Removal of AMR plasmids using a mobile, broad host-range CRISPR-Cas9 delivery tool
dc.contributor.author | Walker-Sünderhauf, D | |
dc.contributor.author | Klümper, U | |
dc.contributor.author | Pursey, E | |
dc.contributor.author | Westra, ER | |
dc.contributor.author | Gaze, WH | |
dc.contributor.author | van Houte, S | |
dc.date.accessioned | 2023-05-30T09:28:59Z | |
dc.date.issued | 2023-05-25 | |
dc.date.updated | 2023-05-29T09:06:32Z | |
dc.description.abstract | Antimicrobial resistance (AMR) genes are widely disseminated on plasmids. Therefore, interventions aimed at blocking plasmid uptake and transfer may curb the spread of AMR. Previous studies have used CRISPR-Cas-based technology to remove plasmids encoding AMR genes from target bacteria, using either phage- or plasmid-based delivery vehicles that typically have narrow host ranges. To make this technology feasible for removal of AMR plasmids from multiple members of complex microbial communities, an efficient, broad host-range delivery vehicle is needed. We engineered the broad host-range IncP1-plasmid pKJK5 to encode cas9 programmed to target an AMR gene. We demonstrate that the resulting plasmid pKJK5::csg has the ability to block the uptake of AMR plasmids and to remove resident plasmids from Escherichia coli. Furthermore, due to its broad host range, pKJK5::csg successfully blocked AMR plasmid uptake in a range of environmental, pig- and human-associated coliform isolates, as well as in isolates of two species of Pseudomonas. This study firmly establishes pKJK5::csg as a promising broad host-range CRISPR-Cas9 delivery tool for AMR plasmid removal, which has the potential to be applied in complex microbial communities to remove AMR genes from a broad range of bacterial species. | en_GB |
dc.description.sponsorship | Medical Research Council (MRC) | en_GB |
dc.description.sponsorship | Biotechnology and Biological Sciences Research Council (BBSRC) | en_GB |
dc.description.sponsorship | Lister Institute for Preventative Medicine | en_GB |
dc.description.sponsorship | JPI-AMR HARISSA programme (MISTAR) | en_GB |
dc.description.sponsorship | Bundesministerium für Bildung und Forschung | en_GB |
dc.description.sponsorship | European Research Council (ERC) | en_GB |
dc.description.sponsorship | College of Life and Environmental Sciences, University of Exeter | en_GB |
dc.format.medium | ||
dc.identifier.citation | Vol. 169(5), article 001334 | en_GB |
dc.identifier.doi | https://doi.org/10.1099/mic.0.001334 | |
dc.identifier.grantnumber | MR/N0137941/1 | en_GB |
dc.identifier.grantnumber | BB/R010781/1 | en_GB |
dc.identifier.grantnumber | BB/S017674/1 | en_GB |
dc.identifier.grantnumber | 01LC1904A | en_GB |
dc.identifier.grantnumber | ERC-STG-2016–714478 | en_GB |
dc.identifier.uri | http://hdl.handle.net/10871/133243 | |
dc.identifier | ORCID: 0000-0002-9345-6204 (Gaze, William H) | |
dc.identifier | ORCID: 0000-0001-7047-1308 (van Houte, Stineke) | |
dc.language.iso | en | en_GB |
dc.publisher | Microbiology Society | en_GB |
dc.relation.url | https://www.ncbi.nlm.nih.gov/pubmed/37226834 | en_GB |
dc.rights | © 2023 The Authors. This is an open-access article distributed under the terms of the Creative Commons Attribution License. | en_GB |
dc.subject | AMR gene removal | en_GB |
dc.subject | CRISPR-Cas plasmids | en_GB |
dc.subject | antibiotic resensitization | en_GB |
dc.subject | antimicrobial resistance | en_GB |
dc.subject | broad host-range plasmids | en_GB |
dc.subject | plasmid curing | en_GB |
dc.title | Removal of AMR plasmids using a mobile, broad host-range CRISPR-Cas9 delivery tool | en_GB |
dc.type | Article | en_GB |
dc.date.available | 2023-05-30T09:28:59Z | |
dc.identifier.issn | 1350-0872 | |
exeter.place-of-publication | England | |
dc.description | This is the final version. Available on open access from the Microbiology Society via the DOI in this record | en_GB |
dc.identifier.eissn | 1465-2080 | |
dc.identifier.journal | Microbiology | en_GB |
dc.relation.ispartof | Microbiology (Reading), 169(5) | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | en_GB |
dcterms.dateAccepted | 2023-04-24 | |
rioxxterms.version | VoR | en_GB |
rioxxterms.licenseref.startdate | 2023-05-25 | |
rioxxterms.type | Journal Article/Review | en_GB |
refterms.dateFCD | 2023-05-30T09:23:52Z | |
refterms.versionFCD | VoR | |
refterms.dateFOA | 2023-05-30T09:29:41Z | |
refterms.panel | A | en_GB |
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