Modulation neural pathways

Abstract

Provided herein are compositions and methods for the design of synthetic regulatory sequences and for subsequent modulation of neural pathways.

1 . A recombinant nucleic acid comprising a nucleic acid encoding a light-sensitive protein operatively linked to a GRM6 regulatory sequence, wherein said GRM6 regulatory sequence is at least 70% identical to SEQ ID NO:1. 
     
     
         2 . (canceled) 
     
     
         3 . A recombinant nucleic acid comprising a nucleic acid encoding a light-sensitive protein operatively linked to a GRM6 regulatory sequence, wherein said GRM6 regulatory sequence comprises a sequence that is at least 95% identical to at least one of the sequences in SEQ ID NOs:2-11. 
     
     
         4 . The recombinant nucleic acid of  claim 1 , wherein, when the recombinant nucleic acid is introduced into the retina of a subject, greater than 80% of cells that express said light-sensitive protein are retinal ON bipolar cells. 
     
     
         5 . (canceled) 
     
     
         6 . The recombinant nucleic acid of  claim 1 , wherein, when the recombinant nucleic acid is introduced into the retina of a subject, less than 20% of cells that express said light-sensitive protein are retinal OFF rod bipolar cells. 
     
     
         7 . The recombinant nucleic acid of  claim 1 , wherein said GRM6 is human GRM6. 
     
     
         8 . The recombinant nucleic acid of  claim 1 , wherein said light-sensitive protein is selected from the group consisting of ChR1, ChR2, VChR1, ChR2 C128A, ChR2 C128S, ChR2 C128T, ChR1-ChR2 hybrids/chimeras, ChD, ChEF, ChF, ChTEF, NpHR, eNpHR, Cheta, ChR65, CIV1, melanopsin, and variants thereof. 
     
     
         9 . The recombinant nucleic acid of  claim 1 , wherein said light-sensitive protein is ChR2 or a light-sensitive protein that is at least 70% identical to ChR2. 
     
     
         10 . (canceled) 
     
     
         11 . (canceled) 
     
     
         12 . The recombinant nucleic acid of  claim 1 , wherein the nucleic acid is encapsidated within a recombinant adeno-associated virus (AAV). 
     
     
         13 . The recombinant nucleic acid of  claim 12  wherein the recombinant adeno-associated virus is of a serotype selected from the group consisting of AAV1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, and hybrids thereof. 
     
     
         14 . (canceled) 
     
     
         15 . The recombinant nucleic acid of  claim 12 , wherein the recombinant AAV virus comprises a mutated capsid protein. 
     
     
         16 . (canceled) 
     
     
         17 . (canceled) 
     
     
         18 . (canceled) 
     
     
         19 . A method of treating a subject suffering from a disease or disorder of the eye comprising introducing into an affected eye a recombinant adeno-associated virus (AAV) comprising the recombinant nucleic acid of  claim 1 . 
     
     
         20 . (canceled) 
     
     
         21 . A method of manufacturing a synthetic regulatory element that targets a specific cell type comprising:
 a. profiling the expression of a plurality of genes in at least one positive target cell and at least one negative target cell, thereby obtaining a set of expression data;   b. analyzing said set of expression data of step (a) in order to identify at least one regulatory motif in said at least one positive target cell that activate gene expression;   c. analyzing said set of expression data of step (a) in order to identify at least one regulatory motif in said at least one negative target cell that inhibits gene expression; and   d. constructing a polynucleotide that comprises said at least one regulatory motif of step (b) or said at least one regulatory motif of step (c).   
     
     
         22 . The method of  claim 21 , wherein said at least one negative target cell is a cell that neighbors said at least one positive target cell. 
     
     
         23 . The method of  claim 21 , wherein said at least one negative target cell is a cell type that is different than said at least one positive target cell. 
     
     
         24 . The method of  claim 21 , wherein said at least one negative target cell is derived from the same tissue type as said at least one positive target cell. 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 .- 46 . (canceled) 
     
     
         47 . The method of  claim 21 , wherein said polynucleotide of step (d) comprises at least one activating regulatory motif and at least one inhibitory regulatory motif. 
     
     
         48 .- 59 . (canceled) 
     
     
         60 . The recombinant nucleic acid of  claim 3 , wherein, when the recombinant nucleic acid is introduced into the retina of a subject, greater than 80% of cells that express said light-sensitive protein are retinal ON bipolar cells. 
     
     
         61 . The recombinant nucleic acid of  claim 3 , wherein, when the recombinant nucleic acid is introduced into the retina of a subject, less than 20% of cells that express said light-sensitive protein are retinal OFF rod bipolar cells. 
     
     
         62 . The recombinant nucleic acid of  claim 3 , wherein said light-sensitive protein is selected from the group consisting of ChR1, ChR2, VChR1, ChR2 C128A, ChR2 C128S, ChR2 C128T, ChR1-ChR2 hybrids/chimeras, ChD, ChEF, ChF, ChTEF, NpHR, eNpHR, Cheta, ChR65, CIV1, melanopsin, and variants thereof. 
     
     
         63 . The recombinant nucleic acid of  claim 3 , wherein said light-sensitive protein is ChR2 or a light-sensitive protein that is at least 70% identical to ChR2. 
     
     
         64 . The recombinant nucleic acid of  claim 3 , wherein the nucleic acid is encapsidated within a recombinant adeno-associated virus (AAV). 
     
     
         65 . The recombinant nucleic acid of  claim 64 , wherein the recombinant adeno-associated virus is of a serotype selected from the group consisting of AAV 1, AAV2, AAV3, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, and hybrids thereof. 
     
     
         66 . The recombinant nucleic acid of  claim 64 , wherein the recombinant AAV virus comprises a mutated capsid protein.