title: A versatile, highly efficient, and potentially safer piggyBac transposon system for mammalian genome manipulations.
abstract: 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.
authors.affiliation: 1S.C.-Y.W., Institute of Molecular Medicine, College of Medicine, Chang Gung University, Tao-Yuan, Taiwan 333. sareinaw@mail.cgu.edu.tw.
authors.forename: Yaa-Jyuhn James
authors.initials: YJ
authors.lastname: Meir
authors.forename: Angelique
authors.initials: A
authors.lastname: Lin
authors.forename: Meng-Fan
authors.initials: MF
authors.lastname: Huang
authors.forename: Jiao-Rung
authors.initials: JR
authors.lastname: Lin
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Weirauch
authors.forename: Hsiang-Chen
authors.initials: HC
authors.lastname: Chou
authors.forename: Siang-Jin Ashley
authors.initials: SJ
authors.lastname: Lin
authors.forename: Sareina Chiung-Yuan
authors.initials: SC
authors.lastname: Wu
mesh_terms: D000818:Animals; D016466:CHO Cells; D065150:Cellular Reprogramming; D002875:Chromosomes / Q000235:genetics; D006224:Cricetinae; D003412:Cricetulus; D004251:DNA Transposable Elements / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods; D016678:Genome; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D019895:Transposases / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; D019895:Transposases
keywords: functional genomics; gene therapy; induced pluripotent stem cell; remobilization