Open AccessA Simple and Efficient System for Regulating Gene Expression in Human Pluripotent Stem Cells and Derivatives
Author(s) -
Qian Kun,
Huang CindyTzuLing,
Chen Hong,
Blackbourn Lisle W.,
Chen Yuejun,
Cao Jingyuan,
Yao Lin,
Sauvey Cornall,
Du Zhongwei,
Zhang SuChun
Publication year - 2014
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1002/stem.1653
Subject(s) - biology , induced pluripotent stem cell , transgene , transcription activator like effector nuclease , stem cell , microbiology and biotechnology , gene silencing , gene , gene expression , effector , cellular differentiation , regulation of gene expression , embryonic stem cell , genetics , genome editing , genome
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. S tem C ells 2014;32:1230–1238