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
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.
