title: PiggyBac mediated multiplex gene transfer in mouse embryonic stem cell.
abstract: PiggyBac system has been shown to have a high efficiency to mediate gene transfer. However, there are no reports on its efficiency to mediate multiplex transgenes in mouse embryonic stem cells. Here we first established an immortalized feeder cell line by introducing four antibiotic resistance genes simultaneously into the original SNL 76/7 feeder cell line utilizing the PiggyBac system. This is the feeder cell line with the most diverse types of antibiotic resistance genes reported so far, which will enable researchers to perform simultaneous multiplex gene transfer or gene targeting experiments in ES cells. With such feeder cell line, we were able to quantitatively characterize the transposition efficiency of PiggyBac system in mouse ES cells using five transposons carrying different inducible fluorescence proteins and antibiotic resistance genes, and the efficiency ranged from about 2% for one transposon to 0.5% for five transposons. The highly efficient multiplex gene transfer mediated by PiggyBac will no doubt provide researchers with more choices in biomedical research and development.
authors.affiliation: Department of Biochemistry, the University of Hong Kong, Hong Kong, China.
authors.forename: Xibin
authors.initials: X
authors.lastname: Lu
authors.affiliation: Department of Biochemistry, the University of Hong Kong, Hong Kong, China; Department of Biology, Shenzhen Key Laboratory of Cell Microenvironment, South University of Science and Technology of China, Shenzhen, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D002999:Clone Cells / Q000166:cytology; D004251:DNA Transposable Elements / Q000235:genetics*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D061252:Feeder Cells / Q000166:cytology; D005786:Gene Expression Regulation; D018390:Gene Targeting; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D014162:Transfection / Q000379:methods*
chemical_list: D015415:Biomarkers; D004251:DNA Transposable Elements; D008164:Luminescent Proteins