J Integr Plant Biol ›› 2024, Vol. 66 ›› Issue (9): 1864-1870.DOI: 10.1111/jipb.13738

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  • 收稿日期:2024-04-25 接受日期:2024-06-20 出版日期:2024-09-01 发布日期:2024-09-06

PE6c greatly enhances prime editing in transgenic rice plants

Zhenghong Cao1†, Wei Sun1†, Dexin Qiao1, Junya Wang1, Siyun Li1, Xiaohan Liu1, Cuiping Xin1, Yu Lu1, Syeda Leeda Gul1, Xue‐Chen Wang1 and Qi‐Jun Chen1,2*   

  1. 1. State Key Laboratory of Plant Environmental Resilience, College of Biological Sciences, China Agricultural University, Beijing100193, China
    2. Center for Crop Functional Genomics and Molecular Breeding, China Agricultural University, Beijing 100193, China
    These authors contributed equally to this work.
    *Correspondence: Qi‐Jun Chen (qjchen@cau.edu.cn)
  • Received:2024-04-25 Accepted:2024-06-20 Online:2024-09-01 Published:2024-09-06
  • Supported by:
    This work was supported by grants from the National Key Research and Development Program of China (2023YFD1202905) and the National Natural Science Foundation of China (U19A2022).

Abstract: Prime editing is a versatile CRISPR/Cas-based precise genome-editing technique for crop breeding. Four new types of prime editors (PEs) named PE6a–d were recently generated using evolved and engineered reverse transcriptase (RT) variants from three different sources. In this study, we tested the editing efficiencies of four PE6 variants and two additional PE6 constructs with double-RT modules in transgenic rice (Oryza sativa) plants. PE6c, with an evolved and engineered RT variant from the yeast Tf1 retrotransposon, yielded the highest prime-editing efficiency. The average fold change in the editing efficiency of PE6c compared with PEmax exceeded 3.5 across 18 agronomically important target sites from 15 genes. We also demonstrated the feasibility of using two RT modules to improve prime-editing efficiency. Our results suggest that PE6c or its derivatives would be an excellent choice for prime editing in monocot plants. In addition, our findings have laid a foundation for prime-editing-based breeding of rice varieties with enhanced agronomically important traits.

Key words: genome editing, PE6, PE6c, prime editing, prime editor, rice

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