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Title: HIT-Cas9: a CRISPR/Cas9 Genome Editing Device Under Tight and Effective Drug Control
Author: Chen Zhao, Yingze Zhao, Jingfang Zhang, Jia Lu, Li Chen, Yue Zhang, Yue Ying, Junjun Xu, Shixian Wei, Yu Wang
Abstract: The CRISPR/Cas9 (clustered, regularly interspaced, short palindromic repeats /CRISPR associated protein9) 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.
Corresponding author: Yu Wang
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PubYear: 2018
Issue: DOI: 10.1016/j.omtn.2018.08.022
Journal: Molecular Therapy - Nucleic Acids
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URL: https://www.sciencedirect.com/science/article/pii/S2162253118302373