Format

Send to

Choose Destination
FEBS Lett. 2018 Jul;592(13):2238-2247. doi: 10.1002/1873-3468.13127. Epub 2018 Jun 15.

iKA-CRISPR hESCs for inducible and multiplex orthogonal gene knockout and activation.

Ma S1, Lv J1, Sun J2,3,4, Tang P5, Li H5, Zhou H5, Zhang Z2,3,4, Lin Y1, Rong Z1.

Author information

1
Cancer Research Institute, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
2
State Key Laboratory of Organ Failure Research, Nanfang Hospital, Southern Medical University, Guangzhou, China.
3
Center for Bioinformatics, School of Basic Medical Sciences, Southern Medical University, Guangzhou, China.
4
Department of Cardiology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
5
Division of Laboratory Medicine, Zhujiang Hospital, Southern Medical University, Guangzhou, China.

Abstract

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.

KEYWORDS:

CRISPR; hESCs; orthogonal gene knockout and activation

PMID:
29869798
DOI:
10.1002/1873-3468.13127

Supplemental Content

Full text links

Icon for Wiley
Loading ...
Support Center