A simple and efficient system for regulating gene expression in human pluripotent stem cells and derivatives.
Regulatable transgene expression in human pluripotent stem cells (hPSCs) and their progenies is often necessary to dissect gene function in a temporal and spatial manner. However, hPSC lines with inducible transgene expression, especially in differentiated progenies, have not been established due to silencing of randomly inserted genes during stem cell expansion and/or differentiation. Here, we report the use of transcription activator-like effector nucleases-mediated targeting to AAVS1 site to generate versatile conditional hPSC lines. Transgene (both green fluorescent protein and a functional gene) expression in hPSCs and their derivatives was not only sustained but also tightly regulated in response to doxycycline both in vitro and in vivo. We modified the donor construct so that any gene of interest can be readily inserted to produce hPSC lines with conditional transgene expression. This technology will substantially improve the way we study human stem cells.
Reproductive Medicine Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China; Waisman Center, School of Medicine and Public Health, University of Wisconsin, Madison, Wisconsin, USA.
Kun
K
Qian
Cindy Tzu-Ling
CT
Huang
CindyTzu-Ling
CL
Huang
Hong
H
Chen
Lisle W
LW
Blackbourn
Yuejun
Y
Chen
Jingyuan
J
Cao
Lin
L
Yao
Cornall
C
Sauvey
Zhongwei
Z
Du
Su-Chun
SC
Zhang
D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000378:metabolism; D015153:Blotting, Western; D004318:Doxycycline / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D016513:Mice, SCID; D018613:Microscopy, Confocal; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D019076:Transgenes / Q000235:genetics*
D049452:Green Fluorescent Proteins; D004318:Doxycycline
Gene editing; Gene regulation; Neural differentiation; Stem cells