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| CsgRNA reduces mutation caused by APOBECs during Cas9-D10A-mediated gene editing |
| REN Rongrong1,CHEN Hongquan2 |
| 1.Department of Medical Laboratory, Zhejiang Hospital, Hangzhou 310013, China;2.Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Medical School of Zhejiang University,Hangzhou 310016, China |
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| Cite this article: |
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REN Rongrong,CHEN Hongquan. CsgRNA reduces mutation caused by APOBECs during Cas9-D10A-mediated gene editing[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2021, 51(9): 724-728.
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Abstract Objective: To investigate whether csgRNA strategy could reduce the indels induced by APOBECs when performing gene editing of Cas9-D10A/sgRNA. Methods: In this study, csgRNA was designed on sgRNA4 and FANCF targeting sites. In order to block on-target indels induced by D10A, csgRNA was optimized finely for its length and space location, with the indels detected in 293FT-APOBEC3B overexpression cell line.Results: CsgRNA dramatically reduced the unwanted mutations caused by APOBECs, which was further improved through optimizing its conditions, and it was playing a better role when 16 bp length and 2 bases extended to the PAM or 20 bp length and 4 bases extended to the PAM. Conclusion: The unexpected mutations caused by APOBECs was significantly lowered using the csgRNA strategy.
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Received: 30 March 2021
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