Methods of in vivo gene transfer using a sleeping beauty transposon system

Abstract

Methods and compositions for introducing a nucleic acid into the genome of at least one cell of a multicellular organism are provided. In the subject methods, a Sleeping Beauty transposon that includes the nucleic acid is administered to the multicellular organism along with a source of a Sleeping Beauty transposase activity. Administration of the transposon and transposase results in integration of the transposon, as well as the nucleic acid present therein, into the genome of at least one cell of the multicellular organism The subject methods find use in a variety of different applications, including the in vivo transfer of genes for use in, among other applications, gene therapy applications.

What is claimed is:  
     
         1 . A method of integrating an exogenous nucleic acid into the genome of at least one cell of a multicellular organism, said method comprising: 
 administering to said multicellular organism: 
 (a) a Sleeping Beauty transposon comprising said exogenous nucleic acid; and  
 (b) a source of Sleeping Beauty transposase activity;  
   whereby said nucleic acid is integrated into said genome.    
     
     
         2 . The method according to  claim 1 , wherein said source of Sleeping Beauty transposase activity comprises a nucleic acid encoding a protein having Sleeping Beauty transposase activity.  
     
     
         3 . The method according to  claim 1 , wherein said transposon and source of transposase activity are present on separate vectors.  
     
     
         4 . The method according to  claim 1 , wherein said transposon and source of transposase activity are present on the same vector.  
     
     
         5 . The method according to  claim 1 , wherein said multicellular organism is a vertebrate.  
     
     
         6 . A method of inserting an exogenous nucleic acid into the genome of at least one cell of a mammal, said method comprising: 
 administering to said mammal: 
 (a) a Sleeping Beauty transposon comprising said exogenous nucleic acid; and  
 (b) a source of a Sleeping Beauty transposase activity;  
   whereby said exogenous nucleic acid is inserted into said genome.    
     
     
         7 . The method according to  claim 6 , wherein said source of Sleeping Beauty transposase activity comprises a nucleic acid encoding a protein having Sleeping Beauty transposase activity.  
     
     
         8 . The method according to  claim 6 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase activity are present on separate vectors.  
     
     
         9 . The method according to  claim 6 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase activity are present on the same vector.  
     
     
         10 . The method according to  claim 6 , wherein said exogenous nucleic acid comprises a gene.  
     
     
         11 . The method according to  claim 10 , wherein said gene encodes a therapeutic protein.  
     
     
         12 . A method for expressing an exogenous gene in at least one cell of a multicellular organism, said method comprising: 
 administering to said organism: 
 (a) a Sleeping Beauty transposon comprising said exogenous gene; and  
 (b) a source of Sleeping Beauty transposase activity,  
   whereby said gene is integrated into the genome of at least one cell of said multicellular organism and expressed therein;    whereby said exogenous gene is expressed in said at least one cell of said multicellular organism.    
     
     
         13 . The method according to  claim 12 , wherein said source of Sleeping Beauty transposase activity comprises a nucleic acid encoding said transposase.  
     
     
         14 . The method according to  claim 12 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase activity are present on separate vectors.  
     
     
         15 . The method according to  claim 12 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase activity are present on the same vector.  
     
     
         16 . The method according to  claim 12 , wherein said multicellular organism is a vertebrate animal.  
     
     
         17 . The method according to  claim 16 , wherein said vertebrate animal is a mammal.  
     
     
         18 . The method according to  claim 12 , wherein said exogenous gene encodes a therapeutic product.  
     
     
         19 . A method for at least enhancing the amount of a protein present in a multicellular organism, said method comprising: 
 administering to said organism: 
 (a) a Sleeping Beauty transposon comprising an exogenous gene encoding said protein; and  
 (b) a source of Sleeping Beauty transposase activity;  
   whereby said gene integrate into the genome of at least one cell of said multicellular organism and is expressed therein;    whereby the amount of said protein in said multicellular organism is at least enhanced.    
     
     
         20 . The method according to  claim 19 , wherein said source of Sleeping Beauty transposase activity comprises a nucleic acid encoding said transposase.  
     
     
         21 . The method according to  claim 19 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase are present on separate vectors.  
     
     
         22 . The method according to  claim 19 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase are present on the same vector.  
     
     
         23 . The method according to  claim 19 , wherein said multicellular organism is a vertebrate animal.  
     
     
         24 . The method according to  claim 23 , wherein said vertebrate animal is a mammal.  
     
     
         25 . The method according to  claim 19 , wherein said multicellular organism comprises substantially no amount of said protein prior to said administering step.  
     
     
         26 . The method according to  claim 19 , wherein said multicellular organism comprises at least some amount of said protein prior to said administering step.  
     
     
         27 . A method of in vivo transfer of a gene into the genome of at least one cell of a multicellular organism, said method comprising: 
 administering to said multicellular organism: 
 (a) a Sleeping Beauty transposon comprising said gene; and  
 (b) a source of a Sleeping Beauty transposase activity;  
   whereby said gene is transferred into said genome.    
     
     
         28 . The method according to  claim 27 , wherein said source of Sleeping Beauty transposase activity comprises a nucleic acid encoding a protein having Sleeping Beauty transposase activity.  
     
     
         29 . The method according to  claim 27 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase activity are present on separate vectors.  
     
     
         30 . The method according to  claim 27 , wherein said Sleeping Beauty transposon and said source of Sleeping Beauty transposase activity are present on the same vector.