CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli

dc.contributor.authorChen, Liang
dc.contributor.authorPeirano, G.
dc.contributor.authorYen, Kelly
dc.contributor.authorWang, Bingjie
dc.contributor.authorTerlecky, Austin
dc.contributor.authorDeVinney, Rebekah
dc.contributor.authorKreiswirth, Barry N.
dc.contributor.authorPitout, Johann D.D.
dc.date.accessioned2025-02-12T13:10:49Z
dc.date.available2025-02-12T13:10:49Z
dc.date.issued2024-01
dc.descriptionDATA AVAILABILITY : Sequences have been deposited at GenBank under BioProject accession number PRJNA549322.en_US
dc.description.abstractIncF plasmids are commonly found in extra-intestinal pathogenic Escherichia coli (ExPEC) strains, serving as reservoirs for antimicrobial resistance (AMR) genes and virulence factors, persistently coexisting with ExPEC lineages. Multidrug-resistant (MDR) high-risk ExPEC clones, particularly ST131, ST1193, and ST410, have acquired diverse IncF plasmids over time, containing various AMR determinants, contributing significantly to their global success. However, the broader roles of these IncF plasmids in the success of MDR ExPEC clones, beyond AMR, remain elusive. In this study, we employed a novel clustered regularly interspaced short palindromic repeats–CRISPR-associated protein-9 nuclease (CRISPR-Cas9)-mediated pCasCure plasmid-curing system to precisely remove specific IncF plasmids among ExPEC clones (ST1193, ST131, and ST410). Antibiotic-resistant parent strains reverted to antibiotic-susceptible states post-curing; however, IncF plasmid curing did not show significant impact on bacterial in vitro growth and had little impact on other in vitro phenotypes, including survival in water, dry environment and biofilm production. In addition, IncF plasmid curing did not affect the conjugation frequency of KPC-producing pKpQIL plasmid. This study represents a pivotal initial step in understanding the precise roles of IncF plasmids in the success of ExPEC. Future research will be crucial in investigating their influence on cell invasion and in vivo fitness, thereby providing a more comprehensive perspective on the functions of IncF plasmids in MDR ExPEC clones. IMPORTANCE : Understanding the role of IncF plasmids in the success of drug-resistant bacteria has far-reaching implications for tackling antibiotic resistance. The study's use of a novel CRISPR-Cas9-mediated plasmid-curing system provides a precision tool for dissecting the specific impact of IncF plasmids on ExPEC clones, especially high-risk, multidrug-resistant strains like ST131, ST1193, and ST410. The study offers a crucial stepping stone for future research into understanding how these plasmids influenceinfluenceinfluencemore complex aspects of bacterial behavior, such as cell invasion and in vivo fitness.en_US
dc.description.departmentMedical Microbiologyen_US
dc.description.librarianam2024en_US
dc.description.sdgSDG-03:Good heatlh and well-beingen_US
dc.description.sponsorshipThe JPIAMR/Canadian Institute Health Research program and the National Institutes of Health. In part supported by the National Institute of Allergy and Infectious Diseases.en_US
dc.description.urihttps://journals.asm.org/journal/spectrumen_US
dc.identifier.citationChen, L., Peirano, G., Yen, K. et al. 2024, 'CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli', Microbiology Spectrum, vol. 12, no. 1, pp. 1-7. DOI: 10.1128/spectrum.03692-23en_US
dc.identifier.issn2165-0497
dc.identifier.other10.1128/spectrum.03692-23
dc.identifier.urihttp://hdl.handle.net/2263/100790
dc.language.isoenen_US
dc.publisherAmerican Society for Microbiologyen_US
dc.rights© 2023 Chen et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.en_US
dc.subjectPlasmiden_US
dc.subjectCRISPRen_US
dc.subjectpCasCureen_US
dc.subjectExtra-intestinal pathogenic Escherichia coli (ExPEC)en_US
dc.subjectAntimicrobial resistance (AMR)en_US
dc.subjectMultidrug-resistant (MDR)en_US
dc.subjectSDG-03: Good health and well-beingen_US
dc.titleCRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia colien_US
dc.typeArticleen_US

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