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Targeted Elimination of bla ~NDM-5~ Gene in Escherichia coli by Conjugative CRISPR-Cas9 System by Peisi Li; Peng Wan; Ruonan Zhao; Jin Chen; Xiaoshen Li; Jie Li; Wenguang Xiong; Zhenling Zeng is a Medicine article available to read on EtoBox.

What is Targeted Elimination of bla ~NDM-5~ Gene in Escherichia coli by Conjugative CRISPR-Cas9 System about?

Plasmid-borne carbapenem resistance gene bla NDM-5 accelerates the dissemination of carbapenem-resistant Enterobacteriaceae. To efficiently eliminate the bla NDM-5 -harboring plasmid and sensitize the antibiotic-resistant bacteria to meropenem, we used the CRISPR-Cas9 system for combating the carbapenem-resistant Escherichia coli (E. coil). Methods: A series of CRISPR-Cas9 plasmids was constructed, and specific guide RNAs(sgRNA) were designed to target the bla NDM-5 gene. We used chemically transformation or conjugation delivery methods, and the elimination efficiency in each recipient strains was evaluated by plate counting, PCR and quantitative real-time PCR (qPCR). Antimicrobial susceptibility test was carried out by using the broth microdilution method. In addition, we assessed the effect of the CRISPR-Cas9 system of adaptive immunity on the prevention of the exogenous resistant plasmids pNDM-5 by introducing the system into E coli J53. Results: The results showed that pCas9, pCas9-oriT and pBAD-Cas9-oriT can effectively eliminate bla NDM-5 in E. coli with >94.00% elimination efficiency. The bla NDM-5 -harboring E. coli successfully restored their susceptibility to meropenem, w

Who reads Targeted Elimination of bla ~NDM-5~ Gene in Escherichia coli by Conjugative CRISPR-Cas9 System?

Author

Peisi Li; Peng Wan; Ruonan Zhao; Jin Chen; Xiaoshen Li; Jie Li; Wenguang Xiong; Zhenling Zeng

Publisher

Dove

Published

2022

Language

EN

Field

Medicine (Health Sciences)