IOCAS-IR
A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy
Wang, Jichao1; Sui, Xinyue1,2; Ding, Yamei3; Fu, Yingxin1; Feng, Xinjun1; Liu, Min1,4; Zhang, Youming4; Xian, Mo1; Zhao, Guang1,4
2021-01-25
发表期刊NUCLEIC ACIDS RESEARCH
ISSN0305-1048
卷号49期号:2页码:15
通讯作者Xian, Mo(xianmo@qibebt.ac.cn) ; Zhao, Guang(zhaoguang@sdu.edu.cn)
摘要The production of optimized strains of a specific phenotype requires the construction and testing of a large number of genome modifications and combinations thereof. Most bacterial iterative genome-editing methods include essential steps to eliminate selection markers, or to cure plasmids. Additionally, the presence of escapers leads to time-consuming separate single clone picking and subsequent cultivation steps. Herein, we report a genome-editing method based on a Rock-Paper-Scissors (RPS) strategy. Each of three constructed sgRNA plasmids can cure, or be cured by, the other two plasmids in the system; plasmids from a previous round of editing can be cured while the current round of editing takes place. Due to the enhanced curing efficiency and embedded double check mechanism, separate steps for plasmid curing or confirmation are not necessary, and only two times of cultivation are needed per genome-editing round. This method was successfully demonstrated in Escherichia toll and Klebsiella pneumoniae with both gene deletions and replacements. To the best of our knowledge, this is the fastest and most robust iterative genome-editing method, with the least times of cultivation decreasing the possibilities of spontaneous genome mutations.
DOI10.1093/nar/gkaa1141
收录类别SCI
语种英语
资助项目National Natural Science Foundation of China[31961133014] ; National Natural Science Foundation of China[31722001] ; National Natural Science Foundation of China[21807101] ; Defence Industrial Technology Development Program[JCKY2018130B005] ; Shandong University
WOS研究方向Biochemistry & Molecular Biology
WOS类目Biochemistry & Molecular Biology
WOS记录号WOS:000626723800006
出版者OXFORD UNIV PRESS
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被引频次:12[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/170288
专题中国科学院海洋研究所
通讯作者Xian, Mo; Zhao, Guang
作者单位1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biobased Mat, Qingdao 266101, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
4.Shandong Univ, State Key Lab Microbial Technol, Qingdao 266237, Peoples R China
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Wang, Jichao,Sui, Xinyue,Ding, Yamei,et al. A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy[J]. NUCLEIC ACIDS RESEARCH,2021,49(2):15.
APA Wang, Jichao.,Sui, Xinyue.,Ding, Yamei.,Fu, Yingxin.,Feng, Xinjun.,...&Zhao, Guang.(2021).A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy.NUCLEIC ACIDS RESEARCH,49(2),15.
MLA Wang, Jichao,et al."A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy".NUCLEIC ACIDS RESEARCH 49.2(2021):15.
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