Targeted genetic screening in bacteria with a Cas12k-guided transposase

doi:10.1016/j.celrep.2021.109635

	Targeted genetic screening in bacteria with a Cas12k-guided transposase
	Chen, Weizhong1 ; Ren, Ze-Hui 2; Tang, Na1,3 ; Chai, Guoshi 2; Zhang, Hongyuan1,3 ; Zhang, Yifei1,3 ; Ma, Jiacheng1,3 ; Wu, Zhaowei1 ; Shen, Xia 2; Huang, Xingxu4,5 ; Luo, Guan-Zheng 2; Ji, Quanjiang1,5
	2021-08-31
发表期刊	CELL REPORTS (IF:7.5[JCR-2023],8.5[5-Year])
ISSN	2211-1247
卷号	36 期号:9
DOI	10.1016/j.celrep.2021.109635
摘要	Microbes employ sophisticated cellular networks encoded by complex genomes to rapidly adapt to changing environments. High-throughput genome engineering methods are valuable tools for functionally profiling genotype-phenotype relationships and understanding the complexity of cellular networks. However, current methods either rely on special homologous recombination systems and are thus applicable in only limited bacterial species or can generate only nonspecific mutations and thus require extensive subsequent screening. Here, we report a site-specific transposon-assisted genome engineering (STAGE) method that allows high-throughput Cas12k-guided mutagenesis in various microorganisms, such as Pseudomonas aeruginosa and Klebsiella pneumoniae. Exploiting the powerful STAGE technique, we construct a site-specific transposon mutant library that focuses on all possible transcription factors (TFs) in P. aeruginosa, enabling the comprehensive identification of essential genes and antibiotic-resistance-related factors. Given its broad host range activity and easy programmability, this method can be widely adapted to diverse microbial species for rapid genome engineering and strain evolution.
收录类别	SCIE
语种	英语
WOS研究方向	Cell Biology
WOS类目	Cell Biology
WOS记录号	WOS:000692168900015
出版者	CELL PRESS
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/128095
专题	物质科学与技术学院_PI研究组_季泉江组生命科学与技术学院_PI研究组_黄行许组物质科学与技术学院_硕士生物质科学与技术学院_博士生
通讯作者	Luo, Guan-Zheng; Ji, Quanjiang
作者单位	1.ShanghaiTech Univ, Sch Phys Sci & Technol, Shanghai 201210, Peoples R China; 2.Sun Yat Sen Univ, Sch Life Sci,State Key Lab Biocontrol, MOE Key Lab Gene Funct & Regulat, Guangdong Prov Key Lab Pharmaceut Funct Genes, Guangzhou 510275, Guangdong, Peoples R China; 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China; 4.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China; 5.Guangzhou Lab, Guangzhou 510120, Peoples R China
第一作者单位	物质科学与技术学院
通讯作者单位	物质科学与技术学院
第一作者的第一单位	物质科学与技术学院
推荐引用方式 GB/T 7714	Chen, Weizhong,Ren, Ze-Hui,Tang, Na,et al. Targeted genetic screening in bacteria with a Cas12k-guided transposase[J]. CELL REPORTS,2021,36(9).
APA	Chen, Weizhong.,Ren, Ze-Hui.,Tang, Na.,Chai, Guoshi.,Zhang, Hongyuan.,...&Ji, Quanjiang.(2021).Targeted genetic screening in bacteria with a Cas12k-guided transposase.CELL REPORTS,36(9).
MLA	Chen, Weizhong,et al."Targeted genetic screening in bacteria with a Cas12k-guided transposase".CELL REPORTS 36.9(2021).