Development of a highly efficient prime editor 2 system in plants

Juan Li; Like Chen; Jing Liang; Rongfang Xu; Yingli Jiang; Yizhen Li; Jian Ding; Min Li; Ruiying Qin; Pengcheng Wei

doi:10.1186/s13059-022-02730-x

Development of a highly efficient prime editor 2 system in plants

Genome Biol. 2022 Jul 25;23(1):161. doi: 10.1186/s13059-022-02730-x.

Authors

Juan Li¹, Like Chen², Jing Liang², Rongfang Xu¹, Yingli Jiang², Yizhen Li², Jian Ding², Min Li², Ruiying Qin¹, Pengcheng Wei^{3

4}

Affiliations

¹ Key Laboratory of Rice Genetic Breeding of Anhui Province, Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, 230031, P.R. China.
² College of Agronomy, Anhui Agricultural University, Hefei, 230036, P.R. China.
³ Key Laboratory of Rice Genetic Breeding of Anhui Province, Rice Research Institute, Anhui Academy of Agricultural Sciences, Hefei, 230031, P.R. China. weipengcheng@gmail.com.
⁴ College of Agronomy, Anhui Agricultural University, Hefei, 230036, P.R. China. weipengcheng@gmail.com.

Abstract

Low efficiency has seriously restricted the application of prime editing (PE) systems in plants. In this study, we develop an enhanced plant prime editor 2 system, enpPE2, by stacking various optimization strategies, including updating the PE architecture to PEmax and expressing engineered pegRNA with a structured motif under the control of a composite promoter. In T₀ rice plants, enpPE2 exhibits editing frequencies of 64.58% to 77.08%, which are much higher than the frequencies with unmodified pPE2. Our results indicate that the enpPE2 system provides a robust and powerful tool for the precise modification of plant genomes.

Keywords: Efficiency; Engineering; Precise editing; Prime editing; Rice.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems
Gene Editing* / methods
Genome, Plant
Oryza* / genetics
Plants / genetics