Assay to determine lrrk2 activity in parkinson's disease

Abstract

Disclosed are novel phosphorylation sites identified in LRRK2 and associated with Parkinson's Disease, antibodies that specifically bind to the novel phosphorylation sites, and laboratory and clinical uses thereof.

What is claimed is: 
     
         1 . A method of editing an LRRK2 gene comprising:
 a) contacting the LRRK2 gene with an engineered nuclease that cleaves the LRRK2 gene at a site in the LRRK2 gene; and   b) producing a base alteration, base deletion or base insertion at the site that results in an amino acid change in an LRRK2 protein encoded by the LRRK2 gene.   
     
     
         2 . The method of  claim 1 , wherein the amino acid change results in reduced aggregation of LRRK2 proteins encoded by the LRRK2 gene. 
     
     
         3 . The method of  claim 1 , wherein the amino acid change results in altering a kinase activity of the LRRK2 protein. 
     
     
         4 . The method of  claim 1 , wherein the amino acid change occurs at a position in the LRRK2 protein selected from amino acid positions 910, 935, 955, 973, 1441, 1699, 2019, and 2020 of the LRRK2 protein. 
     
     
         5 . The method of  claim 1 , wherein producing the amino acid change restores a phosphorylation site in the LRRK2 protein. 
     
     
         6 . The method of  claim 1 , wherein the editing occurs in a neuronal stem cell. 
     
     
         7 . The method of  claim 1 , wherein the editing occurs in an induced pluripotent stem cell. 
     
     
         8 . The method of  claim 1 , wherein the engineered nuclease produces a double-stranded break in the LRRK2 gene. 
     
     
         9 . The method of  claim 8 , wherein the double stranded break is resolved through homology-directed repair or non-homologous end joining. 
     
     
         10 . The method of  claim 8 , wherein the double stranded break is repaired with a donor DNA template. 
     
     
         11 . The method of  claim 1 , wherein the engineered nuclease is a zinc finger nuclease. 
     
     
         12 . The method of  claim 1 , wherein the base alteration, base deletion or base insertion results in correcting a mutation in the LRRK2 protein selected from R1441C, R1441G, Y1699C, 12020T, S910A, S910E, S910D, S955A, S955E, S955D, S973A, S973E, and S973D. 
     
     
         13 . A cell comprising an LRRK2 gene modified by an engineered nuclease. 
     
     
         14 . The cell of  claim 13 , wherein the cell is a neuronal stem cell. 
     
     
         15 . The cell of  claim 13 , wherein the cells is an induced pluripotent stem cell. 
     
     
         16 . The cell of  claim 13 , wherein the LRRK2 gene is modified to correct a genetic mutation, wherein the mutation causes an amino acid alteration in an LRRK2 protein encoded by the LRRK2 gene selected from R1441C, R1441G, Y1699C, 12020T, S910A, S910E, S910D, S955A, S955E, S955D, S973A, S973E, and S973D. 
     
     
         17 . The cell of  claim 13 , wherein the LRRK2 protein comprised a mutation before being modified by the engineered nuclease, and wherein the mutation is selected from R1441C, R1441G, Y1699C, 12020T, S910A, S910E, S910D, S955A, S955E, S955D, S973A, S973E, and S973D. 
     
     
         18 . A method of treating a subject for a condition comprising administering the cell of  claim 13  to the subject. 
     
     
         19 . The method of  claim 18 , wherein the condition is a neurodegenerative disease. 
     
     
         20 . The method of  claim 18 , wherein the condition is Parkinson's disease.