A simple, quick, and efficient CRISPR/Cas9 genome editing method for human induced pluripotent stem cells

Bing-chuan Geng1,2, Kyoung-Han Choi2, Shan-zhi Wang3, Peng Chen2, Xiu-di Pan2, Nian-guo Dong1, Jae-Kyun Ko2, Hua Zhu2
1 Department of Cardiovascular Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
2 Department of Surgery, The Ohio State University Wexner Medical Center, Columbus, OH 43210, USA
3 Chemistry Department, University of Arkansas at Little Rock, Little Rock, AR 72204, USA
Correspondence to: Nian-guo Dong:, Jae-Kyun Ko:, Hua Zhu:,
DOI: 10.1038/s41401-020-0452-0
Received: 8 March 2020
Accepted: 30 May 2020
Advance online: 18 June 2020


Induced pluripotent stem cells (iPSCs) have become an essential research platform to study different human diseases once being discovered by Dr. Shinya Yamanaka in 2006. Another breakthrough in biomedical research is the application of CRISPR/Cas9 system for genome editing in mammalian cells. Although numerous studies have been done to develop methods for gene editing in iPSCs, the current approaches suffer from several limitations, including time and labor consuming, low editing efficiency, and potential off-target effects. In the current study, we report an electroporation-mediated plasmid CRISPR/Cas9 delivery approach for genome editing in iPSCs. With this approach, an edited iPSC cell line could be obtained within 2 weeks. In addition, the transit introducing of CRISPR/Cas9 machinery could minimize genomic integration of Cas9 gene, which avoided potential long-term side effects of Cas9 enzyme. We showed that CRISPR/Cas9-mediated genomic editing did not affect pluripotency and differentiation ability of iPSCs. With the quickly evolving of both iPSC and CRISPR/Cas9-mediated genome editing research fields, we believe that our method can significantly facilitate the application of genome editing in iPSCs research.
Keywords: iPSC; CRISPR; Cas 9; genome editing; cardiomyocytes

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