期刊
FEBS LETTERS
卷 592, 期 13, 页码 2238-2247出版社
WILEY
DOI: 10.1002/1873-3468.13127
关键词
CRISPR; hESCs; orthogonal gene knockout and activation
资金
- National Natural Science Foundation of China [81670093, 81372494]
- Department of Science and Technology of Guangdong Province [2014B020212018]
- Natural Science Foundation of Guangdong Province [2017A030310331]
- Guangzhou Science Technology and Innovation Commission [201508020120]
- Thousand Young Talents Program of China
Human embryonic stem cells (hESCs) have a wide range of applications in early human embryonic development mimics, disease modeling, and cell therapy. To fulfill these applications, we established hESCs for inducible and multiplex orthogonal gene knockout and activation, which we named iKA-CRISPR hESCs. In cells, when complexed with a short guide RNA containing a 14-bp target sequence (14-bp gRNA) or a long 20-bp gRNA, the doxycycline-induced Cas9-p300 protein could activate gene transcription or cleave genomic DNA, respectively. We also demonstrate using iKA-CRISPR hESCs that knockout of OCT4 promoted differentiation, and developmentally relevant microRNAs and transcription factors could be efficiently activated. Thus, iKA-CRISPR hESCs provide a convenient platform to control gene expression networks and, therefore, facilitate the applications of hESCs in basic and translational biomedical research.
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