PE6c greatly enhances prime editing in transgenic rice plants

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 teste...

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Bibliographic Details
Published in:Journal of integrative plant biology 2024-09, Vol.66 (9), p.1864-1870
Main Authors: Cao, Zhenghong, Sun, Wei, Qiao, Dexin, Wang, Junya, Li, Siyun, Liu, Xiaohan, Xin, Cuiping, Lu, Yu, Gul, Syeda Leeda, Wang, Xue‐Chen, Chen, Qi‐Jun
Format: Article
Language:English
Subjects:
Agronomy
CRISPR
Editing
Efficiency
genome editing
Modules
Oryza sativa
PE6
PE6c
Plant breeding
prime editing
prime editor
Rice
RNA-directed DNA polymerase
Transgenic plants
Yeasts
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Summary: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. In rice, the prime editor PE6c, with an evolved and engineered reverse transcriptase variant from the yeast Tf1 retrotransposon, yielded the highest editing efficiency. Moreover, using two reverse transcriptase modules improved prime‐editing efficiency.
ISSN:1672-9072
1744-7909
1744-7909
DOI:10.1111/jipb.13738