Efficient plant genome engineering using a probiotic sourced CRISPR-Cas9 system

文献类型： 外文期刊

第一作者： Zhong, Zhaohui

作者： Zhong, Zhaohui;Tang, Zhongjie;Xiang, Shuyue;Huang, Lan;He, Yao;Fan, Tingting;Liu, Shishi;Zheng, Xuelian;Huang, Jian;Zhang, Yong;Zhong, Zhaohui;Zheng, Xuelian;Zhang, Yong;Liu, Guanqing;Zhang, Tao;Liu, Guanqing;Zhang, Tao;Liu, Guanqing;Zhang, Tao;Yang, Liang;Yang, Liang;Qi, Yiping;Qi, Yiping

作者机构：

期刊名称：NATURE COMMUNICATIONS （ 影响因子：16.6； 五年影响因子：17.0 ）

ISSN：

年卷期： 2023 年 14 卷 1 期

页码：

收录情况： SCI

摘要： Among CRISPR-Cas genome editing systems, Streptococcus pyogenes Cas9 (SpCas9), sourced from a human pathogen, is the most widely used. Here, through in silico data mining, we have established an efficient plant genome engineering system using CRISPR-Cas9 from probiotic Lactobacillus rhamnosus. We have confirmed the predicted 5'-NGAAA-3' PAM via a bacterial PAM depletion assay and showcased its exceptional editing efficiency in rice, wheat, tomato, and Larix cells, surpassing LbCas12a, SpCas9-NG, and SpRY when targeting the identical sequences. In stable rice lines, LrCas9 facilitates multiplexed gene knockout through coding sequence editing and achieves gene knockdown via targeted promoter deletion, demonstrating high specificity. We have also developed LrCas9-derived cytosine and adenine base editors, expanding base editing capabilities. Finally, by harnessing LrCas9's A/T-rich PAM targeting preference, we have created efficient CRISPR interference and activation systems in plants. Together, our work establishes CRISPR-LrCas9 as an efficient and user-friendly genome engineering tool for diverse applications in crops and beyond. In the field of plant genome engineering, new nucleases with improved editing efficiency and alterative PAM requirements are needed. Here, the authors report a probiotic sourced CRISPR-LrCas9 system with similar PAM requirement to Cas12a and show its high efficiencies in various genome editing applications.

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