Elimination of bla_KPC-2-mediated carbapenem resistance in Escherichia coli by CRISPR-Cas9 system.

ABSTRACT:

Objective

The purpose of this study is to re-sensitive bacteria to carbapenemases and reduce the transmission of the bla_KPC-2 gene by curing the bla_KPC-2-harboring plasmid of carbapenem-resistant using the CRISPR-Cas9 system.

Methods

The single guide RNA (sgRNA) specifically targeted to the bla_KPC-2 gene was designed and cloned into plasmid pCas9. The recombinant plasmid pCas9-sgRNA(bla_KPC-2) was transformed into Escherichia coli (E.coli) carrying pET24-bla_KPC-2. The elimination efficiency in strains was evaluated by polymerase chain reaction (PCR) and quantitative real-time PCR (qPCR). Susceptibility testing was performed by broth microdilution assay and by E-test strips (bioMérieux, France) to detect changes in bacteri

SUBMITTER: Tao S 

PROVIDER: S-EPMC10601263 | biostudies-literature | 2023 Oct

REPOSITORIES: biostudies-literature