|Handler, Alfred - Al|
Submitted to: Proceedings of the National Academy of Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/16/2006
Publication Date: 10/10/2006
Citation: Wu, S.C., Meir, Y.J., Coates, C.J., Handler, A.M., Pelczar, P., Moisyadi, S., Kaminski, J.M. 2006. PiggyBac: A flexible and highly active transposon as compared to Sleeping Beauty, Tol2, and Mos1 in mammalian cells. Proceedings of the National Academy of Sciences. 103:15008-15013. Interpretive Summary: The creation of transgenic strains of economically important insects for the development of more effective biological control programs is a major goal of our laboratory at CMAVE. Development of this methodology and strategies to effectively and safely utilize transgenic insects for biological control will depend upon new vector systems that can be targeted to specific genomic integration sites, and by also knowing the potential host range for vector function. This study of transposon vector activity in mammalian cells shows that the piggyBac vector, which is the most commonly used vector in insects, is highly active in mammalian cells. Therefore it is important that piggyBac vectors used in insects be stabilized to limit unintended transmission into mammalian orgainisms. We also show that piggyBac can, uniquely, be coupled to a GAL4-based targeting system for improved transgenesis.
Technical Abstract: A non-viral vector for highly efficient site-specific integration would be desirable for many applications in transgenesis, including gene therapy. In this study, we directly compared the genomic integration efficiencies of piggyBac, hyperactive Sleeping Beauty(SB11), Tol2, and Mos1 in four mammalian cell lines. piggyBac demonstrated significantly higher transposition activity in all cell lines, while Mos1 had no activity. Furthermore, piggyBac transposase coupled to the GAL4 DNA binding domain retains trasposition activity, whereas similarly manipulated gene products of Tol2 and SB11 were inactive. The high transposition activity of piggyBac and the flexibility for molecular modification of its transposase suggest the possibility of utilizing it routinely for mammalian transgenesis.