PiggyBac transposon-based inducible gene expression in vivo after somatic cell gene transfer.

Saridey SK, Liu L, Doherty JE, Kaja A, Galvan DL, Fletcher BS, Wilson MH
Mol Ther. 2009 17 (12): 2115-20

PMID: 19809403 · PMCID: PMC2814386 · DOI:10.1038/mt.2009.234

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.

MeSH Terms (17)

Animals Blotting, Southern Cells, Cultured DNA Transposable Elements Female Genetic Vectors Gene Transfer Techniques Green Fluorescent Proteins Humans Kidney Liver Lung Mice Mice, Inbred C57BL Mice, SCID Transgenes Virus Integration

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