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Stem Cells. 2014 May;32(5):1230-8. doi: 10.1002/stem.1653.

A simple and efficient system for regulating gene expression in human pluripotent stem cells and derivatives.

Author information

1
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.

Erratum in

  • Stem Cells. 2014 Aug;32(8):2290. Huang, CindyTzu-Ling [corrected to Huang, Cindy Tzu–Ling].

Abstract

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.

KEYWORDS:

Gene editing; Gene regulation; Neural differentiation; Stem cells

PMID:
24497442
PMCID:
PMC4121394
DOI:
10.1002/stem.1653
[Indexed for MEDLINE]
Free PMC Article
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