title: Transposition from a gutless adeno-transposon vector stabilizes transgene expression in vivo.
abstract: A major limitation of adenovirus-mediated gene therapy for inherited diseases is the instability of transgene expression in vivo, which originates at least in part from the loss of the linear, extrachromosomal vector genomes. Herein we describe the production of a gene-deleted adenovirus-transposon vector that stably maintains virus-encoded transgenes in vivo through integration into host cell chromosomes. This system utilizes a donor transposon vector that undergoes Flp-mediated recombination and excision of its therapeutic payload in the presence of the Flp and Sleeping Beauty recombinases. Systemic in vivo delivery of this system resulted in efficient generation of transposon circles and stable transposase-mediated integration in mouse liver. Somatic integration was sufficient to maintain therapeutic levels of human coagulation Factor IX for more than six months in mice undergoing extensive liver proliferation. These vectors combine the versatility of adenoviral vectors with the integration capabilities of a eukaryotic DNA transposon and should prove useful in the treatment of genetic diseases.
authors.affiliation: Department of Pediatrics and Genetics, Stanford University School of Medicine, Stanford, CA 94305-5208, USA.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Yant
authors.forename: Anja
authors.initials: A
authors.lastname: Ehrhardt
authors.forename: Jacob Giehm
authors.initials: JG
authors.lastname: Mikkelsen
authors.forename: Leonard
authors.initials: L
authors.lastname: Meuse
authors.forename: Thao
authors.initials: T
authors.lastname: Pham
authors.forename: Mark A
authors.initials: MA
authors.lastname: Kay
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D004251:DNA Transposable Elements / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics; D005786:Gene Expression Regulation; D018014:Gene Transfer Techniques; D006367:HeLa Cells; D006801:Humans; D008099:Liver / Q000201:enzymology*; D051379:Mice; D008810:Mice, Inbred C57BL; D019076:Transgenes / Q000235:genetics*; D019895:Transposases / Q000145:classification / Q000235:genetics
chemical_list: D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D019895:Transposases