Gene Therapy Using Transposon-Based Vectors

Abstract

Methods and compositions are presented for the administration of transposon-based vectors to an animal or human to provide gene therapy to the animal or human.

1 . A method of providing gene therapy to an animal or a human to treat a disease or a condition comprising:
 administering to the animal or the human a transposon-based vector in an acceptable carrier, the transposon-based vector comprising an isolated polynucleotide sequence encoding:   a) a gene operably linked to a first promoter, the gene encoding for a transposase; and,   b) one or more genes of interest operably-linked to one or more additional promoters, wherein the one or more genes of interest and their operably-linked promoters are flanked by transposase insertion sequences recognized by the transposase, and wherein the first promoter and the one or more additional promoters are cell-specific promoters or constitutive promoters.   
     
     
         2 . The method of  claim 1 , wherein the gene of interest codes for production of a protein, peptide or nucleic acid. 
     
     
         3 . The method of  claim 1 , further comprising a polyA sequence located 3′ to the one or more genes of interest. 
     
     
         4 . The method of  claim 1 , wherein the gene therapy comprises production of a protein, peptide or nucleic acid encoded by the one or more genes of interest in the animal or the human. 
     
     
         5 . The method of  claim 1 , wherein the administration is effective to treat a disease or a condition. 
     
     
         6 . The method of  claim 1 , wherein the administration of the transposon-based vector results in a transfection efficiency of at least 40%. 
     
     
         7 . The method of  claim 1 , wherein the administration occurs through the vascular system. 
     
     
         8 . An animal produced by the method of  claim 1 . 
     
     
         9 . The method of  claim 1 , wherein the transposon-based vector comprises at least one of: (a) a Kozak sequence positioned so as to include at least the first codon of the transposase gene; (b) two stop codons operably-linked to the transposase gene; (c) a modified transposase gene sequence, wherein at least one of the first twenty codons of the transposase gene is modified by changing a nucleotide at a third base position of the codon to an adenine or thymine without modifying the amino acid encoded by the codon; or (d) a polyA sequence operably-linked to the transposase gene. 
     
     
         10 . The method of  claim 4 , wherein the nucleic acid is an inhibitory RNA. 
     
     
         11 . The method of  claim 5 , wherein the disease is cancer. 
     
     
         12 . The method of  claim 5 , wherein the condition is fertility. 
     
     
         13 . The method of  claim 5 , wherein the protein is a secreted protein. 
     
     
         14 . The method of  claim 1 , wherein the gene therapy provides cell specific expression or tissue specific expression of the one or more genes of interest. 
     
     
         15 . The method of  claim 7 , wherein the administration through the vascular system comprises administration into the left cardiac ventricle. 
     
     
         16 . The method of  claim 1  further comprising administration of a transfection reagent.