A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR
2019-12-19
发表期刊CELL CHEMICAL BIOLOGY (IF:6.6[JCR-2023],8.2[5-Year])
ISSN2451-9448
卷号26期号:12页码:1732-+
发表状态已发表
DOI10.1016/j.chembiol.2019.09.003
摘要The rapid emergence of extensively drug-resistant A. baumannii has posed a major threat to global public health, emphasizing the desperate need for novel therapeutic strategies. We report the development of a highly efficient genome-engineering platform in A. baumannii by coupling a Cas9 nuclease-mediated genome cleavage system with the RecAb recombination system. We applied the CRISPR-Cas9/RecAb system to dissect the oxidative stress-sensing mechanism of OxyR by performing alanine scanning mutagenesis of 13 residues residing in the H2O2-sensing pocket, pinpointing new vital factors for H2O2 sensing. Moreover, we developed a cytidine base-editing system, enabling programmed C to T conversions. Exploiting this powerful technique, we systematically investigated the drug-resistant mechanisms in a clinically isolated multidrug-resistant A. baumannii strain by generating premature stop codons in the possible resistance genes, unveiling distinct roles of these genes in drug resistance. The development of these genome-engineering methods will facilitate new therapeutic-means development in A. baumannii and related organisms.
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收录类别SCI ; SCIE
语种英语
资助项目China Postdoctoral Science Foundation[2018M632190]
WOS研究方向Biochemistry & Molecular Biology
WOS类目Biochemistry & Molecular Biology
WOS记录号WOS:000503739200013
出版者CELL PRESS
WOS关键词STAPHYLOCOCCUS-AUREUS ; GENE REPLACEMENT ; DNA ; BASE ; RECOMBINATION ; RESISTANCE ; SEQUENCE ; REGULON
原始文献类型Article
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文献类型期刊论文
条目标识符https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/104964
专题物质科学与技术学院_PI研究组_季泉江组
物质科学与技术学院_硕士生
通讯作者Ji, Quanjiang
作者单位
1.ShanghaiTech Univ, Sch Phys Sci & Technol, Shanghai 201210, Peoples R China
2.Zhejiang Univ, Sir Run Run Shaw Hosp, Coll Med, Dept Infect Dis, Hangzhou 310016, Zhejiang, Peoples R China
3.Key Lab Microbial Technol & Bioinformat Zhejiang, Hangzhou 310016, Zhejiang, 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
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Wang, Yu,Wang, Zhipeng,Chen, Yan,et al. A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR[J]. CELL CHEMICAL BIOLOGY,2019,26(12):1732-+.
APA Wang, Yu,Wang, Zhipeng,Chen, Yan,Hua, Xiaoting,Yu, Yunsong,&Ji, Quanjiang.(2019).A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR.CELL CHEMICAL BIOLOGY,26(12),1732-+.
MLA Wang, Yu,et al."A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR".CELL CHEMICAL BIOLOGY 26.12(2019):1732-+.
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