Identification of channelrhodopsin-2 (chr2) mutations and methods of use

Abstract

The invention provides compositions and kits including at least one nucleic acid or polypeptide molecule encoding for a mutant ChR2 protein. Methods of the invention include administering a composition comprising a mutant ChR2 to a subject to preserve, improve, or restore phototransduction. Preferably, the compositions and methods of the invention are provided to a subject having impaired vision, thereby restoring vision to normal levels.

1 .- 50 . (canceled) 
     
     
         51 . An expression vector comprising a polynucleotide encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 26 in which the amino acid at position 132 of SEQ ID NO: 26 is cysteine (C) or alanine (A), and the amino acid at position 159 of SEQ ID NO: 26 is cysteine (C), serine (S), or alanine (A). 
     
     
         52 . The expression vector of  claim 51 , wherein the polypeptide comprises a cysteine (C) at positions 132 and 159 of SEQ ID NO: 26. 
     
     
         53 . The expression vector of  claim 52 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 16 and the polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 15. 
     
     
         54 . The expression vector of  claim 51 , wherein the polypeptide comprises a cysteine (C) at position 132 and a serine (S) at position 159 of SEQ ID NO: 26. 
     
     
         55 . The expression vector of  claim 54 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 19 and the polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 18. 
     
     
         56 . The expression vector of  claim 51 , wherein the polypeptide comprises a cysteine (C) at position 132 and an alanine (A) at position 159 of SEQ ID NO: 26. 
     
     
         57 . The expression vector of  claim 56 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 25 and the polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 24. 
     
     
         58 . The expression vector of  claim 51 , wherein the polypeptide comprises an alanine (A) at position 132 and a cysteine (C) at position 159 of SEQ ID NO: 26. 
     
     
         59 . The expression vector of  claim 58 , wherein the polypeptide comprises the amino acid sequence of SEQ ID NO: 22 and the polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 21. 
     
     
         60 . The expression vector of  claim 51 , wherein the polypeptide comprises an alanine (A) at position 132 and a serine (S) at position 159 of SEQ ID NO: 26. 
     
     
         61 . The expression vector of  claim 51 , wherein the polypeptide comprises an alanine (A) at position 132 and an alanine (A) at position 159 of SEQ ID NO: 26. 
     
     
         62 . The expression vector of  claim 51 , wherein the expression vector is an adeno-associated virus (AAV) vector. 
     
     
         63 . The expression vector of  claim 62 , wherein the AAV vector is an AAV2 vector. 
     
     
         64 . A method of improving or restoring vision in a subject, the method comprising administering to the subject the expression vector of  claim 51 . 
     
     
         65 . A method of improving or restoring vision in a subject, the method comprising administering to the subject the expression vector of  claim 62 . 
     
     
         66 . A method of improving or restoring vision in a subject, the method comprising administering to the subject the expression vector of  claim 63 . 
     
     
         67 . The method of  claim 64 , wherein the subject either has normal vision or the subject is suffering from an ocular disease. 
     
     
         68 . The method of  claim 65 , wherein the subject either has normal vision or the subject is suffering from an ocular disease. 
     
     
         69 . The method of  claim 66 , wherein the subject either has normal vision or the subject is suffering from an ocular disease. 
     
     
         70 . The method of  claim 67 , wherein the subject is suffering from an ocular disease and the ocular disease is macular degeneration or retinitis pigmentosa. 
     
     
         71 . The method of  claim 68 , wherein the subject is suffering from an ocular disease and the ocular disease is macular degeneration or retinitis pigmentosa. 
     
     
         72 . The method of  claim 69 , wherein the subject is suffering from an ocular disease and the ocular disease is macular degeneration or retinitis pigmentosa.