TY - JOUR
T1 - A chemical-inducible CRISPR-Cas9 system for rapid control of genome editing
AU - Liu, Kaiwen Ivy
AU - Ramli, Muhammad Nadzim Bin
AU - Woo, Cheok Wei Ariel
AU - Wang, Yuanming
AU - Zhao, Tianyun
AU - Zhang, Xiujun
AU - Yim, Guo Rong Daniel
AU - Chong, Bao Yi
AU - Gowher, Ali
AU - Chua, Mervyn Zi Hao
AU - Jung, Jonathan
AU - Lee, Jia Hui Jane
AU - Tan, Meng How
N1 - Publisher Copyright:
© 2016 Nature America, Inc. All rights reserved.
PY - 2016/11/1
Y1 - 2016/11/1
N2 - CRISPR-Cas9 has emerged as a powerful technology that enables ready modification of the mammalian genome. The ability to modulate Cas9 activity can reduce off-target cleavage and facilitate precise genome engineering. Here we report the development of a Cas9 variant whose activity can be switched on and off in human cells with 4-hydroxytamoxifen (4-HT) by fusing the Cas9 enzyme with the hormone-binding domain of the estrogen receptor (ERT2). The final optimized variant, termed iCas, showed low endonuclease activity without 4-HT but high editing efficiency at multiple loci with the chemical. We also tuned the duration and concentration of 4-HT treatment to reduce off-target genome modification. Additionally, we benchmarked iCas against other chemical-inducible methods and found that it had the fastest on rate and that its activity could be toggled on and off repeatedly. Collectively, these results highlight the utility of iCas for rapid and reversible control of genome-editing function.
AB - CRISPR-Cas9 has emerged as a powerful technology that enables ready modification of the mammalian genome. The ability to modulate Cas9 activity can reduce off-target cleavage and facilitate precise genome engineering. Here we report the development of a Cas9 variant whose activity can be switched on and off in human cells with 4-hydroxytamoxifen (4-HT) by fusing the Cas9 enzyme with the hormone-binding domain of the estrogen receptor (ERT2). The final optimized variant, termed iCas, showed low endonuclease activity without 4-HT but high editing efficiency at multiple loci with the chemical. We also tuned the duration and concentration of 4-HT treatment to reduce off-target genome modification. Additionally, we benchmarked iCas against other chemical-inducible methods and found that it had the fastest on rate and that its activity could be toggled on and off repeatedly. Collectively, these results highlight the utility of iCas for rapid and reversible control of genome-editing function.
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U2 - 10.1038/nchembio.2179
DO - 10.1038/nchembio.2179
M3 - Article
C2 - 27618190
AN - SCOPUS:84987678200
SN - 1552-4450
VL - 12
SP - 980
EP - 987
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 11
ER -