PiggyBac Transposon-based Inducible Gene Expression In Vivo After Somatic Cell Gene Transfer
| Autor: | Matthew H. Wilson, Sai K. Saridey, Li Liu, Daniel L. Galvan, Aparna Kaja, Bradley S. Fletcher, Joseph E. Doherty |
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| Rok vydání: | 2009 |
| Předmět: |
Transposable element
Somatic cell Transgene Virus Integration Genetic Vectors Green Fluorescent Proteins Mice SCID Biology Kidney Viral vector Mice Plasmid Drug Discovery Genetics Animals Humans Transgenes Gene Lung Molecular Biology Cells Cultured Pharmacology Reporter gene Gene Transfer Techniques Original Articles Molecular biology Mice Inbred C57BL Blotting Southern Liver PiggyBac Transposon System DNA Transposable Elements Molecular Medicine Female |
| Zdroj: | Molecular Therapy. 17(12):2115-2120 |
| ISSN: | 1525-0016 |
| DOI: | 10.1038/mt.2009.234 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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