文献类型： 外文期刊
作者： Zhong, Zhaohui 1 ; Liu, Guanqing 3 ; Tang, Zhongjie 1 ; Xiang, Shuyue 1 ; Yang, Liang 6 ; Huang, Lan 1 ; He, Yao 1 ; Fan, Tingting 1 ; Liu, Shishi 1 ; Zheng, Xuelian 1 ; Zhang, Tao 3 ; Qi, Yiping 8 ; Huang, Jian 1 ; Zhang, Yong 1 ;
作者机构： 1.Univ Elect Sci & Technol China, Ctr Informat Biol, Sch Life Sci & Technol, Dept Biotechnol, Chengdu 610054, Peoples R China
2.Southwest Univ, Integrat Sci Ctr Germplasm Creat Western China Cho, Sch Life Sci, Chongqing Key Lab Plant Resource Conservat & Germp, Chongqing 400715, Peoples R China
3.Yangzhou Univ, Agr Coll, Jiangsu Key Lab Crop Genom & Mol Breeding, Jiangsu Key Lab Crop Genet & Physiol, Yangzhou 225012, Peoples R China
4.Yangzhou Univ, Minist Educ, Minist Educ China, Key Lab Plant Funct Genom,Joint Int Res Lab Agr &, Yangzhou 225012, Peoples R China
5.Yangzhou Univ, Jiangsu Coinnovat Ctr Modern Prod Technol Grain Cr, Jiangsu Key Lab Crop Genet & Physiol, Yangzhou 225012, Peoples R China
6.Sichuan Acad Agr Sci, Hort Res Inst, Chengdu, Sichuan, Peoples R China
7.Vegetable Germplasm Innovat & Variety Improvement, Vegetable Germplasm Innovat & Variety Improvement, Hort Res Inst, Chengdu 610066, Peoples R China
8.Univ Maryland, Dept Plant Sci & Landscape Architecture, College Pk, MD 20742 USA
9.Univ Maryland, Inst Biosci & Biotechnol Res, Rockville, MD 20850 USA
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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