title: Chromosomal transposition of PiggyBac in mouse embryonic stem cells.
abstract: Transposon systems are widely used for generating mutations in various model organisms. PiggyBac (PB) has recently been shown to transpose efficiently in the mouse germ line and other mammalian cell lines. To facilitate PB's application in mammalian genetics, we characterized the properties of the PB transposon in mouse embryonic stem (ES) cells. We first measured the transposition efficiencies of PB transposon in mouse embryonic stem cells. We next constructed a PB/SB hybrid transposon to compare PB and Sleeping Beauty (SB) transposon systems and demonstrated that PB transposition was inhibited by DNA methylation. The excision and reintegration rates of a single PB from two independent genomic loci were measured and its ability to mutate genes with gene trap cassettes was tested. We examined PB's integration site distribution in the mouse genome and found that PB transposition exhibited local hopping. The comprehensive information from this study should facilitate further exploration of the potential of PB and SB DNA transposons in mammalian genetics.
authors.affiliation: The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire CB10 1SA, United Kingdom.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Chengyi
authors.initials: C
authors.lastname: Lin
authors.forename: Dong
authors.initials: D
authors.lastname: Lu
authors.forename: Zeming
authors.initials: Z
authors.lastname: Ning
authors.forename: Tony
authors.initials: T
authors.lastname: Cox
authors.forename: David
authors.initials: D
authors.lastname: Melvin
authors.forename: Xiaozhong
authors.initials: X
authors.lastname: Wang
authors.forename: Allan
authors.initials: A
authors.lastname: Bradley
authors.forename: Pentao
authors.initials: P
authors.lastname: Liu
mesh_terms: D000818:Animals; D001483:Base Sequence; D033481:Chromosomes, Mammalian / Q000378:metabolism*; D019175:DNA Methylation; D004251:DNA Transposable Elements / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000378:metabolism*; D016678:Genome / Q000235:genetics; D051379:Mice; D016296:Mutagenesis; D009153:Mutagens; D011506:Proteins / Q000235:genetics; D022661:RNA, Untranslated; D019895:Transposases / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D009153:Mutagens; D011506:Proteins; D022661:RNA, Untranslated; D019895:Transposases