CRISPR-Cas9-Mediated Carbapenemase Gene and Plasmid Curing in Carbapenem-Resistant <i>Enterobacteriaceae</i>

Abstract

Combating plasmid-mediated carbapenem resistance is essential to control and prevent the dissemination of carbapenem-resistant Enterobacteriaceae (CRE). Here, we conducted a proof-of-concept study to demonstrate that CRISPR-Cas9-mediated resistance gene and plasmid curing can effectively resensitize CRE to carbapenems. A novel CRISPR-Cas9-mediated plasmid-curing system (pCasCure) was developed and electrotransferred into various clinical CRE isolates. The results showed that pCasCure can effectively cure bla KPC , bla NDM , and bla OXA-48 in various Enterobacteriaceae species of Klebsiella pneumoniae , Escherichia coli , Enterobacter hormaechei , Enterobacter xiangfangensis , and Serratia marcescens clinical isolates, with a &gt;94% curing efficiency.