Robustly improved base editing efficiency of Cpf1 base editor using optimized cytidine deaminases

Siyu Chen,Zhanjun Li,Yingqi Jia,Huanhuan Shan,Liangxue Lai,Mao Chen,Zhiquan Liu,Hao Yu

doi:10.1038/s41421-020-00195-5

Abstract

Robustly improved base editing efficiency of Cpf1 base editor using optimized cytidine deaminases

Highlights

1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Dear Editor, Programmable clustered regularly interspaced short palindromic repeats (CRISPR) associated Cpf[1] endonucleases, known as Cas12a, are single RNA-guided effectors[1] that have been commonly utilized in various species to manipulate genome for their remarkable specificity[2–4] and concise structures[1]
We demonstrated that there is no significantly improved base editing frequency observed by using engineering of crRNAs, while the dramatically increased base editing efficiency was perceived by using cytidine deaminase optimized Cpf[1] base editors (BEs)
The results showed that U-rich crRNA slightly improved editing efficiency at all target sites ranging from 1.05- to 1.69-fold and significantly improved base editing efficiency at the EMX1 site

Summary

Introduction

1234567890():,; 1234567890():,; 1234567890():,; 1234567890():,; Dear Editor, Programmable clustered regularly interspaced short palindromic repeats (CRISPR) associated Cpf[1] endonucleases, known as Cas12a, are single RNA-guided (crRNA) effectors[1] that have been commonly utilized in various species to manipulate genome for their remarkable specificity[2–4] and concise structures[1]. We Correspondence: Hao Yu (yu_hao@jlu.edu.cn) or Liangxue Lai (lai_liangxue@gibh.ac.cn) or Zhanjun Li (lizj_1998@jlu.edu.cn) 1Key Laboratory of Zoonosis Research, Ministry of Education, College of Animal Science, Jilin University, 130062 Changchun, China 2CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 510530 Guangzhou, China Full list of author information is available at the end of the article These authors contributed : Siyu Chen, Yingqi Jia, Zhiquan Liu reconstructed dLbCpf1-BE3 (dCpf1-BE3)[9] with three distinctive deaminases (evoAPOBEC1, evoCDA110,11, human APOBEC3A (A3A)12–14) to produce optimized dCpf1based BEs. Here, we demonstrated that there is no significantly improved base editing frequency observed by using engineering of crRNAs, while the dramatically increased base editing efficiency was perceived by using cytidine deaminase optimized Cpf[1] BE (dCpf1-eCDA1).

Results

Conclusion