR) and a second ITR isolated or derived from an AAV vector of serotype AAV8 and a sequence encoding a viral gene isolated or derived from an AAV vector of serotype AAV8. In some embodiments, the delivery vector comprises a sequence encoding a first inverted terminal repeat (ITR) and a second ITR isolated or derived from an AAV vector of serotype AAV2 and a sequence encoding a viral gene isolated or derived from an AAV vector of serotype AAV8. 
     The disclosure provides a pharmaceutical composition comprising a composition of the disclosure and a pharmaceutically-acceptable carrier. In some embodiments, the pharmaceutically-acceptable carrier comprises TMN200. 
     The disclosure provides a pharmaceutical composition comprising a vector of the disclosure a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier comprises TMN200. 
     The disclosure provides a pharmaceutical composition comprising a delivery vector of the disclosure and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier comprises TMN200. 
     The disclosure provides a cell comprising a composition of the disclosure. The disclosure provides a cell comprising a vector of the disclosure. The disclosure provides a cell comprising a delivery vector of the disclosure. The disclosure provides a cell comprising a pharmaceutical composition of the disclosure. In some embodiments, the cell is a mammalian cell. In some embodiments, the mammalian cell is a non-human primate cell, a rodent cell, a mouse cell, a rat cell or a rabbit cell. In some embodiments, the cell is a human cell. In some embodiments, the human cell is a neuronal cell, a glial cell, a retinal cell, a photoreceptor cell, a rod cell, a cone cell or a cuboidal cell of the retinal pigment epithelium (RPE). In some embodiments, the human cell is a photoreceptor cell. In some embodiments, the human cell is an HEK293 cell or an ARPE19 cell. In some embodiments, the human cell is isolated or derived from an RPE of a human retina. In some embodiments, the cell is in vivo, in vitro, ex vivo or in situ. 
     The disclosure provides a method of treating macular dystrophy in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition of the disclosure. 
     The disclosure provides a method of treating macular dystrophy in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising the vector of the disclosure. 
     The disclosure provides a method of treating macular dystrophy in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising the delivery vector of the disclosure. 
     In some embodiments of the methods of the disclosure, the subject is a human. In some embodiments, the subject is a non-human primate, a dog, a cat, a rodent, a mouse, a rat, or a rabbit. In some embodiments, the subject has macular dystrophy. 
     In some embodiments of the methods of the disclosure, the subject has a mutation in one or both copies of a BEST1 gene. In some embodiments, the mutation is heritable as a dominant mutation. In some embodiments, the dominant mutation causes Best Vitelliform Macular Dystrophy (BVMD) in the subject. In some embodiments, the mutation is heritable as a recessive mutation. In some embodiments, the recessive mutation causes Autosomal Recessive Bestrophinopathy (ARB) in the subject. In some embodiments, the mutation occurs in a coding sequence of one or both copies of a BEST1 gene. In some embodiments, the mutation occurs in a non-coding sequence of one or both copies of a BEST1 gene. In some embodiments, the mutation comprises a substitution, an insertion, a deletion, an inversion, a translocation, a frameshift, or a combination thereof in one both copies of a BEST1 gene. 
     In some embodiments of the methods of the disclosure, administering comprises an injection or an infusion via a subretinal, a suprachoroidal or an intravitreal route. In some embodiments, administering comprises an injection or an infusion via a subretinal route. In some embodiments, administering comprises a two-step injection or a two-step infusion via a subretinal route. 
     In some embodiments of the methods of the disclosure, the therapeutically effective amount is formulated in a volume of between 10 and 200 μL, inclusive of the endpoints. In some embodiments, the therapeutically effective amount is formulated in a volume of between 10 and 50 μL, between 50 and 100 μL, between 100 and 150 μL or between 150 and 200 μL, inclusive of the endpoints, for each range. In some embodiments, the therapeutically effective amount is formulated in a volume of between 70 and 120 μL, inclusive of the endpoints, and wherein the administering comprises an injection or an infusion via a subretinal route. In some embodiments, the therapeutically effective amount is formulated in a volume of 100 μL and wherein the administering comprises an injection or an infusion via a subretinal route. 
     In some embodiments of the methods of the disclosure, the therapeutically effective amount comprises a concentration of an AAV delivery vector of at least 1×10 10  DRP/mL, at least 1×10″ DRP/mL, at least 1×10 12  DRP/mL, at least 2×10 12  DRP/mL, at least 5×10 12  DRP/mL or at least 1.5×10″ DRP/mL. In some embodiments, the therapeutically effective amount comprises a concentration of an AAV delivery vector of at least 2×10″ DRP/mL, at least 5×10″ DRP/mL or at least 1.5×10″ DRP/mL. In some embodiments, the therapeutically effective amount comprises a concentration of an AAV delivery vector of at least 5×10 12  DRP/mL. In some embodiments, the therapeutically effective amount comprises a concentration of an AAV delivery vector of at least 1.5×10″ DRP/mL. 
     In some embodiments of the methods of the disclosure, the therapeutically effective amount comprises a dose of 2×10 8  genome particles (gp), 5×10 8  gp, 1.5×10 9  gp, 2×10 9  gp, 5×10 9  gp, 2×10 10  gp, 5×10 10  gp, 6×10 10  gp, 1.2×10 11  gp, 1.5×10 11  gp, 2×10 11  gp, 4.5×10 11  gp, 5×10 11  gp, 1.2×10 12  gp, 1.5×10 12  gp, 2×10 12  gp or 5×10 12  gp. In some embodiments, the subject is a mouse and wherein the therapeutically effective amount comprises a dose of 5×10 8  gp, 1.5×10 9  gp or 5×10 9  gp. In some embodiments, the subject is a non-human primate and wherein the therapeutically effective amount comprises a dose of 1.2×10 11  gp, 4.5×10 11  gp or 1.2×10 12  gp of AAV viral particles. In some embodiments, the subject is human and wherein the therapeutically effective amount comprises a dose of 5×10 10  gp, 1.5×10 11  gp, 1.5×10 12  gp or 1.5×10 12  gp of AAV viral particles. 
     In some embodiments of the methods of the disclosure, the composition further comprises a TMN200 buffer. 
     The disclosure provides a composition of the disclosure for use in treating macular dystrophy in a subject in need thereof. 
     The disclosure provides a vector of the disclosure for use in treating macular dystrophy in a subject in need thereof. 
     The disclosure provides a delivery vector of the disclosure for use in treating macular dystrophy in a subject in need thereof. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee. 
         FIG.  1 A-B  are a pair of maps of a plasmid encoding VMD2. IntEx.BEST1.WPRE.pA construct with AAV2 ITRs. 
         FIG.  2 A-B  are a pair of maps of a plasmid encoding VMD2.BEST1.WPRE.pA construct with AAV2 ITRs. 
         FIG.  3 A-C  are a series of three maps of two plasmids encoding CAG.BEST1.WPRE.pA with AAV2 ITRs.  FIG.  3 A  and  FIG.  3 B  are two maps of a CAG.BEST.WPRE.pA plasmid with an AmpR selectable marker.  FIG.  3 C  is a map of a CAG.BEST.WPRE.pA plasmid with a KanR selectable marker and a stuffer sequence. 
         FIG.  4    is a map of a plasmid encoding VMD2.GFP.WPRE.pA with AAV2 ITRs. 
         FIG.  5    is a map of a plasmid encoding VMD2. Int.Ex.GFP.WPRE.pA with AAV2 ITRs. 
         FIG.  6 A-B  are each a series of images, 6 images and 3 images, respectively, showing BEST1 expression in HEK293 cells transduced with AAV.CAG.BEST1.pA, AAV.CAG.BEST1.WPRE.pA and an untransduced control. Cells are stained with Hoechst blue dye and anti-hBestrophin-1. Bestrophin-1 protein is localized throughout the cytosol 
         FIG.  7 A-B  is a picture of a Western Blot (7A) and a bar graph (7B), respectively, showing the expression of Bestrophin-1 protein and a beta-actin control in HEK293 cells transduced with AAV.CAG.BEST1.pA (sample 1) or AAV.CAG.BEST1.WPRE.pA (sample 2) or a negative control (sample 3). Plasmid-transfected HEK293 cells were used as a positive control.  FIG.  7 B  shows the quantification of Bestrophin-1 protein expression in HEK293 cells transduced with AAV.CAG.BEST1.pA (n=9) or AAV.CAG.BEST1.WPRE.pA (n=9) or an untransduced negative control (n=8). The Y-axis shows the normalized LiCor Value. Error bars are ±SEM. *** indicates p&lt;0.001 when compared to the un-transduced control. 
         FIG.  8 A-B  is a single plot (8A) and a series of four plots (8B), respectively, showing whole-cell patch clamp recording data from HEK293 cells transduced with AAV2/2 CAG.BEST1.pA, AAV2/2 CAG.BEST1.WPRE.pA or AAV2/2 CAG.GFP.WPRE.pA vectors, as well as an untransduced control. In  FIG.  8 A , Current (pA) is plotted on the X-axis from −140 to 500 in increments of 20, while Voltage (in mV) is plotted on the Y-axis from −200 to 500 in units of 100. In  FIG.  8 B , the current waveforms are shown. Current (I)/Voltage (V) plots of HEK293 transduced with the different vectors and an untransduced control are shown, clockwise from the top left: AAV2/2 CAG.BEST1.pA, AAV2/2 CAG.BEST1.WPRE.pA, untransduced control and AAV2/2 CAG.GFP.WPRE.pA. The inset scale bar (center) shows 250 pA on the Y-axis and 100 milliseconds on the X-axis. 
         FIG.  9 A-B  is a pair of plots showing the chord conductance of HEK293 cells transduced with AAV2/2 CAG.BEST1.pA (n=10), AAV2/2 CAG.BEST1.WPRE.pA (n=10), AAV2/2 CAG.GFP.WPRE.pA (n=11) and an untransduced control (n=10). Chord conductance is plotted on the Y-axis from 0 to 10 in units of 2. **** indicates p&lt;0.0001, * indicates p&lt;0.05, ns stands for not significant. 
         FIG.  10 A-B  are a pair of flow charts showing two embodiments of an experimental procedure for assaying BEST1 expression in differentiated ARPE19 cells.  FIG.  10 A  show an experimental procedure for assaying BEST1 expression in transfected differentiated ARPE19 cells.  FIG.  10 B  shows an experimental procedure for assaying BEST1 expression in transfected and/or transduced differentiated ARPE19 cells. 
         FIG.  11 A-B  is a series of 16 images (A) and 6 images (B) showing BEST1 and ZO-1 immunostaining of transfected ARPE19 cells that were differentiated for 1 month. (A) The rows, top to bottom show ARPE19 cells with the following constructs: untransfected control, CAG.BEST1.WPRE, VMD2.BEST1.WPRE and VMD2. IntEx.BEST1.WPRE. The columns from left to right show: nuclei stained with Hoechst in blue, ZO-1 staining in green (ZO-1 is a marker of the cytoplasmic membrane surface of intercellular tight junctions), BEST1 in red, and a merged image (Hoechst, ZO-1, BEST1). Scale bars show 100 microns (μm). (B) Shown in the top row are ARPE19 cells transfected with VMD2.BEST1.WPRE.pA. Shown in the bottom row are ARPE19 cells transfected with VMD2. IntEx.BEST1.WPRE.pA. The images from left to right show ZO-1 (green) and BEST1 (red), and a merged image (Hoechst, ZO-1 and BEST1). The scale bar in the merged images indicates 25 μm. 
         FIG.  12 A-B  is a series of 16 images (A) and 9 images (B) showing BEST1 and ZO-immunostaining of transfected ARPE19 cells that were differentiated for 3 months. (A) The rows, top to bottom show ARPE19 cells with the following constructs: untransfected control, CAG.BEST1.WPRE, VMD2.BEST1.WPRE and VMD2. IntEx.BEST1.WPRE. The columns from left to right show: nuclei stained with Hoechst in blue, ZO-1 staining in green, BEST1 in red, and a merged image (Hoechst, ZO-1, BEST1). Scale bars show 100 microns (μm). (B) Representative images of  FIG.  12 A  at higher magnification. The rows from top to bottom show CAG.BEST1.WPRE, VMD2.BEST1.WPRE and VMD2. IntEx.BEST1.WPRE. The columns from left to right show staining for ZO-1 in green, BEST1 in red and a merged image including Hoechst in blue. The scale bar in the merged images indicates 25 μm. 
         FIG.  13 A-B  shows two series of 8 images each showing GFP fluorescence in ARPE19 cells differentiated for 4 months, pre-treated with 400 nM doxorubicin and transduced with ( FIG.  13 A ) AAV2/2.CAG.GFP.WPRE or ( FIG.  13 B ) AAV2/2.VMD2. InEx.GFP.WPRE at 3 different multiplicities of infection (MOI). The MOIs used were 2, 4 and 8×10 4  genome particles (gp)/cell. The scale bars in the negative control (untransduced and untreated cells) indicates 50 pin. The top row in each panel indicates untreated control cells, the bottom row are cells pre-treated with 400 nM doxorubicin. 
         FIG.  14    is a series of 20 images showing BEST1 and ZO-1 immunostaining of ARPE19 cells differentiated for 4 months, pre-treated with 400 nM doxorubicin and transduced with AAV2/2.CAG.BEST1.WPRE and AAV2/2.VMD2. InEx.BEST1.WPRE at two different MOIs: 1 and 4×10 4  gp/cell. The rows, top to bottom show ARPE19 cells with the following viral vectors: untransduced control, AAV2/2.CAG.BEST1.WPRE at a MOI 10,000 gp/cell, AAV2/2.CAG.BEST1.WPRE at a MOI 40,000 gp/cell, AAV2/2.VMD2. InEx.BEST1.WPRE at a MOI 10,000 gp/cell and AAV2/2.VMD2. InEx.BEST1.WPRE at a MOI 40,000 gp/cell. The columns from left to right show: nuclei stained with Hoechst in blue, ZO-1 staining in green, BEST1 in red, and a merged image (Hoechst, ZO-1, BEST1). Scale bars show 50 μm. 
         FIG.  15    is a table outlining a 4/8 week in vivo pilot study protocol in mice. 
         FIG.  16    is a series of 6 optical coherence tomography (OCT) images of mouse eyes four weeks after being injected with sham, VMD2.BEST1.WPRE or VMD2. IntEx.BEST1.WPRE AAV constructs. The columns, from left to right, show mice injected with a sham, with VMD2.BEST1.WPRE and with VMD2. IntEx.BEST1.WPRE AAV constructs. 
         FIG.  17    is a series of 3 OCT images of mouse eyes four weeks after being injected with, from left to right: sham, VMD2.BEST1.WPRE or VMD2. IntEx.BEST1.WPRE AAV constructs. Indicated morphological structures are the retinal ganglion cell (RGC), inner plexiform layer (IPL), the inner nuclear layer (INL), the outer plexiform layer (OPL), the outer nuclear layer (ONL), the retinal pigmented epithelium (RPE). Blue and red arrows indicate retinal thicknesses. 
         FIG.  18    is a series of 12 OCT images of mouse eyes four and eight weeks after being injected with sham, VMD2.BEST1.WPRE or VMD2. IntEx.BEST1.WPRE AAV constructs (columns, from left to right). Mid-sagittal and off-center views are shown in alternating rows. The top two rows are animals imaged at 4 weeks post injection, and the bottom two rows are animals imaged at 8 weeks post injection. 
         FIG.  19    is a series of 12 fluorescent microscopy images of mouse eyes four weeks after being injected with sham, VMD2.BEST1.WPRE or VMD2. IntEx.BEST1.WPRE AAV constructs and stained with anti BEST1 (green), anti Rhodopsin (red) and DAPI (blue). The rows show, from top to bottom, sham injected eyes, eyes injected VMD2.BEST1.WPRE or VMD2. IntEx.BEST1.WPRE AAV particles. The columns, from left to right, show anti BEST1 (green), anti Rhodopsin (red), DAPI (blue) and a merged image. The retinal pigment epithelium (RPE), photoreceptors (PR) and retinal ganglion cells (RGC) are indicated at bottom. 
         FIG.  20    is a series of 12 images of mouse eyes eight weeks after being injected with, in columns from left to right: sham, VMD2.BEST1.WPRE or VMD2. IntEx.BEST1.WPRE AAV particles and stained for BEST1 (green), Rhodopsin (red) and DAPI (blue). The rows, from top to bottom, are a merged image, anti-BEST1 (also called huBEST1), anti-Rhodopsin and a bright field image. 
         FIG.  21    is an image of a western blot showing BEST1 protein expression in dissected mouse RPE and choroid complex four weeks after injection with sham, CAG.BEST1.WPRE, VMD2.BEST1.WPRE or VMD2. IntEx.BEST1.WPRE AAV constructs. The blue arrow indicates a recombinant human Bestrophin-1 protein, while the red arrow indicates the suggested size of the BEST1 protein. CAG.BEST1.WPRE was used as a control. CAG is a strong promoter with a constitutive expression in mammalian cells. It is an hybrid between the cytomegalovirus (CMV) enhancer element, the chicken beta-actin promoter (CBA) and the splice acceptor of the rabbit beta-globin gene. 
         FIG.  22    is a table outlining a 4/13 week in vivo proof of concept (PoC) study protocol in mice. 
         FIG.  23    is a series of 20 OCT images of mouse eyes four weeks and 13 weeks after being injected with sham, VMD2. IntEx.BEST1.WPRE or VMD2.BEST1.WPRE AAV constructs at two different dosages (1×10 8  GC/μL/eye and 1×10 9  GC/μL/eye). Mid-sagittal (top row) and off-center (bottom row) views are shown in alternating rows. 
         FIG.  24    is a series of 20 microscopy images of mouse eyes four weeks after being injected with sham, VMD2. IntEx.BEST1.WPRE or VMD2.BEST1.WPRE AAV constructs at two different dosages (1×10 8  GC/μL/eye and 1×10 9  GC/μL/eye), and stained with anti-BEST1 (huBEST1, green), anti-Rhodopsin (red) and DAPI (blue). Also shown are bright field images (bottom row). The columns, from left to right, show VMD2. IntEx.BEST1.WPRE at 1×10 8  GC/μL/eye, VMD2. IntEx.BEST1.WPRE at 1×10 9  GC/μL/eye, VMD2.BEST1.WPRE at 1×10 8  GC/μL/eye and VMD2.BEST1.WPRE at 1×10 9  GC/μL/eye. Rows, from top to bottom show: a merged image, anti-BEST1, anti-Rhodopsin and bright field. Anatomical structures indicated in the upper left image are the inner nuclear layer (INL), the outer nuclear layer (ONL), the outer segment (OS), the retinal pigment epithelium (RPE) and the choroid. 
         FIG.  25 A-B  are a pair of images of western blots looking at BEST1 protein expression in cells or injected mouse RPE and choroid complex.  FIG.  25 A  is a western blot showing the expression of Bestrophin-1 protein and a beta-actin control in HEK293 and ARPE-19 cells transfected with pCAG.BEST1.WPRE, pVMD2.BEST1.WPRE and pVMD2. InEx.BEST1.WPRE or an untransfected sample as negative control.  FIG.  25 B  is a western blot showing BEST1 protein in isolated RPE and choroid samples from mice injected with either a high does (1×10 9  GC/μL/eye) or low dose (1×10 8  GC/μL/eye) of either VMD2. IntEx.BEST1.WPRE or VMD2.BEST1.WPRE AAV particles. 
         FIG.  26    is a table showing a study design for assaying human BEST1 expression by immunohistochemistry and western blot in mice injected with AAV2/2.VMD2. InEx.BEST1.WPRE. 
         FIG.  27    is a table showing a protocol for a proposed good laboratory practice (GLP) study to assess potential toxicity in mice. 
         FIG.  28    is a table showing a protocol for the evaluation of toxicity assessment study materials at 4 weeks. 
         FIG.  29    is a table showing a protocol for a proposed good laboratory practice (GLP) study to assess potential toxicity in non-human primates. 
         FIG.  30    is a table showing a dosing regimen in mouse, non-human primate and human equivalent doses in genome particles (gp) using a BEST1 AAV viral vector of the disclosure. The BEST1 AAV viral vector for the proposed doses is at a concentration of 2×10 12  DRP/mL and made according to current good manufacturing practice (GMP) standards. 
         FIG.  31    is a table showing a dosing regimen and the required concentrations of DNAse resistant particles (DRP) and number of genome particles (gp) per dose in mouse, non-human primate and human of a BEST1 AAV viral vector of the disclosure. 
     
    
    
     DETAILED DESCRIPTION 
     The disclosure relates to the finding that in many cases macular degeneration may be caused by mutations in or the abnormal function of the protein Bestrophin-1 (BEST1, also known as VMD2). The macula is a region near the center of the retina, and is responsible for central, high-resolution color vision. The fovea, located near the center of the macula, contains the largest concentration of cone cell photoreceptors in the eye. Mutations in a gene called Bestrophin-1 (BEST1, or human BEST1 (hBEST1), also known as VMD2) are associated with at least five distinct retinal degeneration diseases, called bestrinopathies. Bestrinopathies comprise best vitelliform macular dystrophy (BVMD), autosomal recessive bestrophinopathy, adult-onset vitelliform macular dystrophy, autosomal dominant vitreoretinochoroidopathy and retinitis pigmentosa. These mutations can be either dominant (for example, BVMD) or recessive. Best Vitelliform Macular Dystrophy (BVMD) and Autosomal Recessive Bestrophinopathy may cause macular degeneration with an onset in late childhood or adolescence. However, in some cases, macular degeneration begins in adulthood. However, regardless of age of onset, bestrinopathies can have a devastating effect on vision, and there is currently no known effective treatment. Given the key role that BEST1 function plays in bestrophinopathies, one approach to the treatment of bestophinopathy is to deliver a functional BEST1 protein to the affected cells of the patient. 
     Bestrophin-1 (BEST1) 
     Bestrophin-1 (BEST1) is an integral membrane protein found primarily in the retinal pigment epithelium of the eye (RPE) and predominantly localizes to the basolateral plasma membrane. BEST1 protein is thought to function as an ion channel and a regulator of intracellular calcium signaling. Human BEST1 can be found in the NCBI database with accession numbers NP_004174.1 and NM_004183.3, the contents of which are incorporated by reference in their entirety herein. 
     In some embodiments of the compositions of the disclosure, a sequence encoding a BEST1 protein of the disclosure comprises or consists of an amino acid sequence having at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to the sequence of: 
     
       
         
           
               
            
               
                 (SEQ ID NO: 4) 
               
            
           
           
               
               
            
               
                 1 
                 MTITYTSQVA NARLGSFSRL LLCWRGSIYK 
               
               
                   
               
               
                   
                 LLYGEFLIFL LCYYIIRFIY RLALTEEQQL 
               
               
                   
               
               
                 61 
                 MFEKLTLYCD SYIQLIPISF VLGFYVTLVV 
               
               
                   
               
               
                   
                 TRWWNQYENL PWPDRLMSLV SGFVEGKDEQ 
               
               
                   
               
               
                 121 
                 GRLLRRTLIR YANLGNVLIL RSVSTAVYKR 
               
               
                   
               
               
                   
                 FPSAQHLVQA GFMTPAEHKQ LEKLSLPHNM 
               
               
                   
               
               
                 181 
                 FWVPWVWFAN LSMKAWLGGR IRDPILLQSL 
               
               
                   
               
               
                   
                 LNEMNTLRTQ CGHLYAYDWI SIPLVYTQVV 
               
               
                   
               
               
                 241 
                 TVAVYSFFLT CLVGRQFLNP AKAYPGHELD 
               
               
                   
               
               
                   
                 LVVPVFTFLQ FFFYVGWLKV AEQLINPFGE 
               
               
                   
               
               
                 301 
                 DDDDFETNWI VDRNLQVSLL AVDEMHQDLP 
               
               
                   
               
               
                   
                 RMEPDMYWNK PEPQPPYTAA SAQFRRASFM 
               
               
                   
               
               
                 361 
                 GSTFNISLNK EEMEFQPNQE DEEDAHAGII 
               
               
                   
               
               
                   
                 GRFLGLQSHD HHPPRANSRT KLLWPKRESL 
               
               
                   
               
               
                 421 
                 LHEGLPKNHK AAKQNVRGQE DNKAWKLKAV 
               
               
                   
               
               
                   
                 DAFKSAPLYQ RPGYYSAPQT PLSPTPMFFP 
               
               
                   
               
               
                 481 
                 LEPSAPSKLH SVTGIDTKDK SLKTVSSGAK 
               
               
                   
               
               
                   
                 KSFELLSESD GALMEHPEVS QVRRKTVEFN 
               
               
                   
               
               
                 541 
                 LTDMPEIPEN HLKEPLEQSP TNIHTTLKDH 
               
               
                   
               
               
                   
                 MDPYWALENR DEAHS. 
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, a sequence encoding a BEST1 protein of the disclosure comprises or consists of the amino acid sequence: 
     
       
         
           
               
            
               
                 (SEQ ID NO: 4) 
               
            
           
           
               
               
            
               
                 1 
                 MTITYTSQVA NARLGSFSRL LLCWRGSIYK 
               
               
                   
               
               
                   
                 LLYGEFLIFL LCYYIIRFIY RLALTEEQQL 
               
               
                   
               
               
                 61 
                 MFEKLTLYCD SYIQLIPISF VLGFYVTLVV 
               
               
                   
               
               
                   
                 TRWWNQYENL PWPDRLMSLV SGFVEGKDEQ 
               
               
                   
               
               
                 121 
                 GRLLRRTLIR YANLGNVLIL RSVSTAVYKR 
               
               
                   
               
               
                   
                 FPSAQHLVQA GFMTPAEHKQ LEKLSLPHNM 
               
               
                   
               
               
                 181 
                 FWVPWVWFAN LSMKAWLGGR IRDPILLQSL 
               
               
                   
               
               
                   
                 LNEMNTLRTQ CGHLYAYDWI SIPLVYTQVV 
               
               
                   
               
               
                 241 
                 TVAVYSFFLT CLVGRQFLNP AKAYPGHELD 
               
               
                   
               
               
                   
                 LVVPVFTFLQ FFFYVGWLKV AEQLINPFGE 
               
               
                   
               
               
                 301 
                 DDDDFETNWI VDRNLQVSLL AVDEMHQDLP 
               
               
                   
               
               
                   
                 RMEPDMYWNK PEPQPPYTAA SAQFRRASFM 
               
               
                   
               
               
                 361 
                 GSTFNISLNK EEMEFQPNQE DEEDAHAGII 
               
               
                   
               
               
                   
                 GRFLGLQSHD HHPPRANSRT KLLWPKRESL 
               
               
                   
               
               
                 421 
                 LHEGLPKNHK AAKQNVRGQE DNKAWKLKAV 
               
               
                   
               
               
                   
                 DAFKSAPLYQ RPGYYSAPQT PLSPTPMFFP 
               
               
                   
               
               
                 481 
                 LEPSAPSKLH SVTGIDTKDK SLKTVSSGAK 
               
               
                   
               
               
                   
                 KSFELLSESD GALMEHPEVS QVRRKTVEFN 
               
               
                   
               
               
                 541 
                 LTDMPEIPEN HLKEPLEQSP TNIHTTLKDH 
               
               
                   
               
               
                   
                 MDPYWALENR DEAHS. 
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, a nucleic acid sequence encoding a BEST1 protein of the disclosure comprises or consists of a nucleic acid having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to the nucleic acid sequence of: 
     
       
         
           
               
            
               
                 (SEQ ID NO: 3) 
               
            
           
           
               
               
            
               
                 1 
                 ATGACCATCA CTTACACAAG CCAAGTGGCT 
               
               
                   
               
               
                   
                 AATGCCCGCT TAGGCTCCTT CTCCCGCCTG 
               
               
                   
               
               
                 61 
                 CTGCTGTGCT GGCGGGGCAG CATCTACAAG 
               
               
                   
               
               
                   
                 CTGCTATATG GCGAGTTCTT AATCTTCCTG 
               
               
                   
               
               
                 121 
                 CTCTGCTACT ACATCATCCG CTTTATTTAT 
               
               
                   
               
               
                   
                 AGGCTGGCCC TCACGGAAGA ACAACAGCTG 
               
               
                   
               
               
                 181 
                 ATGTTTGAGA AACTGACTCT GTATTGCGAC 
               
               
                   
               
               
                   
                 AGCTACATCC AGCTCATCCC CATTTCCTTC 
               
               
                   
               
               
                 241 
                 GTGCTGGGCT TCTACGTGAC GCTGGTCGTG 
               
               
                   
               
               
                   
                 ACCCGCTGGT GGAACCAGTA CGAGAACCTG 
               
               
                   
               
               
                 301 
                 CCGTGGCCCG ACCGCCTCAT GAGCCTGGTG 
               
               
                   
               
               
                   
                 TCGGGCTTCG TCGAAGGCAA GGACGAGCAA 
               
               
                   
               
               
                 361 
                 GGCCGGCTGC TGCGGCGCAC GCTCATCCGC 
               
               
                   
               
               
                   
                 TACGCCAACC TGGGCAACGT GCTCATCCTG 
               
               
                   
               
               
                 421 
                 CGCAGCGTCA GCACCGCAGT CTACAAGCGC 
               
               
                   
               
               
                   
                 TTCCCCAGCG CCCAGCACCT GGTGCAAGCA 
               
               
                   
               
               
                 481 
                 GGCTTTATGA CTCCGGCAGA ACACAAGCAG 
               
               
                   
               
               
                   
                 TTGGAGAAAC TGAGCCTACC ACACAACATG 
               
               
                   
               
               
                 541 
                 TTCTGGGTGC CCTGGGTGTG GTTTGCCAAC 
               
               
                   
               
               
                   
                 CTGTCAATGA AGGCGTGGCT TGGAGGTCGA 
               
               
                   
               
               
                 601 
                 ATCCGGGACC CTATCCTGCT CCAGAGCCTG 
               
               
                   
               
               
                   
                 CTGAACGAGA TGAACACCTT GCGTACTCAG 
               
               
                   
               
               
                 661 
                 TGTGGACACC TGTATGCCTA CGACTGGATT 
               
               
                   
               
               
                   
                 AGTATCCCAC TGGTGTATAC ACAGGTGGTG 
               
               
                   
               
               
                 721 
                 ACTGTGGCGG TGTACAGCTT CTTCCTGACT 
               
               
                   
               
               
                   
                 TGTCTAGTTG GGCGGCAGTT TCTGAACCCA 
               
               
                   
               
               
                 781 
                 GCCAAGGCCT ACCCTGGCCA TGAGCTGGAC 
               
               
                   
               
               
                   
                 CTCGTTGTGC CCGTCTTCAC GTTCCTGCAG 
               
               
                   
               
               
                 841 
                 TTCTTCTTCT ATGTTGGCTG GCTGAAGGTG 
               
               
                   
               
               
                   
                 GCAGAGCAGC TCATCAACCC CTTTGGAGAG 
               
               
                   
               
               
                 901 
                 GATGATGATG ATTTTGAGAC CAACTGGATT 
               
               
                   
               
               
                   
                 GTCGACAGGA ATTTGCAGGT GTCCCTGTTG 
               
               
                   
               
               
                 961 
                 GCTGTGGATG AGATGCACCA GGACCTGCCT 
               
               
                   
               
               
                   
                 CGGATGGAGC CGGACATGTA CTGGAATAAG 
               
               
                   
               
               
                 1021 
                 CCCGAGCCAC AGCCCCCCTA CACAGCTGCT 
               
               
                   
               
               
                   
                 TCCGCCCAGT TCCGTCGAGC CTCCTTTATG 
               
               
                   
               
               
                 1081 
                 GGCTCCACCT TCAACATCAG CCTGAACAAA 
               
               
                   
               
               
                   
                 GAGGAGATGG AGTTCCAGCC CAATCAGGAG 
               
               
                   
               
               
                 1141 
                 GACGAGGAGG ATGCTCACGC TGGCATCATT 
               
               
                   
               
               
                   
                 GGCCGCTTCC TAGGCCTGCA GTCCCATGAT 
               
               
                   
               
               
                 1201 
                 CACCATCCTC CCAGGGCAAA CTCAAGGACC 
               
               
                   
               
               
                   
                 AAACTACTGT GGCCCAAGAG GGAATCCCTT 
               
               
                   
               
               
                 1261 
                 CTCCACGAGG GCCTGCCCAA AAACCACAAG 
               
               
                   
               
               
                   
                 GCAGCCAAAC AGAACGTTAG GGGCCAGGAA 
               
               
                   
               
               
                 1321 
                 GACAACAAGG CCTGGAAGCT TAAGGCTGTG 
               
               
                   
               
               
                   
                 GACGCCTTCA AGTCTGCCCC ACTGTATCAG 
               
               
                   
               
               
                 1381 
                 AGGCCAGGCT ACTACAGTGC CCCACAGACG 
               
               
                   
               
               
                   
                 CCCCTCAGCC CCACTCCCAT GTTCTTCCCC 
               
               
                   
               
               
                 1441 
                 CTAGAACCAT CAGCGCCGTC AAAGCTTCAC 
               
               
                   
               
               
                   
                 AGTGTCACAG GCATAGACAC CAAAGACAAA 
               
               
                   
               
               
                 1501 
                 AGCTTAAAGA CTGTGAGTTC TGGGGCCAAG 
               
               
                   
               
               
                   
                 AAAAGTTTTG AATTGCTCTC AGAGAGCGAT 
               
               
                   
               
               
                 1561 
                 GGGGCCTTGA TGGAGCACCC AGAAGTATCT 
               
               
                   
               
               
                   
                 CAAGTGAGGA GGAAAACTGT GGAGTTTAAC 
               
               
                   
               
               
                 1621 
                 CTGACGGATA TGCCAGAGAT CCCCGAAAAT 
               
               
                   
               
               
                   
                 CACCTCAAAG AACCTTTGGA ACAATCACCA 
               
               
                   
               
               
                 1681 
                 ACCAACATAC ACACTACACT CAAAGATCAC 
               
               
                   
               
               
                   
                 ATGGATCCTT ATTGGGCCTT GGAAAACAGG 
               
               
                   
               
               
                 1741 
                 GATGAAGCAC ATTCCTAA. 
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, a nucleic acid sequence encoding a BEST1 protein of the disclosure comprises or consists of the nucleic acid sequence: 
     
       
         
           
               
            
               
                 (SEQ ID NO: 3) 
               
            
           
           
               
               
            
               
                 1 
                 ATGACCATCA CTTACACAAG CCAAGTGGCT 
               
               
                   
               
               
                   
                 AATGCCCGCT TAGGCTCCTT CTCCCGCCTG 
               
               
                   
               
               
                 61 
                 CTGCTGTGCT GGCGGGGCAG CATCTACAAG 
               
               
                   
               
               
                   
                 CTGCTATATG GCGAGTTCTT AATCTTCCTG 
               
               
                   
               
               
                 121 
                 CTCTGCTACT ACATCATCCG CTTTATTTAT 
               
               
                   
               
               
                   
                 AGGCTGGCCC TCACGGAAGA ACAACAGGTG 
               
               
                   
               
               
                 181 
                 ATGTTTGAGA AACTGACTCT GTATTGCGAC 
               
               
                   
               
               
                   
                 AGCTACATCC AGCTCATCCC CATTTCCTTC 
               
               
                   
               
               
                 241 
                 GTGCTGGGCT TCTACGTGAC GCTGGTCGTG 
               
               
                   
               
               
                   
                 ACCCGCTGGT GGAACCAGTA CGAGAACCTG 
               
               
                   
               
               
                 301 
                 CCGTGGCCCG ACCGCCTCAT GAGCCTGGTG 
               
               
                   
               
               
                   
                 TCGGGCTTCG TCGAAGGCAA GGACGAGCAA 
               
               
                   
               
               
                 361 
                 GGCCGGCTGC TGCGGCGCAC GCTCATCCGC 
               
               
                   
               
               
                   
                 TACGCCAACC TGGGCAACGT GCTCATCCTG 
               
               
                   
               
               
                 421 
                 CGCAGCGTCA GCACCGCAGT CTACAAGCGC 
               
               
                   
               
               
                   
                 TTCCCCAGCG CCCAGCACCT GGTGCAAGCA 
               
               
                   
               
               
                 481 
                 GGCTTTATGA CTCCGGCAGA ACACAAGCAG 
               
               
                   
               
               
                   
                 TTGGAGAAAC TGAGCCTACC ACACAACATG 
               
               
                   
               
               
                 541 
                 TTCTGGGTGC CCTGGGTGTG GTTTGCCAAC 
               
               
                   
               
               
                   
                 CTGTCAATGA AGGCGTGGCT TGGAGGTCGA 
               
               
                   
               
               
                 601 
                 ATCCGGGACC CTATCCTGCT CCAGAGCCTG 
               
               
                   
               
               
                   
                 CTGAACGAGA TGAACACCTT GCGTACTCAG 
               
               
                   
               
               
                 661 
                 TGTGGACACC TGTATGCCTA CGACTGGATT 
               
               
                   
               
               
                   
                 AGTATCCCAC TGGTGTATAC ACAGGTGGTG 
               
               
                   
               
               
                 721 
                 ACTGTGGCGG TGTACAGCTT CTTCCTGACT 
               
               
                   
               
               
                   
                 TGTCTAGTTG GGCGGCAGTT TCTGAACCCA 
               
               
                   
               
               
                 781 
                 GCCAAGGCCT ACCCTGGCCA TGAGCTGGAC 
               
               
                   
               
               
                   
                 CTCGTTGTGC CCGTCTTCAC GTTCCTGCAG 
               
               
                   
               
               
                 841 
                 TTCTTCTTCT ATGTTGGCTG GCTGAAGGTG 
               
               
                   
               
               
                   
                 GCAGAGCAGC TCATCAACCC CTTTGGAGAG 
               
               
                   
               
               
                 901 
                 GATGATGATG ATTTTGAGAC CAACTGGATT 
               
               
                   
               
               
                   
                 GTCGACAGGA ATTTGCAGGT GTCCCTGTTG 
               
               
                   
               
               
                 961 
                 GCTGTGGATG AGATGCACCA GGACCTGCCT 
               
               
                   
               
               
                   
                 CGGATGGAGC CGGACATGTA CTGGAATAAG 
               
               
                   
               
               
                 1021 
                 CCCGAGCCAC AGCCCCCCTA CACAGCTGCT 
               
               
                   
               
               
                   
                 TCCGCCCAGT TCCGTCGAGC CTCCTTTATG 
               
               
                   
               
               
                 1081 
                 GGCTCCACCT TCAACATCAG CCTGAACAAA 
               
               
                   
               
               
                   
                 GAGGAGATGG AGTTCCAGCC CAATCAGGAG 
               
               
                   
               
               
                 1141 
                 GACGAGGAGG ATGCTCACGC TGGCATCATT 
               
               
                   
               
               
                   
                 GGCCGCTTCC TAGGCCTGCA GTCCCATGAT 
               
               
                   
               
               
                 1201 
                 CACCATCCTC CCAGGGCAAA CTCAAGGACC 
               
               
                   
               
               
                   
                 AAACTACTGT GGCCCAAGAG GGAATCCCTT 
               
               
                   
               
               
                 1261 
                 CTCCACGAGG GCCTGCCCAA AAACCACAAG 
               
               
                   
               
               
                   
                 GCAGCCAAAC AGAAGGTTAG GGGCCAGGAA 
               
               
                   
               
               
                 1321 
                 GACAACAAGG CCTGGAAGCT TAAGGCTGTG 
               
               
                   
               
               
                   
                 GACGCCTTCA AGTCTGCCCC ACTGTATCAG 
               
               
                   
               
               
                 1381 
                 AGGCCAGGCT ACTACAGTGC CCCACAGACG 
               
               
                   
               
               
                   
                 CCCCTCAGCC CCACTCCCAT GTTCTTCCCC 
               
               
                   
               
               
                 1441 
                 CTAGAACCAT CAGCGCCGTC AAAGCTTCAC 
               
               
                   
               
               
                   
                 AGTGTCACAG GCATAGACAC CAAAGACAAA 
               
               
                   
               
               
                 1501 
                 AGCTTAAAGA CTGTGAGTTC TGGGGCCAAG 
               
               
                   
               
               
                   
                 AAAAGTTTTG AATTGCTCTC AGAGAGGGAT 
               
               
                   
               
               
                 1561 
                 GGGGCCTTGA TGGAGCACCC AGAAGTATCT 
               
               
                   
               
               
                   
                 CAAGTGAGGA GGAAAACTGT GGAGTTTAAC 
               
               
                   
               
               
                 1621 
                 CTGACGGATA TGCCAGAGAT CCCCGAAAAT 
               
               
                   
               
               
                   
                 CACCTCAAAG AACCTTTGGA ACAATCACCA 
               
               
                   
               
               
                 1681 
                 ACCAACATAC ACACTACACT CAAAGATCAC 
               
               
                   
               
               
                   
                 ATGGATCCTT ATTGGGCCTT GGAAAACAGG 
               
               
                   
               
               
                 1741 
                 GATGAAGCAC ATTCCTAA. 
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, a nucleic acid sequence encoding a BEST1 protein of the disclosure comprises a codon optimized sequence. In some embodiments, the sequence has been codon optimized for expression in a mammalian cell. In some embodiments, the sequence has been codon optimized for expression in a human cell. 
     BEST1 Expression 
     In some embodiments of the compositions of the disclosure, a nucleic acid sequence encoding a BEST1 protein of the disclosure further comprises a sequence encoding a regulatory element that enhances or increases BEST1 transcript or BEST1 protein expression. Exemplary regulatory element that enhances or increases BEST1 transcript or BEST1 protein expression include, but are not limited to, a promoter, an enhancer, a superenhancer, an intron, an exon, a combination of an intron and exon, a sequence encoding an untranslated region (e.g. a 5′ untranslated region (UTR) or a 3′ UTR), a sequence comprising a polyadenylation (polyA) signal, and a posttranscriptional regulatory element (PRE). 
     Exemplary promoters of the disclosure include, but are not limited to, those promoters capable of expressing a sequence encoding a BEST1 protein or a BEST1 protein in a mammalian cell. Exemplary promoters of the disclosure include, but are not limited to, those promoters capable of expressing a sequence encoding a BEST1 protein or a BEST1 protein in a human cell. In some embodiments, the mammalian or the human cell may be in vivo, ex vivo, in vitro or in situ. In some embodiments, the promoter may be constitutively active. In some embodiments, the promoter may be cell-type specific. In some embodiments, the promoter may be inducible. 
     Exemplary constitutively active promoters of the disclosure include, but are not limited to, a viral promoter. Viral promoters of the disclosure may include, but are not limited to, a simian virus 40 (SV40) promoter, a cytomegalovirus (CMV) promoter, ubiquitin C (UBC) promoter, elongation factor-1 alpha (EF1A) promoter, phosphoglycerate kinase 1 (PGK) promoter and a CAG promoter (a combination of a (C) the cytomegalovirus (CMV) early enhancer element, (A) the promoter comprising the first exon and the first intron of chicken beta-actin gene, and (G) the splice acceptor of the rabbit beta-globin gene). In some embodiments, a CMV promoter is used to control expression of a nucleic acid sequence encoding a BEST1 protein of the disclosure. In some embodiments, a CAG promoter is used to control expression of a nucleic acid sequence encoding a BEST1 protein of the disclosure. Non-viral promoters of the disclosure may include, but are not limited to, a chicken beta actin (CBA) promoter. In some embodiments, the CBA promoter comprises the chicken beta actin the first exon and intron of the CBA gene. In some embodiments, the promoter comprises the chicken beta actin promoter and the cytomegalovirus early enhancer elements. In some embodiments, the promoter further comprises a rabbit beta globin splice acceptor sequence (the CAG promoter). In some embodiments, the CAG promoter comprises or consists of a nucleic acid sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identity to the nucleic acid sequence of: 
     
       
         
           
               
            
               
                 (SEQ ID NO: 2) 
               
            
           
           
               
               
            
               
                 1 
                 CCATTGACGT CAATAATGAC GTATGTTCCC 
               
               
                   
               
               
                   
                 ATAGTAACGC CAATAGGGAC TTTCCATTGA 
               
               
                   
               
               
                 61 
                 CGTCAATGGG TGGAGTATTT ACGGTAAACT 
               
               
                   
               
               
                   
                 GCCCACTTGG CAGTACATGA AGTGTATCAT 
               
               
                   
               
               
                 121 
                 ATGCCAAGTA CGCCCCCTAT TGACGTCAAT 
               
               
                   
               
               
                   
                 GACGGTAAAT GGCCCGCCTG GCATTATGCC 
               
               
                   
               
               
                 181 
                 CAGTACATGA CCTTATGGGA CTTTCCTACT 
               
               
                   
               
               
                   
                 TGGCAGTACA TCTACGTATT AGTCATCGCT 
               
               
                   
               
               
                 241 
                 ATTACCATGG TCGAGGTGAG CCCCACGTTC 
               
               
                   
               
               
                   
                 TGCTTCACTC TCCCCATCTC CCCCCCCTCC 
               
               
                   
               
               
                 301 
                 CCACCCCCAA TTTTGTATTT ATTTATTTTT 
               
               
                   
               
               
                   
                 TAATTATTTT GTGCAGCGAT GGGGGGGGGG 
               
               
                   
               
               
                 361 
                 GGGGGGGGGG GGCGCGCGCC AGGCGGGGCG 
               
               
                   
               
               
                   
                 GGGGGGGGGG AGGGGCGGGG CGGGGCGAGG 
               
               
                   
               
               
                 421 
                 CGGAGAGGTG CGGCGGCAGC CAATCAGAGC 
               
               
                   
               
               
                   
                 GGCGCGCTCC GAAAGTTTCC TTTTATGGCG 
               
               
                   
               
               
                 481 
                 AGGCGGCGGC GGCGGCGGCC CTATAAAAAG 
               
               
                   
               
               
                   
                 CGAAGCGCGC GGGGGGGGGG AGTCGCTGCG 
               
               
                   
               
               
                 541 
                 CGCTGCCTTC GCCCCGTGCC CCGCTCCGCC 
               
               
                   
               
               
                   
                 GCCGCCTCGC GCCGCCCGCC CCGGCTCTGA 
               
               
                   
               
               
                 601 
                 CTGACCGCGT TACTCCCACA GGTGAGCGGG 
               
               
                   
               
               
                   
                 CGGGACGGCC CTTCTCCTCC GGGCTGTAAT 
               
               
                   
               
               
                 661 
                 TAGCGCTTGG TTTAATGACG GCTTGTTTCT 
               
               
                   
               
               
                   
                 TTTCTGTGGC TGCGTGAAAG CCTTGAGGGG 
               
               
                   
               
               
                 721 
                 CTCCGGGAGG GCCCTTTGTG CGGGGGGAGC 
               
               
                   
               
               
                   
                 GGCTCGGGGC TGTCCGCGGG GGGACGGCTG 
               
               
                   
               
               
                 781 
                 CCTTCGGGGG GGACGGGGCA GGGCGGGGTT 
               
               
                   
               
               
                   
                 CGGCTTCTGG CGTGTGACCG GCGGCTCTAG 
               
               
                   
               
               
                 841 
                 AGCCTCTGCT AACCATGTTC ATGCCTTCTT 
               
               
                   
               
               
                   
                 CTTTTTCCTA CAGCTCCTGG GCAACGTGCT 
               
               
                   
               
               
                 901 
                 GGTTATTGTG CTGTCTCATC ATTTTGGCAA 
               
               
                   
               
               
                   
                 AGAATTGGAT C. 
               
            
           
         
       
     
     Exemplary cell-type specific promoters of the disclosure include, but are not limited to, a promoter capable of expressing a nucleic acid or a protein in a neuron, a promoter capable of expressing a nucleic acid or a protein in a retinal cell, a promoter capable of expressing a nucleic acid or a protein in a photoreceptor, a promoter capable of expressing a nucleic acid or a protein in a rod cell, and a promoter capable of expressing a nucleic acid or a protein in a cone cell. In some embodiments, a sequence encoding a tissue specific promoter comprises a sequence encoding a human VMD2 gene (also known as Bestrophin-1). In some embodiments, a tissue specific promoter comprises a human VMD2 promoter (also known as Bestrophin-1). In some embodiments, the human VMD2 promoter comprises or consists of a nucleic acid sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to the nucleic acid sequence of: 
     
       
         
           
               
            
               
                 (SEQ ID NO: 1) 
               
            
           
           
               
               
            
               
                 1 
                 AATTCTGTCA TTTTACTAGG GTGATGAAAT 
               
               
                   
               
               
                   
                 TCCCAAGCAA CACCATCCTT TTCAGATAAG 
               
               
                   
               
               
                 61 
                 GGCACTGAGG CTGAGAGAGG AGCTGAAACC 
               
               
                   
               
               
                   
                 TACCCGGGGT CACCACACAC AGGTGGCAAG 
               
               
                   
               
               
                 121 
                 GCTGGGACCA GAAACCAGGA CTGTTGACTG 
               
               
                   
               
               
                   
                 CAGCCCGGTA TTCATTCTTT CCATAGCCCA 
               
               
                   
               
               
                 181 
                 CAGGGCTGTC AAAGACCCCA GGGCCTAGTC 
               
               
                   
               
               
                   
                 AGAGGCTCCT CCTTCCTGGA GAGTTCCTGG 
               
               
                   
               
               
                 241 
                 CACAGAAGTT GAAGCTCAGC ACAGCCCCCT 
               
               
                   
               
               
                   
                 AACCCCCAAC TCTCTCTGCA AGGCCTCAGG 
               
               
                   
               
               
                 301 
                 GGTCAGAACA CTGGTGGAGC AGATCCTTTA 
               
               
                   
               
               
                   
                 GCCTCTGGAT TTTAGGGCCA TGGTAGAGGG 
               
               
                   
               
               
                 361 
                 GGTGTTGCCC TAAATTCCAG CCCTGGTCTC 
               
               
                   
               
               
                   
                 AGCCCAACAC CCTCCAAGAA GAAATTAGAG 
               
               
                   
               
               
                 421 
                 GGGCCATGGC CAGGCTGTGC TAGCCGTTGC 
               
               
                   
               
               
                   
                 TTCTGAGCAG ATTACAAGAA GGGACTAAGA 
               
               
                   
               
               
                 481 
                 CAAGGACTCC TTTGTGGAGG TCCTGGCTTA 
               
               
                   
               
               
                   
                 GGGAGTCAAG TGACGGCGGC TCAGCACTCA 
               
               
                   
               
               
                 541 
                 CGTGGGCAGT GCCAGCCTCT AAGAGTGGGC 
               
               
                   
               
               
                   
                 AGGGGCACTG GCCACAGAGT CCCAGGGAGT 
               
               
                   
               
               
                 601 
                 CCCACCAGCC TAGTCGCCAG ACC. 
               
            
           
         
       
     
     In some embodiments, the human VMD2 promoter comprises or consists of a nucleic acid sequence having 100% identity to the nucleic acid sequence of: 
     
       
         
           
               
            
               
                 (SEQ ID NO: 1) 
               
            
           
           
               
               
            
               
                 1 
                 AATTCTGTCA TTTTACTAGG GTGATGAAAT 
               
               
                   
               
               
                   
                 TCCCAAGCAA CACCATCCTT TTCAGATAAG 
               
               
                   
               
               
                 61 
                 GGCACTGAGG CTGAGAGAGG AGCTGAAACC 
               
               
                   
               
               
                   
                 TACCCGGGGT CACCACACAC AGGTGGCAAG 
               
               
                   
               
               
                 121 
                 GCTGGGACCA GAAACCAGGA CTGTTGACTG 
               
               
                   
               
               
                   
                 CAGCCCGGTA TTCATTCTTT CCATAGCCCA 
               
               
                   
               
               
                 181 
                 CAGGGCTGTC AAAGACCCCA GGGCCTAGTC 
               
               
                   
               
               
                   
                 AGAGGCTCCT CCTTCCTGGA GAGTTCCTGG 
               
               
                   
               
               
                 241 
                 CACAGAAGTT GAAGCTCAGC ACAGCCCCCT 
               
               
                   
               
               
                   
                 AACCCCCAAC TCTCTCTGCA AGGCCTCAGG 
               
               
                   
               
               
                 301 
                 GGTCAGAACA CTGGTGGAGC AGATCCTTTA 
               
               
                   
               
               
                   
                 GCCTCTGGAT TTTAGGGCCA TGGTAGAGGG 
               
               
                   
               
               
                 361 
                 GGTGTTGCCC TAAATTCCAG CCCTGGTCTC 
               
               
                   
               
               
                   
                 AGCCCAACAC CCTCCAAGAA GAAATTAGAG 
               
               
                   
               
               
                 421 
                 GGGCCATGGC CAGGCTGTGC TAGCCGTTGC 
               
               
                   
               
               
                   
                 TTCTGAGCAG ATTACAAGAA GGGACTAAGA 
               
               
                   
               
               
                 481 
                 CAAGGACTCC TTTGTGGAGG TCCTGGCTTA 
               
               
                   
               
               
                   
                 GGGAGTCAAG TGACGGCGGC TCAGCACTCA 
               
               
                   
               
               
                 541 
                 CGTGGGCAGT GCCAGCCTCT AAGAGTGGGC 
               
               
                   
               
               
                   
                 AGGGGCACTG GCCACAGAGT CCCAGGGAGT 
               
               
                   
               
               
                 601 
                 CCCACCAGCC TAGTCGCCAG ACC. 
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, the nucleic acid sequence comprising a sequence encoding a BEST1 protein and a sequence encoding a promoter, further comprises an intron and an exon. The presence of an intron and an exon increases levels of protein expression. In some embodiments, the intron is positioned between the VMD2 promoter and the exon. In some embodiments, including those embodiments wherein the intron is positioned between the VMD2 promoter and the exon, the exon is positioned 5′ of the BEST coding sequence. 
     The exon may comprise a coding sequence, a non-coding sequence, or a combination of both. In some embodiments, the exon comprises non-coding sequence. In some embodiments, the exon is isolated or derived from a mammalian gene. In embodiments, the mammal is a rabbit (Oryctolagus  cuniculus ). In some embodiments, the mammalian gene comprises a rabbit beta globin gene. In some embodiments, the exon comprises a nucleic acid sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                   
                 (SEQ ID NO: 6) 
               
               
                   
                 CTCCTGGGCA ACGTGCTGGT TATTGTGCTG 
               
               
                   
                   
               
               
                   
                 TCTCATCATT TTGGCAAAGA ATT 
               
            
           
         
       
     
     In some embodiments, the exon comprises a nucleic acid sequence having 100% identify to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                   
                 (SEQ ID NO: 6) 
               
               
                   
                 CTCCTGGGCA ACGTGCTGGT TATTGTGCTG 
               
               
                   
                   
               
               
                   
                 TCTCATCATT TTGGCAAAGA ATT 
               
            
           
         
       
     
     Introns may comprise a splice donor site, a splice acceptor site or a branch point. Introns may comprise a splice donor site, a splice acceptor site and a branch point. Exemplary splice acceptor sites comprise nucleotides “GT” (“GU” in the pre-mRNA) at the 5′ end of the intron. Exemplary splice acceptor sites comprise an “AG” at the 3′ end of the intron. In some embodiments, the branch point comprises an adenosine (A) between 20 and 40 nucleotides, inclusive of the endpoints, upstream of the 3′ end of the intron. The intron may be an artificial or non-naturally occurring sequence. Alternatively, the intron may be isolated or derived from a vertebrate gene. The intron may comprise a sequence encoding a fusion of two sequences, each of which may be isolated or derived from a plurality of vertebrate genes. In some embodiments, a vertebrate gene contributing to the intron nucleic acid sequence comprises a chicken ( Gallus gallus ) gene. In some embodiments, the chicken gene comprises the chicken beta actin gene. In some embodiments, a vertebrate gene contributing to the intron nucleic acid sequence comprises a rabbit (Oryctolagus  cuniculus ) gene. In some embodiments, the rabbit gene comprises the rabbit beta globin gene. In some embodiments, the intron comprises a nucleic acid sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 7) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 GTGCCGCAGG GGGACGGCTG CCTTCGGGGG GGACGGGGCA GGGCGGGGTT CGGCTTCTGG 
                   
               
               
                   
               
               
                 61 
                 CGTGTGACCG GCGGCTCTAG AGCCTCTGCT AACCATGTTC ATGCCTTCTT CTTTTTCCTA 
               
               
                   
               
               
                 121 
                 CAG. 
               
            
           
         
       
     
     In some embodiments, the intron comprises a nucleic acid sequence having 100% identify to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 7) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 GTGCCGCAGG GGGACGGCTG CCTTCGGGGG GGACGGGGCA GGGCGGGGTT CGGCTTCTGG 
                   
               
               
                   
               
               
                 61 
                 CGTGTGACCG GCGGCTCTAG AGCCTCTGCT AACCATGTTC ATGCCTTCTT CTTTTTCCTA 
               
               
                   
               
               
                 121 
                 CAG. 
               
            
           
         
       
     
     Kozak sequences are short sequence motifs that are recognized by the ribosome as the translation start site. Kozak sequences may be positioned immediately upstream, or surrounding the translational start site. In vertebrates, the Kozak consensus sequence comprises a sequence of having at least 50% identity to the consensus sequence of gccRccATGG, where R represents an A or G, and the ATG encoding the start methionine is bolded. An exemplary Kozak sequence of the disclosure comprises a sequence of GGCACCATGA. In some embodiments, the nucleic acid comprising a nucleic acid sequence encoding BEST1, further comprises a sequence encoding a 5′ untranslated sequence (5′ UTR). In some embodiments, the 5′ UTR comprises a Kozak sequence. In some embodiments, the 5′ UTR comprises a portion of a Kozak sequence. In some embodiments, the 5′ UTR comprises at least 50%, at least 60%, at least 70% or at least 80% of a Kozak sequence. 
     In some embodiments, the nucleic acid comprising a nucleic acid sequence encoding BEST1, further comprises a nucleic acid sequence encoding transcriptional response element (PRE). Exemplary PREs comprise a Woodchuck PRE (WPRE), which is derived from the Woodchuck hepatitis virus. In some embodiments, a sequence encoding a WPRE is positioned 3′ of the nucleic acid sequence encoding BEST1. In some embodiments, a sequence encoding a WPRE is positioned between the nucleic acid sequence encoding BEST1 and the sequence encoding a polyA signal. In some embodiments, a sequence encoding a WPRE comprises a nucleic acid sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 8) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 ATCGATAATC AACCTCTGGA TTACAAAATT TGTGAAAGAT TGACTGGTAT TCTTAACTAT 
                   
               
               
                   
               
               
                 61 
                 GTTGCTCCTT TTACGCTATG TGGATACGCT GCTTTAATGC CTTTGTATCA TGCTATTGCT 
               
               
                   
               
               
                 121 
                 TCCCGTATGG CTTTCATTTT CTCCTCCTTG TATAAATCCT GGTTGCTGTC TCTTTATGAG 
               
               
                   
               
               
                 181 
                 GAGTTGTGGC CCGTTGTCAG GCAACGTGGC GTGGTGTGCA CTGTGTTTGC TGACGCAACC 
               
               
                   
               
               
                 241 
                 CCCACTGGTT GGGGCATTGC CACCACCTGT CAGCTCCTTT CCGGGACTTT CGCTTTCCCC 
               
               
                   
               
               
                 301 
                 CTCCCTATTG CCACGGCGGA ACTCATCGCC GCCTGCCTTG CCCGCTGCTG GACAGGGGCT 
               
               
                   
               
               
                 361 
                 CGGCTGTTGG GCACTGACAA TTCCGTGGTG TTGTCGGGGA AATCATCGTC CTTTCCTTGG 
               
               
                   
               
               
                 421 
                 CTGCTCGCCT GTGTTGCCAC CTGGATTCTG CGCGGGACGT CCTTCTGCTA CGTCCCTTCG 
               
               
                   
               
               
                 481 
                 GCCCTCAATC CAGCGGACCT TCCTTCCCGC GGCCTGCTGC CGGCTCTGCG GCCTCTTCCG 
               
               
                   
               
               
                 541 
                 CGTCTTCGCC TTCGCCCTCA GACGAGTCGG ATCTCCCTTT GGGCCGCCTC CCC. 
               
            
           
         
       
     
     In some embodiments, a sequence encoding a WPRE comprises a nucleic acid sequence having 100% identity to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 8) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 ATCGATAATC AACCTCTGGA TTACAAAATT TGTGAAAGAT TGACTGGTAT TCTTAACTAT 
                   
               
               
                   
               
               
                 61 
                 GTTGCTCCTT TTACGCTATG TGGATACGCT GCTTTAATGC CTTTGTATCA TGCTATTGCT 
               
               
                   
               
               
                 121 
                 TCCCGTATGG CTTTCATTTT CTCCTCCTTG TATAAATCCT GGTTGCTGTC TCTTTATGAG 
               
               
                   
               
               
                 181 
                 GAGTTGTGGC CCGTTGTCAG GCAACGTGGC GTGGTGTGCA CTGTGTTTGC TGACGCAACC 
               
               
                   
               
               
                 241 
                 CCCACTGGTT GGGGCATTGC CACCACCTGT CAGCTCCTTT CCGGGACTTT CGCTTTCCCC 
               
               
                   
               
               
                 301 
                 CTCCCTATTG CCACGGCGGA ACTCATCGCC GCCTGCCTTG CCCGCTGCTG GACAGGGGCT 
               
               
                   
               
               
                 361 
                 CGGCTGTTGG GCACTGACAA TTCCGTGGTG TTGTCGGGGA AATCATCGTC CTTTCCTTGG 
               
               
                   
               
               
                 421 
                 CTGCTCGCCT GTGTTGCCAC CTGGATTCTG CGCGGGACGT CCTTCTGCTA CGTCCCTTCG 
               
               
                   
               
               
                 481 
                 GCCCTCAATC CAGCGGACCT TCCTTCCCGC GGCCTGCTGC CGGCTCTGCG GCCTCTTCCG 
               
               
                   
               
               
                 541 
                 CGTCTTCGCC TTCGCCCTCA GACGAGTCGG ATCTCCCTTT GGGCCGCCTC CCC. 
               
            
           
         
       
     
     In some embodiments, the nucleic acid comprising a nucleic acid sequence encoding BEST1, further comprises a sequence encoding a polyadenylation (polyA) signal. The polyA signal facilitates nuclear export, enhances translation and increases mRNA stability. In some embodiments, the sequence encoding the polyA signal comprises a synthetic or an artificial sequence. In some embodiments, the sequence encoding the polyA signal comprises a sequence isolated or derived from a mammalian gene. In some embodiments, the mammalian gene is a human gene. In some embodiments, the mammalian gene is a bovine growth hormone gene 
     (BGH). In some embodiments, the sequence encoding the polyA signal comprises a nucleic acid sequence having at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% identity to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 9) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 CGCTGATCAG CCTCGACTGT GCCTTCTAGT TGCCAGCCAT CTGTTGTTTG CCCCTCCCCC 
                   
               
               
                   
               
               
                 61 
                 GTGCCTTCCT TGACCCTGGA AGGTGCCACT CCCACTGTCC TTTCCTAATA AAATGAGGAA 
               
               
                   
               
               
                 121 
                 ATTGCATCGC ATTGTCTGAG TAGGTGTCAT TCTATTCTGG GGGGTGGGGT GGGGCAGGAC 
               
               
                   
               
               
                 181 
                 AGCAAGGGGG AGGATTGGGA AGACAATAGC AGGCATGCTG GGGATGCGGT GGGCTCTATG 
               
               
                   
               
               
                 241 
                 GCTTCTGAGG CGGAAAGAAC CAGCTGGGG. 
               
            
           
         
       
     
     In some embodiments, the sequence encoding the polyA signal comprises a nucleic acid sequence having 100% identity to the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 9) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 CGCTGATCAG CCTCGACTGT GCCTTCTAGT TGCCAGCCAT CTGTTGTTTG CCCCTCCCCC 
                   
               
               
                   
               
               
                 61 
                 GTGCCTTCCT TGACCCTGGA AGGTGCCACT CCCACTGTCC TTTCCTAATA AAATGAGGAA 
               
               
                   
               
               
                 121 
                 ATTGCATCGC ATTGTCTGAG TAGGTGTCAT TCTATTCTGG GGGGTGGGGT GGGGCAGGAC 
               
               
                   
               
               
                 181 
                 AGCAAGGGGG AGGATTGGGA AGACAATAGC AGGCATGCTG GGGATGCGGT GGGCTCTATG 
               
               
                   
               
               
                 241 
                 GCTTCTGAGG CGGAAAGAAC CAGCTGGGG. 
               
            
           
         
       
     
     AAV Vectors 
     A vector may comprise the nucleic acid comprising a nucleic acid sequence encoding BEST1. In some embodiments of the compositions of the disclosure, the vector may be a viral delivery vector. Viral delivery vectors of the disclosure may contain sequences necessary for packaging a nucleic acid sequence of the disclosure into a viral delivery system for delivery to a target cell or tissue. Typical viral delivery vectors of the disclosure include, but are not limited to, lentiviral, retroviral or adeno-associated viral (AAV) vectors. 
     An AAV viral delivery system of the disclosure may be in the form of a mature AAV particle or virion, i.e. nucleic acid surrounded by an AAV protein capsid. In some embodiments, the AAV viral delivery vector may comprise an AAV genome or a derivative thereof. 
     An AAV genome is a nucleic acid sequence which encodes functions needed for production of an AAV particle. These functions include those operating in the replication and packaging cycle of AAV in a host cell, including encapsidation of the AAV genome into an AAV particle. Naturally occurring AAVs are replication-deficient and rely on the provision of helper functions in trans for completion of a replication and packaging cycle. In preferred embodiments, an AAV genome of a vector of the disclosure is replication-deficient. 
     The AAV genome may be in single-stranded form, either positive or negative-sense, or alternatively in double-stranded form. The use of a double-stranded form allows bypass of the DNA replication step in the target cell and so can accelerate transgene expression. The AAV genome of a vector of the disclosure may be single-stranded form. 
     The AAV genome may be from any naturally derived serotype, isolate or Glade of AAV. Thus, the AAV genome may be the full genome of a naturally occurring AAV. As is known to the person skilled in the art, AAVs occurring in nature may be classified according to various biological systems. 
     AAVs are referred to in terms of their serotype. A serotype corresponds to a variant subspecies of AAV which, owing to its profile of expression of capsid surface antigens, has a distinctive reactivity which can be used to distinguish it from other variant subspecies. A virus having a particular AAV serotype does not efficiently cross-react with neutralizing antibodies specific for any other AAV serotype. AAV serotypes include AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10 and AAV11, and also recombinant serotypes, such as Rec2 and Rec3, recently identified from primate brain. Any of these AAV serotypes may be used in the invention. Thus, in some embodiments, an AAV vector of the invention may be derived from an AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, Rec2 or Rec3 AAV. 
     Reviews of AAV serotypes may be found in Choi et al. (2005) Cur. Gene There. 5: 299-310 and Wu et al. (2006) Molecular Therapy 14: 316-27. The sequences of AAV genomes or of elements of AAV genomes including ITR sequences, rep or cap genes may be derived from the following accession numbers for AAV whole genome sequences: Adeno-associated virus 1 NC_002077, AF063497; Adeno-associated virus 2 NC_001401; Adeno-associated virus 3 NC_001729; Adeno-associated virus 3B NC_001863; Adeno-associated virus 4 NC_001829; Adeno-associated virus 5 Y18065, AF085716; Adeno-associated virus 6 NC_001862; Avian AAV ATCC VR-865 AY186198, AY629583, NC_004828; Avian AAV strain DA-1 NC_006263, AY629583; Bovine AAV NC_005889, AY388617. 
     AAV may also be referred to in terms of clades or clones. This refers to the phylogenetic relationship of naturally derived AAVs, as well as to a phylogenetic group of AAVs which can be traced back to a common ancestor, and includes all descendants thereof. Additionally, AAVs may be referred to in terms of a specific isolate, i.e. a genetic isolate of a specific AAV found in nature. The term genetic isolate describes a population of AAVs which has undergone limited genetic mixing with other naturally occurring AAVs, thereby defining a recognizably distinct population at a genetic level. 
     The AAV serotype determines the tissue specificity of infection (or tropism) of an AAV virus. Accordingly, preferred AAV serotypes for use in AAVs administered to patients in accordance with the invention are those which have natural tropism for or a high efficiency of infection of target cells within the eye. In one embodiment, AAV serotypes for use in the invention are those which infect cells of the neurosensory retina, retinal pigment epithelium and/or macula. 
     The AAV genome of a naturally derived serotype, isolate or Glade of AAV comprises at least one inverted terminal repeat sequence (ITR). An ITR sequence acts in cis to provide a functional origin of replication and allows for integration and excision of the vector from the genome of a cell. 
     An AAV viral delivery vector may include at least one inverted terminal repeat sequence (ITR), preferably more than one ITR, such as two ITRs or more. One or more of the ITRs may be derived from AAV genomes having different serotypes, or may be a chimeric or mutant ITR. A preferred mutant ITR is one having a deletion of a trs (terminal resolution site). This deletion allows for continued replication of the genome to generate a single-stranded genome which contains both coding and complementary sequences, i.e. a self-complementary AAV genome. This allows for bypass of DNA replication in the target cell, and so enables accelerated transgene expression. 
     The inclusion of one or more ITRs is preferred to aid concatamer formation of a viral delivery vector of the invention in the nucleus of a host cell, for example following the conversion of single-stranded vector DNA into double-stranded DNA by the action of host cell DNA polymerases. The formation of such episomal concatamers protects the vector construct during the life of the host cell, thereby allowing for prolonged expression of the transgene in vivo. 
     In some embodiments, ITR elements are the only sequences retained from the native AAV genome in the viral delivery vector. Thus, in some embodiments, a viral delivery vector does not include either the rep or cap genes of the native genome and, furthermore, lacks any other sequences of the native genome. This is preferred for the reasons described above, and also to reduce the possibility of integration of the vector into the host cell genome. Additionally, reducing the size of the AAV genome allows for increased flexibility in incorporating other sequence elements (such as regulatory elements) within the vector in addition to the transgene. In some embodiments, the viral delivery vector of the disclosure comprises sequences encoding AAV2 ITRs. In some embodiments, the sequences encoding the two AAV2 ITRs may comprise or consist of a nucleic acid sequence of: 
                        (SEQ ID NO: 10)                             1   ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt                   61   ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact               121   aggggttcct tgtagttaat gatt.            
and/or
 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 11) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg 
                   
               
               
                   
               
               
                 61 
                 cccgggcggc ctcagtgagc gagcgagcgc gcagagcttt ttgcaaaagc ctaggcctcc 
               
               
                   
               
               
                 121 
                 aaaaaagcct cctcactact tctgg. 
               
            
           
         
       
     
     The AAV genome may comprise a nucleic acid sequence of about 4.7 kb in length. Thus, in those embodiments where the nucleic acid sequence to be delivered by an AAV viral vector is less than 4.7 kb in length, a stuffer or filler sequence may be used. The presence of a stuffer sequence can, in some embodiments, aid in AAV viral vector packaging into the viral particle. In some embodiments, the stuffer sequence comprises a random sequence. An exemplary stuffer sequence of the disclosure may comprise or consist of the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 12) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 GATGTAACCA TATACTTAGG CTGGATCTTC TCCCGCGAAT TTTAACCCTC ACCAACTACG 
                   
               
               
                   
               
               
                 61 
                 AGATATGAGG TAAGCCAAAA AAGCACGTAG TGGCGCTCTC CGACTGTTCC CAAATTGTAA 
               
               
                   
               
               
                 121 
                 CTTATCGTTC CGTGAAGGCC AGAGTTACTT CCCGGCCCTT TCCATGCGCG CACCATACCC 
               
               
                   
               
               
                 181 
                 TCCTAGTTCC CCGGTTATCT TTCCGAAGTG GGAGTGAGCG AACCTCCGTT TACGTCTTGT 
               
               
                   
               
               
                 241 
                 TACCAATGAT GTAGCTATGC ACTTTGTACA GGGTGCCAAC GGGTTTCACA ATTCACAGAT 
               
               
                   
               
               
                 301 
                 AGTGGGGATC CCGGCAAAGG GCCTATATTT GCGGTCCAAC TTAGGCGTAA ACCTCGATGC 
               
               
                   
               
               
                 361 
                 TACCTACTCA GACCCACCTC GCGCGGGGTA AATAAGGCAC TCATCCCAGC TGGTTCTTGG 
               
               
                   
               
               
                 421 
                 CGTTCTACGC AGCGACATGT TTATTAACAG TTGTCTGGCA GCACAAAACT TTTACCATGG 
               
               
                   
               
               
                 481 
                 TCGTAGAAGC CCCCCAGAGT TAGTTCATAC CTAATGCCAC AAATGTGACA GGACGCCGAT 
               
               
                   
               
               
                 541 
                 GGGTACCGGA CTTTAGGTCG AGCACAGTTC GGTAACGGAG AGACCCTGCG GCGTACTTCA 
               
               
                   
               
               
                 601 
                 TTATGTATAT GGAACGTGCC CAAGTGACGC CAGGCAAGTC TCAGCTGGTT CCTGTGTTAG 
               
               
                   
               
               
                 661 
                 CTCGAGGGTA GACATACGAG CTGATTGAAC ATGGGTTGGG GGCCTCGAAC CGTCGAGGAC 
               
               
                   
               
               
                 721 
                 CCCATAGTAC CTCGGAGACC AAGTAGGGCA GCCTATAGTT TGAAGCAGAA CTATTTCGGG 
               
               
                   
               
               
                 781 
                 GGGCGAGCCC TCATCGTCTC TTCTGCGGAT GACTCAACAC GCTAGGGACG TGAAGTCGAT 
               
               
                   
               
               
                 841 
                 TCCTTCGATG GTTATAAATC AAAGACTCAG AGTGCTGTCT GGAGCGTGAA TCTAACGGTA 
               
               
                   
               
               
                 901 
                 CGTATCTCGA TTGCTCGGTC GCTTTTCGCA CTCCGCGAAA GTTCGTACCG CTCATTCACT 
               
               
                   
               
               
                 961 
                 AGGTTGCGAA GCCTATGCTG ATATATGAAT CCAAACTAGA GCAGGGCTCT TAAGATTCGG 
               
               
                   
               
               
                 1021 
                 AGTTGTAAAT ACTTAATACT CCAATCGGCT TTTACGTGCA CCACCGCGGG CGGCTGACAA 
               
               
                   
               
               
                 1081 
                 GGGTCTCACA TCGAGAAACA AGACAGTTCC GGGCTGGAAG TAGCGCCGGC TAAGGAAGAC 
               
               
                   
               
               
                 1141 
                 GCCTGGTACG GCAGGACTAT GAAACCAGTA CAAAGGCAAC ATCCTCACTT GGGTGAACGG 
               
               
                   
               
               
                 1201 
                 AAACGCAGTA TTATGGTTAC TTTTTGGATA CGTGAAACAT ATCCCATGGT AGTCCTTAGA 
               
               
                   
               
               
                 1261 
                 CTTGGGAGTC TATCACCCCT AGGGCCCATA TCTGGAAATA GACGCCAGGT TGAATCCGTA 
               
               
                   
               
               
                 1321 
                 TTTGGAGGTA CGATGGAACA GTCTGGGTGG GACGTGCTTC ATTTATAGCC TGCGCAGGCT 
               
               
                   
               
               
                 1381 
                 GGACCGAGGA CCGCAAGGTG CGGCGGTGCA CAAGCAATTG ACAACTAACC ACCGTGTATT 
               
               
                   
               
               
                 1441 
                 CATTATGGTA CCAGGAACTT TAAGCCGAGT CAATGAAGCT CGCATTACAG TGTTTACCGC 
               
               
                   
               
               
                 1501 
                 ATCTTGCCGT TACTCACAAA CTGTGATCCA CCACAAGTCA AGCCATTGCC TCTCTGACAC 
               
               
                   
               
               
                 1561 
                 GCCGTAAGAA TTAATATGTA AACTTTGCGC GGGTTGACTG CGATCCGTTC AGTCTCGTCC 
               
               
                   
               
               
                 1621 
                 GAGGGCACAA TCCTATTCCC ATTTGTATGT TCAGCTAACT TCTACCCATC CCCCGAAGTT 
               
               
                   
               
               
                 1681 
                 AAGTAGGTCG TGAGATGCCA TGGAGGCTCT CGTTCATCCC GTGGGACATC AAGCTTCCCC 
               
               
                   
               
               
                 1741 
                 TTGATAAAGC ACCCCGCTCG GGTGTAGCAG AGAAGACGCC TTCTGAATTG TGCAATCCCT 
               
               
                   
               
               
                 1801 
                 CCACCTTATC TAAGCTTGCT ACCAATAATT AGCATTTTTG CCTTGCGACA GACCTCCTAC 
               
               
                   
               
               
                 1861 
                 TTAGATTGCC ACACATTGAG CTAGTCAGTG AGCGATAAGC TTGACGCGCT TTCAAGGGTC 
               
               
                   
               
               
                 1921 
                 GCGAGTACGT GAACTAAGGC TCCGGACAGG ACTATATACT TGGGTTTGAT CTCGCCCCGA 
               
               
                   
               
               
                 1981 
                 CAACTGCAAA CCTCAACTTT TTTAGATTAT ATGGTTAGCC GAAGTTGCAC GAGGTGGCGT 
               
               
                   
               
               
                 2041 
                 CCGCGGACTG CTCCCCGAGT GTGGCTCTTT CATCTGACAA CGTGCAACCC CTATCGCGGC 
               
               
                   
               
               
                 2101 
                 CGATTGTTTC TGCGGACGAT GTTGTCCTCA TAGTTTGGGC ATGTTTCCCT TGTAGGTGTG 
               
               
                   
               
               
                 2161 
                 AAACCACTTA GCTTCGCGCC GTAGTCCCAA TGAAAAACCT ATGGACTTTG TTTTGGGTAG 
               
               
                   
               
               
                 2221 
                 CACCAGGAAT CTGAACCGTG TGAATGTGGA CGTCGCGCGC GTAGACCTTT ATCTCCGGTT 
               
               
                   
               
               
                 2281 
                 CAAGCTAGGG ATGTGGCTGC ATGCTACGTT GTCACACCTA CACTGCTCGA AGTAAATATG 
               
               
                   
               
               
                 2341 
                 CGAAGCGCGC GGCCTGGCCG GAGGCGTTCC GCGCCGCCAC GTGTTCGTTA ACTGTTGATT 
               
               
                   
               
               
                 2401 
                 GGTGGCACAT AAGCAATATC GTAGTCCGTC AAATTCAGCT CTGTTATCCC GGGCGTTATG 
               
               
                   
               
               
                 2461 
                 TGTCAAATGG CGTAGAACGG GATTGACTGT TTGACGGTAG. 
               
            
           
         
       
     
     In some embodiments, the AAV viral delivery vector comprises a nucleic acid sequence comprising a sequence encoding a VMD2 promoter, a sequence encoding a BEST1 protein, and a sequence encoding a WPRE. An exemplary AAV viral delivery vector of the disclosure comprising this nucleic acid sequence (VMD2.BEST1.WPRE.pA) comprises or consists of the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 13) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 TAGCTGCGCG CTCGCTCGCT CACTGAGGCC GCCCGGGCAA AGCCCGGGCG TCGGGCGACC 
                   
               
               
                   
               
               
                 61 
                 TTTGGTCGCC CGGCCTCAGT GAGCGAGCGA GCGCGCAGAG AGGGAGTGGC CAACTCCATC 
               
               
                   
               
               
                 121 
                 ACTAGGGGTT CCTTGTAGTT AATGATTAAC CCGCCATGCT ACTTATCTAC GTAGCCATGC 
               
               
                   
               
               
                 181 
                 TCTAGGTAAA TTCTGTCATT TTACTAGGGT GATGAAATTC CCAAGCAACA CCATCCTTTT 
               
               
                   
               
               
                 241 
                 CAGATAAGGG CACTGAGGCT GAGAGAGGAG CTGAAACCTA CCCGGGGTCA CCACACACAG 
               
               
                   
               
               
                 301 
                 GTGGCAAGGC TGGGACCAGA AACCAGGACT GTTGACTGCA GCCCGGTATT CATTCTTTCC 
               
               
                   
               
               
                 361 
                 ATAGCCCACA GGGCTGTCAA AGACCCCAGG GCCTAGTCAG AGGCTCCTCC TTCCTGGAGA 
               
               
                   
               
               
                 421 
                 GTTCCTGGCA CAGAAGTTGA AGCTCAGCAC AGCCCCCTAA CCCCCAACTC TCTCTGCAAG 
               
               
                   
               
               
                 481 
                 GCCTCAGGGG TCAGAACACT GGTGGAGCAG ATCCTTTAGC CTCTGGATTT TAGGGCCATG 
               
               
                   
               
               
                 541 
                 GTAGAGGGGG TGTTGCCCTA AATTCCAGCC CTGGTCTCAG CCCAACACCC TCCAAGAAGA 
               
               
                   
               
               
                 601 
                 AATTAGAGGG GCCATGGCCA GGCTGTGCTA GCCGTTGCTT CTGAGCAGAT TACAAGAAGG 
               
               
                   
               
               
                 661 
                 GACTAAGACA AGGACTCCTT TGTGGAGGTC CTGGCTTAGG GAGTCAAGTG ACGGCGGCTC 
               
               
                   
               
               
                 721 
                 AGCACTCACG TGGGCAGTGC CAGCCTCTAA GAGTGGGCAG GGGCACTGGC CACAGAGTCC 
               
               
                   
               
               
                 781 
                 CAGGGAGTCC CACCAGCCTA GTCGCCAGAC CGGCACCATG ACCATCACTT ACACAAGCCA 
               
               
                   
               
               
                 841 
                 AGTGGCTAAT GCCCGCTTAG GCTCCTTCTC CCGCCTGCTG CTGTGCTGGC GGGGCAGCAT 
               
               
                   
               
               
                 901 
                 CTACAAGGTG CTATATGGCG AGTTCTTAAT CTTCCTGCTC TGCTACTACA TCATCCGCTT 
               
               
                   
               
               
                 961 
                 TATTTATAGG CTGGCCCTCA CGGAAGAACA ACAGCTGATG TTTGAGAAAC TGACTCTGTA 
               
               
                   
               
               
                 1021 
                 TTGCGACAGC TACATCCAGC TCATCCCCAT TTCCTTCGTG CTGGGCTTCT ACGTGACGCT 
               
               
                   
               
               
                 1081 
                 GGTCGTGACC CGCTGGTGGA ACCAGTACGA GAACCTGCCG TGGCCCGACC GCCTCATGAG 
               
               
                   
               
               
                 1141 
                 CCTGGTGTCG GGCTTCGTCG AAGGCAAGGA CGAGCAAGGC CGGCTGCTGC GGCGCACGCT 
               
               
                   
               
               
                 1201 
                 CATCCGCTAC GCCAACCTGG GCAACGTGCT CATCCTGCGC AGCGTCAGCA CCGCAGTCTA 
               
               
                   
               
               
                 1261 
                 CAAGCGCTTC CCCAGCGCCC AGCACCTGGT GCAAGCAGGC TTTATGACTC CGGCAGAACA 
               
               
                   
               
               
                 1321 
                 CAAGCAGTTG GAGAAACTGA GCCTACCACA CAACATGTTC TGGGTGCCCT GGGTGTGGTT 
               
               
                   
               
               
                 1381 
                 TGCCAACCTG TCAATGAAGG CGTGGCTTGG AGGTCGAATC CGGGACCCTA TCCTGCTCCA 
               
               
                   
               
               
                 1441 
                 GAGCCTGCTG AACGAGATGA ACACCTTGCG TACTCAGTGT GGACACCTGT ATGCCTACGA 
               
               
                   
               
               
                 1501 
                 CTGGATTAGT ATCCCACTGG TGTATACACA GGTGGTGACT GTGGCGGTGT ACAGCTTCTT 
               
               
                   
               
               
                 1561 
                 CCTGACTTGT CTAGTTGGGC GGCAGTTTCT GAACCCAGCC AAGGCCTACC CTGGCCATGA 
               
               
                   
               
               
                 1621 
                 GCTGGACCTC GTTGTGCCCG TCTTCACGTT CCTGCAGTTC TTCTTCTATG TTGGCTGGCT 
               
               
                   
               
               
                 1681 
                 GAAGGTGGCA GAGCAGCTCA TCAACCCCTT TGGAGAGGAT GATGATGATT TTGAGACCAA 
               
               
                   
               
               
                 1741 
                 CTGGATTGTC GACAGGAATT TGCAGGTGTC CCTGTTGGCT GTGGATGAGA TGCACCAGGA 
               
               
                   
               
               
                 1801 
                 CCTGCCTCGG ATGGAGCCGG ACATGTACTG GAATAAGCCC GAGCCACAGC CCCCCTACAC 
               
               
                   
               
               
                 1861 
                 AGCTGCTTCC GCCCAGTTCC GTCGAGCCTC CTTTATGGGC TCCACCTTCA ACATCAGCCT 
               
               
                   
               
               
                 1921 
                 GAACAAAGAG GAGATGGAGT TCCAGCCCAA TCAGGAGGAC GAGGAGGATG CTCACGCTGG 
               
               
                   
               
               
                 1981 
                 CATCATTGGC CGCTTCCTAG GCCTGCAGTC CCATGATCAC CATCCTCCCA GGGCAAACTC 
               
               
                   
               
               
                 2041 
                 AAGGACCAAA CTACTGTGGC CCAAGAGGGA ATCCCTTCTC CACGAGGGCC TGCCCAAAAA 
               
               
                   
               
               
                 2101 
                 CCACAAGGCA GCCAAACAGA ACGTTAGGGG CCAGGAAGAC AACAAGGCCT GGAAGCTTAA 
               
               
                   
               
               
                 2161 
                 GGCTGTGGAC GCCTTCAAGT CTGCCCCACT GTATCAGAGG CCAGGCTACT ACAGTGCCCC 
               
               
                   
               
               
                 2221 
                 ACAGACGCCC CTCAGCCCCA CTCCCATGTT CTTCCCCCTA GAACCATCAG CGCCGTCAAA 
               
               
                   
               
               
                 2281 
                 GCTTCACAGT GTCACAGGCA TAGACACCAA AGACAAAAGC TTAAAGACTG TGAGTTCTGG 
               
               
                   
               
               
                 2341 
                 GGCCAAGAAA AGTTTTGAAT TGCTCTCAGA GAGCGATGGG GCCTTGATGG AGCACCCAGA 
               
               
                   
               
               
                 2401 
                 AGTATCTCAA GTGAGGAGGA AAACTGTGGA GTTTAACCTG ACGGATATGC CAGAGATCCC 
               
               
                   
               
               
                 2461 
                 CGAAAATCAC CTCAAAGAAC CTTTGGAACA ATCACCAACC AACATACACA CTACACTCAA 
               
               
                   
               
               
                 2521 
                 AGATCACATG GATCCTTATT GGGCCTTGGA AAACAGGGAT GAAGCACATT CCTAATCTAG 
               
               
                   
               
               
                 2581 
                 CGGCCGCGAA TTCGATATCA AGCTTATCGA TAATCAACCT CTGGATTACA AAATTTGTGA 
               
               
                   
               
               
                 2641 
                 AAGATTGACT GGTATTCTTA ACTATGTTGC TCCTTTTACG CTATGTGGAT ACGCTGCTTT 
               
               
                   
               
               
                 2701 
                 AATGCCTTTG TATCATGCTA TTGCTTCCCG TATGGCTTTC ATTTTCTCCT CCTTGTATAA 
               
               
                   
               
               
                 2761 
                 ATCCTGGTTG CTGTCTCTTT ATGAGGAGTT GTGGCCCGTT GTCAGGCAAC GTGGCGTGGT 
               
               
                   
               
               
                 2821 
                 GTGCACTGTG TTTGCTGACG CAACCCCCAC TGGTTGGGGC ATTGCCACCA CCTGTCAGCT 
               
               
                   
               
               
                 2881 
                 CCTTTCCGGG ACTTTCGCTT TCCCCCTCCC TATTGCCACG GCGGAACTCA TCGCCGCCTG 
               
               
                   
               
               
                 2941 
                 CCTTGCCCGC TGCTGGACAG GGGCTCGGCT GTTGGGCACT GACAATTCCG TGGTGTTGTC 
               
               
                   
               
               
                 3001 
                 GGGGAAATCA TCGTCCTTTC CTTGGCTGCT CGCCTGTGTT GCCACCTGGA TTCTGCGCGG 
               
               
                   
               
               
                 3061 
                 GACGTCCTTC TGCTACGTCC CTTCGGCCCT CAATCCAGCG GACCTTCCTT CCCGCGGCCT 
               
               
                   
               
               
                 3121 
                 GCTGCCGGCT CTGCGGCCTC TTCCGCGTCT TCGCCTTCGC CCTCAGACGA GTCGGATCTC 
               
               
                   
               
               
                 3181 
                 CCTTTGGGCC GCCTCCCCGG CGGCCGCGCA CCGTCGACTC GCTGATCAGC CTCGACTGTG 
               
               
                   
               
               
                 3241 
                 CCTTCTAGTT GCCAGCCATC TGTTGTTTGC CCCTCCCCCG TGCCTTCCTT GACCCTGGAA 
               
               
                   
               
               
                 3301 
                 GGTGCCACTC CCACTGTCCT TTCCTAATAA AATGAGGAAA TTGCATCGCA TTGTCTGAGT 
               
               
                   
               
               
                 3361 
                 AGGTGTCATT CTATTCTGGG GGGTGGGGTG GGGCAGGACA GCAAGGGGGA GGATTGGGAA 
               
               
                   
               
               
                 3421 
                 GACAATAGGA GGCATGCTGG GGATGCGGTG GGCTCTATGG CTTCTGAGGC GGAAAGAACC 
               
               
                   
               
               
                 3481 
                 AGCTGGGGCT CGACTAGAGC ATGGCTACGT AGATAAGTAG CATGGCGGGT TAATCATTAA 
               
               
                   
               
               
                 3541 
                 CTACAAGGAA CCCCTAGTGA TGGAGTTGGC CACTCCCTCT CTGCGCGCTC GCTCGCTCAC 
               
               
                   
               
               
                 3601 
                 TGAGGCCGGG CGACCAAAGG TCGCCCGACG CCCGGGCGGC CTCAGTGAGC GAGCGAGCGC 
               
               
                   
               
               
                 3661 
                 GCAGAGCTTT TTGCAAAAGC CTAGGCCTCC AAAAAAGCCT CCTCACTACT TCTGGAATAG 
               
               
                   
               
               
                 3721 
                 CTCAGAGGCC GAGGCGGCCT CGGCCTCTGC ATAAATAAAA AAAATTAGTC AGCCATGGGG 
               
               
                   
               
               
                 3781 
                 CGGAGAATGG GCGGAACTGG GCGGAGTTAG GGGCGGGATG GGCGGAGTTA GGGGCGGGAC 
               
               
                   
               
               
                 3841 
                 TATGGTTGCT GACTAATTGA GATGCATGCT TTGCATACTT CTGCCTGCTG GGGAGCCTGG 
               
               
                   
               
               
                 3901 
                 GGACTTTCCA CACCTGGTTG CTGACTAATT GAGATGCATG CTTTGCATAC TTCTGCCTGC 
               
               
                   
               
               
                 3961 
                 TGGGGAGCCT GGGGACTTTC CACACCCTAA CTGACACACA TTCCACAGCT GCATTAATGA 
               
               
                   
               
               
                 4021 
                 ATCGGCCAAC GCGCGGGGAG AGGCGGTTTG CGTATTGGGC GCTCTTCCGC TTCCTCGCTC 
               
               
                   
               
               
                 4081 
                 ACTGACTCGC TGCGCTCGGT CGTTCGGCTG CGGCGAGCGG TATCAGCTCA CTCAAAGGCG 
               
               
                   
               
               
                 4141 
                 GTAATACGGT TATCCACAGA ATCAGGGGAT AACGCAGGAA AGAACATGTG AGCAAAAGGC 
               
               
                   
               
               
                 4201 
                 CAGCAAAAGG CCAGGAACCG TAAAAAGGCC GCGTTGCTGG CGTTTTTCCA TAGGCTCCGC 
               
               
                   
               
               
                 4261 
                 CCCCCTGACG AGCATCACAA AAATCGACGC TCAAGTCAGA GGTGGCGAAA CCCGACAGGA 
               
               
                   
               
               
                 4321 
                 CTATAAAGAT ACCAGGCGTT TCCCCCTGGA AGCTCCCTCG TGCGCTCTCC TGTTCCGACC 
               
               
                   
               
               
                 4381 
                 CTGCCGCTTA CCGGATACCT GTCCGCCTTT CTCCCTTCGG GAAGCGTGGC GCTTTCTCAT 
               
               
                   
               
               
                 4441 
                 AGCTCACGCT GTAGGTATCT CAGTTCGGTG TAGGTCGTTC GCTCCAAGCT GGGCTGTGTG 
               
               
                   
               
               
                 4501 
                 CACGAACCCC CCGTTCAGCC CGACCGCTGC GCCTTATCCG GTAACTATCG TCTTGAGTCC 
               
               
                   
               
               
                 4561 
                 AACCCGGTAA GACACGACTT ATCGCCACTG GCAGCAGCCA CTGGTAACAG GATTAGCAGA 
               
               
                   
               
               
                 4621 
                 GCGAGGTATG TAGGCGGTGC TACAGAGTTC TTGAAGTGGT GGCCTAACTA CGGCTACACT 
               
               
                   
               
               
                 4681 
                 AGAAGAACAG TATTTGGTAT CTGCGCTCTG CTGAAGCCAG TTACCTTCGG AAAAAGAGTT 
               
               
                   
               
               
                 4741 
                 GGTAGCTCTT GATCCGGCAA ACAAACCACC GCTGGTAGCG GTGGTTTTTT TGTTTGCAAG 
               
               
                   
               
               
                 4801 
                 CAGCAGATTA CGCGCAGAAA AAAAGGATCT CAAGAAGATC CTTTGATCTT TTCTACGGGG 
               
               
                   
               
               
                 4861 
                 TCTGACGCTC AGTGGAACGA AAACTCACGT TAAGGGATTT TGGTCATGAG ATTATCAAAA 
               
               
                   
               
               
                 4921 
                 AGGATCTTCA CCTAGATCCT TTTAAATTAA AAATGAAGTT TTAAATCAAT CTAAAGTATA 
               
               
                   
               
               
                 4981 
                 TATGAGTAAA CTTGGTCTGA CAGTTACCAA TGCTTAATCA GTGAGGCACC TATCTCAGCG 
               
               
                   
               
               
                 5041 
                 ATCTGTCTAT TTCGTTCATC CATAGTTGCC TGACTCCTGC AAACCACGTT GTGTCTCAAA 
               
               
                   
               
               
                 5101 
                 ATCTCTGATG TTACATTGCA CAAGATAAAA ATATATCATC ATGAACAATA AAACTGTCTG 
               
               
                   
               
               
                 5161 
                 CTTACATAAA CAGTAATACA AGGGGTGTTA TGAGCCATAT TCAACGGGAA ACGTCTTGCT 
               
               
                   
               
               
                 5221 
                 CGAGGCCGCG ATTAAATTCC AACATGGATG CTGATTTATA TGGGTATAAA TGGGCTCGCG 
               
               
                   
               
               
                 5281 
                 ATAATGTCGG GCAATCAGGT GCGACAATCT ATCGATTGTA TGGGAAGCCC GATGCGCCAG 
               
               
                   
               
               
                 5341 
                 AGTTGTTTCT GAAACATGGC AAAGGTAGCG TTGCCAATGA TGTTACAGAT GAGATGGTCA 
               
               
                   
               
               
                 5401 
                 GACTAAACTG GCTGACGGAA TTTATGCCTC TTCCGACCAT CAAGCATTTT ATCCGTACTC 
               
               
                   
               
               
                 5461 
                 CTGATGATGC ATGGTTACTC ACCACTGCGA TCCCCGGGAA AACAGCATTC CAGGTATTAG 
               
               
                   
               
               
                 5521 
                 AAGAATATCC TGATTCAGGT GAAAATATTG TTGATGCGCT GGCAGTGTTC CTGCGCCGGT 
               
               
                   
               
               
                 5581 
                 TGCATTCGAT TCCTGTTTGT AATTGTCCTT TTAACAGCGA TCGCGTATTT CGTCTCGCTC 
               
               
                   
               
               
                 5641 
                 AGGCGCAATC ACGAATGAAT AACGGTTTGG TTGATGCGAG TGATTTTGAT GACGAGCGTA 
               
               
                   
               
               
                 5701 
                 ATGGCTGGCC TGTTGAACAA GTCTGGAAAG AAATGCATAA GCTTTTGCCA TTCTCACCGG 
               
               
                   
               
               
                 5761 
                 ATTCAGTCGT CACTCATGGT GATTTCTCAC TTGATAACCT TATTTTTGAC GAGGGGAAAT 
               
               
                   
               
               
                 5821 
                 TAATAGGTTG TATTGATGTT GGACGAGTCG GAATCGCAGA CCGATACCAG GATCTTGCCA 
               
               
                   
               
               
                 5881 
                 TCCTATGGAA CTGCCTCGGT GAGTTTTCTC CTTCATTACA GAAACGGCTT TTTCAAAAAT 
               
               
                   
               
               
                 5941 
                 ATGGTATTGA TAATCCTGAT ATGAATAAAT TGCAGTTTCA TTTGATGCTC GATGAGTTTT 
               
               
                   
               
               
                 6001 
                 TCTAAGGGCG GCCTGCCACC ATACCCACGC CGAAACAAGC GCTCATGAGC CCGAAGTGGC 
               
               
                   
               
               
                 6061 
                 GAGCCCGATC TTCCCCATCG GTGATGTCGG CGATATAGGC GCCAGCAACC GCACCTGTGG 
               
               
                   
               
               
                 6121 
                 CGCCGGTGAT GCCGGCCACG ATGCGTCCGG CGTAGAGGAT CTGGCTAGCG ATGACCCTGC 
               
               
                   
               
               
                 6181 
                 TGATTGGTTC GCTGACCATT TCCGGGTGCG GGACGGCGTT ACCAGAAACT CAGAAGGTTC 
               
               
                   
               
               
                 6241 
                 GTCCAACCAA ACCGACTCTG ACGGCAGTTT ACGAGAGAGA TGATAGGGTC TGCTTCAGGG 
               
               
                   
               
               
                 6301 
                 TGAGCGATGT AACCATATAC TTAGGCTGGA TCTTCTCCCG CGAATTTTAA CCCTCACCAA 
               
               
                   
               
               
                 6361 
                 CTACGAGATA TGAGGTAAGC CAAAAAAGCA CGTAGTGGCG CTCTCCGACT GTTCCCAAAT 
               
               
                   
               
               
                 6421 
                 TGTAACTTAT CGTTCCGTGA AGGCCAGAGT TACTTCCCGG CCCTTTCCAT GCGCGCACCA 
               
               
                   
               
               
                 6481 
                 TACCCTCCTA GTTCCCCGGT TATCTTTCCG AAGTGGGAGT GAGCGAACCT CCGTTTACGT 
               
               
                   
               
               
                 6541 
                 CTTGTTACCA ATGATGTAGC TATGCACTTT GTACAGGGTG CCAACGGGTT TCACAATTCA 
               
               
                   
               
               
                 6601 
                 CAGATAGTGG GGATCCCGGC AAAGGGCCTA TATTTGCGGT CCAACTTAGG CGTAAACCTC 
               
               
                   
               
               
                 6661 
                 GATGCTACCT ACTCAGACCC ACCTCGCGCG GGGTAAATAA GGCACTCATC CCAGCTGGTT 
               
               
                   
               
               
                 6721 
                 CTTGGCGTTC TACGCAGCGA CATGTTTATT AACAGTTGTC TGGCAGCACA AAACTTTTAC 
               
               
                   
               
               
                 6781 
                 CATGGTCGTA GAAGCCCCCC AGAGTTAGTT CATACCTAAT GCCACAAATG TGACAGGACG 
               
               
                   
               
               
                 6841 
                 CCGATGGGTA CCGGACTTTA GGTCGAGCAC AGTTCGGTAA CGGAGAGACC CTGCGGCGTA 
               
               
                   
               
               
                 6901 
                 CTTCATTATG TATATGGAAC GTGCCCAAGT GACGCCAGGC AAGTCTCAGC TGGTTCCTGT 
               
               
                   
               
               
                 6961 
                 GTTAGCTCGA GGGTAGACAT ACGAGCTGAT TGAACATGGG TTGGGGGCCT CGAACCGTCG 
               
               
                   
               
               
                 7021 
                 AGGACCCCAT AGTACCTCGG AGACCAAGTA GGGCAGCCTA TAGTTTGAAG CAGAACTATT 
               
               
                   
               
               
                 7081 
                 TCGGGGGGCG AGCCCTCATC GTCTCTTCTG CGGATGACTC AACACGCTAG GGACGTGAAG 
               
               
                   
               
               
                 7141 
                 TCGATTCCTT CGATGGTTAT AAATCAAAGA CTCAGAGTGC TGTCTGGAGC GTGAATCTAA 
               
               
                   
               
               
                 7201 
                 CGGTACGTAT CTCGATTGCT CGGTCGCTTT TCGCACTCCG CGAAAGTTCG TACCGCTCAT 
               
               
                   
               
               
                 7261 
                 TCACTAGGTT GCGAAGCCTA TGCTGATATA TGAATCCAAA CTAGAGCAGG GCTCTTAAGA 
               
               
                   
               
               
                 7321 
                 TTCGGAGTTG TAAATACTTA ATACTCCAAT CGGCTTTTAC GTGCACCACC GCGGGCGGCT 
               
               
                   
               
               
                 7381 
                 GACAAGGGTC TCACATCGAG AAACAAGACA GTTCCGGGCT GGAAGTAGCG CCGGCTAAGG 
               
               
                   
               
               
                 7441 
                 AAGACGCCTG GTACGGCAGG ACTATGAAAC CAGTACAAAG GCAACATCCT CACTTGGGTG 
               
               
                   
               
               
                 7501 
                 AACGGAAACG CAGTATTATG GTTACTTTTT GGATACGTGA AACATATCCC ATGGTAGTCC 
               
               
                   
               
               
                 7561 
                 TTAGACTTGG GAGTCTATCA CCCCTAGGGC CCATATCTGG AAATAGACGC CAGGTTGAAT 
               
               
                   
               
               
                 7621 
                 CCGTATTTGG AGGTACGATG GAACAGTCTG GGTGGGACGT GCTTCATTTA TACCCTGCGC 
               
               
                   
               
               
                 7681 
                 AGGCTGGACC GAGGACCGCA AGGTGCGGCG GTGCACAAGC AATTGACAAC TAACCACCGT 
               
               
                   
               
               
                 7741 
                 GTATTCATTA TGGTACCAGG AACTTTAAGC CGAGTCAATG AAGCTCGCAT TACAGTGTTT 
               
               
                   
               
               
                 7801 
                 ACCGCATCTT GCCGTTACTC ACAAACTGTG ATCCACCACA AGTCAAGCCA TTGCCTCTCT 
               
               
                   
               
               
                 7861 
                 GACACGCCGT AAGAATTAAT ATGTAAACTT TGCGCGGGTT GACTGCGATC CGTTCAGTCT 
               
               
                   
               
               
                 7921 
                 CGTCCGAGGG CACAATCCTA TTCCCATTTG TATGTTCAGC TAACTTCTAC CCATCCCCCG 
               
               
                   
               
               
                 7981 
                 AAGTTAAGTA GGTCGTGAGA TGCCATGGAG GCTCTCGTTC ATCCCGTGGG ACATCAAGCT 
               
               
                   
               
               
                 8041 
                 TCCCCTTGAT AAAGCACCCC GCTCGGGTGT AGCAGAGAAG ACGCCTTCTG AATTGTGCAA 
               
               
                   
               
               
                 8101 
                 TCCCTCCACC TTATCTAAGC TTGCTACCAA TAATTAGCAT TTTTGCCTTG CGACAGACCT 
               
               
                   
               
               
                 8161 
                 CCTACTTAGA TTGCCACACA TTGAGCTAGT CAGTGAGCGA TAAGCTTGAC GCGCTTTCAA 
               
               
                   
               
               
                 8221 
                 GGGTCGCGAG TACGTGAACT AAGGCTCCGG ACAGGACTAT ATACTTGGGT TTGATCTCGC 
               
               
                   
               
               
                 8281 
                 CCCGACAACT GCAAACCTCA ACTTTTTTAG ATTATATGGT TAGCCGAAGT TGCACGAGGT 
               
               
                   
               
               
                 8341 
                 GGCGTCCGCG GACTGCTCCC CGAGTGTGGC TCTTTCATCT GACAACGTGC AACCCCTATC 
               
               
                   
               
               
                 8401 
                 GCGGCCGATT GTTTCTGCGG ACGATGTTGT CCTCATAGTT TGGGCATGTT TCCCTTGTAG 
               
               
                   
               
               
                 8461 
                 GTGTGAAACC ACTTAGCTTC GCGCCGTAGT CCCAATGAAA AACCTATGGA CTTTGTTTTG 
               
               
                   
               
               
                 8521 
                 GGTAGCACCA GGAATCTGAA CCGTGTGAAT GTGGACGTCG CGCGCGTAGA CCTTTATCTC 
               
               
                   
               
               
                 8581 
                 CGGTTCAAGC TAGGGATGTG GCTGCATGCT ACGTTGTCAC ACCTACACTG CTCGAAGTAA 
               
               
                   
               
               
                 8641 
                 ATATGCGAAG CGCGCGGCCT GGCCGGAGGC GTTCCGCGCC GCCACGTGTT CGTTAACTGT 
               
               
                   
               
               
                 8701 
                 TGATTGGTGG CACATAAGCA ATATCGTAGT CCGTCAAATT CAGCTCTGTT ATCCCGGGCG 
               
               
                   
               
               
                 8761 
                 TTATGTGTGA AATGGCGTAG AACGGGATTG ACTGTTTGAC GGTAGGGTGA CCTAAGCCAG 
               
               
                   
               
               
                 8821 
                 ATGCTACACA ATTAGGCTTG TACATATTGT CGTTAGAACG CGGCTACAAT TAATACATAA 
               
               
                   
               
               
                 8881 
                 CCTTATGTAT CATACACATA CGATTTAGGT GACACTATAG AATACACGGA ATTAATTC. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 1 
               
             
            
               
                   
               
               
                 Features of the VMD2.BEST1.WPRE.pA plasmid sequence 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                 Mini- 
                 Maxi- 
                   
                 Direc- 
               
               
                 Name 
                 Type 
                 mum 
                 mum 
                 Length 
                 tion 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 130 bp AAV2 
                 LTR 
                 4 
                 133 
                 130 
                 forward 
               
               
                 5′ITR 
               
               
                 VMD2 promoter 
                 promoter 
                 189 
                 811 
                 623 
                 forward 
               
               
                 Kozak 
                 Kozak 
                 812 
                 821 
                 10 
                 forward 
               
               
                 BEST1 
                 CDS 
                 818 
                 2,575 
                 1,758 
                 forward 
               
               
                 WPRE 
                 WPRE 
                 2,606 
                 3,198 
                 593 
                 forward 
               
               
                 bGH pA 
                 polyA_signal 
                 3,220 
                 3,488 
                 269 
                 forward 
               
               
                 112 bp AAV2 
                 LTR 
                 3,546 
                 3,657 
                 112 
                 reverse 
               
               
                 3′ITR 
               
               
                 pBR322 rep 
                 rep_origin 
                 4,230 
                 4,849 
                 620 
                 reverse 
               
               
                 origin 
               
               
                 AphR (KanR) 
                 CDS 
                 5,190 
                 6,005 
                 816 
                 forward 
               
               
                 Randomly 
                 Stuffer 
                 6,306 
                 8,805 
                 2,500 
                 none 
               
               
                 generated 
               
               
                 stuffer sequence 
               
               
                   
               
            
           
         
       
     
     In some embodiments, the AAV viral delivery vector comprising a nucleic acid sequence comprising a sequence encoding a VMD2 promoter, a sequence encoding a BEST1 protein, a sequence encoding an intron, a sequence encoding an exon and a sequence encoding a WPRE. An exemplary AAV viral delivery vector of the disclosure comprises a nucleic acid sequence encoding a VMD2. IntEx.BEST1.WPRE.pA sequence comprising or consisting of the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 14) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 TAGCTGCGCG CTCGCTCGCT CACTGAGGCC GCCCGGGCAA AGCCCGGGCG TCGGGCGACC 
                   
               
               
                   
               
               
                 61 
                 TTTGGTCGCC CGGCCTCAGT GAGCGAGCGA GCGCGCAGAG AGGGAGTGGC CAACTCCATC 
               
               
                   
               
               
                 121 
                 ACTAGGGGTT CCTTGTAGTT AATGATTAAC CCGCCATGCT ACTTATCTAC GTAGCCATGC 
               
               
                   
               
               
                 181 
                 TCTAGGTAAA TTCTGTCATT TTACTAGGGT GATGAAATTC CCAAGCAACA CCATCCTTTT 
               
               
                   
               
               
                 241 
                 CAGATAAGGG CACTGAGGCT GAGAGAGGAG CTGAAACCTA CCCGGGGTCA CCACACACAG 
               
               
                   
               
               
                 301 
                 GTGGCAAGGC TGGGACCAGA AACCAGGACT GTTGACTGCA GCCCGGTATT CATTCTTTCC 
               
               
                   
               
               
                 361 
                 ATAGCCCACA GGGCTGTCAA AGACCCCAGG GCCTAGTCAG AGGCTCCTCC TTCCTGGAGA 
               
               
                   
               
               
                 421 
                 GTTCCTGGCA CAGAAGTTGA AGCTCAGCAC AGCCCCCTAA CCCCCAACTC TCTCTGCAAG 
               
               
                   
               
               
                 481 
                 GCCTCAGGGG TCAGAACACT GGTGGAGCAG ATCCTTTAGC CTCTGGATTT TAGGGCCATG 
               
               
                   
               
               
                 541 
                 GTAGAGGGGG TGTTGCCCTA AATTCCAGCC CTGGTCTCAG CCCAACACCC TCCAAGAAGA 
               
               
                   
               
               
                 601 
                 AATTAGAGGG GCCATGGCCA GGCTGTGCTA GCCGTTGCTT CTGAGCAGAT TACAAGAAGG 
               
               
                   
               
               
                 661 
                 GACTAAGACA AGGACTCCTT TGTGGAGGTC CTGGCTTAGG GAGTCAAGTG ACGGCGGCTC 
               
               
                   
               
               
                 721 
                 AGCACTCACG TGGGCAGTGC CAGCCTCTAA GAGTGGGCAG GGGCACTGGC CACAGAGTCC 
               
               
                   
               
               
                 781 
                 CAGGGAGTCC CACCAGCCTA GTCGCCAGAC CGGGTGCCGC AGGGGGACGG CTGCCTTCGG 
               
               
                   
               
               
                 841 
                 GGGGGACGGG GCAGGGCGGG GTTCGGCTTC TGGCGTGTGA CCGGCGGCTC TAGAGCCTCT 
               
               
                   
               
               
                 901 
                 GCTAACCATG TTCATGCCTT CTTCTTTTTC CTACAGCTCC TGGGCAACGT GCTGGTTATT 
               
               
                   
               
               
                 961 
                 GTGCTGTCTC ATCATTTTGG CAAAGAATTG GCACCATGAC CATCACTTAC ACAAGCCAAG 
               
               
                   
               
               
                 1021 
                 TGGCTAATGC CCGCTTAGGC TCCTTCTCCC GCCTGCTGCT GTGCTGGCGG GGCAGCATCT 
               
               
                   
               
               
                 1081 
                 ACAAGCTGCT ATATGGCGAG TTCTTAATCT TCCTGCTCTG CTACTACATC ATCCGCTTTA 
               
               
                   
               
               
                 1141 
                 TTTATAGGCT GGCCCTCACG GAAGAACAAC AGCTGATGTT TGAGAAACTG ACTCTGTATT 
               
               
                   
               
               
                 1201 
                 GCGACAGCTA CATCCAGCTC ATCCCCATTT CCTTCGTGCT GGGCTTCTAC GTGACGCTGG 
               
               
                   
               
               
                 1261 
                 TCGTGACCCG CTGGTGGAAC CAGTACGAGA ACCTGCCGTG GCCCGACCGC CTCATGAGCC 
               
               
                   
               
               
                 1321 
                 TGGTGTCGGG CTTCGTCGAA GGCAAGGACG AGCAAGGCCG GCTGCTGCGG CGCACGCTCA 
               
               
                   
               
               
                 1381 
                 TCCGCTACGC CAACCTGGGC AACGTGCTCA TCCTGCGCAG CGTCAGCACC GCAGTCTACA 
               
               
                   
               
               
                 1441 
                 AGCGCTTCCC CAGCGCCCAG CACCTGGTGC AAGCAGGCTT TATGACTCCG GCAGAACACA 
               
               
                   
               
               
                 1501 
                 AGCAGTTGGA GAAACTGAGC CTACCACACA ACATGTTCTG GGTGCCCTGG GTGTGGTTTG 
               
               
                   
               
               
                 1561 
                 CCAACCTGTC AATGAAGGCG TGGCTTGGAG GTCGAATCCG GGACCCTATC CTGCTCCAGA 
               
               
                   
               
               
                 1621 
                 GCCTGCTGAA CGAGATGAAC ACCTTGCGTA CTCAGTGTGG ACACCTGTAT GCCTACGACT 
               
               
                   
               
               
                 1681 
                 GGATTAGTAT CCCACTGGTG TATACACAGG TGGTGACTGT GGCGGTGTAC AGCTTCTTCC 
               
               
                   
               
               
                 1741 
                 TGACTTGTCT AGTTGGGCGG CAGTTTCTGA ACCCAGCCAA GGCCTACCCT GGCCATGAGC 
               
               
                   
               
               
                 1801 
                 TGGACCTCGT TGTGCCCGTC TTCACGTTCC TGCAGTTCTT CTTCTATGTT GGCTGGCTGA 
               
               
                   
               
               
                 1861 
                 AGGTGGCAGA GCAGCTCATC AACCCCTTTG GAGAGGATGA TGATGATTTT GAGACCAACT 
               
               
                   
               
               
                 1921 
                 GGATTGTCGA CAGGAATTTG CAGGTGTCCC TGTTGGCTGT GGATGAGATG CACCAGGACC 
               
               
                   
               
               
                 1981 
                 TGCCTCGGAT GGAGCCGGAC ATGTACTGGA ATAAGCCCGA GCCACAGCCC CCCTACACAG 
               
               
                   
               
               
                 2041 
                 CTGCTTCCGC CCAGTTCCGT CGAGCCTCCT TTATGGGCTC CACCTTCAAC ATCAGCCTGA 
               
               
                   
               
               
                 2101 
                 ACAAAGAGGA GATGGAGTTC CAGCCCAATC AGGAGGACGA GGAGGATGCT CACGCTGGCA 
               
               
                   
               
               
                 2161 
                 TCATTGGCCG CTTCCTAGGC CTGCAGTCCC ATGATCACCA TCCTCCCAGG GCAAACTCAA 
               
               
                   
               
               
                 2221 
                 GGACCAAACT ACTGTGGCCC AAGAGGGAAT CCCTTCTCCA CGAGGGCCTG CCCAAAAACC 
               
               
                   
               
               
                 2281 
                 ACAAGGCAGC CAAACAGAAC GTTAGGGGCC AGGAAGACAA CAAGGCCTGG AAGCTTAAGG 
               
               
                   
               
               
                 2341 
                 CTGTGGACGC CTTCAAGTCT GCCCCACTGT ATCAGAGGCC AGGCTACTAC AGTGCCCCAC 
               
               
                   
               
               
                 2401 
                 AGACGCCCCT CAGCCCCACT CCCATGTTCT TCCCCCTAGA ACCATCAGCG CCGTCAAAGC 
               
               
                   
               
               
                 2461 
                 TTCACAGTGT CACAGGCATA GACACCAAAG ACAAAAGCTT AAAGACTGTG AGTTCTGGGG 
               
               
                   
               
               
                 2521 
                 CCAAGAAAAG TTTTGAATTG CTCTCAGAGA GCGATGGGGC CTTGATGGAG CACCCAGAAG 
               
               
                   
               
               
                 2581 
                 TATCTCAAGT GAGGAGGAAA ACTGTGGAGT TTAACCTGAC GGATATGCCA GAGATCCCCG 
               
               
                   
               
               
                 2641 
                 AAAATCACCT CAAAGAACCT TTGGAACAAT CACCAACCAA CATACACACT ACACTCAAAG 
               
               
                   
               
               
                 2701 
                 ATCACATGGA TCCTTATTGG GCCTTGGAAA ACAGGGATGA AGCACATTCC TAATCTAGCG 
               
               
                   
               
               
                 2761 
                 GCCGCGAATT CGATATCAAG CTTATCGATA ATCAACCTCT GGATTACAAA ATTTGTGAAA 
               
               
                   
               
               
                 2821 
                 GATTGACTGG TATTCTTAAC TATGTTGCTC CTTTTACGCT ATGTGGATAC GCTGCTTTAA 
               
               
                   
               
               
                 2881 
                 TGCCTTTGTA TCATGCTATT GCTTCCCGTA TGGCTTTCAT TTTCTCCTCC TTGTATAAAT 
               
               
                   
               
               
                 2941 
                 CCTGGTTGCT GTCTCTTTAT GAGGAGTTGT GGCCCGTTGT CAGGCAACGT GGCGTGGTGT 
               
               
                   
               
               
                 3001 
                 GCACTGTGTT TGCTGACGCA ACCCCCACTG GTTGGGGCAT TGCCACCACC TGTCAGCTCC 
               
               
                   
               
               
                 3061 
                 TTTCCGGGAC TTTCGCTTTC CCCCTCCCTA TTGCCACGGC GGAACTCATC GCCGCCTGCC 
               
               
                   
               
               
                 3121 
                 TTGCCCGCTG CTGGACAGGG GCTCGGCTGT TGGGCACTGA CAATTCCGTG GTGTTGTCGG 
               
               
                   
               
               
                 3181 
                 GGAAATCATC GTCCTTTCCT TGGCTGCTCG CCTGTGTTGC CACCTGGATT CTGCGCGGGA 
               
               
                   
               
               
                 3241 
                 CGTCCTTCTG CTACGTCCCT TCGGCCCTCA ATCCAGCGGA CCTTCCTTCC CGCGGCCTGC 
               
               
                   
               
               
                 3301 
                 TGCCGGCTCT GCGGCCTCTT CCGCGTCTTC GCCTTCGCCC TCAGACGAGT CGGATCTCCC 
               
               
                   
               
               
                 3361 
                 TTTGGGCCGC CTCCCCGGCG GCCGCGCACC GTCGACTCGC TGATCAGCCT CGACTGTGCC 
               
               
                   
               
               
                 3421 
                 TTCTAGTTGC CAGCCATCTG TTGTTTGCCC CTCCCCCGTG CCTTCCTTGA CCCTGGAAGG 
               
               
                   
               
               
                 3481 
                 TGCCACTCCC ACTGTCCTTT CCTAATAAAA TGAGGAAATT GCATCGCATT GTCTGAGTAG 
               
               
                   
               
               
                 3541 
                 GTGTCATTCT ATTCTGGGGG GTGGGGTGGG GCAGGACAGC AAGGGGGAGG ATTGGGAAGA 
               
               
                   
               
               
                 3601 
                 CAATAGCAGG CATGCTGGGG ATGCGGTGGG CTCTATGGCT TCTGAGGCGG AAAGAACCAG 
               
               
                   
               
               
                 3661 
                 CTGGGGCTCG ACTAGAGCAT GGCTACGTAG ATAAGTAGCA TGGCGGGTTA ATCATTAACT 
               
               
                   
               
               
                 3721 
                 ACAAGGAACC CCTAGTGATG GAGTTGGCCA CTCCCTCTCT GCGCGCTCGC TCGCTCACTG 
               
               
                   
               
               
                 3781 
                 AGGCCGGGCG ACCAAAGGTC GCCCGACGCC CGGGCGGCCT CAGTGAGCGA GCGAGCGCGC 
               
               
                   
               
               
                 3841 
                 AGAGCTTTTT GCAAAAGCCT AGGCCTCCAA AAAAGCCTCC TCACTACTTC TGGAATAGCT 
               
               
                   
               
               
                 3901 
                 CAGAGGCCGA GGCGGCCTCG GCCTCTGCAT AAATAAAAAA AATTAGTCAG CCATGGGGCG 
               
               
                   
               
               
                 3961 
                 GAGAATGGGC GGAACTGGGC GGAGTTAGGG GCGGGATGGG CGGAGTTAGG GGCGGGACTA 
               
               
                   
               
               
                 4021 
                 TGGTTGCTGA CTAATTGAGA TGCATGCTTT GCATACTTCT GCCTGCTGGG GAGCCTGGGG 
               
               
                   
               
               
                 4081 
                 ACTTTCCACA CCTGGTTGCT GACTAATTGA GATGCATGCT TTGCATACTT CTGCCTGCTG 
               
               
                   
               
               
                 4141 
                 GGGAGCCTGG GGACTTTCCA CACCCTAACT GACACACATT CCACAGCTGC ATTAATGAAT 
               
               
                   
               
               
                 4201 
                 CGGCCAACGC GCGGGGAGAG GCGGTTTGCG TATTGGGCGC TCTTCCGCTT CCTCGCTCAC 
               
               
                   
               
               
                 4261 
                 TGACTCGCTG CGCTCGGTCG TTCGGCTGCG GCGAGCGGTA TCAGCTCACT CAAAGGCGGT 
               
               
                   
               
               
                 4321 
                 AATACGGTTA TCCACAGAAT CAGGGGATAA CGCAGGAAAG AACATGTGAG CAAAAGGCCA 
               
               
                   
               
               
                 4381 
                 GCAAAAGGCC AGGAACCGTA AAAAGGCCGC GTTGCTGGCG TTTTTCCATA GGCTCCGCCC 
               
               
                   
               
               
                 4441 
                 CCCTGACGAG CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC CGACAGGACT 
               
               
                   
               
               
                 4501 
                 ATAAAGATAC CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG TTCCGACCCT 
               
               
                   
               
               
                 4561 
                 GCCGCTTACC GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC TTTCTCATAG 
               
               
                   
               
               
                 4621 
                 CTCACGCTGT AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA 
               
               
                   
               
               
                 4681 
                 CGAACCCCCC GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC TTGAGTCCAA 
               
               
                   
               
               
                 4741 
                 CCCGGTAAGA CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA TTAGCAGAGC 
               
               
                   
               
               
                 4801 
                 GAGGTATGTA GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG GCTACACTAG 
               
               
                   
               
               
                 4861 
                 AAGAACAGTA TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG 
               
               
                   
               
               
                 4921 
                 TAGCTCTTGA TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA 
               
               
                   
               
               
                 4981 
                 GCAGATTACG CGCAGAAAAA AAGGATCTCA AGAAGATCCT TTGATCTTTT CTACGGGGTC 
               
               
                   
               
               
                 5041 
                 TGACGCTCAG TGGAACGAAA ACTCACGTTA AGGGATTTTG GTCATGAGAT TATCAAAAAG 
               
               
                   
               
               
                 5101 
                 GATCTTCACC TAGATCCTTT TAAATTAAAA ATGAAGTTTT AAATCAATCT AAAGTATATA 
               
               
                   
               
               
                 5161 
                 TGAGTAAACT TGGTCTGACA GTTACCAATG CTTAATCAGT GAGGCACCTA TCTCAGCGAT 
               
               
                   
               
               
                 5221 
                 CTGTCTATTT CGTTCATCCA TAGTTGCCTG ACTCCTGCAA ACCACGTTGT GTCTCAAAAT 
               
               
                   
               
               
                 5281 
                 CTCTGATGTT ACATTGCACA AGATAAAAAT ATATCATCAT GAACAATAAA ACTGTCTGCT 
               
               
                   
               
               
                 5341 
                 TACATAAACA GTAATACAAG GGGTGTTATG AGCCATATTC AACGGGAAAC GTCTTGCTCG 
               
               
                   
               
               
                 5401 
                 AGGCCGCGAT TAAATTCCAA CATGGATGCT GATTTATATG GGTATAAATG GGCTCGCGAT 
               
               
                   
               
               
                 5461 
                 AATGTCGGGC AATCAGGTGC GACAATCTAT CGATTGTATG GGAAGCCCGA TGCGCCAGAG 
               
               
                   
               
               
                 5521 
                 TTGTTTCTGA AACATGGCAA AGGTAGCGTT GCCAATGATG TTACAGATGA GATGGTCAGA 
               
               
                   
               
               
                 5581 
                 CTAAACTGGC TGACGGAATT TATGCCTCTT CCGACCATCA AGCATTTTAT CCGTACTCCT 
               
               
                   
               
               
                 5641 
                 GATGATGCAT GGTTACTCAC CACTGCGATC CCCGGGAAAA CAGCATTCCA GGTATTAGAA 
               
               
                   
               
               
                 5701 
                 GAATATCCTG ATTCAGGTGA AAATATTGTT GATGCGCTGG CAGTGTTCCT GCGCCGGTTG 
               
               
                   
               
               
                 5761 
                 CATTCGATTC CTGTTTGTAA TTGTCCTTTT AACAGCGATC GCGTATTTCG TCTCGCTCAG 
               
               
                   
               
               
                 5821 
                 GCGCAATCAC GAATGAATAA CGGTTTGGTT GATGCGAGTG ATTTTGATGA CGAGCGTAAT 
               
               
                   
               
               
                 5881 
                 GGCTGGCCTG TTGAACAAGT CTGGAAAGAA ATGCATAAGC TTTTGCCATT CTCACCGGAT 
               
               
                   
               
               
                 5941 
                 TCAGTCGTCA CTCATGGTGA TTTCTCACTT GATAACCTTA TTTTTGACGA GGGGAAATTA 
               
               
                   
               
               
                 6001 
                 ATAGGTTGTA TTGATGTTGG ACGAGTCGGA ATCGCAGACC GATACCAGGA TCTTGCCATC 
               
               
                   
               
               
                 6061 
                 CTATGGAACT GCCTCGGTGA GTTTTCTCCT TCATTAGAGA AACGGCTTTT TCAAAAATAT 
               
               
                   
               
               
                 6121 
                 GGTATTGATA ATCCTGATAT GAATAAATTG CAGTTTCATT TGATGCTCGA TGAGTTTTTC 
               
               
                   
               
               
                 6181 
                 TAAGGGCGGC CTGCCACCAT ACCCACGCCG AAACAAGCGC TCATGAGCCC GAAGTGGCGA 
               
               
                   
               
               
                 6241 
                 GCCCGATCTT CCCCATCGGT GATGTCGGCG ATATAGGCGC CAGCAACCGC ACCTGTGGCG 
               
               
                   
               
               
                 6301 
                 CCGGTGATGC CGGCCACGAT GCGTCCGGCG TAGAGGATCT GGCTAGCGAT GACCCTGCTG 
               
               
                   
               
               
                 6361 
                 ATTGGTTCGC TGACCATTTC CGGGTGCGGG ACGGCGTTAC CAGAAACTCA GAAGGTTCGT 
               
               
                   
               
               
                 6421 
                 CCAACCAAAC CGACTCTGAC GGCAGTTTAC GAGAGAGATG ATAGGGTCTG CTTCAGGGTG 
               
               
                   
               
               
                 6481 
                 ACCGATGTAA CCATATACTT AGGCTGGATC TTCTCCCGCG AATTTTAACC CTCACCAACT 
               
               
                   
               
               
                 6541 
                 ACGAGATATG AGGTAAGCCA AAAAAGCACG TAGTGGCGCT CTCCGACTGT TCCCAAATTG 
               
               
                   
               
               
                 6601 
                 TAACTTATCG TTCCGTGAAG GCCAGAGTTA CTTCCCGGCC CTTTCCATGC GCGCACCATA 
               
               
                   
               
               
                 6661 
                 CCCTCCTAGT TCCCCGGTTA TCTTTCCGAA GTGGGAGTGA GCGAACCTCC GTTTACGTCT 
               
               
                   
               
               
                 6721 
                 TGTTACCAAT GATGTAGCTA TGCACTTTGT ACAGGGTGCC AACGGGTTTC ACAATTCACA 
               
               
                   
               
               
                 6781 
                 GATAGTGGGG ATCCCGGCAA AGGGCCTATA TTTGCGGTCC AACTTAGGCG TAAACCTCGA 
               
               
                   
               
               
                 6841 
                 TGCTACCTAC TCAGACCCAC CTCGCGCGGG GTAAATAAGG CACTCATCCC AGCTGGTTCT 
               
               
                   
               
               
                 6901 
                 TGGCGTTCTA CGCAGCGACA TGTTTATTAA CAGTTGTCTG GCAGCACAAA ACTTTTACCA 
               
               
                   
               
               
                 6961 
                 TGGTCGTAGA AGCCCCCCAG AGTTAGTTCA TACCTAATGC CACAAATGTG ACAGGACGCC 
               
               
                   
               
               
                 7021 
                 GATGGGTACC GGACTTTAGG TCGAGCACAG TTCGGTAACG GAGAGACCCT GCGGCGTACT 
               
               
                   
               
               
                 7081 
                 TCATTATGTA TATGGAACGT GCCCAAGTGA CGCCAGGCAA GTCTCAGCTG GTTCCTGTGT 
               
               
                   
               
               
                 7141 
                 TAGCTCGAGG GTAGACATAC GAGCTGATTG AACATGGGTT GGGGGCCTCG AACCGTCGAG 
               
               
                   
               
               
                 7201 
                 GACCCCATAG TACCTCGGAG ACCAAGTAGG GCAGCCTATA GTTTGAAGCA GAACTATTTC 
               
               
                   
               
               
                 7261 
                 GGGGGGCGAG CCCTCATCGT CTCTTCTGCG GATGACTCAA CACGCTAGGG ACGTGAAGTC 
               
               
                   
               
               
                 7321 
                 GATTCCTTCG ATGGTTATAA ATCAAAGACT CAGAGTGCTG TCTGGAGCGT GAATCTAACG 
               
               
                   
               
               
                 7381 
                 GTACGTATCT CGATTGCTCG GTCGCTTTTC GCACTCCGCG AAAGTTCGTA CCGCTCATTC 
               
               
                   
               
               
                 7441 
                 ACTAGGTTGC GAAGCCTATG CTGATATATG AATCCAAACT AGAGCAGGGC TCTTAAGATT 
               
               
                   
               
               
                 7501 
                 CGGAGTTGTA AATACTTAAT ACTCCAATCG GCTTTTACGT GCACCACCGC GGGCGGCTGA 
               
               
                   
               
               
                 7561 
                 CAAGGGTCTC ACATCGAGAA ACAAGACAGT TCCGGGCTGG AAGTAGCGCC GGCTAAGGAA 
               
               
                   
               
               
                 7621 
                 GACGCCTGGT ACGGCAGGAC TATGAAACCA GTACAAAGGC AACATCCTCA CTTGGGTGAA 
               
               
                   
               
               
                 7681 
                 CGGAAACGCA GTATTATGGT TACTTTTTGG ATACGTGAAA CATATCCCAT GGTAGTCCTT 
               
               
                   
               
               
                 7741 
                 AGACTTGGGA GTCTATCACC CCTAGGGCCC ATATCTGGAA ATAGACGCCA GGTTGAATCC 
               
               
                   
               
               
                 7801 
                 GTATTTGGAG GTACGATGGA ACAGTCTGGG TGGGACGTGC TTCATTTATA CCCTGCGCAG 
               
               
                   
               
               
                 7861 
                 GCTGGACCGA GGACCGCAAG GTGCGGCGGT GCACAAGCAA TTGACAACTA ACCACCGTGT 
               
               
                   
               
               
                 7921 
                 ATTCATTATG GTACCAGGAA CTTTAAGCCG AGTCAATGAA GCTCGCATTA CAGTGTTTAC 
               
               
                   
               
               
                 7981 
                 CGCATCTTGC GGTTACTCAC AAACTGTGAT CCACCACAAG TCAAGCCATT GCCTCTCTGA 
               
               
                   
               
               
                 8041 
                 CACGCCGTAA GAATTAATAT GTAAACTTTG CGCGGGTTGA CTGCGATCCG TTCAGTCTCG 
               
               
                   
               
               
                 8101 
                 TCCGAGGGCA CAATCCTATT CCCATTTGTA TGTTCAGCTA ACTTCTACCC ATCCCCCGAA 
               
               
                   
               
               
                 8161 
                 GTTAAGTAGG TCGTGAGATG CCATGGAGGC TCTCGTTCAT CCCGTGGGAC ATCAAGCTTC 
               
               
                   
               
               
                 8221 
                 CCCTTGATAA AGCACCCCGC TCGGGTGTAG CAGAGAAGAC GCCTTCTGAA TTGTGCAATC 
               
               
                   
               
               
                 8281 
                 CCTCCACCTT ATCTAAGCTT GCTACCAATA ATTAGCATTT TTGCCTTGCG ACAGACCTCC 
               
               
                   
               
               
                 8341 
                 TACTTAGATT GCCACACATT GAGCTAGTCA GTGAGCGATA AGCTTGACGC GCTTTCAAGG 
               
               
                   
               
               
                 8401 
                 GTCGCGAGTA CGTGAACTAA GGCTCCGGAC AGGACTATAT ACTTGGGTTT GATCTCGCCC 
               
               
                   
               
               
                 8461 
                 CGACAACTGC AAACCTCAAC TTTTTTAGAT TATATGGTTA GCCGAAGTTG CACGAGGTGG 
               
               
                   
               
               
                 8521 
                 CGTCCGCGGA CTGCTCCCCG AGTGTGGCTC TTTCATCTGA CAACGTGCAA CCCCTATCGC 
               
               
                   
               
               
                 8581 
                 GGCCGATTGT TTCTGCGGAC GATGTTGTCC TCATAGTTTG GGCATGTTTC CCTTGTAGGT 
               
               
                   
               
               
                 8641 
                 GTGAAACCAC TTAGCTTCGC GCCGTAGTCC CAATGAAAAA CCTATGGACT TTGTTTTGGG 
               
               
                   
               
               
                 8701 
                 TAGCACCAGG AATCTGAACC GTGTGAATGT GGACGTCGCG CGCGTAGACC TTTATCTCCG 
               
               
                   
               
               
                 8761 
                 GTTCAAGCTA GGGATGTGGC TGCATGCTAC GTTGTCACAC CTACACTGCT CGAAGTAAAT 
               
               
                   
               
               
                 8821 
                 ATGCGAAGCG CGCGGCCTGG CCGGAGGCGT TCCGCGCCGC CACGTGTTCG TTAACTGTTG 
               
               
                   
               
               
                 8881 
                 ATTGGTGGCA CATAAGCAAT ATCGTAGTCC GTCAAATTCA GCTCTGTTAT CCCGGGCGTT 
               
               
                   
               
               
                 8941 
                 ATGTGTCAAA TGGCGTAGAA CGGGATTGAC TGTTTGACGG TAGGGTGACC TAAGCCAGAT 
               
               
                   
               
               
                 9001 
                 GCTACACAAT TAGGCTTGTA CATATTGTCG TTAGAACGCG GCTACAATTA ATACATAACC 
               
               
                   
               
               
                 9061 
                 TTATGTATGA TACACATACG ATTTAGGTGA CACTATAGAA TACACGGAAT TAATTC. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 2 
               
             
            
               
                   
               
               
                 Features of the VMD2.IntEx.BEST1.WPRE.pA plasmid sequence 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                 Mini- 
                 Maxi- 
                   
                 Direc- 
               
               
                 Name 
                 Type 
                 mum 
                 mum 
                 Length 
                 tion 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 AAV2 ITR 
                 LTR 
                 4 
                 133 
                 130 
                 forward 
               
               
                 −585 to +38 
                 promoter 
                 189 
                 811 
                 623 
                 forward 
               
               
                 VMD2 promoter 
               
               
                 Intron 
                 intron 
                 814 
                 936 
                 123 
                 forward 
               
               
                 Exon 
                 exon 
                 937 
                 989 
                 53 
                 forward 
               
               
                 Kozak 
                 Kozak 
                 990 
                 999 
                 10 
                 forward 
               
               
                 BEST1 
                 CDS 
                 996 
                 2753 
                 1758 
                 forward 
               
               
                 NotI 
                 RBS 
                 2758 
                 2765 
                 8 
                 none 
               
               
                 WPRE 
                 WPRE 
                 2784 
                 3376 
                 593 
                 forward 
               
               
                 NotI 
                 RBS 
                 3378 
                 3385 
                 8 
                 none 
               
               
                 bGH pA 
                 polyA_signal 
                 3398 
                 3666 
                 269 
                 forward 
               
               
                 AAV2 ITR 
                 LTR 
                 3724 
                 3844 
                 121 
                 reverse 
               
               
                 pBR322 rep origin 
                 rep_origin 
                 4408 
                 5027 
                 620 
                 reverse 
               
               
                 AphR (KanR) 
                 CDS 
                 5368 
                 6183 
                 816 
                 forward 
               
               
                 BstEII 
                 RBS 
                 6477 
                 6483 
                 7 
                 none 
               
               
                 Randomly generated 
                 Stuffer 
                 6484 
                 8983 
                 2500 
                 none 
               
               
                 stuffer sequence 
               
               
                 BstEII 
                 RBS 
                 8984 
                 8990 
                 7 
                 none 
               
               
                   
               
            
           
         
       
     
     In some embodiments, the AAV viral delivery vector comprises a nucleic acid sequence comprising a sequence encoding a CAG promoter, a sequence encoding a BEST1 protein and a sequence encoding a WPRE. An exemplary AAV viral delivery vector of the disclosure comprising a nucleic acid sequence encoding a CAG.BEST1.WPRE.pA sequence comprises or consists of the nucleic acid sequence of: 
     
       
         
           
               
               
            
               
                 (SEO Id NO: 15) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 TAGCTGCGCG CTCGCTCGCT CACTGAGGCC GCCCGGGCAA AGCCCGGGCG TCGGGCGACC 
                   
               
               
                   
               
               
                 61 
                 TTTGGTCGCC CGGCCTCAGT GAGCGAGCGA GCGCGCAGAG AGGGAGTGGC CAACTCCATC 
               
               
                   
               
               
                 121 
                 ACTAGGGGTT CCTTGTAGTT AATGATTAAC CCGCCATGCT ACTTATCTAC GTAGCCATGC 
               
               
                   
               
               
                 181 
                 TCTAGGTACC ATTGAGGTGA ATAATGACGT ATGTTCCCAT AGTAACGCCA ATAGGGACTT 
               
               
                   
               
               
                 241 
                 TCCATTGACG TCAATGGGTG GAGTATTTAC GGTAAACTGC CCACTTGGCA GTACATCAAG 
               
               
                   
               
               
                 301 
                 TGTATCATAT GCCAAGTACG CCCCCTATTG ACGTCAATGA CGGTAAATGG CCCGCCTGGC 
               
               
                   
               
               
                 361 
                 ATTATGCCCA GTACATGACC TTATGGGACT TTCCTACTTG GCAGTACATC TAGGTATTAG 
               
               
                   
               
               
                 421 
                 TCATCGCTAT TACCATGGTC GAGGTGAGCC CCACGTTCTG CTTCAGTCTC CCCATCTCCC 
               
               
                   
               
               
                 481 
                 CCCCCTCCCC ACCCCCAATT TTGTATTTAT TTATTTTTTA ATTATTTTGT GCAGCGATGG 
               
               
                   
               
               
                 541 
                 GGGCGGGGGG GGGGGGGGGG CGCGCGCCAG GGGGGGGGGG GCGGGGCGAG GGGGGGGGGG 
               
               
                   
               
               
                 601 
                 GGGCGAGGCG GAGAGGTGCG GCGGCAGCCA ATCAGAGCGG CGCGCTCCGA AAGTTTCCTT 
               
               
                   
               
               
                 661 
                 TTATGGCGAG GGGGGGGGGG CGGCGGCCCT ATAAAAAGCG AAGCGCGCGG CGGGCGGGAG 
               
               
                   
               
               
                 721 
                 TCGCTGCGCG CTGCCTTCGC CCCGTGCCCC GCTCCGCCGC CGCCTCGCGC CGCCCGCCCC 
               
               
                   
               
               
                 781 
                 GGCTCTGACT GACCGCGTTA CTCCCACAGG TGAGCGGGCG GGACGGCCCT TCTCCTCCGG 
               
               
                   
               
               
                 841 
                 GCTGTAATTA GCGCTTGGTT TAATGACGGC TTGTTTCTTT TCTGTGGCTG CGTGAAAGCC 
               
               
                   
               
               
                 901 
                 TTGAGGGGCT CCGGGAGGGC CCTTTGTGCG GGGGGAGCGG CTCGGGGCTG TCCGCGGGGG 
               
               
                   
               
               
                 961 
                 GACGGCTGCC TTCGGGGGGG ACGGGGCAGG GCGGGGTTCG GCTTCTGGCG TGTGACCGGC 
               
               
                   
               
               
                 1021 
                 GGCTCTAGAG CCTCTGCTAA CCATGTTCAT GCCTTCTTCT TTTTCCTACA GCTCCTGGGC 
               
               
                   
               
               
                 1081 
                 AACGTGCTGG TTATTGTGCT GTCTCATCAT TTTGGCAAAG AATTGGATCC GCGGCCGCAG 
               
               
                   
               
               
                 1141 
                 CTTGGTACCG CCACCATGAC CATCACTTAC ACAAGCCAAG TGGCTAATGC CCGCTTAGGC 
               
               
                   
               
               
                 1201 
                 TCCTTCTCCC GCCTGCTGCT GTGCTGGCGG GGCAGCATCT ACAAGCTGCT ATATGGCGAG 
               
               
                   
               
               
                 1261 
                 TTCTTAATCT TCCTGCTCTG CTACTACATC ATCCGCTTTA TTTATAGGCT GGCCCTCACG 
               
               
                   
               
               
                 1321 
                 GAAGAACAAC AGCTGATGTT TGAGAAACTG ACTCTGTATT GCGACAGTTA CATCCAGCTC 
               
               
                   
               
               
                 1381 
                 ATCCCCATTT CCTTCGTGCT GGGCTTCTAC GTGACGCTGG TCGTGACCCG CTGGTGGAAC 
               
               
                   
               
               
                 1441 
                 CAGTACGAGA ACCTGCCGTG GCCCGACCGC CTCATGAGCC TGGTGTCGGG CTTCGTCGAA 
               
               
                   
               
               
                 1501 
                 GGCAAGGACG AGCAAGGCCG GCTGCTGCGG CGCACGCTCA TCCGCTACGC CAACCTGGGC 
               
               
                   
               
               
                 1561 
                 AACGTGCTCA TCCTGCGCAG CGTCAGCACC GCAGTCTACA AGCGCTTCCC CAGCGCCCAG 
               
               
                   
               
               
                 1621 
                 CACCTGGTGC AAGCAGGCTT TATGACTCCG GCAGAACACA AGCAGTTGGA GAAACTGAGC 
               
               
                   
               
               
                 1681 
                 CTACCACACA ACATGTTCTG GGTGCCCTGG GTGTGGTTTG CCAACCTGTC AATGAAGGCG 
               
               
                   
               
               
                 1741 
                 TGGCTTGGAG GTCGAATCCG GGACCCTATC CTGCTCCAGA GCCTGCTGAA CGAGATGAAC 
               
               
                   
               
               
                 1801 
                 ACCTTGCGTA CTCAGTGTGG ACACCTGTAT GCCTACGACT GGATTAGTAT CCCACTGGTG 
               
               
                   
               
               
                 1861 
                 TATACACAGG TGGTGACTGT GGCGGTGTAC AGCTTCTTCC TGACTTGTCT AGTTGGGCGG 
               
               
                   
               
               
                 1921 
                 CAGTTTCTGA ACCCAGCCAA GGCCTACCCT GGCCATGAGC TGGACCTCGT TGTGCCCGTC 
               
               
                   
               
               
                 1981 
                 TTCACGTTCC TGCAGTTCTT CTTCTATGTT GGCTGGCTGA AGGTGGCAGA GCAGCTCATC 
               
               
                   
               
               
                 2041 
                 AACCCCTTTG GAGAGGATGA TGATGATTTT GAGACCAACT GGATTGTCGA CAGGAATTTG 
               
               
                   
               
               
                 2101 
                 CAGGTGTCCC TGTTGGCTGT GGATGAGATG CACCAGGACC TGCCTCGGAT GGAGCCGGAC 
               
               
                   
               
               
                 2161 
                 ATGTACTGGA ATAAGCCCGA GCCACAGCCC CCCTACACAG CTGCTTCCGC CCAGTTCCGT 
               
               
                   
               
               
                 2221 
                 CGAGCCTCCT TTATGGGCTC CACCTTCAAC ATCAGCCTGA ACAAAGAGGA GATGGAGTTC 
               
               
                   
               
               
                 2281 
                 CAGCCCAATC AGGAGGACGA GGAGGATGCT CACGCTGGCA TCATTGGCCG CTTCCTAGGC 
               
               
                   
               
               
                 2341 
                 CTGCAGTCCC ATGATCACCA TCCTCCCAGG GCAAACTCAA GGACCAAACT ACTGTGGCCC 
               
               
                   
               
               
                 2401 
                 AAGAGGGAAT CCCTTCTCCA CGAGGGCCTG CCCAAAAACC ACAAGGCAGC CAAACAGAAC 
               
               
                   
               
               
                 2461 
                 GTTAGGGGCC AGGAAGACAA CAAGGCCTGG AAGCTTAAGG CTGTGGACGC CTTCAAGTCT 
               
               
                   
               
               
                 2521 
                 GCCCCACTGT ATCAGAGGCC AGGCTACTAC AGTGCCCCAC AGACGCCCCT CAGCCCCACT 
               
               
                   
               
               
                 2581 
                 CCCATGTTCT TCCCCCTAGA ACCATCAGCG CCGTCAAAGC TTCACAGTGT CACAGGCATA 
               
               
                   
               
               
                 2641 
                 GACACCAAAG ACAAAAGCTT AAAGACTGTG AGTTCTGGGG CCAAGAAAAG TTTTGAATTG 
               
               
                   
               
               
                 2701 
                 CTCTCAGAGA GCGATGGGGC CTTGATGGAG CACCCAGAAG TATCTCAAGT GAGGAGGAAA 
               
               
                   
               
               
                 2761 
                 ACTGTGGAGT TTAACCTGAC GGATATGCCA GAGATCCCCG AAAATCACCT CAAAGAACCT 
               
               
                   
               
               
                 2821 
                 TTGGAACAAT CACCAACCAA CATACACACT ACACTCAAAG ATCACATGGA TCCTTATTGG 
               
               
                   
               
               
                 2881 
                 GCCTTGGAAA ACAGGGATGA AGCACATTCC TAAGAGCTCA AGCTTATCGA TAATCAACCT 
               
               
                   
               
               
                 2941 
                 CTGGATTACA AAATTTGTGA AAGATTGACT GGTATTCTTA ACTATGTTGC TCCTTTTACG 
               
               
                   
               
               
                 3001 
                 CTATGTGGAT ACGCTGCTTT AATGCCTTTG TATCATGCTA TTGCTTCCCG TATGGCTTTC 
               
               
                   
               
               
                 3061 
                 ATTTTCTCCT CCTTGTATAA ATCCTGGTTG CTGTCTCTTT ATGAGGAGTT GTGGCCCGTT 
               
               
                   
               
               
                 3121 
                 GTCAGGCAAC GTGGCGTGGT GTGCACTGTG TTTGCTGACG CAACCCCCAC TGGTTGGGGC 
               
               
                   
               
               
                 3181 
                 ATTGCCACCA CCTGTCAGCT CCTTTCCGGG ACTTTCGCTT TCCCCCTCCC TATTGCCACG 
               
               
                   
               
               
                 3241 
                 GCGGAACTCA TCGCCGCCTG CCTTGCCCGC TGCTGGACAG GGGCTCGGCT GTTGGGCACT 
               
               
                   
               
               
                 3301 
                 GACAATTCCG TGGTGTTGTC GGGGAAATCA TCGTCCTTTC CTTGGCTGCT CGCCTGTGTT 
               
               
                   
               
               
                 3361 
                 GCCACCTGGA TTCTGCGCGG GACGTCCTTC TGCTACGTCC CTTCGGCCCT CAATCCAGCG 
               
               
                   
               
               
                 3421 
                 GACCTTCCTT CCCGCGGCCT GCTGCCGGCT CTGCGGCCTC TTCCGCGTCT TCGCCTTCGC 
               
               
                   
               
               
                 3481 
                 CCTCAGACGA GTCGGATCTC CCTTTGGGCC GCCTCCCCGC ATCGATACCG TCGACTCGCT 
               
               
                   
               
               
                 3541 
                 GATCAGCCTC GACTGTGCCT TCTAGTTGCC AGCCATCTGT TGTTTGCCCC TCCCCCGTGC 
               
               
                   
               
               
                 3601 
                 CTTCCTTGAC CCTGGAAGGT GCCACTCCCA CTGTCCTTTC CTAATAAAAT GAGGAAATTG 
               
               
                   
               
               
                 3661 
                 CATCGCATTG TCTGAGTAGG TGTCATTCTA TTCTGGGGGG TGGGGTGGGG CAGGACAGCA 
               
               
                   
               
               
                 3721 
                 AGGGGGAGGA TTGGGAAGAC AATAGCAGGC ATGCTGGGGA TGCGGTGGGC TCTATGGCTT 
               
               
                   
               
               
                 3781 
                 CTGAGGCGGA AAGAACCAGC TGGGGCTCGA CTAGAGCATG GCTACGTAGA TAAGTAGCAT 
               
               
                   
               
               
                 3841 
                 GGCGGGTTAA TCATTAACTA CAAGGAACCC CTAGTGATGG AGTTGGCCAC TCCCTCTCTG 
               
               
                   
               
               
                 3901 
                 CGCGCTCGCT CGCTCACTGA GGCCGGGCGA CCAAAGGTCG CCCGACGCCC GGGCGGCCTC 
               
               
                   
               
               
                 3961 
                 AGTGAGCGAG CGAGCGCGCA GAGCTTTTTG CAAAAGCCTA GGCCTCCAAA AAAGCCTCCT 
               
               
                   
               
               
                 4021 
                 CACTACTTCT GGAATAGCTC AGAGGCCGAG GCGGCCTCGG CCTCTGCATA AATAAAAAAA 
               
               
                   
               
               
                 4081 
                 ATTAGTCAGC CATGGGGCGG AGAATGGGCG GAACTGGGCG GAGTTAGGGG CGGGATGGGC 
               
               
                   
               
               
                 4141 
                 GGAGTTAGGG GCGGGACTAT GGTTGCTGAC TAATTGAGAT GCATGCTTTG CATACTTCTG 
               
               
                   
               
               
                 4201 
                 CCTGCTGGGG AGCCTGGGGA CTTTCCACAC CTGGTTGCTG ACTAATTGAG ATGCATGCTT 
               
               
                   
               
               
                 4261 
                 TGCATACTTC TGCCTGCTGG GGAGCCTGGG GACTTTCCAC ACCCTAACTG ACACACATTC 
               
               
                   
               
               
                 4321 
                 CACAGCTGCA TTAATGAATC GGCCAACGCG CGGGGAGAGG CGGTTTGCGT ATTGGGCGCT 
               
               
                   
               
               
                 4381 
                 CTTCCGCTTC CTCGCTCACT GACTCGCTGC GCTCGGTCGT TCGGCTGCGG CGAGCGGTAT 
               
               
                   
               
               
                 4441 
                 CAGCTCACTC AAAGGCGGTA ATACGGTTAT CCACAGAATC AGGGGATAAC GCAGGAAAGA 
               
               
                   
               
               
                 4501 
                 ACATGTGAGC AAAAGGCCAG CAAAAGGCCA GGAACCGTAA AAAGGCCGCG TTGCTGGCGT 
               
               
                   
               
               
                 4561 
                 TTTTCCATAG GCTCCGCCCC CCTGACGAGC ATCACAAAAA TCGACGCTCA AGTCAGAGGT 
               
               
                   
               
               
                 4621 
                 GGCGAAACCC GACAGGACTA TAAAGATACC AGGCGTTTCC CCCTGGAAGC TCCCTCGTGC 
               
               
                   
               
               
                 4681 
                 GCTCTCCTGT TCCGACCCTG CCGCTTACCG GATACCTGTC CGCCTTTCTC CCTTCGGGAA 
               
               
                   
               
               
                 4741 
                 GCGTGGCGCT TTCTCATAGC TCACGCTGTA GGTATCTCAG TTCGGTGTAG GTCGTTCGCT 
               
               
                   
               
               
                 4801 
                 CCAAGCTGGG CTGTGTGCAC GAACCCCCCG TTCAGCCCGA CCGCTGCGCC TTATCCGGTA 
               
               
                   
               
               
                 4861 
                 ACTATCGTCT TGAGTCCAAC CCGGTAAGAC ACGACTTATC GCCACTGGCA GCAGCCACTG 
               
               
                   
               
               
                 4921 
                 GTAACAGGAT TAGCAGAGCG AGGTATGTAG GCGGTGCTAC AGAGTTCTTG AAGTGGTGGC 
               
               
                   
               
               
                 4981 
                 CTAACTACGG CTACACTAGA AGAACAGTAT TTGGTATCTG CGCTCTGCTG AAGCCAGTTA 
               
               
                   
               
               
                 5041 
                 CCTTCGGAAA AAGAGTTGGT AGCTCTTGAT CCGGCAAACA AACCACCGCT GGTAGCGGTG 
               
               
                   
               
               
                 5101 
                 GTTTTTTTGT TTGCAAGCAG CAGATTACGC GCAGAAAAAA AGGATCTCAA GAAGATCCTT 
               
               
                   
               
               
                 5161 
                 TGATCTTTTC TACGGGGTCT GACGCTCAGT GGAACGAAAA CTCACGTTAA GGGATTTTGG 
               
               
                   
               
               
                 5221 
                 TCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT AAATTAAAAA TGAAGTTTTA 
               
               
                   
               
               
                 5281 
                 AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG TTACCAATGC TTAATCAGTG 
               
               
                   
               
               
                 5341 
                 AGGCACCTAT CTCAGCGATC TGTCTATTTC GTTCATCCAT AGTTGCCTGA CTCCCCGTCG 
               
               
                   
               
               
                 5401 
                 TGTAGATAAC TACGATACGG GAGGGCTTAC CATCTGGCCC CAGTGCTGCA ATGATACCGC 
               
               
                   
               
               
                 5461 
                 GAGACCCACG CTCACCGGCT CCAGATTTAT CAGCAATAAA CCAGCCAGCC GGAAGGGCCG 
               
               
                   
               
               
                 5521 
                 AGCGCAGAAG TGGTCCTGCA ACTTTATCCG CCTCCATCCA GTCTATTAAT TGTTGCCGGG 
               
               
                   
               
               
                 5581 
                 AAGCTAGAGT AAGTAGTTCG CCAGTTAATA GTTTGCGCAA CGTTGTTGCC ATTGCTACAG 
               
               
                   
               
               
                 5641 
                 GCATCGTGGT GTCACGCTCG TCGTTTGGTA TGGCTTCATT CAGCTCCGGT TCCCAACGAT 
               
               
                   
               
               
                 5701 
                 CAAGGCGAGT TACATGATCC CCCATGTTGT GCAAAAAAGC GGTTAGCTCC TTCGGTCCTC 
               
               
                   
               
               
                 5761 
                 CGATCGTTGT CAGAAGTAAG TTGGCCGCAG TGTTATCACT CATGGTTATG GCAGCACTGC 
               
               
                   
               
               
                 5821 
                 ATAATTCTCT TACTGTCATG CCATCCGTAA GATGCTTTTC TGTGACTGGT GAGTACTCAA 
               
               
                   
               
               
                 5881 
                 CCAAGTCATT CTGAGAATAG TGTATGCGGC GACCGAGTTG CTCTTGCCCG GCGTCAATAC 
               
               
                   
               
               
                 5941 
                 GGGATAATAC CGCGCCACAT AGCAGAACTT TAAAAGTGCT CATCATTGGA AAACGTTCTT 
               
               
                   
               
               
                 6001 
                 CGGGGCGAAA ACTCTCAAGG ATCTTACCGC TGTTGAGATC CAGTTCGATG TAACCCACTC 
               
               
                   
               
               
                 6061 
                 GTGCACCCAA CTGATCTTCA GCATCTTTTA CTTTCACCAG CGTTTCTGGG TGAGCAAAAA 
               
               
                   
               
               
                 6121 
                 CAGGAAGGCA AAATGCCGCA AAAAAGGGAA TAAGGGCGAC ACGGAAATGT TGAATACTCA 
               
               
                   
               
               
                 6181 
                 TACTCTTCCT TTTTCAATAT TATTGAAGCA TTTATCAGGG TTATTGTCTC ATGAGCGGAT 
               
               
                   
               
               
                 6241 
                 ACATATTTGA ATGTATTTAG AAAAATAAAC AAATAGGGGT TCCGCGCACA TTTCCCCGAA 
               
               
                   
               
               
                 6301 
                 AAGTGCCACC TGACGTCTAA GAAACCATTA TTATCATGAC ATTAACCTAT AAAAATAGGC 
               
               
                   
               
               
                 6361 
                 GTATCACGAG GCCCTTTCGT CTCGCGCGTT TCGGTGATGA CGGTGAAAAC CTCTGACACA 
               
               
                   
               
               
                 6421 
                 TGCAGCTCCC GGAGACGGTC ACAGCTTGTC TGTAAGCGGA TGCCGGGAGC AGACAAGCCC 
               
               
                   
               
               
                 6481 
                 GTCAGGGCGC GTCAGCGGGT GTTGGCGGGT GTCGGGGCTG GCTTAACTAT GCGGCATCAG 
               
               
                   
               
               
                 6541 
                 AGCAGATTGT ACTGAGAGTG CACCATTCGA CGCTCTCCCT TATGCGACTC CTGCATTAGG 
               
               
                   
               
               
                 6601 
                 AAGCAGCCCA GTAGTAGGTT GAGGCCGTTG AGCACCGCCG CCGCAAGGAA TGGTGCATGC 
               
               
                   
               
               
                 6661 
                 AAGGAGATGG CGCCCAACAG TCCCCCGGCC ACGGGGCCTG CCACCATACC CACGCCGAAA 
               
               
                   
               
               
                 6721 
                 CAAGCGCTCA TGAGCCCGAA GTGGCGAGCC CGATCTTCCC CATCGGTGAT GTCGGCGATA 
               
               
                   
               
               
                 6781 
                 TAGGCGCCAG CAACCGCACC TGTGGCGCCG GTGATGCCGG CCACGATGCG TCCGGCGTAG 
               
               
                   
               
               
                 6841 
                 AGGATCTGGC TAGCGATGAC CCTGCTGATT GGTTCGCTGA CCATTTCCGG GTGCGGGACG 
               
               
                   
               
               
                 6901 
                 GCGTTACCAG AAACTCAGAA GGTTCGTCCA ACCAAACCGA CTCTGACGGC AGTTTACGAG 
               
               
                   
               
               
                 6961 
                 AGAGATGATA GGGTCTGCTT CAGTAAGCCA GATGCTACAC AATTAGGCTT GTACATATTG 
               
               
                   
               
               
                 7021 
                 TCGTTAGAAC GCGGCTACAA TTAATACATA ACCTTATGTA TCATACACAT ACGATTTAGG 
               
               
                   
               
               
                 7081 
                 TGACACTATA GAATACACGG AATTAATTC. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 3 
               
             
            
               
                   
               
               
                 Features of the CAG.BEST1.WPRE.PA plasmid sequence 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                 Mini- 
                 Maxi- 
                   
                 Direc- 
               
               
                 Name 
                 Type 
                 mum 
                 mum 
                 Length 
                 tion 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 AAV2 ITR 
                 repeat_region 
                 7,066 
                 133 
                 177 
                 forward 
               
               
                 amp prom 
                 promoter 
                 6,223 
                 6,251 
                 29 
                 reverse 
               
               
                 AmpR gene 
                 gene 
                 5,321 
                 6,181 
                 861 
                 reverse 
               
               
                 Bla gene 
                 gene 
                 5,321 
                 5,983 
                 663 
                 reverse 
               
               
                 ColE1 origin 
                 rep_origin 
                 4,503 
                 5,176 
                 674 
                 forward 
               
               
                 rep origin 
               
               
                 SV40 origin 
                 origin of 
                 4,059 
                 4,136 
                 78 
                 reverse 
               
               
                   
                 replication 
               
               
                 AAV2 ITR 
                 repeat_region 
                 3,864 
                 4,000 
                 137 
                 reverse 
               
               
                 bGH_PA term 
                 terminator 
                 3,550 
                 3,777 
                 228 
                 forward 
               
               
                 WPRE 
                 misc_feature 
                 2,932 
                 3,520 
                 589 
                 forward 
               
               
                 Exon 10 
                 exon 
                 2,895 
                 2,913 
                 19 
                 forward 
               
               
                 Exon 9 
                 exon 
                 2,256 
                 2,894 
                 639 
                 forward 
               
               
                 Exon 8 
                 exon 
                 2,104 
                 2,255 
                 152 
                 forward 
               
               
                 Exon 7 
                 exon 
                 2,023 
                 2,103 
                 81 
                 forward 
               
               
                 Exon 6 
                 exon 
                 1,870 
                 2,022 
                 153 
                 forward 
               
               
                 Exon 5 
                 exon 
                 1,792 
                 1,869 
                 78 
                 forward 
               
               
                 Exon 4 
                 exon 
                 1,637 
                 1,791 
                 155 
                 forward 
               
               
                 Exon 3 
                 exon 
                 1,403 
                 1,636 
                 234 
                 forward 
               
               
                 C &gt; T 
                 modified_base 
                 1,368 
                 1,368 
                 1 
                 forward 
               
               
                 Exon 2 
                 exon 
                 1,308 
                 1,402 
                 95 
                 forward 
               
               
                 hBEST1 CDS 
                 CDS 
                 1,156 
                 2,913 
                 1,758 
                 forward 
               
               
                 Exon 1 
                 exon 
                 1,156 
                 1,307 
                 152 
                 forward 
               
               
                 Kozak 
                 unsure 
                 1,147 
                 1,155 
                 9 
                 forward 
               
               
                   
                 Editing History 
                 &lt;1133 
                 1,138 
                 &gt;6 
                 none 
               
               
                   
                 Insertion 
               
               
                 CAG promoter 
                 promoter 
                 189 
                 1,129 
                 941 
                 forward 
               
               
                 5′ITR on REP1 
                 LTR 
                 64 
                 183 
                 120 
                 forward 
               
               
                 official 
               
               
                 sequence file 
               
               
                   
               
            
           
         
       
     
     In some embodiments of the compositions of the disclosure, a vector may comprise a sequence encoding a marker, which may be expressed in a cell when the cell is either in vitro or in vivo. For example, in a vector or nucleic acid sequence of the disclosure, a sequence encoding a marker may be used in place of or may replace a sequence encoding a BEST1 protein of the disclosure (e.g. a sequence comprising a coding sequence of a BEST1 gene). Exemplary markers of the disclosure include, but are not limited to, fluorophore proteins such as GFP, YFP or dsRED as well as various epitope tags such as FLAG, HA, His or Myc. The fluorophore or epitope tag may be fused to the BEST1 coding sequence, for example as an N or C terminal fusion, or may be used in place of BEST1 to characterize a vector of the disclosure. Exemplary uses for a vector containing a marker of the disclosure include, but are not limited to characterizing gene expression, for example levels of expression, or characterizing the cell type specificity of a vector of the disclosure. 
     An exemplary a vector of the disclosure comprising a marker includes VMD2.GFP.WPRE.pA. A nucleic acid sequence encoding a VMD2.GFP.WPRE.pA construct comprises or consists of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 16) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 TAGCTGCGCG CTCGCTCGCT CACTGAGGCC GCCCGGGCAA AGCCCGGGCG TCGGGCGACC 
                   
               
               
                   
               
               
                 61 
                 TTTGGTCGCC CGGCCTCAGT GAGCGAGCGA GCGCGCAGAG AGGGAGTGGC CAACTCCATC 
               
               
                   
               
               
                 121 
                 ACTAGGGGTT CCTTGTAGTT AATGATTAAC CCGCCATGCT ACTTATCTAC GTAGCCATGC 
               
               
                   
               
               
                 181 
                 TCTAGGTAAA TTCTGTCATT TTACTAGGGT GATGAAATTC CCAAGCAACA CCATCCTTTT 
               
               
                   
               
               
                 241 
                 CAGATAAGGG CACTGAGGCT GAGAGAGGAG CTGAAACCTA CCCGGGGTCA CCACACACAG 
               
               
                   
               
               
                 301 
                 GTGGCAAGGC TGGGACCAGA AACCAGGACT GTTGACTGCA GCCCGGTATT CATTCTTTCC 
               
               
                   
               
               
                 361 
                 ATAGCCCACA GGGCTGTCAA AGACCCCAGG GCCTAGTCAG AGGCTCCTCC TTCCTGGAGA 
               
               
                   
               
               
                 421 
                 GTTCCTGGCA CAGAAGTTGA AGCTCAGCAC AGCCCCCTAA CCCCCAACTC TCTCTGCAAG 
               
               
                   
               
               
                 481 
                 GCCTCAGGGG TCAGAACACT GGTGGAGCAG ATCCTTTAGC CTCTGGATTT TAGGGCCATG 
               
               
                   
               
               
                 541 
                 GTAGAGGGGG TGTTGCCCTA AATTCCAGCC CTGGTCTCAG CCCAACACCC TCCAAGAAGA 
               
               
                   
               
               
                 601 
                 AATTAGAGGG GCCATGGCCA GGCTGTGCTA GCCGTTGCTT CTGAGCAGAT TACAAGAAGG 
               
               
                   
               
               
                 661 
                 GACTAAGACA AGGACTCCTT TGTGGAGGTC CTGGCTTAGG GAGTCAAGTG ACGGCGGCTC 
               
               
                   
               
               
                 721 
                 AGCACTCACG TGGGCAGTGC CAGCCTCTAA GAGTGGGCAG GGGCACTGGC CACAGAGTCC 
               
               
                   
               
               
                 781 
                 CAGGGAGTCC CACCAGCCTA GTCGCCAGAC CGGCACCATG AGCAAGGGCG AGGAACTGTT 
               
               
                   
               
               
                 841 
                 CACTGGCGTG GTCCCAATTC TCGTGGAACT GGATGGCGAT GTGAATGGGC ACAAATTTTC 
               
               
                   
               
               
                 901 
                 TGTCAGCGGA GAGGGTGAAG GTGATGCCAC ATACGGAAAG CTCACCCTGA AATTCATCTG 
               
               
                   
               
               
                 961 
                 CACCACTGGA AAGCTCCCTG TGCCATGGCC AACACTGGTC ACTACCCTGA CCTATGGCGT 
               
               
                   
               
               
                 1021 
                 GCAGTGCTTT TCCAGATACC CAGACCATAT GAAGCAGCAT GACTTTTTCA AGAGCGCCAT 
               
               
                   
               
               
                 1081 
                 GCCCGAGGGC TATGTGCAGG AGAGAACCAT CTTTTTCAAA GATGACGGGA ACTACAAGAC 
               
               
                   
               
               
                 1141 
                 CCGCGCTGAA GTCAAGTTCG AAGGTGACAC CCTGGTGAAT AGAATCGAGC TGAAGGGCAT 
               
               
                   
               
               
                 1201 
                 TGACTTTAAG GAGGATGGAA ACATTCTCGG CCACAAGCTG GAATACAACT ATAACTCCCA 
               
               
                   
               
               
                 1261 
                 CAATGTGTAC ATCATGGCCG ACAAGCAAAA GAATGGCATC AAGGTCAACT TCAAGATCAG 
               
               
                   
               
               
                 1321 
                 ACACAACATT GAGGATGGAT CCGTGCAGCT GGCCGACCAT TATCAACAGA ACACTCCAAT 
               
               
                   
               
               
                 1381 
                 CGGCGACGGC CCTGTGCTCC TCCCAGACAA CCATTACCTG TCCACCCAGT CTGCCCTGTC 
               
               
                   
               
               
                 1441 
                 TAAAGATCCC AACGAAAAGA GAGACCACAT GGTCCTGCTG GAGTTTGTGA CCGCTGCTGG 
               
               
                   
               
               
                 1501 
                 GATCACACAT GGCATGGACG AGCTGTACAA GTGAAAGCTT ATCGATAATC AACCTCTGGA 
               
               
                   
               
               
                 1561 
                 TTACAAAATT TGTGAAAGAT TGACTGGTAT TCTTAACTAT GTTGCTCCTT TTACGCTATG 
               
               
                   
               
               
                 1621 
                 TGGATACGCT GCTTTAATGC CTTTGTATCA TGCTATTGCT TCCCGTATGG CTTTCATTTT 
               
               
                   
               
               
                 1681 
                 CTCCTCCTTG TATAAATCCT GGTTGCTGTC TCTTTATGAG GAGTTGTGGC CCGTTGTCAG 
               
               
                   
               
               
                 1741 
                 GCAACGTGGC GTGGTGTGCA CTGTGTTTGC TGACGCAACC CCCACTGGTT GGGGCATTGC 
               
               
                   
               
               
                 1801 
                 CACCACCTGT CAGCTCCTTT CCGGGACTTT CGCTTTCCCC CTCCCTATTG CCACGGCGGA 
               
               
                   
               
               
                 1861 
                 ACTCATCGCC GCCTGCCTTG CCCGCTGCTG GACAGGGGCT CGGCTGTTGG GCACTGACAA 
               
               
                   
               
               
                 1921 
                 TTCCGTGGTG TTGTCGGGGA AATCATCGTC CTTTCCTTGG CTGCTCGCCT GTGTTGCCAC 
               
               
                   
               
               
                 1981 
                 CTGGATTCTG CGCGGGACGT CCTTCTGCTA CGTCCCTTCG GCCCTCAATC CAGCGGACCT 
               
               
                   
               
               
                 2041 
                 TCCTTCCCGC GGCCTGCTGC CGGCTCTGCG GCCTCTTCCG CGTCTTCGCC TTCGCCCTCA 
               
               
                   
               
               
                 2101 
                 GACGAGTCGG ATCTCCCTTT GGGCCGCCTC CCCGGCGGCC GCGCACCGTC GACTCGCTGA 
               
               
                   
               
               
                 2161 
                 TCAGCCTCGA CTGTGCCTTC TAGTTGCCAG CCATCTGTTG TTTGCCCCTC CCCCGTGCCT 
               
               
                   
               
               
                 2221 
                 TCCTTGACCC TGGAAGGTGC CACTCCCACT GTCCTTTCCT AATAAAATGA GGAAATTGCA 
               
               
                   
               
               
                 2281 
                 TCGCATTGTC TGAGTAGGTG TCATTCTATT CTGGGGGGTG GGGTGGGGCA GGACAGCAAG 
               
               
                   
               
               
                 2341 
                 GGGGAGGATT GGGAAGACAA TAGCAGGCAT GCTGGGGATG CGGTGGGCTC TATGGCTTCT 
               
               
                   
               
               
                 2401 
                 GAGGCGGAAA GAACCAGCTG GGGCTCGACT AGAGCATGGC TACGTAGATA AGTAGCATGG 
               
               
                   
               
               
                 2461 
                 CGGGTTAATC ATTAACTACA AGGAACCCCT AGTGATGGAG TTGGCCACTC CCTCTCTGCG 
               
               
                   
               
               
                 2521 
                 CGCTCGCTCG CTCACTGAGG CCGGGCGACC AAAGGTCGCC CGACGCCCGG GCGGCCTCAG 
               
               
                   
               
               
                 2581 
                 TGAGCGAGCG AGCGCGCAGA GCTTTTTGCA AAAGCCTAGG CCTCCAAAAA AGCCTCCTCA 
               
               
                   
               
               
                 2641 
                 CTACTTCTGG AATAGCTCAG AGGCCGAGGC GGCCTCGGCC TCTGCATAAA TAAAAAAAAT 
               
               
                   
               
               
                 2701 
                 TAGTCAGCCA TGGGGCGGAG AATGGGCGGA ACTGGGCGGA GTTAGGGGCG GGATGGGCGG 
               
               
                   
               
               
                 2761 
                 AGTTAGGGGC GGGACTATGG TTGCTGACTA ATTGAGATGC ATGCTTTGCA TACTTCTGCC 
               
               
                   
               
               
                 2821 
                 TGCTGGGGAG CCTGGGGACT TTCCACACCT GGTTGCTGAC TAATTGAGAT GCATGCTTTG 
               
               
                   
               
               
                 2881 
                 CATACTTCTG CCTGCTGGGG AGCCTGGGGA CTTTCCACAC CCTAACTGAC ACACATTCCA 
               
               
                   
               
               
                 2941 
                 CAGCTGCATT AATGAATCGG CCAACGCGCG GGGAGAGGCG GTTTGCGTAT TGGGCGCTCT 
               
               
                   
               
               
                 3001 
                 TCCGCTTCCT CGCTCACTGA CTCGCTGCGC TCGGTCGTTC GGCTGCGGCG AGCGGTATCA 
               
               
                   
               
               
                 3061 
                 GCTCACTCAA AGGCGGTAAT ACGGTTATCC ACAGAATCAG GGGATAACGC AGGAAAGAAC 
               
               
                   
               
               
                 3121 
                 ATGTGAGCAA AAGGCCAGCA AAAGGCCAGG AACCGTAAAA AGGCCGCGTT GCTGGCGTTT 
               
               
                   
               
               
                 3181 
                 TTCCATAGGC TCCGCCCCCC TGACGAGCAT CACAAAAATC GACGCTCAAG TCAGAGGTGG 
               
               
                   
               
               
                 3241 
                 CGAAACCCGA CAGGACTATA AAGATACCAG GCGTTTCCCC CTGGAAGCTC CCTCGTGCGC 
               
               
                   
               
               
                 3301 
                 TCTCCTGTTC CGACCCTGCC GCTTACCGGA TACCTGTCCG CCTTTCTCCC TTCGGGAAGC 
               
               
                   
               
               
                 3361 
                 GTGGCGCTTT CTCATAGCTC ACGCTGTAGG TATCTCAGTT CGGTGTAGGT CGTTCGCTCC 
               
               
                   
               
               
                 3421 
                 AAGCTGGGCT GTGTGCACGA ACCCCCCGTT CAGCCCGACC GCTGCGCCTT ATCCGGTAAC 
               
               
                   
               
               
                 3481 
                 TATCGTCTTG AGTCCAACCC GGTAAGACAC GACTTATCGC CACTGGCAGC AGCCACTGGT 
               
               
                   
               
               
                 3541 
                 AACAGGATTA GCAGAGCGAG GTATGTAGGC GGTGCTACAG AGTTCTTGAA GTGGTGGCCT 
               
               
                   
               
               
                 3601 
                 AACTACGGCT ACACTAGAAG AACAGTATTT GGTATCTGCG CTCTGCTGAA GCCAGTTACC 
               
               
                   
               
               
                 3661 
                 TTCGGAAAAA GAGTTGGTAG CTCTTGATCC GGCAAACAAA CCACCGCTGG TAGCGGTGGT 
               
               
                   
               
               
                 3721 
                 TTTTTTGTTT GCAAGCAGCA GATTACGCGC AGAAAAAAAG GATCTCAAGA AGATCCTTTG 
               
               
                   
               
               
                 3781 
                 ATCTTTTCTA CGGGGTCTGA CGCTCAGTGG AACGAAAACT CACGTTAAGG GATTTTGGTC 
               
               
                   
               
               
                 3841 
                 ATGAGATTAT CAAAAAGGAT CTTCACCTAG ATCCTTTTAA ATTAAAAATG AAGTTTTAAA 
               
               
                   
               
               
                 3901 
                 TCAATCTAAA GTATATATGA GTAAACTTGG TCTGACAGTT ACCAATGCTT AATCAGTGAG 
               
               
                   
               
               
                 3961 
                 GCACCTATCT CAGCGATCTG TCTATTTCGT TCATCCATAG TTGCCTGACT CCTGCAAACC 
               
               
                   
               
               
                 4021 
                 ACGTTGTGTC TCAAAATCTC TGATGTTACA TTGCACAAGA TAAAAATATA TCATCATGAA 
               
               
                   
               
               
                 4081 
                 CAATAAAACT GTCTGCTTAC ATAAACAGTA ATACAAGGGG TGTTATGAGC CATATTCAAC 
               
               
                   
               
               
                 4141 
                 GGGAAACGTC TTGCTCGAGG CCGCGATTAA ATTCCAACAT GGATGCTGAT TTATATGGGT 
               
               
                   
               
               
                 4201 
                 ATAAATGGGC TCGCGATAAT GTCGGGCAAT CAGGTGCGAC AATCTATCGA TTGTATGGGA 
               
               
                   
               
               
                 4261 
                 AGCCCGATGC GCCAGAGTTG TTTCTGAAAC ATGGCAAAGG TAGCGTTGCC AATGATGTTA 
               
               
                   
               
               
                 4321 
                 CAGATGAGAT GGTCAGACTA AACTGGCTGA CGGAATTTAT GCCTCTTCCG ACCATCAAGC 
               
               
                   
               
               
                 4381 
                 ATTTTATCCG TACTCCTGAT GATGCATGGT TACTCACCAC TGCGATCCCC GGGAAAACAG 
               
               
                   
               
               
                 4441 
                 CATTCCAGGT ATTAGAAGAA TATCCTGATT CAGGTGAAAA TATTGTTGAT GCGCTGGCAG 
               
               
                   
               
               
                 4501 
                 TGTTCCTGCG CCGGTTGCAT TCGATTCCTG TTTGTAATTG TCCTTTTAAC AGCGATCGCG 
               
               
                   
               
               
                 4561 
                 TATTTCGTCT CGCTCAGGCG CAATCACGAA TGAATAACGG TTTGGTTGAT GCGAGTGATT 
               
               
                   
               
               
                 4621 
                 TTGATGACGA GCGTAATGGC TGGCCTGTTG AACAAGTCTG GAAAGAAATG CATAAGCTTT 
               
               
                   
               
               
                 4681 
                 TGCCATTCTC ACCGGATTCA GTCGTCACTC ATGGTGATTT CTCACTTGAT AACCTTATTT 
               
               
                   
               
               
                 4741 
                 TTGACGAGGG GAAATTAATA GGTTGTATTG ATGTTGGACG AGTCGGAATC GCAGACCGAT 
               
               
                   
               
               
                 4801 
                 ACCAGGATCT TGCCATCCTA TGGAACTGCC TCGGTGAGTT TTCTCCTTCA TTACAGAAAC 
               
               
                   
               
               
                 4861 
                 GGCTTTTTCA AAAATATGGT ATTGATAATC CTGATATGAA TAAATTGCAG TTTCATTTGA 
               
               
                   
               
               
                 4921 
                 TGCTCGATGA GTTTTTCTAA GGGCGGCCTG CCACCATACC CACGCCGAAA CAAGCGCTCA 
               
               
                   
               
               
                 4981 
                 TGAGCCCGAA GTGGCGAGCC CGATCTTCCC CATCGGTGAT GTCGGCGATA TAGGCGCCAG 
               
               
                   
               
               
                 5041 
                 CAACCGCACC TGTGGCGCCG GTGATGCCGG CCACGATGCG TCCGGCGTAG AGGATCTGGC 
               
               
                   
               
               
                 5101 
                 TAGCGATGAC CCTGCTGATT GGTTCGCTGA CCATTTCCGG GTGCGGGACG GCGTTACCAG 
               
               
                   
               
               
                 5161 
                 AAACTCAGAA GGTTCGTCCA ACCAAACCGA CTCTGACGGC AGTTTACGAG AGAGATGATA 
               
               
                   
               
               
                 5221 
                 GGGTCTGCTT CAGGGTGACC GATGTAACCA TATACTTAGG CTGGATCTTC TCCCGCGAAT 
               
               
                   
               
               
                 5281 
                 TTTAACCCTC ACCAACTACG AGATATGAGG TAAGCCAAAA AAGCACGTAG TGGCGCTCTC 
               
               
                   
               
               
                 5341 
                 CGACTGTTCC CAAATTGTAA CTTATCGTTC CGTGAAGGCC AGAGTTACTT CCCGGCCCTT 
               
               
                   
               
               
                 5401 
                 TCCATGCGCG CACCATACCC TCCTAGTTCC CCGGTTATCT TTCCGAAGTG GGAGTGAGCG 
               
               
                   
               
               
                 5461 
                 AACCTCCGTT TACGTCTTGT TACCAATGAT GTAGCTATGC ACTTTGTACA GGGTGCCAAC 
               
               
                   
               
               
                 5521 
                 GGGTTTCACA ATTCACAGAT AGTGGGGATC CCGGCAAAGG GCCTATATTT GCGGTCCAAC 
               
               
                   
               
               
                 5581 
                 TTAGGCGTAA ACCTCGATGC TACCTACTCA GACCCACCTC GCGCGGGGTA AATAAGGCAC 
               
               
                   
               
               
                 5641 
                 TCATCCCAGC TGGTTCTTGG CGTTCTACGC AGCGACATGT TTATTAACAG TTGTCTGGCA 
               
               
                   
               
               
                 5701 
                 GCACAAAACT TTTACCATGG TCGTAGAAGC CCCCCAGAGT TAGTTCATAC CTAATGCCAC 
               
               
                   
               
               
                 5761 
                 AAATGTGACA GGACGCCGAT GGGTACCGGA CTTTAGGTCG AGCACAGTTC GGTAACGGAG 
               
               
                   
               
               
                 5821 
                 AGACCCTGCG GCGTACTTCA TTATGTATAT GGAACGTGCC CAAGTGACGC CAGGCAAGTC 
               
               
                   
               
               
                 5881 
                 TCAGCTGGTT CCTGTGTTAG CTCGAGGGTA GACATACGAG CTGATTGAAC ATGGGTTGGG 
               
               
                   
               
               
                 5941 
                 GGCCTCGAAC CGTCGAGGAC CCCATAGTAC CTCGGAGACC AAGTAGGGCA GCCTATAGTT 
               
               
                   
               
               
                 6001 
                 TGAAGCAGAA CTATTTCGGG GGGCGAGCCC TCATCGTCTC TTCTGCGGAT GACTCAACAC 
               
               
                   
               
               
                 6061 
                 GCTAGGGACG TGAAGTCGAT TCCTTCGATG GTTATAAATC AAAGACTCAG AGTGCTGTCT 
               
               
                   
               
               
                 6121 
                 GGAGCGTGAA TCTAACGGTA CGTATCTCGA TTGCTCGGTC GCTTTTCGCA CTCCGCGAAA 
               
               
                   
               
               
                 6181 
                 GTTCGTACCG CTCATTCACT AGGTTGCGAA GCCTATGCTG ATATATGAAT CCAAACTAGA 
               
               
                   
               
               
                 6241 
                 GCAGGGCTCT TAAGATTCGG AGTTGTAAAT ACTTAATACT CCAATCGGCT TTTACGTGCA 
               
               
                   
               
               
                 6301 
                 CCACCGCGGG CGGCTGACAA GGGTCTCACA TCGAGAAACA AGACAGTTCC GGGCTGGAAG 
               
               
                   
               
               
                 6361 
                 TAGCGCCGGC TAAGGAAGAC GCCTGGTACG GCAGGACTAT GAAACCAGTA CAAAGGCAAC 
               
               
                   
               
               
                 6421 
                 ATCCTCACTT GGGTGAACGG AAACGCAGTA TTATGGTTAC TTTTTGGATA CGTGAAACAT 
               
               
                   
               
               
                 6481 
                 ATCCCATGGT AGTCCTTAGA CTTGGGAGTC TATCACCCCT AGGGCCCATA TCTGGAAATA 
               
               
                   
               
               
                 6541 
                 GACGCCAGGT TGAATCCGTA TTTGGAGGTA CGATGGAACA GTCTGGGTGG GACGTGCTTC 
               
               
                   
               
               
                 6601 
                 ATTTATACCC TGCGCAGGCT GGACCGAGGA CCGCAAGGTG CGGCGGTGCA CAAGCAATTG 
               
               
                   
               
               
                 6661 
                 ACAACTAACC ACCGTGTATT CATTATGGTA CCAGGAACTT TAAGCCGAGT CAATGAAGCT 
               
               
                   
               
               
                 6721 
                 CGCATTACAG TGTTTACCGC ATCTTGCCGT TACTCACAAA CTGTGATCCA CCACAAGTCA 
               
               
                   
               
               
                 6781 
                 AGCCATTGCC TCTCTGACAC GCCGTAAGAA TTAATATGTA AACTTTGCGC GGGTTGACTG 
               
               
                   
               
               
                 6841 
                 CGATCCGTTC AGTCTCGTCC GAGGGCACAA TCCTATTCCC ATTTGTATGT TCAGCTAACT 
               
               
                   
               
               
                 6901 
                 TCTACCCATC CCCCGAAGTT AAGTAGGTCG TGAGATGCCA TGGAGGCTCT CGTTCATCCC 
               
               
                   
               
               
                 6961 
                 GTGGGACATC AAGCTTCCCC TTGATAAAGC ACCCCGCTCG GGTGTAGCAG AGAAGACGCC 
               
               
                   
               
               
                 7021 
                 TTCTGAATTG TGCAATCCCT CCACCTTATC TAAGCTTGCT ACCAATAATT AGCATTTTTG 
               
               
                   
               
               
                 7081 
                 CCTTGCGACA GACCTCCTAC TTAGATTGCC ACACATTGAG CTAGTCAGTG AGCGATAAGC 
               
               
                   
               
               
                 7141 
                 TTGACGCGCT TTCAAGGGTC GCGAGTACGT GAACTAAGGC TCCGGACAGG ACTATATACT 
               
               
                   
               
               
                 7201 
                 TGGGTTTGAT CTCGCCCCGA CAACTGCAAA CCTCAACTTT TTTAGATTAT ATGGTTAGCC 
               
               
                   
               
               
                 7261 
                 GAAGTTGCAC GAGGTGGCGT CCGCGGACTG CTCCCCGAGT GTGGCTCTTT CATCTGACAA 
               
               
                   
               
               
                 7321 
                 CGTGCAACCC CTATCGCGGC CGATTGTTTC TGCGGACGAT GTTGTCCTCA TAGTTTGGGC 
               
               
                   
               
               
                 7381 
                 ATGTTTCCCT TGTAGGTGTG AAACCACTTA GCTTCGCGCC GTAGTCCCAA TGAAAAACCT 
               
               
                   
               
               
                 7441 
                 ATGGACTTTG TTTTGGGTAG CACCAGGAAT CTGAACCGTG TGAATGTGGA CGTCGCGCGC 
               
               
                   
               
               
                 7501 
                 GTAGACCTTT ATCTCCGGTT CAAGCTAGGG ATGTGGCTGC ATGCTACGTT GTCACACCTA 
               
               
                   
               
               
                 7561 
                 CACTGCTCGA AGTAAATATG CGAAGCGCGC GGCCTGGCCG GAGGCGTTCC GCGCCGCCAC 
               
               
                   
               
               
                 7621 
                 GTGTTCGTTA ACTGTTGATT GGTGGCACAT AAGCAATATC GTAGTCCGTC AAATTCAGCT 
               
               
                   
               
               
                 7681 
                 CTGTTATCCC GGGCGTTATG TGTCAAATGG CGTAGAACGG GATTGACTGT TTGACGGTAG 
               
               
                   
               
               
                 7741 
                 GGTGACCTAA GCCAGATGCT ACACAATTAG GCTTGTACAT ATTGTCGTTA GAACGCGGCT 
               
               
                   
               
               
                 7801 
                 ACAATTAATA CATAACCTTA TGTATCATAC ACATACGATT TAGGTGACAC TATAGAATAC 
               
               
                   
               
               
                 7861 
                 ACGGAATTAA TTC. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 4 
               
             
            
               
                   
               
               
                 Features of the VMD2.GFP.WPRE.pA plasmid sequence 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                 Mini- 
                 Maxi- 
                   
                 Direc- 
               
               
                 Name 
                 Type 
                 mum 
                 mum 
                 Length 
                 tion 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Randomly 
                 Stuffer 
                 5,241 
                 7,740 
                 2,500 
                 none 
               
               
                 generated 
               
               
                 stuffer sequence 
               
               
                 AphR (KanR) 
                 CDS 
                 4,125 
                 4,940 
                 816 
                 forward 
               
               
                 pBR322 rep 
                 rep_origin 
                 3,165 
                 3,784 
                 620 
                 reverse 
               
               
                 origin 
               
               
                 AAV2 ITR 
                 LTR 
                 2,481 
                 2,601 
                 121 
                 reverse 
               
               
                 bGH pA 
                 polyA_signal 
                 2,155 
                 2,423 
                 269 
                 forward 
               
               
                 WPRE 
                 WPRE 
                 1,547 
                 2,136 
                 590 
                 forward 
               
               
                 GFP 
                 misc_feature 
                 818 
                 1,534 
                 717 
                 forward 
               
               
                 Kozak 
                 Kozak 
                 812 
                 817 
                 6 
                 forward 
               
               
                 −585 to +38 
                 promoter 
                 189 
                 811 
                 623 
                 forward 
               
               
                 VMD2 promoter 
               
               
                 AAV2 ITR 
                 LTR 
                 4 
                 133 
                 130 
                 forward 
               
               
                   
               
            
           
         
       
     
     An exemplary a vector of the disclosure comprising a marker includes VMD. IntEx.GFP.WPRE.pA. A nucleic acid sequence encoding a VMD. IntEx.GFP.WPRE.pA construct comprises or consists of: 
     
       
         
           
               
               
            
               
                 (SEQ ID NO: 17) 
                   
               
            
           
           
               
               
               
            
               
                 1 
                 TAGCTGCGCG CTCGCTCGCT CACTGAGGCC GCCCGGGCAA AGCCCGGGCG TCGGGCGACC 
                   
               
               
                   
               
               
                 61 
                 TTTGGTCGCC CGGCCTCAGT GAGCGAGCGA GCGCGCAGAG AGGGAGTGGC CAACTCCATC 
               
               
                   
               
               
                 121 
                 ACTAGGGGTT CCTTGTAGTT AATGATTAAC CCGCCATGCT ACTTATCTAC GTAGCCATGC 
               
               
                   
               
               
                 181 
                 TCTAGGTAAA TTCTGTCATT TTACTAGGGT GATGAAATTC CCAAGCAACA CCATCCTTTT 
               
               
                   
               
               
                 241 
                 CAGATAAGGG CACTGAGGCT GAGAGAGGAG CTGAAACCTA CCCGGGGTCA CGAGACACAG 
               
               
                   
               
               
                 301 
                 GTGGCAAGGC TGGGACCAGA AACCAGGACT GTTGACTGCA GCCCGGTATT CATTCTTTCC 
               
               
                   
               
               
                 361 
                 ATAGCCCACA GGGCTGTCAA AGACCCCAGG GCCTAGTCAG AGGCTCCTCC TTCCTGGAGA 
               
               
                   
               
               
                 421 
                 GTTCCTGGCA CAGAAGTTGA AGCTCAGCAC AGCCCCCTAA CCCCCAACTC TCTCTGCAAG 
               
               
                   
               
               
                 481 
                 GCCTCAGGGG TCAGAACACT GGTGGAGCAG ATCCTTTAGC CTCTGGATTT TAGGGCCATG 
               
               
                   
               
               
                 541 
                 GTAGAGGGGG TGTTGCCCTA AATTCCAGCC CTGGTCTCAG CCCAACACCC TCCAAGAAGA 
               
               
                   
               
               
                 601 
                 AATTAGAGGG GCCATGGCCA GGCTGTGCTA GCCGTTGCTT CTGAGCAGAT TACAAGAAGG 
               
               
                   
               
               
                 661 
                 GACTAAGACA AGGACTCCTT TGTGGAGGTC CTGGCTTAGG GAGTCAAGTG ACGGCGGCTC 
               
               
                   
               
               
                 721 
                 AGCACTCACG TGGGCAGTGC CAGCCTCTAA GAGTGGGCAG GGGCACTGGC CACAGAGTCC 
               
               
                   
               
               
                 781 
                 CAGGGAGTCC CACCAGCCTA GTCGCCAGAC CGGGTGCCGC AGGGGGACGG CTGCCTTCGG 
               
               
                   
               
               
                 841 
                 GGGGGACGGG GCAGGGCGGG GTTCGGCTTC TGGCGTGTGA CCGGCGGCTC TAGAGCCTCT 
               
               
                   
               
               
                 901 
                 GCTAACCATG TTCATGCCTT CTTCTTTTTC CTACAGCTCC TGGGCAACGT GCTGGTTATT 
               
               
                   
               
               
                 961 
                 GTGCTGTCTC ATCATTTTGG CAAAGAATTG GCACCATGAG CAAGGGCGAG GAACTGTTCA 
               
               
                   
               
               
                 1021 
                 CTGGCGTGGT CCCAATTCTC GTGGAACTGG ATGGCGATGT GAATGGGCAC AAATTTTCTG 
               
               
                   
               
               
                 1081 
                 TCAGCGGAGA GGGTGAAGGT GATGCCACAT ACGGAAAGCT CACCCTGAAA TTCATCTGCA 
               
               
                   
               
               
                 1141 
                 CCACTGGAAA GCTCCCTGTG CCATGGCCAA CACTGGTCAC TACCCTGACC TATGGCGTGC 
               
               
                   
               
               
                 1201 
                 AGTGCTTTTC CAGATACCCA GAGCATATGA AGCAGCATGA CTTTTTCAAG AGCGCCATGC 
               
               
                   
               
               
                 1261 
                 CCGAGGGCTA TGTGCAGGAG AGAACCATCT TTTTCAAAGA TGACGGGAAC TACAAGACCC 
               
               
                   
               
               
                 1321 
                 GCGCTGAAGT CAAGTTCGAA GGTGACACCC TGGTGAATAG AATCGAGCTG AAGGGCATTG 
               
               
                   
               
               
                 1381 
                 ACTTTAAGGA GGATGGAAAC ATTCTCGGCC ACAAGCTGGA ATACAACTAT AACTCCCACA 
               
               
                   
               
               
                 1441 
                 ATGTGTACAT CATGGCCGAC AAGCAAAAGA ATGGCATCAA GGTCAACTTC AAGATCAGAC 
               
               
                   
               
               
                 1501 
                 ACAACATTGA GGATGGATCC GTGCAGCTGG CCGACCATTA TCAACAGAAC ACTCCAATCG 
               
               
                   
               
               
                 1561 
                 GCGACGGCCC TGTGCTCCTC CCAGACAACC ATTACCTGTC CACCCAGTCT GCCCTGTCTA 
               
               
                   
               
               
                 1621 
                 AAGATCCCAA CGAAAAGAGA GACCACATGG TCCTGCTGGA GTTTGTGACC GCTGCTGGGA 
               
               
                   
               
               
                 1681 
                 TCACACATGG CATGGACGAG CTGTACAAGT GAAAGCTTAT CGATAATCAA CCTCTGGATT 
               
               
                   
               
               
                 1741 
                 ACAAAATTTG TGAAAGATTG ACTGGTATTC TTAACTATGT TGCTCCTTTT ACGCTATGTG 
               
               
                   
               
               
                 1801 
                 GATACGCTGC TTTAATGCCT TTGTATCATG CTATTGCTTC CCGTATGGCT TTCATTTTCT 
               
               
                   
               
               
                 1861 
                 CCTCCTTGTA TAAATCCTGG TTGCTGTCTC TTTATGAGGA GTTGTGGCCC GTTGTCAGGC 
               
               
                   
               
               
                 1921 
                 AACGTGGCGT GGTGTGCACT GTGTTTGCTG ACGCAACCCC CACTGGTTGG GGCATTGCCA 
               
               
                   
               
               
                 1981 
                 CCACCTGTCA GCTCCTTTCC GGGACTTTCG CTTTCCCCCT CCCTATTGCC ACGGCGGAAC 
               
               
                   
               
               
                 2041 
                 TCATCGCCGC CTGCCTTGCC CGCTGCTGGA CAGGGGCTCG GCTGTTGGGC ACTGACAATT 
               
               
                   
               
               
                 2101 
                 CCGTGGTGTT GTCGGGGAAA TCATCGTCCT TTCCTTGGCT GCTCGCCTGT GTTGCCACCT 
               
               
                   
               
               
                 2161 
                 GGATTCTGCG CGGGACGTCC TTCTGCTACG TCCCTTCGGC CCTCAATCCA GCGGACCTTC 
               
               
                   
               
               
                 2221 
                 CTTCCCGCGG CCTGCTGCCG GCTCTGCGGC CTCTTCCGCG TCTTCGCCTT CGCCCTCAGA 
               
               
                   
               
               
                 2281 
                 CGAGTCGGAT CTCCCTTTGG GCCGCCTCCC CGGCGGCCGC GCACCGTCGA CTCGCTGATC 
               
               
                   
               
               
                 2341 
                 AGCCTCGACT GTGCCTTCTA GTTGCCAGCC ATCTGTTGTT TGCCCCTCCC CCGTGCCTTC 
               
               
                   
               
               
                 2401 
                 CTTGACCCTG GAAGGTGCCA CTCCCACTGT CCTTTCCTAA TAAAATGAGG AAATTGCATC 
               
               
                   
               
               
                 2461 
                 GCATTGTCTG AGTAGGTGTC ATTCTATTCT GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG 
               
               
                   
               
               
                 2521 
                 GGAGGATTGG GAAGACAATA GCAGGCATGC TGGGGATGCG GTGGGCTCTA TGGCTTCTGA 
               
               
                   
               
               
                 2581 
                 GGCGGAAAGA ACCAGCTGGG GCTCGACTAG AGCATGGCTA CGTAGATAAG TAGCATGGCG 
               
               
                   
               
               
                 2641 
                 GGTTAATCAT TAACTACAAG GAACCCCTAG TGATGGAGTT GGCCACTCCC TCTCTGCGCG 
               
               
                   
               
               
                 2701 
                 CTCGCTCGCT CACTGAGGCC GGGCGACCAA AGGTCGCCCG ACGCCCGGGC GGCCTCAGTG 
               
               
                   
               
               
                 2761 
                 AGCGAGCGAG CGCGCAGAGC TTTTTGCAAA AGCCTAGGCC TCCAAAAAAG CCTCCTCACT 
               
               
                   
               
               
                 2821 
                 ACTTCTGGAA TAGCTCAGAG GCCGAGGCGG CCTCGGCCTC TGCATAAATA AAAAAAATTA 
               
               
                   
               
               
                 2881 
                 GTCAGCCATG GGGCGGAGAA TGGGCGGAAC TGGGCGGAGT TAGGGGCGGG ATGGGCGGAG 
               
               
                   
               
               
                 2941 
                 TTAGGGGCGG GACTATGGTT GCTGACTAAT TGAGATGCAT GCTTTGCATA CTTCTGCCTG 
               
               
                   
               
               
                 3001 
                 CTGGGGAGCC TGGGGACTTT CCACACCTGG TTGCTGACTA ATTGAGATGC ATGCTTTGCA 
               
               
                   
               
               
                 3061 
                 TACTTCTGCC TGCTGGGGAG CCTGGGGACT TTCCACACCC TAACTGACAC ACATTCCACA 
               
               
                   
               
               
                 3121 
                 GCTGCATTAA TGAATCGGCC AACGCGCGGG GAGAGGCGGT TTGCGTATTG GGCGCTCTTC 
               
               
                   
               
               
                 3181 
                 CGCTTCCTCG CTCACTGACT CGCTGCGCTC GGTCGTTCGG CTGCGGCGAG CGGTATCAGC 
               
               
                   
               
               
                 3241 
                 TCACTCAAAG GCGGTAATAC GGTTATCCAC AGAATCAGGG GATAACGCAG GAAAGAACAT 
               
               
                   
               
               
                 3301 
                 GTGAGCAAAA GGCCAGCAAA AGGCCAGGAA CCGTAAAAAG GCCGCGTTGC TGGCGTTTTT 
               
               
                   
               
               
                 3361 
                 CCATAGGCTC CGCCCCCCTG ACGAGCATCA CAAAAATCGA CGCTCAAGTC AGAGGTGGCG 
               
               
                   
               
               
                 3421 
                 AAACCCGACA GGACTATAAA GATACCAGGC GTTTCCCCCT GGAAGCTCCC TCGTGCGCTC 
               
               
                   
               
               
                 3481 
                 TCCTGTTCCG ACCCTGCCGC TTACCGGATA CCTGTCCGCC TTTCTCCCTT CGGGAAGCGT 
               
               
                   
               
               
                 3541 
                 GGCGCTTTCT CATAGCTCAC GCTGTAGGTA TCTCAGTTCG GTGTAGGTCG TTCGCTCCAA 
               
               
                   
               
               
                 3601 
                 GCTGGGCTGT GTGCACGAAC CCCCCGTTCA GCCCGACCGC TGCGCCTTAT CCGGTAACTA 
               
               
                   
               
               
                 3661 
                 TCGTCTTGAG TCCAACCCGG TAAGACACGA CTTATCGCCA CTGGCAGCAG CCACTGGTAA 
               
               
                   
               
               
                 3721 
                 CAGGATTAGC AGAGCGAGGT ATGTAGGCGG TGCTACAGAG TTCTTGAAGT GGTGGCCTAA 
               
               
                   
               
               
                 3781 
                 CTACGGCTAC ACTAGAAGAA CAGTATTTGG TATCTGCGCT CTGCTGAAGC CAGTTACCTT 
               
               
                   
               
               
                 3841 
                 CGGAAAAAGA GTTGGTAGCT CTTGATCCGG CAAACAAACC ACCGCTGGTA GCGGTGGTTT 
               
               
                   
               
               
                 3901 
                 TTTTGTTTGC AAGCAGCAGA TTACGCGCAG AAAAAAAGGA TCTCAAGAAG ATCCTTTGAT 
               
               
                   
               
               
                 3961 
                 CTTTTCTACG GGGTCTGACG CTCAGTGGAA CGAAAACTCA CGTTAAGGGA TTTTGGTCAT 
               
               
                   
               
               
                 4021 
                 GAGATTATCA AAAAGGATCT TCACCTAGAT CCTTTTAAAT TAAAAATGAA GTTTTAAATC 
               
               
                   
               
               
                 4081 
                 AATCTAAAGT ATATATGAGT AAACTTGGTC TGACAGTTAC CAATGCTTAA TCAGTGAGGC 
               
               
                   
               
               
                 4141 
                 ACCTATCTCA GCGATCTGTC TATTTCGTTC ATCCATAGTT GCCTGACTCC TGCAAACCAC 
               
               
                   
               
               
                 4201 
                 GTTGTGTCTC AAAATCTCTG ATGTTACATT GCACAAGATA AAAATATATC ATCATGAACA 
               
               
                   
               
               
                 4261 
                 ATAAAACTGT CTGCTTACAT AAACAGTAAT ACAAGGGGTG TTATGAGCCA TATTCAACGG 
               
               
                   
               
               
                 4321 
                 GAAACGTCTT GCTCGAGGCC GCGATTAAAT TCCAACATGG ATGCTGATTT ATATGGGTAT 
               
               
                   
               
               
                 4381 
                 AAATGGGCTC GCGATAATGT CGGGCAATCA GGTGCGACAA TCTATCGATT GTATGGGAAG 
               
               
                   
               
               
                 4441 
                 CCCGATGCGC CAGAGTTGTT TCTGAAACAT GGCAAAGGTA GCGTTGCCAA TGATGTTACA 
               
               
                   
               
               
                 4501 
                 GATGAGATGG TCAGACTAAA CTGGCTGACG GAATTTATGC CTCTTCCGAC CATCAAGCAT 
               
               
                   
               
               
                 4561 
                 TTTATCCGTA CTCCTGATGA TGCATGGTTA CTCACCACTG CGATCCCCGG GAAAACAGCA 
               
               
                   
               
               
                 4621 
                 TTCCAGGTAT TAGAAGAATA TCCTGATTCA GGTGAAAATA TTGTTGATGC GCTGGCAGTG 
               
               
                   
               
               
                 4681 
                 TTCCTGCGCC GGTTGCATTC GATTCCTGTT TGTAATTGTC CTTTTAACAG CGATCGCGTA 
               
               
                   
               
               
                 4741 
                 TTTCGTCTCG CTCAGGCGCA ATCACGAATG AATAACGGTT TGGTTGATGC GAGTGATTTT 
               
               
                   
               
               
                 4801 
                 GATGACGAGC GTAATGGCTG GCCTGTTGAA CAAGTCTGGA AAGAAATGCA TAAGCTTTTG 
               
               
                   
               
               
                 4861 
                 CCATTCTCAC CGGATTCAGT CGTCACTCAT GGTGATTTCT CACTTGATAA CCTTATTTTT 
               
               
                   
               
               
                 4921 
                 GACGAGGGGA AATTAATAGG TTGTATTGAT GTTGGACGAG TCGGAATCGC AGACCGATAC 
               
               
                   
               
               
                 4981 
                 CAGGATCTTG CCATCCTATG GAACTGCCTC GGTGAGTTTT CTCCTTCATT ACAGAAACGG 
               
               
                   
               
               
                 5041 
                 CTTTTTCAAA AATATGGTAT TGATAATCCT GATATGAATA AATTGCAGTT TCATTTGATG 
               
               
                   
               
               
                 5101 
                 CTCGATGAGT TTTTCTAAGG GCGGCCTGCC ACCATACCCA CGCCGAAACA AGCGCTCATG 
               
               
                   
               
               
                 5161 
                 AGCCCGAAGT GGCGAGCCCG ATCTTCCCCA TCGGTGATGT CGGCGATATA GGCGCCAGCA 
               
               
                   
               
               
                 5221 
                 ACCGCACCTG TGGCGCCGGT GATGCCGGCC ACGATGCGTC CGGCGTAGAG GATCTGGCTA 
               
               
                   
               
               
                 5281 
                 GCGATGACCC TGCTGATTGG TTCGCTGACC ATTTCCGGGT GCGGGACGGC GTTACCAGAA 
               
               
                   
               
               
                 5341 
                 ACTCAGAAGG TTCGTCCAAC CAAACCGACT CTGACGGCAG TTTACGAGAG AGATGATAGG 
               
               
                   
               
               
                 5401 
                 GTCTGCTTCA GGGTGACCGA TGTAACCATA TACTTAGGCT GGATCTTCTC CCGCGAATTT 
               
               
                   
               
               
                 5461 
                 TAACCCTCAC CAACTACGAG ATATGAGGTA AGCCAAAAAA GCACGTAGTG GCGCTCTCCG 
               
               
                   
               
               
                 5521 
                 ACTGTTCCCA AATTGTAACT TATCGTTCCG TGAAGGCCAG AGTTACTTCC CGGCCCTTTC 
               
               
                   
               
               
                 5581 
                 CATGCGCGCA CCATACCCTC CTAGTTCCCC GGTTATCTTT CCGAAGTGGG AGTGAGCGAA 
               
               
                   
               
               
                 5641 
                 CCTCCGTTTA CGTCTTGTTA CCAATGATGT AGCTATGCAC TTTGTACAGG GTGCCAACGG 
               
               
                   
               
               
                 5701 
                 GTTTCACAAT TCACAGATAG TGGGGATCCC GGCAAAGGGC CTATATTTGC GGTCCAACTT 
               
               
                   
               
               
                 5761 
                 AGGCGTAAAC CTCGATGCTA CCTACTCAGA CCCACCTCGC GCGGGGTAAA TAAGGCACTC 
               
               
                   
               
               
                 5821 
                 ATCCCAGCTG GTTCTTGGCG TTCTACGCAG CGACATGTTT ATTAACAGTT GTCTGGCAGC 
               
               
                   
               
               
                 5881 
                 ACAAAACTTT TACCATGGTC GTAGAAGCCC CCCAGAGTTA GTTCATACCT AATGCCACAA 
               
               
                   
               
               
                 5941 
                 ATGTGACAGG ACGCCGATGG GTACCGGACT TTAGGTCGAG CACAGTTCGG TAACGGAGAG 
               
               
                   
               
               
                 6001 
                 ACCCTGCGGC GTACTTCATT ATGTATATGG AACGTGCCCA AGTGACGCCA GGCAAGTCTC 
               
               
                   
               
               
                 6061 
                 AGCTGGTTCC TGTGTTAGCT CGAGGGTAGA CATACGAGCT GATTGAACAT GGGTTGGGGG 
               
               
                   
               
               
                 6121 
                 CCTCGAACCG TCGAGGACCC CATAGTACCT CGGAGACCAA GTAGGGCAGC CTATAGTTTG 
               
               
                   
               
               
                 6181 
                 AAGCAGAACT ATTTCGGGGG GCGAGCCCTC ATCGTCTCTT CTGCGGATGA CTCAACACGC 
               
               
                   
               
               
                 6241 
                 TAGGGACGTG AAGTCGATTC CTTCGATGGT TATAAATCAA AGACTCAGAG TGCTGTCTGG 
               
               
                   
               
               
                 6301 
                 AGCGTGAATC TAACGGTACG TATCTCGATT GCTCGGTCGC TTTTCGCACT CCGCGAAAGT 
               
               
                   
               
               
                 6361 
                 TCGTACCGCT CATTCACTAG GTTGCGAAGC CTATGCTGAT ATATGAATCC AAACTAGAGC 
               
               
                   
               
               
                 6421 
                 AGGGCTCTTA AGATTCGGAG TTGTAAATAC TTAATACTCC AATCGGCTTT TACGTGCACC 
               
               
                   
               
               
                 6481 
                 ACCGCGGGCG GCTGACAAGG GTCTCACATC GAGAAACAAG ACAGTTCCGG GCTGGAAGTA 
               
               
                   
               
               
                 6541 
                 GCGCCGGCTA AGGAAGACGC CTGGTACGGC AGGACTATGA AACCAGTACA AAGGCAACAT 
               
               
                   
               
               
                 6601 
                 CCTCACTTGG GTGAACGGAA ACGCAGTATT ATGGTTACTT TTTGGATACG TGAAACATAT 
               
               
                   
               
               
                 6661 
                 CCCATGGTAG TCCTTAGACT TGGGAGTCTA TCACCCCTAG GGCCCATATC TGGAAATAGA 
               
               
                   
               
               
                 6721 
                 CGCCAGGTTG AATCCGTATT TGGAGGTACG ATGGAACAGT CTGGGTGGGA CGTGCTTCAT 
               
               
                   
               
               
                 6781 
                 TTATACCCTG CGCAGGCTGG ACCGAGGACC GCAAGGTGCG GCGGTGCACA AGCAATTGAC 
               
               
                   
               
               
                 6841 
                 AACTAACCAC CGTGTATTCA TTATGGTACC AGGAACTTTA AGCCGAGTCA ATGAAGCTCG 
               
               
                   
               
               
                 6901 
                 CATTACAGTG TTTACCGCAT CTTGCCGTTA CTCACAAACT GTGATCCACC ACAAGTCAAG 
               
               
                   
               
               
                 6961 
                 CCATTGCCTC TCTGACACGC CGTAAGAATT AATATGTAAA CTTTGCGCGG GTTGACTGCG 
               
               
                   
               
               
                 7021 
                 ATCCGTTCAG TCTCGTCCGA GGGCACAATC CTATTCCCAT TTGTATGTTC AGCTAACTTC 
               
               
                   
               
               
                 7081 
                 TACCCATCCC CCGAAGTTAA GTAGGTCGTG AGATGCCATG GAGGCTCTCG TTCATCCCGT 
               
               
                   
               
               
                 7141 
                 GGGACATCAA GCTTCCCCTT GATAAAGCAC CCCGCTCGGG TGTAGCAGAG AAGACGCCTT 
               
               
                   
               
               
                 7201 
                 CTGAATTGTG CAATCCCTCC ACCTTATCTA AGCTTGCTAC CAATAATTAG CATTTTTGCC 
               
               
                   
               
               
                 7261 
                 TTGCGACAGA CCTCCTACTT AGATTGCCAC ACATTGAGCT AGTCAGTGAG CGATAAGCTT 
               
               
                   
               
               
                 7321 
                 GACGCGCTTT CAAGGGTCGC GAGTACGTGA ACTAAGGCTC CGGACAGGAC TATATACTTG 
               
               
                   
               
               
                 7381 
                 GGTTTGATCT CGCCCCGACA ACTGCAAACC TCAACTTTTT TAGATTATAT GGTTAGCCGA 
               
               
                   
               
               
                 7441 
                 AGTTGCACGA GGTGGCGTCC GCGGACTGCT CCCCGAGTGT GGCTCTTTCA TCTGACAACG 
               
               
                   
               
               
                 7501 
                 TGCAACCCCT ATCGCGGCCG ATTGTTTCTG CGGACGATGT TGTCCTCATA GTTTGGGCAT 
               
               
                   
               
               
                 7561 
                 GTTTCCCTTG TAGGTGTGAA ACCACTTAGC TTCGCGCCGT AGTCCCAATG AAAAACCTAT 
               
               
                   
               
               
                 7621 
                 GGACTTTGTT TTGGGTAGCA CCAGGAATCT GAACCGTGTG AATGTGGACG TCGCGCGCGT 
               
               
                   
               
               
                 7681 
                 AGACCTTTAT CTCCGGTTCA AGGTAGGGAT GTGGCTGCAT GCTACGTTGT CACACCTACA 
               
               
                   
               
               
                 7741 
                 CTGCTCGAAG TAAATATGCG AAGCGCGCGG CCTGGCCGGA GGCGTTCCGC GCCGCCACGT 
               
               
                   
               
               
                 7801 
                 GTTCGTTAAC TGTTGATTGG TGGCACATAA GCAATATCGT AGTCCGTCAA ATTCAGCTCT 
               
               
                   
               
               
                 7861 
                 GTTATCCCGG GCGTTATGTG TCAAATGGCG TAGAACGGGA TTGACTGTTT GACGGTAGGG 
               
               
                   
               
               
                 7921 
                 TGACCTAAGC CAGATGCTAC ACAATTAGGC TTGTACATAT TGTCGTTAGA ACGCGGCTAC 
               
               
                   
               
               
                 7981 
                 AATTAATACA TAACCTTATG TATCATACAC ATACGATTTA GGTGACACTA TAGAATACAC 
               
               
                   
               
               
                 8041 
                 GGAATTAATT C. 
               
            
           
         
       
     
     
       
         
           
               
             
               
                 TABLE 5 
               
             
            
               
                   
               
               
                 Features of the VMD2.IntEx.GFP.WPRE.pA plasmid sequence 
               
            
           
           
               
               
               
               
               
               
            
               
                   
                   
                 Mini- 
                 Maxi- 
                   
                 Direc- 
               
               
                 Name 
                 Type 
                 mum 
                 mum 
                 Length 
                 tion 
               
               
                   
               
            
           
           
               
               
               
               
               
               
            
               
                 Randomly 
                 Stuffer 
                 5,419 
                 7,918 
                 2,500 
                 none 
               
               
                 generated 
               
               
                 stuffer sequence 
               
               
                 AphR (KanR) 
                 CDS 
                 4,303 
                 5,118 
                 816 
                 forward 
               
               
                 pBR322 rep 
                 rep_origin 
                 3,343 
                 3,962 
                 620 
                 reverse 
               
               
                 origin 
               
               
                 AAV2 ITR 
                 LTR 
                 2,659 
                 2,779 
                 121 
                 reverse 
               
               
                 bGH pA 
                 polyA_signal 
                 2,333 
                 2,601 
                 269 
                 forward 
               
               
                 WPRE 
                 WPRE 
                 1,725 
                 2,314 
                 590 
                 forward 
               
               
                 GFP 
                 misc_feature 
                 996 
                 1,712 
                 717 
                 forward 
               
               
                 Kozak 
                 Kozak 
                 990 
                 995 
                 6 
                 forward 
               
               
                 Exon 
                 exon 
                 937 
                 989 
                 53 
                 forward 
               
               
                 Intron 
                 intron 
                 814 
                 936 
                 123 
                 forward 
               
               
                 −585 to +38 
                 promoter 
                 189 
                 811 
                 623 
                 forward 
               
               
                 VMD2 promoter 
               
               
                 AAV2 ITR 
                 LTR 
                 4 
                 133 
                 130 
                 forward 
               
               
                   
               
            
           
         
       
     
     AAV Particles 
     The AAV vectors of the disclosure contain an AAV genome that has been derivatized for the purpose of administration to patients. Such derivatization is standard in the art and the invention encompasses the use of any known derivative of an AAV genome, and derivatives which could be generated by applying techniques known in the art. Derivatization of the AAV genome and of the AAV capsid are reviewed in Coura and Nardi (2007) Virology Journal 4: 99, and in Choi et al. and Wu et al., referenced above. 
     Derivatives of an AAV genome include any truncated or modified forms of an AAV genome which allow for expression of a transgene from a vector of the invention in vivo. It is possible to truncate the AAV genome significantly to include minimal viral sequence yet retain the above function. This is preferred for safety reasons to reduce the risk of recombination of the vector with wild-type virus, and also to avoid triggering a cellular immune response by the presence of viral gene proteins in the target cell. 
     The following portions could therefore be removed in a derivative of the invention: one inverted terminal repeat (ITR) sequence, the replication (rep) and capsid (cap) genes. However, in some embodiments, derivatives may additionally include one or more rep and/or cap genes or other viral sequences of an AAV genome. Naturally occurring AAV integrates with a high frequency at a specific site on human chromosome 19, and shows a negligible frequency of random integration, such that retention of an integrative capacity in the vector may be tolerated in a therapeutic setting. 
     The AAV genome comprises packaging genes, such as rep and/or cap genes which encode packaging functions for an AAV particle. The rep gene encodes one or more of the proteins Rep78, Rep68, Rep52 and Rep40 or variants thereof. The cap gene encodes one or more capsid proteins such as VP1, VP2 and VP3 or variants thereof. These proteins make up the capsid of an AAV particle. 
     Where a derivative comprises capsid proteins i.e. VP1, VP2 and/or VP3, the derivative may be a chimeric, shuffled or capsid-modified derivative of one or more naturally occurring AAVs. In particular, the invention encompasses the provision of capsid protein sequences from different serotypes, clades, clones, or isolates of AAV within the same vector (i.e. a pseudotyped vector). 
     Chimeric, shuffled or capsid-modified derivatives are selected to provide one or more desired functionalities for the viral vector. Thus, these derivatives may display increased efficiency of gene delivery, decreased immunogenicity (humoral or cellular), an altered tropism range and/or improved targeting of a particular cell type compared to an AAV vector comprising a naturally occurring AAV genome, such as that of AAV2. Increased efficiency of gene delivery may be effected by improved receptor or co-receptor binding at the cell surface, improved internalization, improved trafficking within the cell and into the nucleus, improved uncoating of the viral particle and improved conversion of a single-stranded genome to double-stranded form. Increased efficiency may also relate to an altered tropism range or targeting of a specific cell population, such that the vector dose is not diluted by administration to tissues where it is not needed. 
     Chimeric capsid proteins include those generated by recombination between two or more capsid coding sequences of naturally occurring AAV serotypes. This may be performed for example by a marker rescue approach in which non-infectious capsid sequences of one serotype are co-transfected with capsid sequences of a different serotype, and directed selection is used to select for capsid sequences having desired properties. The capsid sequences of the different serotypes can be altered by homologous recombination within the cell to produce novel chimeric capsid proteins. 
     Chimeric capsid proteins also include those generated by engineering of capsid protein sequences to transfer specific capsid protein domains, surface loops or specific amino acid residues between two or more capsid proteins, for example between two or more capsid proteins of different serotypes. 
     Shuffled or chimeric capsid proteins may also be generated by DNA shuffling or by error-prone PCR. Hybrid AAV capsid genes can be created by randomly fragmenting the sequences of related AAV genes e.g. those encoding capsid proteins of multiple different serotypes and then subsequently reassembling the fragments in a self-priming polymerase reaction, which may also cause crossovers in regions of sequence homology. A library of hybrid AAV genes created in this way by shuffling the capsid genes of several serotypes can be screened to identify viral clones having a desired functionality. Similarly, error prone PCR may be used to randomly mutate AAV capsid genes to create a diverse library of variants which may then be selected for a desired property. 
     The sequences of the capsid genes may also be genetically modified to introduce specific deletions, substitutions or insertions with respect to the native wild-type sequence. In particular, capsid genes may be modified by the insertion of a sequence of an unrelated protein or peptide within an open reading frame of a capsid coding sequence, or at the N- and/or C-terminus of a capsid coding sequence. The unrelated protein or peptide may advantageously be one which acts as a ligand for a particular cell type, thereby conferring improved binding to a target cell or improving the specificity of targeting of the vector to a particular cell population. The unrelated protein may also be one which assists purification of the viral particle as part of the production process, i.e. an epitope or affinity tag. The site of insertion will is selected so as not to interfere with other functions of the viral particle e.g. internalization, trafficking of the viral particle. The skilled person can identify suitable sites for insertion based on their common general knowledge. Particular sites are disclosed in Choi et al., referenced above. 
     The invention additionally encompasses the provision of sequences of an AAV genome in a different order and configuration to that of a native AAV genome. The invention also encompasses the replacement of one or more AAV sequences or genes with sequences from another virus or with chimeric genes composed of sequences from more than one virus. Such chimeric genes may be composed of sequences from two or more related viral proteins of different viral species. 
     AAV vectors of the invention include transcapsidated forms wherein an AAV genome or derivative having an ITR of one serotype is packaged in the capsid of a different serotype. AAV vectors of the invention also include mosaic forms wherein a mixture of unmodified capsid proteins from two or more different serotypes makes up the viral capsid. An AAV vector may also include chemically modified forms bearing ligands adsorbed to the capsid surface. For example, such ligands may include antibodies for targeting a particular cell surface receptor. 
     Thus, for example, AAV vectors of the invention include those with an AAV2 genome and AAV2 capsid proteins (AAV2/2), those with an AAV2 genome and AAV5 capsid proteins (AAV2/5) and those with an AAV2 genome and AAV8 capsid proteins (AAV2/8). An AAV vector of the invention may comprise a mutant AAV capsid protein. In one embodiment, an AAV vector of the invention comprises a mutant AAV8 capsid protein. Preferably the mutant AAV8 capsid protein is an AAV8 Y733F capsid protein. 
     Methods of making AAV viral particles of the disclosure will be known to one of skill in the art. An exemplary, but non-limiting method of preparing AAV viral particles of the disclosure is described below. For generation of a given AAV vector, three plasmids are required: one comprising the viral delivery vector encoding the nucleic acid sequence of interest to be delivered (i.e. the nucleic acid sequence encoding BEST1), a plasmid encoding the rep and cap genes, and a third helper plasmid that contains the required adenoviral genes necessary for successful AAV generation. A promoter may be operably linked to each of the packaging genes. Specific examples of such promoters include the p5, p19 and p40 promoters (Laughlin et al. (1979) Proc. Natl. Acad. Sci. USA 76: 5567-5571). For example, the p5 and p19 promoters are generally used to express the rep gene, while the p40 promoter is generally used to express the cap gene. The plasmids are used to transfect suitable cells that are capable of replicating the AAV viral vector, transcribing and translating the AAV protein, and packaging the AAV viral vector into an AAV viral particle. Exemplary suitable cells comprise HEK293 cells. Post-transfection, the cells are collected and lysed. AAV particles can then be purified from the lysate through a variety of methods. Alternatively, AAV particles can be purified from the supernatant. For example, the lysate can be treated with Benzonase and clarified before applying to an iodixanol gradient comprised of 15%, 25%, 40% and 60% phases. The gradients can spun at 59,000 rpm for 1 hour 30 minutes and the 40% fraction then withdrawn. This AAV phase can then purified and concentrated using an Amicon Ultra-15 100K filter unit. 
     Pharmaceutical Compositions 
     The AAV vectors of the invention may be formulated into pharmaceutical compositions. These compositions may comprise, in addition to the medicament, a pharmaceutically acceptable carrier, diluent, excipient, buffer, stabilizer or other materials well known in the art. Such materials should be non-toxic and should not interfere with the efficacy of the active ingredient. The precise nature of the carrier or other material may be determined by the skilled person according to the route of administration, e.g. subretinal, direct retinal or intravitreal injection. 
     The pharmaceutical composition may be formulated as a liquid. Liquid pharmaceutical compositions may include a liquid carrier such as water, petroleum, animal or vegetable oils, mineral oil or synthetic oil. Physiological saline solution, magnesium chloride, dextrose or other saccharide solution, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol may be included. In some cases, a surfactant, such as pluronic acid (PF68) 0.001% may be used. 
     For injection at the site of affliction, the active ingredient may be in the form of an aqueous solution which is pyrogen-free, and has suitable pH, isotonicity and stability. The skilled person is well able to prepare suitable solutions using, for example, isotonic vehicles such as Sodium Chloride Injection, Ringer&#39;s Injection or Lactated Ringer&#39;s Injection. Preservatives, stabilisers, buffers, antioxidants and/or other additives may be included as required. 
     Buffers may have an effect on the stability and biocompatibity of the viral vectors and vector particles of the disclosure following storage and passage through injection devices for AAV gene therapy. In some embodiments, the viral vectors and vector particles of the disclosure may be diluted in TMN 200 buffer to maintain biocompatibility and stability. TMN 200 buffer comprises 20 mM Tris (pH adjusted to 8.0), 1 mM MgCl 2  and 200 mM NaCl. 
     The determination of the physical viral genome titer comprises part of the characterization of the viral vector or viral particle. In some embodiments, determination of the physical viral genome titre comprises a step in ensuring the potency and safety of viral vectors and viral particles during gene therapy. In some embodiments, a method to determine the AAV titer comprises quantitative PCR (qPCR). There are different variables that can influence the results, such as the conformation of the DNA used as standard or the enzymatic digestion during the sample preparation. The viral vector or particle preparation whose titer may be measured may be compared against a standard dilution curve generated using a plasmid. In some embodiments, the plasmid DNA used in the standard curve is in the supercoiled conformation. In some embodiments, the plasmid DNA used in the standard curve is in the linear conformation. Linearized plasmid can be prepared, for example by digestion with HindIII restriction enzyme, visualized by agarose gel electrophoresis and purified using the QIAquick Gel Extraction Kit (Qiagen) following manufacturer&#39;s instructions. Other restriction enzymes that cut within the plasmid used to generate the standard curve may also be appropriate. In some embodiments, the use of supercoiled plasmid as the standard increased the titre of the AAV vector compared to the use of linearized plasmid. 
     To extract the DNA from purified AAV vectors for quantification of AAV genome titer, two enzymatic methods can be used. In some embodiments, the AAV vector may be singly digested with DNase I. In some embodiments, the AAV vector may be double digested with DNase I and an additional proteinase K treatment. QPCR can then performed with the CFX Connect Real-Time PCR Detection System (BioRad) using primers and Taqman probe specific to the transgene sequence. 
     For delayed release, the medicament may be included in a pharmaceutical composition which is formulated for slow release, such as in microcapsules formed from biocompatible polymers or in liposomal carrier systems according to methods known in the art. 
     Dosages 
     As used herein, the term “Dnase resistant particle (DRP)” refers to AAV particles that are resistant to Dnase digestion, and are therefore thought to completely encapsulate and protect the AAV vector of the disclosure from Dnase digestion. AAV particles may also be quantified in terms of the total numbers of genome particles (gp) administered in a dose, or gp/mL, the number genome particles per milliliter (mL) of solution. As used herein, genome particle (gp) refers to AAV particles containing a copy of an AAV delivery vector (or AAV genome) of the disclosure. As used herein, the term genome content (GC) per mL refers to the number of viral genomes per mL of solution, and may be determined, for example, by qPCR as described above. The terms GC and VG (viral genomes) may be used synonymously to characterize AAV dosages and concentrations of the disclosure. 
     In some embodiments of the compositions of the disclosure, a composition comprising an AAV vector or an AAV vector is administered to a subject as a single dose. 
     In some embodiments of the compositions of the disclosure, a composition comprising an AAV vector or an AAV vector may be formulated as a liquid suspension wherein the AAV vectors are suspended in a pharmaceutically-acceptable carrier. In some embodiments, compositions of the disclosure may comprise a plurality of AAV vectors at a concentration of 1−2×10 9 , 1−2×10 10 , 1−2×10 11 , 1−2×10 12  or 1−2×10 13  genome particles (gp) per mL. In some embodiments, compositions of the disclosure may comprise a plurality of AAV vectors at a concentration of 5×10 11  DRP/mL, 1.5×10 12  DRP/mL, 5×10 12  DRP/mL, 1.2×10 12  DRP/mL, 4.5×10 12  DRP/mL, 1.2×10 13  DRP/mL, 1.5×10 13  DRP/mL or 5×10 13  DRP/1.2×10 12  DRP/mL. In some embodiments, compositions of the disclosure may comprise a plurality of AAV vectors at a concentration of 5×10 12  DRP per mL. In some embodiments, compositions of the disclosure may comprise a plurality of AAV vectors at a concentration of 1.5×10 13  DRP per mL. Thus, to administer a dose of AAV vector of about 2×10 10  gp, for example, a single injection of about 10 microliters of a pharmaceutical composition having a concentration of about 2×10 12  gp per mL will achieve the desired dose in vivo. 
     In some embodiments of the compositions of the disclosure, a composition comprising an AAV vector or an AAV vector may comprise a volume of between 1 and 500 μl, inclusive of the endpoints. In some embodiments of the compositions of the disclosure, a composition comprising an AAV vector or an AAV vector may comprise a volume of between 10-500, 50-500, 100-500, 200-500, 300-500, 400-500, 50-250, 100-250, 200-250, 50-150, 1-100 or 1-10 μl, inclusive of the endpoints for each range. In some embodiments of the compositions of the disclosure, a composition comprising an AAV vector or an AAV vector may comprise a volume of 1, 2, 5, 10, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500 μl or any number of microliters in between. In some embodiments, a composition comprising an AAV vector or an AAV vector may comprise 100 μl. 
     In some embodiments of the compositions of the disclosure, an entire volume of a composition comprising an AAV vector or an AAV vector may be injected in a single injection. In some embodiments, a portion of a volume of a composition comprising an AAV vector or an AAV vector may be injected in a single injection. In some embodiments, a first portion of a volume of a composition comprising an AAV vector or an AAV vector may be injected in a first single injection and a second portion of a volume of a composition comprising an AAV vector or an AAV vector may be injected in a second single injection 
     In some embodiments of the compositions of the disclosure, a composition comprising an AAV vector or an AAV vector is administered at a dosage of at least 2×10 7 , 2×10 8 , 5×10 8 , 1.5×10 9 , 2×10 9 , 5×10 9 , 2×10 10 , 5×10 10 , 6×10 10 , 1.2×10 11 , 2×10 11 , 4.5×10 11 , 5×10 11 , 1.2×10 12 , 1.5×10 12 , 2×10 12  or 5×10 12  gp per eye. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 5×10 10 , 1.5×10 11 , 5×10 11  or 1.5×10 11  gp per eye. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 5×10 11  DRP per eye, by subretinal injection. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 2×10 10  gp per eye, by subretinal injection. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 5×10 10  gp per eye, by subretinal injection. In some embodiments, the AAV vector is administered at a dosage of about 6×10 10  gp per eye, by subretinal injection. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 1.5×10 11  gp per eye, by subretinal injection. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 2×10 11  gp per eye, by subretinal injection. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 5×10 11  gp per eye, by subretinal injection. In some embodiments, a composition comprising an AAV vector or an AAV vector is administered at a dosage of about 1.5×10 12  gp per eye, by subretinal injection. 
     Dosages or volumes may be calculated based on allometric scaling between species based on vitreal volume. “Allometry”, as used herein, refers to the changes in organisms with respect to body size. Some factors to take into account when comparing species include body volume, surface area, metabolic rate, and unique anatomical, physiological or biochemical processes. The human equivalent dose can be normalized to body surface area, body weight or a combination of surface area and weight. Other factors may also be taken into account. 
     Delivery 
     The viral vectors of the invention may be administered to the eye of a subject by subretinal, direct retinal, suprachoroidal or intravitreal injection. A skilled person will be familiar with and well able to carry out individual subretinal, direct retinal or intravitreal injections. 
     Subretinal injections are injections into the subretinal space, i.e. underneath the neurosensory retina. During a subretinal injection, the injected material is directed into, and creates a space between, the photoreceptor cell and retinal pigment epithelial (RPE) layers. When the injection is carried out through a small retinotomy, a retinal detachment may be created. The detached, raised layer of the retina that is generated by the injected material is referred to as a “bleb”. The hole created by the subretinal injection must be sufficiently small that the injected solution does not significantly reflux back into the vitreous cavity after administration. Such reflux would be particularly problematic when a medicament is injected, because the effects of the medicament would be directed away from the target zone. Preferably, the injection creates a self-sealing entry point in the neurosensory retina, i.e. once the injection needle is removed, the hole created by the needle reseals such that very little or substantially no injected material is released through the hole. 
     To facilitate this process, specialist subretinal injection needles are commercially available (e.g. DORC 41G Teflon subretinal injection needle, Dutch Ophthalmic Research Center International BV, Zuidland, The Netherlands). These are needles designed to carry out subretinal injections. 
     Alternatively, subretinal injections can be performed by delivering the composition comprising AAV particles under direct visual guidance using an operating microscope (Leica Microsystems, Germany). One exemplary approach is that of using a scleral tunnel approach through the posterior pole to the superior retina with a Hamilton syringe and 34-gauge needle (ESS labs, UK). Alternatively, sub-retinal injections can be performed using an anterior chamber paracentesis with a 33G needle prior to the subretinal injection using a WPI syringe and a beveled 35G-needle system (World Precision Instruments, UK). An additional alternative is a WPI Nanofil Syringe (WPI, part #NANOFIL) and a 34 gauge WBI Nanofil needle (WPI, part # NF34BL-2). 
     Vectors or compositions of the disclosure may be administered via suprachoroidal injection. Any means of suprachoroidal injection is envisaged as a potential delivery system for a vector or a composition of the disclosure. Suprachoroidal injections are injections into the suprachoroidal space, which is the space between the choroid and the sclera. Injection into the suprachoroidal space is thus a potential route of administration for the delivery of compositions to proximate eye structures such as the retina, retinal pigment epithelium (RPE) or macula. In some embodiments, injection into the suprachoroidal space is done in an anterior portion of the eye using a microneedle, microcannula, or microcatheter. An anterior portion of the eye may comprise or consist of an area anterior to the equator of the eye. The vector composition or AAV viral particles may diffuse posteriorly from an injection site via a suprachoroidal route. In some embodiments, the suprachoroidal space in the posterior eye is injected directly using a catheter system. In this embodiment, the suprachoroidal space may be catheterized via an incision in the pars plana. In some embodiments, an injection or an infusion via a suprachoroidal route traverses the choroid, Bruch&#39;s membrane and/or RPE layer to deliver a vector or a composition of the disclosure to a subretinal space. In some embodiments, including those in which a vector or a composition of the disclosure is delivered to a subretinal space via a suprachoroidal route, one or more injections is made into at least one of the sclera, the pars plana, the choroid, the Bruch&#39;s membrane, and the RPE layer. In some embodiments, including those in which a vector or a composition of the disclosure is delivered to a subretinal space via a suprachoroidal route, a two-step procedure is used to create a bleb in a suprachoroidal or a subretinal space prior to delivery of a vector or a composition of the disclosure. 
     In those embodiments where mice are injected, animals can be anaesthetized by intraperitoneal injection containing ketamine (40-80 mg/kg) and xylazine (1-10 mg/kg) and pupils fully dilated with tropicamide eye drops (Mydriaticum 1%, Bausch &amp; Lomb, UK) and phenylephrine eye drops (phenylephrine hydrochloride 2.5%, Bausch &amp; Lomb, UK). Proxymetacaine eye drops (proxymetacaine hydrochloride 0.5%, Bausch &amp; Lomb, UK) can also applied prior to sub-retinal injection. Post-injection, chloramphenicol eye drops can applied (chloramphenicol 0.5%, Bausch &amp; Lomb, UK) and anaesthesia reversed with atipamezole (2 mg/kg) and carbomer gel applied (Viscotears, Novartis, UK) to prevent cataract formation. 
     Unless damage to the retina occurs during the injection, and as long as a sufficiently small needle is used, substantially all injected material remains localized between the detached neurosensory retina and the RPE at the site of the localized retinal detachment (i.e. does not reflux into the vitreous cavity). Indeed, the typical persistence of the bleb over a short time frame indicates that there is usually little escape of the injected material into the vitreous. The bleb may dissipate over a longer time frame as the injected material is absorbed. 
     Visualizations of the eye, in particular the retina, for example using optical coherence tomography, may be made pre-operatively. 
     The AAV vectors of the invention may be delivered with increased accuracy and safety by using a two-step method in which a localized retinal detachment is created by the subretinal injection of a first solution. The first solution does not comprise the vector. A second subretinal injection is then used to deliver the medicament comprising the vector into the subretinal fluid of the bleb created by the first subretinal injection. Because the injection delivering the medicament is not being used to detach the retina, a specific volume of solution may be injected in this second step. An AAV vector of the invention may be delivered by: (a) administering a solution to the subject by subretinal injection in an amount effective to at least partially detach the retina to form a subretinal bleb, wherein the solution does not comprise the vector; and (b) administering a medicament composition by subretinal injection into the bleb formed by step (a), wherein the medicament comprises the vector. 
     EXAMPLES 
     Example 1: Bestrophin-1 Protein in HEK293 Cells Using the CAG Promoter 
     HEK293 cells were transduced with an AAV2/2 vector containing the CAG promoter driving Best1 expression with a WPRE (AAV2/2 CAG.BEST1.WPRE.pA,  FIG.  3   ) and without a WPRE (AAV2/2 CAG.BEST1.pA), and the expression and localization of Bestrophin-1 protein was examined. In  FIG.  6   , transduced HEK293 cells were stained with Hoechst and an anti-human Bestrophin-1 (hBEST1 or huBEST1) antibody. Bestrophin-1 protein was found throughout the cytosol when compared to untransduced control cells. 
     Bestrophin-1 expression in HEK293 cells was quantified from Western Blot ( FIG.  7   ). In  FIG.  7 A , sample 1 was the AAV2/2 CAG.hBEST1.pA vector; sample 2 was the AAV2/2 CAG.hBEST1.WPRE.pA vector and sample 3 was a negative control. Plasmid-transfected HEK293 cells were used as a positive control. In  FIG.  7 B , quantification showed that AAV2/2 CAG.BEST1.WPRE.pA (n=9) showed an approximately 4-fold increase in Bestrophin-1 expression over AAV2/2 CAG.BEST1.pA (n=9) (p&lt;0.01 by One-way ANOVA with Tukey&#39;s Multiple Comparisons Test) and a statistically significant increase over un-transduced control cells (n=8) (p&lt;0.001) was seen. Although Bestrophin-1 expression was seen in the AAV2/2 CAG.BEST1.pA cells, this was not statistically significant over un-transduced cells. Error bars=±SEM, and *** indicates p&lt;0.001 when compared to un-transduced control. 
     HEK293 cells expressing Bestrophin-1 were additionally assayed with whole-cell patch clamp recording.  FIG.  8 A  shows the Current (I)/Voltage (V) plots of HEK293 cells transduced with AAV2/2 CAG.BEST1.pA, AAV2/2 CAG.BEST1.WPRE.pA and AAV2/2 CAG.GFP.WPRE.pA vectors as well as an untransduced control.  FIG.  8 B  shows the current waveforms, and chord conductance is shown in  FIG.  9   . 
     Example 2: Bestrophin-1 Protein in Cultured ARPE19 Cells Using the VMD2 Promoter 
     Appropriately differentiated ARPE19 are known to have gene expression profiles similar to those of native retinal pigment epithelium (RPE) cells, and can be used as an alternative to native RPE cells to test gene expression. Differentiated ARPE19 cells were used to test the ability of the VMD2 and CAG promoters to drive BEST1 expression in RPE cells, and to test the effect of the intron-exon (IntEx) sequence on expression from the VMD2 promoter. 
     ARPE19 cells were transfected and assayed for BEST1 expression using the protocol outlined in  FIG.  10 B . ARPE19 cells were grown in differentiation medium (DMEM with 4.5 g/l glucose, L-glutamine, and 1 mM sodium pyruvate supplemented with 1% fetal bovine serum (FBS) for 1-4 months at 37° C. and 5% CO 2  in 96 well plates. Differentiated ARPE19 cells were then transfected with either pCAG.BEST1.WPRE (CAG promoter), pVMD.BEST1.WPRE (VMD2 promoter), or pVMD2. IntEx.BEST1.WPRE (VMD2 promoter and an intron-exon construct) at 3.8×10 10  number of copies of each plasmid per well. Cells treated with TranslT-LT1 reagent alone and cells without transfection reagent or plasmid served as negative controls. Cells were then cultured for 2 days at 37° C., before being fixed and stained with Anti-hBest1 and Anti-ZO1 (also called ZO-1, or zona occludens-1, or tight junction protein 1, a protein located on the cytoplasmic membrane surface of intercellular tight junctions).  FIGS.  11 - 13    show BEST1 expression in differentiated ARPE19 cells transfected with the three vectors encoding BEST1 and an untransfected control. 
     In ARPE19 cells that were differentiated for one month before transfection, the untransfected cells showed no expression. In contrast, both the pCAG.BEST1.WPRE and pVMD2. IntEx.BEST1.WPRE were able to drive the expression of BEST1 protein in differentiated ARPE19 cells (see  FIG.  11 A , contrast the first, second and fourth rows). pVMD2.BEST1.WPRE (no exon-intron) was also able to drive the expression of BEST1 in 1 month differentiated ARPE19 cells ( FIG.  11 B ), although this construct seemed to express BEST1 at lower levels than the construct with the intron-exon sequence (pVMD2. IntEx.BEST1.WPRE). In ARPE19 cells that were differentiated for three months, similar results were obtained: pCAG.BEST1.WPRE, pVMD2. IntEx.BEST1.WPRE and pVMD2.BEST1.WPRE were all able to drive expression of BEST1 protein, although the expression with the CAG promoter was higher than with the VMD2 promoter and, with the VMD2 promoter, the intron-exon sequence improves the expression (contrast the first row of  FIG.  12 A , the untransfected control, with  FIG.  12 B ). 
     ARPE19 cells were transduced and assayed for BEST1 expression using the protocol outlined in  FIG.  10   . Differentiated ARPE19 cells were pre-treated with 400 nM doxorubicin before transduction. This drug has been proved to improve AAV2 transduction efficiency in several in vitro models. Four hours after the treatment, cells were transduced with the different viral constructs at different multiplicities of infection (MOIs). 
     ARPE19 cells differentiated for 4 months, pre-treated with 400 nM doxorubicin and transduced with AAV2/2.CAG.GFP.WPRE and AAV2/2.VMD2. InEx.GFP.WPRE at 2, 4 and 8×10 4  gp/cell showed higher GFP fluorescence compared to transduced cells without pre-treatment with doxorubicin 10 days after transduction (contrast top and bottom row of each panel of  FIG.  13 A ) AAV2/2.CAG.GFP.WPRE and B) AAV2/2.VMD2. InEx.GFP.WPRE). GFP fluorescence was not detected in untransduced cells, used as negative controls (first column of  FIG.  13 A  and B). 
     In ARPE19 cells differentiated for 4 months, pre-treated with 400 nM doxorubicin and transduced with AAV2/2.CAG.BEST1.WPRE and AAV2/2.VMD2. InEx.BEST1.WPRE at 1 and 4×10 4  gp/cell, BEST1 expression could be detected by immunostaining with anti-hBEST1 (red, third column, second to fifth row of  FIG.  14   ) compared to the untransduced control (first row,  FIG.  14   ) 10 days after transduction. 
     Example 3: 4/8 Week In Vivo Pilot Study in Mice 
     The ability of the VMD2.BEST1.WPRE and VMD2. IntEx.BEST1.WPRE constructs to drive the expression BEST1 was assayed in vivo. The protocol of the 4/8 week in vivo pilot study is shown in  FIG.  15   . C57BL/6 mice (6 per group) were injected bilaterally with either a sham injection, AAV2/2 VMD2.BEST1.WPRE or AAV2/2 VMD2. IntEx.BEST1.WPRE AAV viral particles. 1 μL of AAV solution was injected subretinally with a 34 gauge Nanofil needle (WPI #NF34BL-2) at 1×10 9  GC/μL/eye. Eyes were imaged using optical coherence tomography at 4 and 8 weeks to assess for retinal thinning (toxicity), and 3 animals were sacrificed at each time point to assay BEST1 protein expression by immunohistochemistry and Western Blot. 
     OCT imaging at 4 and 8 weeks showed that neither VMD2 construct showed photoreceptor toxicity when compared to the sham treatment ( FIGS.  16 - 18   ). 
     Three animals were sacrificed at both the 4- and 8-week time points, and BEST1 protein expression was further characterized by western blot ( FIG.  21   ) and immunohistochemistry ( FIG.  19   ).  FIG.  19    shows immunohistochemistry results for eyes four weeks post injection, while  FIG.  20    shows immunohistochemistry results for eyes eight weeks post injection. Eyes were stained with anti-BEST1 (green) and anti-Rhodopsin (red), which marks photoreceptor cells, and DAPI ( FIGS.  19  and  20   ). BEST1 protein expression was observed from VMD2.BEST1.WPRE.pA and VMD2. IntEx.BEST1.WPRE.pA. VMD2 promoter driven BEST1 expression localized to the conjunction of the RPE layer and photoreceptor outer layer. Western Blot on dissected RPE/choroid complex tissue from four week injected eyes shows protein expression ( FIG.  21   ). 
     Example 4: 4/13 Week In Vivo Proof of Concept Study in Mice 
     An additional 4 and 13 week in vivo proof of concept (PoC) study was carried out in mice to confirm the results of the pilot study, assay the effect of AAV viral particle dosage, and look at the effects at later time points post AAV injection. An outline of the protocol for the 4/13 week Proof of Concept study is set forth in  FIG.  22   . C57BL/6 mice (12 per cohort) were bilaterally injected with VMD2. IntEx.BEST1.WPRE or VMD2.BEST1.WPRE.pA AAV particles at either 1×10 8  GC/μL/eye or 1×10 9  GC/μL/eye, or with a sham injection. 1 μL of AAV solution was injected subretinally with a 34 gauge Nanofil needle (WPI #NF34BL-2). Eyes were imaged with OCT at 4 and 13 weeks post injection. Four mice were sacrificed four weeks post injection, and the remaining eight at 13 weeks post injection, and BEST1 expression was characterized by immunohistochemistry and Western blot. 
     OCT imaging at 4 weeks and 13 weeks showed that neither VMD2 construct (with or without the intron-exon sequence) at either the high dose (1×10 9  GC/eye) or the low dose (1×10 8  GC/eye) showed toxicity as evidenced by retinal thinning when compared to the sham control ( FIG.  23   ). Staining with anti-BEST1 (huBEST1 in  FIG.  24   ) and anti-Rhodopsin showed that VMD2 driven BEST1 localized to the RPE layer, with a trend of more BEST1 expression in VMD2. IntEx.BEST1.WPRE injected eyes. Western Blot on pooled dissected RPE/choroid complex tissue from four week injected eyes (4) shows protein expression ( FIG.  25 B ). 
     Example 5: Good Laboratory Practice (GLP) Toxicity Assessment Study in Mice 
     The safety and expression of BEST1 AAV over longer periods of time is verified in mice with a Good Laboratory Practice (GLP) toxicity study in mice. An outline of the study is set forth in  FIG.  27   . Cohorts of 8 male and 8 female mice are injected subretinally and bilaterally with a low (5.0×10 8  GC/eye), medium (1.5×10 9  GC/eye) or high (5.0×10 9  GC/eye) dose of VMD2. IntEx.BEST1.WPRE AAV particles. Using allometric volume scaling, the high mouse dose is equivalent to a dose of 100 μL at 5×10 12  GC/mL/eye in humans. Mice are evaluated and sacrificed at 4 weeks and 26 weeks. Eyes are assessed with an ophthalmic examination, tonometry to measure intraocular pressure (TOP), OCT for retinal thickness (predose, and the end of 4 and 13 weeks). Post sacrifice, necropsies assess organ weights and tissues such as the left eye, brain, heart, skeletal muscle, lung, liver, kidney, testes and ovary are collected for qPCR. Histopathological evaluations are carried out, and tissues are reserved, .e.g. by storage in formalin, for additional immunohistochemistry. Alternatively, or in addition, groups of 4 mice are injected with dosages of 2×10 9  GC/eye and 5×10 9  GC/eye of VMD2. IntEx.BEST1.WPRE AAV particles and evaluated at 4 weeks to optimize protocols for the larger toxicity study (see  FIG.  28    for an outline). 
     INCORPORATION BY REFERENCE 
     Every document cited herein, including any cross referenced or related patent or application is hereby incorporated herein by reference in its entirety unless expressly excluded or otherwise limited. The citation of any document is not an admission that it is prior art with respect to any invention disclosed or claimed herein or that it alone, or in any combination with any other reference or references, teaches, suggests or discloses any such invention. Further, to the extent that any meaning or definition of a term in this document conflicts with any meaning or definition of the same term in a document incorporated by reference, the meaning or definition assigned to that term in this document shall govern. 
     OTHER EMBODIMENTS 
     While particular embodiments of the disclosure have been illustrated and described, various other changes and modifications can be made without departing from the spirit and scope of the disclosure. The scope of the appended claims includes all such changes and modifications that are within the scope of this disclosure.