title: PiggyBac transposon-based inducible gene expression in vivo after somatic cell gene transfer.
abstract: Somatic cell gene transfer has permitted inducible gene expression in vivo through coinfection of multiple viruses. We hypothesized that the highly efficient plasmid-based piggyBac transposon system would enable long-term inducible gene expression in mice in vivo. We used a multiple-transposon delivery strategy to create a tetracycline-inducible expression system in vitro in human cells by delivering the two genes on separate transposons for inducible reporter gene expression along with a separate selectable transposon marker. Evaluation of stable cell lines revealed 100% of selected clones exhibited inducible expression via stable expression from three separate transposons simultaneously. We next tested and found that piggyBac-mediated gene transfer to liver or lung could achieve stable reporter gene expression in mice in vivo in either immunocompetent or immune deficient animals. A single injection of piggyBac transposons could achieve long-term inducible gene expression in the livers of mice in vivo, confirming our multiple-transposon strategy used in cultured cells. The plasmid-based piggyBac transposon system enables constitutive or inducible gene expression in vivo for potential therapeutic and biological applications without using viral vectors.
authors.affiliation: Department of Medicine, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: Sai K
authors.initials: SK
authors.lastname: Saridey
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Doherty
authors.forename: Aparna
authors.initials: A
authors.lastname: Kaja
authors.forename: Daniel L
authors.initials: DL
authors.lastname: Galvan
authors.forename: Bradley S
authors.initials: BS
authors.lastname: Fletcher
authors.forename: Matthew H
authors.initials: MH
authors.lastname: Wilson
mesh_terms: D000818:Animals; D015139:Blotting, Southern; D002478:Cells, Cultured; D004251:DNA Transposable Elements / Q000235:genetics*; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000378:metabolism / Q000821:virology; D008168:Lung / Q000166:cytology / Q000378:metabolism / Q000821:virology; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D019076:Transgenes / Q000235:genetics*; D016662:Virus Integration
chemical_list: D004251:DNA Transposable Elements; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins