title: Somatic integration and long-term transgene expression in normal and haemophilic mice using a DNA transposon system.
abstract: The development of non-viral gene-transfer technologies that can support stable chromosomal integration and persistent gene expression in vivo is desirable. Here we describe the successful use of transposon technology for the nonhomologous insertion of foreign genes into the genomes of adult mammals using naked DNA. We show that the Sleeping Beauty transposase can efficiently insert transposon DNA into the mouse genome in approximately 5-6% of transfected mouse liver cells. Chromosomal transposition resulted in long-term expression (>5 months) of human blood coagulation factor IX at levels that were therapeutic in a mouse model of haemophilia B. Our results establish DNA-mediated transposition as a new genetic tool for mammals, and provide new strategies to improve existing non-viral and viral vectors for human gene therapy applications.
authors.affiliation: Departments of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, California, USA.
authors.forename: S R
authors.initials: SR
authors.lastname: Yant
authors.forename: L
authors.initials: L
authors.lastname: Meuse
authors.forename: W
authors.initials: W
authors.lastname: Chiu
authors.forename: Z
authors.initials: Z
authors.lastname: Ivics
authors.forename: Z
authors.initials: Z
authors.lastname: Izsvak
authors.forename: M A
authors.initials: MA
authors.lastname: Kay
mesh_terms: D000818:Animals; D001483:Base Sequence; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D004198:Disease Susceptibility; D018628:Gene Dosage; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D002836:Hemophilia B / Q000235:genetics* / Q000628:therapy; D006801:Humans; D007275:Injections, Intravenous; D008099:Liver / Q000166:cytology / Q000201:enzymology / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D008969:Molecular Sequence Data; D010957:Plasmids / Q000008:administration & dosage / Q000627:therapeutic use; D019076:Transgenes / Q000235:genetics*; D019895:Transposases / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; C111652:mariner transposases; C081454:Tc1 transposase; D019895:Transposases