CRISPR-Cas9-Mediated Carbapenemase Gene and Plasmid Curing in Carbapenem-Resistant Enterobacteriaceae

Author:

Hao Mingju1,He Yuzhang23,Zhang Haifang4,Liao Xiao-Ping23,Liu Ya-Hong235,Sun Jian23ORCID,Du Hong4,Kreiswirth Barry N.6,Chen Liang67ORCID

Affiliation:

1. Department of Clinical Laboratory Medicine, The First Affiliated Hospital of Shandong First Medical University, Jinan, Shandong, China

2. National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, South China Agricultural University, Guangzhou, China

3. Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, South China Agricultural University, Guangzhou, China

4. Department of Clinical Laboratory, Second Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China

5. Jiangsu Co-Innovation Centre for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu, China

6. Center for Discovery and Innovation, Hackensack-Meridian Health, Nutley, New Jersey, USA

7. Hackensack Meridian School of Medicine, Nutley, New Jersey, USA

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 >94% curing efficiency.

Funder

HHS | NIH | National Institute of Allergy and Infectious Diseases

Publisher

American Society for Microbiology

Subject

Infectious Diseases,Pharmacology (medical),Pharmacology

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