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A versatile, highly efficient, and potentially safer piggyBac transposon system for mammalian genome manipulations
Author(s) -
Meir YaaJyuhn James,
Lin Angelique,
Huang MengFan,
Lin JiaoRung,
Weirauch Matthew T.,
Chou HsiangChen,
Lin SiangJin Ashley,
Wu Sareina ChiungYuan
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fj.12-223586
Subject(s) - transposase , transposable element , biology , genetics , hek 293 cells , genome , zebrafish , transposition (logic) , embryonic stem cell , p element , microbiology and biotechnology , computational biology , gene , linguistics , philosophy
The piggyBac transposon is one of the most attractive nonviral tools for mammalian genome manipulations. Given that piggybac mobilizes in a “cut‐and‐paste” fashion, integrant remobilization could potentially damage the host genome. Here, we report a novel piggyBac transposon system with a series of recombinant transposases. We found that the transposition activity of wild‐type (PBase) and hyperactive (hyPBase) piggyBac transposases can be significantly increased by peptide fusions in a cell‐type dependent fashion, with the greatest change typically seen in mouse embryonic stem (ES) cells. The two most potent recombinant transposases, TPLGMH and ThyPLGMH, give a 9‐ and 7‐fold increase, respectively, in the number of integrants in HEK293 compared with Myc‐tagged PBase (MycPBase), and both display 4‐fold increase in generating induced pluripotential stem cells. Interestingly, ThyPLGMH but not TPLGMH shows improved chromosomal excision activity (2.5‐fold). This unique feature of TPLGMH provides the first evidence that integration activity of a transposase can be drastically improved without increasing its remobilization activity. Transposition catalyzed by ThyPLGMH is more random and occurs further from CpG islands than that catalyzed by MycPBase or TPLGMH. Our transposon system diversifies the mammalian genetic toolbox and provides a spectrum of piggyBac transposases that is better suited to different experimental purposes.—Meir, Y.‐J. J., Lin, A., Huang, M.‐F., Lin, J.‐R., Weirauch, M. T., Chou, H.‐C, Lin, S.‐J., Wu, S. C. A versatile, highly efficient, and potentially safer piggyBac transposon system for mammalian genome manipulations. FASEB J . 27, 4429–4443 (2013). www.fasebj.org