Determination of Acr-mediated immunosuppression in Pseudomonas aeruginosa
dc.contributor.author | Pons, BJ | |
dc.contributor.author | Westra, ER | |
dc.contributor.author | van Houte, S | |
dc.date.accessioned | 2023-08-15T14:05:42Z | |
dc.date.issued | 2022-11-28 | |
dc.date.updated | 2023-08-15T13:21:01Z | |
dc.description.abstract | Bacteria have a broad array of defence mechanisms to fight bacteria-specific viruses (bacteriophages, phages) and other invading mobile genetic elements. Among those mechanisms, the 'CRISPR-Cas' (Clustered Regularly Interspaced Short Palindromic Repeats - CRISPR-associated) system keeps record of previous infections to prevent re-infection and thus provides acquired immunity. However, phages are not defenceless against CRISPR-based bacterial immunity. Indeed, they can escape CRISPR systems by encoding one or several anti-CRISPR (Acr) proteins. Acr proteins are among the earliest proteins produced upon phage infection, as they need to quickly inhibit CRISPR-Cas system before it can destroy phage genetic material. As a result, Acrs do not perfectly protect phage from the CRISPR-Cas system, and infection often fails. However, even if the infection fails, Acr can induce a lasting inactivation of the CRISPR-Cas system. The method presented here aims to assess the lasting CRISPR-Cas inhibition in Pseudomonas aeruginosa induced by Acr proteins by:•Infecting the P. aeruginosa strain with a phage carrying an acr gene.•Making the cell electrocompetent while eliminating the phage•Transforming the cells with a plasmid targeted by the CRISPR-Cas system and a non-targeted one to measure the relative transformation efficiency of the plasmids. This method can be adapted to measure which parameters influence Acr-induced immunosuppression in different culture conditions. | en_GB |
dc.description.sponsorship | Biotechnology and Biological Sciences Research Council (BBSRC) | en_GB |
dc.description.sponsorship | Engineering and Physical Sciences Research Council (EPSRC) | en_GB |
dc.description.sponsorship | European Union Horizon 2020 | en_GB |
dc.format.extent | 101941- | |
dc.format.medium | Electronic-eCollection | |
dc.identifier.citation | Vol. 10, article 101941 | en_GB |
dc.identifier.doi | https://doi.org/10.1016/j.mex.2022.101941 | |
dc.identifier.grantnumber | BB/S017674/1 | en_GB |
dc.identifier.grantnumber | BB/R010781/1 | en_GB |
dc.identifier.grantnumber | EP/X026507/1 | en_GB |
dc.identifier.grantnumber | ERC-STG-2016-714478 | en_GB |
dc.identifier.uri | http://hdl.handle.net/10871/133782 | |
dc.identifier | ORCID: 0000-0003-4396-0354 (Westra, Edze R) | |
dc.identifier | ORCID: 0000-0001-7047-1308 (van Houte, Stineke) | |
dc.language.iso | en | en_GB |
dc.publisher | Elsevier | en_GB |
dc.relation.url | https://www.ncbi.nlm.nih.gov/pubmed/36504499 | en_GB |
dc.rights | © 2022 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/) | en_GB |
dc.subject | Anti-CRISPR | en_GB |
dc.subject | CRISPR-Cas | en_GB |
dc.subject | Transformation efficiency | en_GB |
dc.title | Determination of Acr-mediated immunosuppression in Pseudomonas aeruginosa | en_GB |
dc.type | Article | en_GB |
dc.date.available | 2023-08-15T14:05:42Z | |
dc.identifier.issn | 2215-0161 | |
exeter.article-number | 101941 | |
exeter.place-of-publication | Netherlands | |
dc.description | This is the final version. Available on open access from Elsevier via the DOI in this record | en_GB |
dc.description | Data availability: Data will be made available on request. | en_GB |
dc.identifier.eissn | 2215-0161 | |
dc.identifier.journal | MethodsX | en_GB |
dc.relation.ispartof | MethodsX, 10 | |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | en_GB |
dcterms.dateAccepted | 2022-11-22 | |
dc.rights.license | CC BY | |
rioxxterms.version | VoR | en_GB |
rioxxterms.licenseref.startdate | 2022-11-28 | |
rioxxterms.type | Journal Article/Review | en_GB |
refterms.dateFCD | 2023-08-15T14:03:12Z | |
refterms.versionFCD | VoR | |
refterms.dateFOA | 2023-08-15T14:05:45Z | |
refterms.panel | A | en_GB |
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