Crispr/cas-related methods and compositions for treating cystic fibrosis

Abstract

CRISPR/CAS-related compositions and methods for treatment of Cystic Fibrosis (CF).

What is claimed is: 
     
         1 . A method of altering a cystic fibrosis transmembrane conductance regulator (CFTR) gene in a cell, comprising administering to said cell:
 (i) a CRISPR/Cas system comprising: (a) a gRNA molecule comprising a targeting domain that is complementary with a target sequence of a CFTR gene and (b) a Cas9 molecule; or   (ii) a nucleic acid composition comprising: (a) a first nucleotide sequence encoding a gRNA molecule comprising a targeting domain that is complementary with a target sequence of a CFTR gene and (b) a second nucleotide sequence encoding a Cas9 molecule.   
     
     
         2 . The method of  claim 1 , wherein said alteration comprises correction of one or more mutations in said CFTR gene. 
     
     
         3 . The method of  claim 1 , wherein said alteration of said CFTR gene results in restoration of a mutant CFTR gene to its wild type state thereby providing a functional CFTR protein. 
     
     
         4 . The method of  claim 1 , wherein said cell is from a subject suffering from or at risk for Cystic Fibrosis (CF) and CF-like disease. 
     
     
         5 . The method of  claim 1 , wherein said cell is from a subject having a mutation at a CF target mutant position. 
     
     
         6 . The method of  claim 1 , wherein said cell is selected from the group consisting of an airway cell, an epithelial cell, and a stem cell. 
     
     
         7 . The method of  claim 6 , wherein said epithelial cell is selected from the group consisting of a lung epithelial cell, a gastrointestinal epithelial cell, a biliary epithelial cell, a pancreatic ductal epithelial cell, a gastrointestinal cell, a hepatobiliary epithelial cell, a gallbladder epithelial cell, a large intestine epithelial cell, a small intestine epithelial cell, a jejunum epithelial cell, a epithelial cell of the ileum, a duodenum epithelial cell, and a reproductive epithelial cell; and said stem cell is selected from the group consisting of an embryonic stem cell, an induced pluripotent stem cell, a pulmonary stem cell, a gastrointestinal stem cell, and a mesenchymal stem cell. 
     
     
         8 . The method of  claim 1 , wherein said targeting domain that is complementary with a target sequence of said CFTR gene comprises or consists of a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a nucleotide sequence set forth in SEQ ID NOs: 15101-27265. 
     
     
         9 . A method of altering a sodium channel epithelial 1 alpha (SCNN1A) gene in a cell, comprising administering to said cell:
 (i) a CRISPR/Cas system comprising: (a) a gRNA molecule comprising a targeting domain that is complementary with a target sequence of a SCNN1A gene and (b) a Cas9 molecule; or   (ii) a nucleic acid composition comprising: (a) a first nucleotide sequence encoding a gRNA molecule comprising a targeting domain that is complementary with a target sequence of a SCNN1A gene and (b) a second nucleotide sequence encoding a Cas9 molecule.   
     
     
         10 . The method of  claim 9 , wherein said alteration comprises correction of one or more mutations in said SCNN1A gene, knockout of said SCNN1A gene, or knockdown of said SCNN1A gene. 
     
     
         11 . The method of  claim 9 , wherein the alteration of said SCNN1A gene results in a reduction or elimination of expression of said SCNN1A gene. 
     
     
         12 . The method of  claim 9 , wherein said cell is from a subject suffering from or at risk for Cystic Fibrosis (CF) and CF-like disease. 
     
     
         13 . The method of  claim 9 , wherein said cell is from a subject having a mutation at a CF target mutant position, or from a subject who would benefit from having a mutation in said SCNN1A gene. 
     
     
         14 . The method of  claim 9 , wherein the cell is selected from the group consisting of an airway cell, an epithelial cell, and a stem cell. 
     
     
         15 . The method of  claim 14 , wherein said epithelial cell is selected from the group consisting of a lung epithelial cell, a gastrointestinal epithelial cell, a biliary epithelial cell, a pancreatic ductal epithelial cell, a gastrointestinal cell, a hepatobiliary epithelial cell, a gallbladder epithelial cell, a large intestine epithelial cell, a small intestine epithelial cell, a jejunum epithelial cell, a epithelial cell of the ileum, a duodenum epithelial cell, and a reproductive epithelial cell; and said stem cell is selected from the group consisting of an embryonic stem cell, an induced pluripotent stem cell, a pulmonary stem cell, a gastrointestinal stem cell, and a mesenchymal stem cell. 
     
     
         16 . The method of  claim 9 , wherein said targeting domain that is complementary to a target sequence of said SCNN1A gene comprises or consists of a nucleotide sequence that is the same as, or differs by no more than 3 nucleotides from, a nucleotide sequence set forth in SEQ ID NOs: 497-654, 803-916, and 4176-15100.