首页|Versatile plant genome engineering using anti-CRISPR-Cas12a systems

Versatile plant genome engineering using anti-CRISPR-Cas12a systems

扫码查看
CRISPR-Cas12a genome engineering systems have been widely used in plant research and crop breeding.To date,the performance and use of anti-CRISPR-Cas 12a systems have not been fully established in plants.Here,we conduct in silico analysis to identify putative anti-CRISPR systems for Cas12a.These putative anti-CRISPR proteins,along with known anti-CRISPR proteins,are assessed for their ability to inhibit Cas12a cleavage activity in vivo and in planta.Among all anti-CRISPR proteins tested,AcrVA1 shows robust inhibition of Mb2Cas12a and LbCas12a in E.coli.Further tests show that AcrVA1 inhibits LbCas12a mediated genome editing in rice protoplasts and stable transgenic lines.Impressively,co-expression of AcrVA1 mitigates off-target effects by CRISPR-LbCas12a,as revealed by whole genome sequencing.In addition,transgenic plants expressing AcrVA1 exhibit different levels of inhibition to LbCas12a mediated genome editing,representing a novel way of fine-tuning genome editing efficiency.By controlling temporal and spatial expression of AcrVA1,we show that inducible and tissue specific genome editing can be achieved in plants.Furthermore,we demonstrate that AcrVA1 also inhibits LbCas12a-based CRISPR activation(CRISPRa)and based on this principle we build logic gates to turn on and off target genes in plant cells.Together,we have established an efficient anti-CRISPR-Cas12a system in plants and demonstrate its versatile applications in mitigating off-target effects,fine-tuning genome editing efficiency,achieving spatial-temporal control of genome editing,and generating synthetic logic gates for controlling target gene expression in plant cells.

anti-CRISPRCas12aAcrVA1off-target effectsfine-tuning genome editinginducible and tissue specific genome editingCRISPRasynthetic logic circuit

Yao He、Shishi Liu、Long Chen、Dongkai Pu、Zhaohui Zhong、Tang Xu、Qiurong Ren、Chuan Dong、Yawei Wang、Danning Wang、Xuelian Zheng、Fengbiao Guo、Tao Zhang、Yiping Qi、Yong Zhang

展开 >

Integrative Science Center of Germplasm Creation in Western China(Chongqing)Science City,Chongqing Key Laboratory of Tree Germplasm Innovation and Utilization,School of Life Sciences,Southwest University,Chongqing 400715,China

Department of Biotechnology,School of Life Sciences and Technology,Center for Informational Biology,University of Electronic Science and Technology of China,Chengdu 610054,China

Sichuan Institute of Edible Fungi,Sichuan Academy of Agricultural Sciences,Chengdu 610066,China

Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Zhongshan Biological Breeding Laboratory/Key Laboratory of Plant Functional Genomics of the Ministry of Education,College of Agriculture,Yangzhou University,Yangzhou 225009,China

Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops/Jiangsu Key Laboratory of Crop Genetics and Physiology,Yangzhou University,Yangzhou 225009,China

Department of Respiratory and Critical Care Medicine,Zhongnan Hospital of Wuhan University,Wuhan 430017,China

Key Laboratory of Combinatorial Biosynthesis and Drug Discovery,Ministry of Education and School of Pharmaceutical Sciences,Wuhan University,Wuhan 430072,China

Department of Plant Science and Landscape Architecture,University of Maryland,College Park 20742,USA

Institute for Bioscience and Biotechnology Research,University of Maryland,Rockville 20850,USA

展开 >

2024

中国科学:生命科学(英文版)
中国科学院

中国科学:生命科学(英文版)

CSTPCD
影响因子:0.806
ISSN:1674-7305
年,卷(期):2024.67(12)