Promoters, expression cassettes, vectors, kits, and methods for the treatment of achromatopsia and other diseases

Abstract

The present invention provides isolated promoters, transgene expression cassettes, vectors, kits, and methods for treatment of genetic diseases that affect the cone cells of the retina.

1 . A nucleic acid comprising a portion of the cone cell specific promoter PR 2.1. 
     
     
         2 . The nucleic acid of  claim 1 , comprising the sequence SEQ ID NO: 4. 
     
     
         3 . The nucleic acid of  claim 1 , wherein PR2.1 is truncated at the 5′ or the 3′ end. 
     
     
         4 . The nucleic acid of  claim 3 , wherein the truncation is between about 100 base pairs to 1,500 base pairs. 
     
     
         5 . (canceled) 
     
     
         6 . (canceled) 
     
     
         7 . (canceled) 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . A nucleic acid comprising a nucleotide sequence selected from a nucleotide sequence of SEQ ID NO:1, SEQ ID NO: 2 or SEQ ID NO: 3. 
     
     
         15 . (canceled) 
     
     
         16 . (canceled) 
     
     
         17 . (canceled) 
     
     
         18 . The nucleic acid of  claim 1 , wherein the promoter is capable of promoting CNGB3 expression in S-cone cells, M-cone cells, and L-cone cells. 
     
     
         19 . The nucleic acid of  claim 1 , wherein the promoter is capable of promoting CNGA3 expression in S-cone cells, M-cone cells, and L-cone cells. 
     
     
         20 . The nucleic acid of  claim 1 , wherein the promoter is capable of promoting GNAT2 expression in S-cone cells, M-cone cells, and L-cone cells. 
     
     
         21 . A recombinant adeno-associated (rAAV) expression vector comprising a target nucleic acid sequence operably linked to the nucleic acid of  claim 1 . 
     
     
         22 . (canceled) 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . The expression vector of  claim 21 , wherein the target nucleic acid sequence encodes a cyclic nucleotide-gated channel subunit B (CNGB3) polypeptide. 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . The expression vector of  claim 21 , wherein the target nucleic acid sequence encodes a cyclic nucleotide-gated channel subunit A (CNGA3) polypeptide. 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . The expression vector of  claim 21 , wherein the target nucleic acid sequence encodes a Guanine nucleotide-binding protein G(t) subunit alpha-2 (GNAT-2) polypeptide. 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . A mammalian cell comprising the expression vector of  claim 21 . 
     
     
         39 . A transgene expression cassette comprising:
 (a) the nucleic acid of  claim 1 ;   (b) a nucleic acid selected from the group consisting of a CNGB3 nucleic acid, a CNGA3 nucleic acid, and a GNAT2 nucleic acid; and   (c) minimal regulatory elements.   
     
     
         40 . A nucleic acid vector comprising the expression cassette of  claim 39 . 
     
     
         41 . The vector of  claim 40  wherein the vector is an adeno-associated viral (AAV) vector. 
     
     
         42 . A kit comprising the expression vector of  claim 21  and instructions for use. 
     
     
         43 . A method of treating an eye disease comprising administering to a subject in need thereof the expression vector of  claim 21 , thereby treating the subject. 
     
     
         44 . A method of promoting CNGA3 or CNGB3 expression in the cone cells of a subject comprising administering to the subject the expression vector of  claim 21 , thereby promoting CNGA3 or CNGB3 expression. 
     
     
         45 . The method of  claim 43 , wherein the eye disease is associated with a genetic mutation, substitution, or deletion that affects retinal cone cells. 
     
     
         46 . (canceled) 
     
     
         47 . (canceled) 
     
     
         48 . (canceled) 
     
     
         49 . (canceled) 
     
     
         50 . (canceled) 
     
     
         51 . (canceled)