Methods and compositions for nucleic acid transfer into cells

Abstract

The present invention provides compositions and methods for increasing the transfer of nucleic acids into cells. In particular, the present invention provides for the use of inhibitors of HDAC6, a cytoplasmic histone deacetylase present in mammalian cells by, for example, small molecules or siRNA treatment, in increasing gene transfer and/or expression in cells in vitro and in vivo for research and gene therapy applications.

1 . A composition for increasing gene transfer into cells comprising an inhibitor of major tubulin deacetylase (HDAC6) and a gene transfer reagent. 
     
     
         2 . The composition of  claim 1 , wherein said gene transfer reagent is a lipid composition. 
     
     
         3 . The composition of  claim 1 , wherein said gene transfer reagent is calcium phosphate. 
     
     
         4 . The composition of  claim 1 , wherein said gene transfer reagent is DEAE/dextran. 
     
     
         5 . The composition of  claim 1 , wherein said inhibitor is trichostatin A or compound 17b. 
     
     
         6 . The composition of  claim 1 , wherein said inhibitor is a siRNA or shRNA to HDAC6. 
     
     
         7 . A composition comprising an inhibitor of major tubulin deacetylase (HDAC6) and a vector. 
     
     
         8 . The composition of  claim 7 , wherein said inhibitor is trichostatin A or compound 17b. 
     
     
         9 . The composition of  claim 7 , wherein said inhibitor is a siRNA or shRNA to HDAC6. 
     
     
         10 . A method for increasing nucleic acid expression in cells comprising:
 a) providing:
 i) a gene transfer system 
 ii) an inhibitor of major tubulin deacetylase (HDAC6), 
 iii) mammalian cells, and 
 iv) a vector, 
   b) contacting said cells with said vector and said inhibitor of major tubulin deacteylase in the presence of said gene transfer system such that gene expression from said vector is increased in the presence of said inhibitor compared to expression when said inhibitor is absent.   
     
     
         11 . The method of  claim 10 , wherein the gene transfer system comprises an electroporation system. 
     
     
         12 . The method of  claim 10 , wherein the gene transfer system is lipids. 
     
     
         13 . The method of  claim 10 , wherein the gene transfer system is calcium chloride. 
     
     
         14 . The method of  claim 10 , wherein the gene transfer system is DEAE/dextran. 
     
     
         15 . The method of  claim 10 , wherein said inhibitor is trichostatin A or compound 17b. 
     
     
         16 . The method of  claim 10 , wherein said inhibitor is a siRNA or shRNA to HDAC6. 
     
     
         17 . The method of  claim 10 , wherein said cell is in vitro or in vivo. 
     
     
         18 . A method for transfecting lung cells comprising: contacting lung cells with a gene transfer system and a vector under conditions of cyclic stretch such that gene expression from said vector is increased by employing said conditions of cyclic stretch compared to expression when cyclic stretch is not employed. 
     
     
         19 . The method of  claim 18 , wherein said contacting is in vivo in a subject. 
     
     
         20 . The method of  claim 19 , wherein said condition of cyclic stretch comprise ventilating said subject.