A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy

doi:10.1093/nar/gkaa1141

CORC > 海洋研究所 > 中国科学院海洋研究所

	A fast and robust iterative genome-editing method based on a Rock-Paper-Scissors strategy
	Wang, Jichao 3; Sui, Xinyue 2,3; Ding, Yamei 4; Fu, Yingxin 3; Feng, Xinjun 3; Liu, Min 1,3; Zhang, Youming 1; Xian, Mo 3; Zhao, Guang 1,3
刊名	NUCLEIC ACIDS RESEARCH
	2021-01-25
卷号	49 期号:2 页码:15
ISSN号	0305-1048
DOI	10.1093/nar/gkaa1141
通讯作者	Xian, Mo(xianmo@qibebt.ac.cn) ; Zhao, Guang(zhaoguang@sdu.edu.cn)
英文摘要	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.
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
语种	英语
出版者	OXFORD UNIV PRESS
WOS记录号	WOS:000626723800006
内容类型	期刊论文
源URL	[http://ir.qdio.ac.cn/handle/337002/170287]
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.Shandong Univ, State Key Lab Microbial Technol, Qingdao 266237, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, CAS Key Lab Biobased Mat, Qingdao 266101, Peoples R China 4.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
推荐引用方式 GB/T 7714	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.