Small molecules promote CRISPR-Cpf1-mediated genome editing in human pluripotent stem cells

  • Nat Commun. 2018 Apr 3;9(1):1303. doi: 10.1038/s41467-018-03760-5.
Xiaojie Ma  1 ,  Xi Chen  1 ,  Yan Jin  1 ,  Wenyan Ge  1 ,  Weiyun Wang  1 ,  Linghao Kong  1 ,  Junfang Ji  1 ,  Xing Guo  1 ,  Jun Huang  1 ,  Xin-Hua Feng  1 ,  Junfen Fu  2 ,  Saiyong Zhu  3  4
Affiliations
  • 1. Life Sciences Institute, Zhejiang University, 310058, Hangzhou, China.
  • 2. The Children's Hospital, Zhejiang University School of Medicine, 310003, Hangzhou, China.
  • 3. Life Sciences Institute, Zhejiang University, 310058, Hangzhou, China. [email protected].
  • 4. Stem Cell Institute, Zhejiang University, 310058, Hangzhou, China. [email protected].
PMID: 29610531 DOI: 10.1038/s41467-018-03760-5
Abstract

Human pluripotent stem cells (hPSCs) have potential applications in biological studies and regenerative medicine. However, precise genome editing in hPSCs remains time-consuming and labor-intensive. Here we demonstrate that the recently identified CRISPR-Cpf1 can be used to efficiently generate knockout and knockin hPSC lines. The unique properties of CRISPR-Cpf1, including shorter crRNA length and low off-target activity, are very attractive for many applications. In particular, we develop an unbiased drug-selection-based platform feasible for high-throughput screening in hPSCs and this screening system enables us to identify small molecules VE-822 and AZD-7762 that can promote CRISPR-Cpf1-mediated precise genome editing. Significantly, the combination of CRISPR-Cpf1 and small molecules provides a simple and efficient strategy for precise genome engineering.

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