Volume 31, No 10, Oct 2021

ISSN: 1001-0602 
EISSN: 1748-7838 2018 
impact factor 17.848* 
(Clarivate Analytics, 2019)

Volume 31 Issue 10, October 2021: 1134-1136   |  Open Access

LETTERS TO THE EDITOR

Enhancing prime editing by Csy4-mediated processing of pegRNA

Yao Liu¹ , Guang Yang² , Shuhong Huang¹ , Xiangyang Li² , Xin Wang² , Guanglei Li² , Tian Chi² , Yulin Chen¹ , Xingxu Huang^2,3,* , Xiaolong Wang^1,*

¹Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, China
²School of Life Science and Technology, ShanghaiTech University, Shanghai, China
³CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China
Correspondence: Xingxu Huang(huangxx@shanghaitech.edu.cn)Xiaolong Wang(xiaolongwang@nwafu.edu.cn)

Dear Editor,

The most advanced prime editor 3 (PE3) system comprises the editor, a fusion protein of Cas9 H840A nickase and mutant reverse transcriptase (RTase) (hereafter termed NMRT), a prime editing guide RNA (pegRNA) and an alternative single-guide RNA (sgRNA).1 The pegRNA contains a primer binding site (PBS) and a reverse transcription (RT) template for introducing new genetic information1 (Fig. 1a; Supplementary information, Fig. S1a). We noted that the PBS, which is generally 10–16 nt at the 3′ end of pegRNA, is complementary to part of the spacer at the 5′ end of pegRNA, and their annealing is expected to cause pegRNA circularization, which can potentially hamper editing (Fig. 1a; Supplementary information, Fig. S1b, c).

https://doi.org/10.1038/s41422-021-00520-x

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