CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli
ABSTRACT IncF 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...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2024-01-01
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| Series: | Microbiology Spectrum |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.03692-23 |
| _version_ | 1797357369497223168 |
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| author | Liang Chen Gisele Peirano Kelly Yen Bingjie Wang Austin Terlecky Rebekah DeVinney Barry N. Kreiswirth Johann D. D. Pitout |
| author_facet | Liang Chen Gisele Peirano Kelly Yen Bingjie Wang Austin Terlecky Rebekah DeVinney Barry N. Kreiswirth Johann D. D. Pitout |
| author_sort | Liang Chen |
| collection | DOAJ |
| description | ABSTRACT IncF 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 influence more complex aspects of bacterial behavior, such as cell invasion and in vivo fitness. |
| first_indexed | 2024-03-08T14:44:08Z |
| format | Article |
| id | doaj.art-9a1a24ed801d4298aa6fafe427db4ba0 |
| institution | Directory Open Access Journal |
| issn | 2165-0497 |
| language | English |
| last_indexed | 2024-03-08T14:44:08Z |
| publishDate | 2024-01-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj.art-9a1a24ed801d4298aa6fafe427db4ba02024-01-11T14:04:38ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972024-01-0112110.1128/spectrum.03692-23CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coliLiang Chen0Gisele Peirano1Kelly Yen2Bingjie Wang3Austin Terlecky4Rebekah DeVinney5Barry N. Kreiswirth6Johann D. D. Pitout7Center for Discovery and Innovation, Hackensack-Meridian Health , Nutley, New Jersey, USACummings School of Medicine, University of Calgary , Calgary, Alberta, CanadaCenter for Discovery and Innovation, Hackensack-Meridian Health , Nutley, New Jersey, USACenter for Discovery and Innovation, Hackensack-Meridian Health , Nutley, New Jersey, USACenter for Discovery and Innovation, Hackensack-Meridian Health , Nutley, New Jersey, USACummings School of Medicine, University of Calgary , Calgary, Alberta, CanadaCenter for Discovery and Innovation, Hackensack-Meridian Health , Nutley, New Jersey, USACummings School of Medicine, University of Calgary , Calgary, Alberta, CanadaABSTRACT IncF 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 influence more complex aspects of bacterial behavior, such as cell invasion and in vivo fitness.https://journals.asm.org/doi/10.1128/spectrum.03692-23extraintestinal pathogenic Escherichia coliplasmidCRISPRpCasCure |
| spellingShingle | Liang Chen Gisele Peirano Kelly Yen Bingjie Wang Austin Terlecky Rebekah DeVinney Barry N. Kreiswirth Johann D. D. Pitout CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli Microbiology Spectrum extraintestinal pathogenic Escherichia coli plasmid CRISPR pCasCure |
| title | CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli |
| title_full | CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli |
| title_fullStr | CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli |
| title_full_unstemmed | CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli |
| title_short | CRISPR-Cas9-mediated IncF plasmid curing in extraintestinal pathogenic Escherichia coli |
| title_sort | crispr cas9 mediated incf plasmid curing in extraintestinal pathogenic escherichia coli |
| topic | extraintestinal pathogenic Escherichia coli plasmid CRISPR pCasCure |
| url | https://journals.asm.org/doi/10.1128/spectrum.03692-23 |
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