title: PiggyBac transposon-mediated, reversible gene transfer in human embryonic stem cells.
abstract: Permanent and reversible genetic modifications are important approaches to study gene function in different cell types. They are also important for stem cell researchers to explore and test the therapeutic potential of stem cells. The piggyBac transposon from insects is a rising nonviral system that efficiently mutagenizes and mediates gene transfer into the mammalian genome. It is also characterized by its precise excision, leaving no trace sequence behind so that the genomic integrity of the mutated cell can be restored. Here, we use an optimized piggyBac transposon system to mediate gene transfer and expression of a bifunctional fluorescent reporter in human embryonic stem (ES) cells. We provide molecular evidence for transposase-mediated piggyBac integration events and functional evidence for successful expression of a transferred fluorescent protein genes in human ES cells and their in vitro differentiated derivatives. We also demonstrate that the integrated piggyBac transposon can be removed and an undisrupted insertion site can be restored, which implies potential applications for its use in gene therapy and genetics studies.
authors.affiliation: Graduate Institute of Clinical Genomics and Graduate Institute of Clinical Medicine, Stem Cell Core Laboratory, NTU Research Center for Medical Excellence, Taipei, Taiwan. youtzungchen@ntu.edu.tw
authors.forename: You-Tzung
authors.initials: YT
authors.lastname: Chen
authors.forename: Kenryo
authors.initials: K
authors.lastname: Furushima
authors.forename: Pei-Shan
authors.initials: PS
authors.lastname: Hou
authors.forename: Amy T
authors.initials: AT
authors.lastname: Ku
authors.forename: Jian Min
authors.initials: JM
authors.lastname: Deng
authors.forename: Chuan-Wei
authors.initials: CW
authors.lastname: Jang
authors.forename: Haotian
authors.initials: H
authors.lastname: Fang
authors.forename: Henry P
authors.initials: HP
authors.lastname: Adams
authors.forename: Min-Liang
authors.initials: ML
authors.lastname: Kuo
authors.forename: Hong-Nerng
authors.initials: HN
authors.lastname: Ho
authors.forename: Chung-Liang
authors.initials: CL
authors.lastname: Chien
authors.forename: Richard R
authors.initials: RR
authors.lastname: Behringer
mesh_terms: D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D048430:Cell Shape; D002645:Chickens; D004251:DNA Transposable Elements / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D018014:Gene Transfer Techniques; D006801:Humans; D008969:Molecular Sequence Data; D019076:Transgenes / Q000235:genetics; D019895:Transposases / Q000378:metabolism
chemical_list: D004251:DNA Transposable Elements; D019895:Transposases