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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 Huang2,3,* , Xiaolong Wang1,*
1Key Laboratory of Animal Genetics, Breeding and Reproduction of Shaanxi Province, College of Animal Science and Technology, Northwest A&F University, Yangling, Shaanxi, ChinaDear 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