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 . An isolated promoter comprising approximately 1.8 kb of the 5′-NTR of the cyclic nucleotide-gated ion channel beta 3 (CNGB3) gene. 
     
     
         2 . The promoter of  claim 1  comprising the sequence SEQ ID NO: 1 
     
     
         3 . An isolated promoter comprising approximately 1.6 kb of the 5′-NTR of the CNGB3 gene. 
     
     
         4 . The promoter of  claim 2  comprising SEQ ID NO:2. 
     
     
         5 . An isolated promoter comprising
 approximately 400 bp of the cytomegalovirus (CMV) enhancer and   approximately 1.4 kb of the 5′-NTR of the CNGB3 gene.   
     
     
         6 . The promoter of  claim 5  comprising the following sequences:
 cytomegalovirus (CMV) enhancer set forth as SEQ ID NO: 3 and the 5′-NTR of the CNGB3 gene set forth as SEQ ID NO: 4. 
 
     
     
         7 . The promoter of  claim 1 , wherein the CNGB3 gene is the human CNGB3 gene. 
     
     
         8 . The promoter of  claim 1 , wherein said promoter is capable of promoting CNGB3 expression in S-cone cells, M-cone cells, and L-cone cells. 
     
     
         9 . The promoter of  claim 1 , wherein said promoter is capable of promoting cyclic nucleotide-gated ion channel alpha 3 (CNGA3) expression in S-cone cells, M-cone cells, and L-cone cells. 
     
     
         10 . The promoter of  claim 1 , wherein said promoter is capable of promoting guanine nucleotide binding protein subunit alpha transducin 2 (GNAT2) expression in S-cone cells, M-cone cells, and L-cone cells. 
     
     
         11 . A transgene expression cassette comprising
 (a) the promoter 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.   
     
     
         12 . A transgene expression cassette comprising
 (a) the promoter of  claim 1 ,   (b) a CNGB3 nucleic acid, and   (c) minimal regulatory elements.   
     
     
         13 . (canceled) 
     
     
         14 . A nucleic acid vector comprising the expression cassette of  claim 1 . 
     
     
         15 . The vector of  claim 14 , wherein the vector is an adeno-associated vital (AAV) vector. 
     
     
         16 . The vector of  claim 15 , wherein the serotype of the capsid sequence and the serotype of the ITRs of said AAV vector are independently selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, and AAV12. 
     
     
         17 . (canceled) 
     
     
         18 . A method for treating a disease associated with a genetic mutation, substitution, or deletion that affects retinal cone cells, wherein the method comprises administering to a subject in need of such treatment a vector that comprises the promoter of  claim 1 , thereby treating the subject. 
     
     
         19 . (canceled) 
     
     
         20 . A method for treating achromatopsia comprising administering the vector of  claim 14  to a subject in need of such treatment, thereby treating the subject. 
     
     
         21 . (canceled) 
     
     
         22 . (canceled) 
     
     
         23 . A kit comprising
 (a) a vector that comprises the promoter of  claim 1 , and   (b) instructions for use thereof.   
     
     
         24 . A kit comprising
 (a) the nucleic acid vector of  claim 14 , and   (b) instructions for use thereof.   
     
     
         25 . A method of making a recombinant adeno-associated viral (rAAV) vector comprising inserting into an adeno-associated viral vector any one of the promoters of  claim 1  and a nucleic acid selected from the group consisting of a CNGB3 nucleic acid, a CNGA3 nucleic acid, and a GNAT2 nucleic acid. 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . (canceled)