Molecular Therapy - Nucleic Acids
Volume 13, 7 December 2018, Pages 208-219
Journal home page for Molecular Therapy - Nucleic Acids

Original Article
HIT-Cas9: A CRISPR/Cas9 Genome-Editing Device under Tight and Effective Drug Control

https://doi.org/10.1016/j.omtn.2018.08.022Get rights and content
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The CRISPR/Cas9 enabled efficient gene editing in an easy and programmable manner. Controlling its activity in greater precision is desired for biomedical research and potential therapeutic translation. Here, we engrafted the CRISPR/Cas9 system with a mutated human estrogen receptor (ERT2), which renders it 4-hydroxytamoxifen (4-OHT) inducible for the access of genome, and a nuclear export signal (NES), which lowers the background activity. Tight and efficient drug-inducible genome editing was achieved across several human cell types, including embryonic stem cells (ESCs) and mesenchymal stem cells (MSCs), upon vigorous optimization. Optimized terminal device, which we named hybrid drug inducible CRISPR/Cas9 technology (HIT-Cas9), delivered advantageous performances over several existing designs. Such architecture was also successfully applied to an orthogonal Cas9. The HIT-Cas9 system developed in this study will find broad utility in controlled editing of potentially any genomic loci.

Keywords

CRISPR/Cas9
genome editing
drug inducible
stem cells

Cited by (0)

5

These authors contributed equally to this work.

6

Present address: National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.

7

Present address: School of Life Sciences, Beijing University of Chinese Medicine, Beijing 100029, China.