中文版 | English
Title

High-throughput genome editing in rice with a virus-based surrogate system

Author
Corresponding AuthorZhu Jian-kang; Lu Yuming
Joint first authorTian Yifu; Zhong Dating; Li Xinbo
Publication Years
2022-10-11
DOI
Source Title
ISSN
1672-9072
EISSN
1744-7909
Abstract

With the widespread use of CRISPR/Cas technologies in plants, large-scale genome editing is increasingly needed. Here, we developed a geminivirus-mediated surrogate system, called WDV-Gate (Wheat Dwarf Virus-surrogate), to facilitate high-throughput genome editing. WDV-Gate has two parts: one is the recipient callus from a transgenic rice line expressing Cas9 and a mutated hygromycin-resistant gene (HygM) for surrogate selection; the other is a WDV-based construct expressing two sgRNAs targeting HygM and a gene of interest, respectively. We evaluated WDV-Gate on six rice loci by producing a total of 874 T0 plants. Compared with the conventional method, the WDV-Gate system, which was characterized by a transient and high level of sgRNA expression, significantly increased editing frequency (66.8% vs. 90.1%), plantlet regeneration efficiency (2.31-fold increase), and numbers of homozygous-edited plants (36.3% vs. 70.7%). Large-scale editing using pooled sgRNAs targeting the SLR1 gene resulted in a high editing frequency of 94.4%, further demonstrating its feasibility. We also tested WDV-Gate on sequence knock-in for protein tagging. By co-delivering a chemically modified donor DNA with the WDV-Gate plasmid, 3xFLAG peptides were successfully fused to three loci with an efficiency of up to 13%. Thus, by combining transiently expressed sgRNAs and a surrogate selection system, WDV-Gate could be useful for high-throughput gene knock-out and sequence knock-in. This article is protected by copyright. All rights reserved.

Keywords
URL[Source Record]
Indexed By
Language
English
SUSTech Authorship
Corresponding
Funding Project
[2021YFD1201300] ; [2021YFA1300404] ; [32070396] ; [32188102] ; [BX20220098]
WOS Research Area
Biochemistry & Molecular Biology ; Plant Sciences
WOS Subject
Biochemistry & Molecular Biology ; Plant Sciences
WOS Accession No
WOS:000905248100001
Publisher
Data Source
人工提交
Publication Status
在线出版
Citation statistics
Cited Times [WOS]:2
Document TypeJournal Article
Identifierhttp://kc.sustech.edu.cn/handle/2SGJ60CL/416233
DepartmentInstitute of Advanced Biotechnology
生命科学学院
Affiliation
1.Shanghai Center for Plant Stress Biology, Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences
2.Center for Advanced Bioindustry Technologies, and Institute of Crop Sciences, Chinese Academy of Agricultural Sciences
3.Hainan Yazhou Bay Seed Lab
4.Shanghai Collaborative Innovation Center of Agri-Seeds, Joint Center for Single Cell Biology, School of Agriculture and Biology, Shanghai Jiao Tong University
5.Institute of Advanced Biotechnology, and School of Life Sciences, Southern University of Science and Technology
Corresponding Author AffilicationInstitute of Advanced Biotechnology;  School of Life Sciences
Recommended Citation
GB/T 7714
Tian Yifu,Zhong Dating,Li Xinbo,et al. High-throughput genome editing in rice with a virus-based surrogate system[J]. Journal of Integrative Plant Biology,2022.
APA
Tian Yifu.,Zhong Dating.,Li Xinbo.,Shen Rundong.,Han Han.,...&Lu Yuming.(2022).High-throughput genome editing in rice with a virus-based surrogate system.Journal of Integrative Plant Biology.
MLA
Tian Yifu,et al."High-throughput genome editing in rice with a virus-based surrogate system".Journal of Integrative Plant Biology (2022).
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