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Cas9-PE: a robust multiplex gene editing tool for simultaneous precise editing and site-specific random mutation in rice

文献类型： 外文期刊

作者： Zou, Jinpeng 1 ; Meng, Xiangbing 2 ; Hong, Zhengyuan 1 ; Rao, Yuchun 3 ; Wang, Kejian 1 ; Li, Jiayang 2 ; Yu, Hong 2 ; Wang, Chun 1 ;

作者机构： 1.Chinese Acad Agr Sci, China Natl Rice Res Inst, State Key Lab Rice Biol & Breeding, Hangzhou 310006, Peoples R China

2.Chinese Acad Sci, Inst Genet & Dev Biol, Key Lab Seed Innovat, Beijing 100101, Peoples R China

3.Zhejiang Normal Univ, Coll Life Sci, Jinhua 321004, Peoples R China

4.Minist Agr & Rural Affairs, Key Lab Gene Editing Technol Hainan, Sanya 572025, Peoples R China

5.Yazhouwan Natl Lab, Sanya 572024, Peoples R China

期刊名称：TRENDS IN BIOTECHNOLOGY （ 影响因子：14.9； 五年影响因子：16.6 ）

ISSN： 0167-7799

年卷期： 2025 年 43 卷 2 期

页码：

收录情况： SCI

摘要： In molecular design breeding, the simultaneous introduction of desired functional genes through specific nucleotide modifications and the elimination of genes regu-lating undesired phenotypic traits or agronomic components require advanced gene editing tools. Due to limited editing efficiency, even with the use of highly precise editing tools, such as prime editing (PE), simultaneous editing of multiple mutation types poses a challenge. Here, we replaced Cas9 nickase (nCas9) with Cas9 to con-struct a Cas9-mediated PE (Cas9-PE) system in rice. This system not only enables precise editing, but also allows for site-specific random mutation. Moreover, leveraging the precision of Cas9-PE, we established a transgene-free multiplex gene editing system using a co-editing strategy. This strategy involved the Agrobacterium-mediated transient expression of the precise editing rice endoge-nous acetolactate synthase gene ALSS627I to confer herbicide bispyribac-sodium (BS) resistance as a selection marker. This study provides a versatile and efficient multiplex gene editing tool for molecular design breeding.

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