Generation of bacteriophage-insensitive mutants of Streptococcus thermophilus using an antisense RNA CRISPR-Cas silencing approach

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dc.contributor.author McDonnell, Brian
dc.contributor.author Mahony, Jennifer
dc.contributor.author Hanemaaijer, Laurens
dc.contributor.author Kouwen, Thijs R.H.M.
dc.contributor.author van Sinderen, Douwe
dc.date.accessioned 2018-01-09T10:12:18Z
dc.date.available 2018-01-09T10:12:18Z
dc.date.issued 2017-11-27
dc.identifier.citation McDonnell, B., Mahony, J., Hanemaaijer, L., Kouwen, T. R. H. M. and van Sinderen, D. (2017) 'Generation of bacteriophage-insensitive mutants of Streptococcus thermophilus using an antisense RNA CRISPR-Cas silencing approach', Applied and Environmental Microbiology, 84(4), e01733-17 (14pp). doi: 10.1128/aem.01733-17 en
dc.identifier.volume 84
dc.identifier.issued 4
dc.identifier.startpage 1 en
dc.identifier.endpage 14 en
dc.identifier.issn 0099-2240
dc.identifier.issn 1098-5336
dc.identifier.uri http://hdl.handle.net/10468/5246
dc.identifier.doi 10.1128/aem.01733-17
dc.description.abstract Predation of starter lactic acid bacteria such as Streptococcus thermophilus by bacteriophages is a persistent and costly problem in the dairy industry. CRISPR-mediated Bacteriophage Insensitive Mutants (BIMs), while straightforward to generate and verify, can quickly be overcome by mutant phages. The aim of this study was to develop a tool allowing the generation of derivatives of commercial S. thermophilus strains which are resistant to phage attack through a non-CRISPR-mediated mechanism, with the objective of generating BIMs exhibiting stable resistance against a range of isolated lytic S. thermophilus phages. To achieve this, standard BIM generation was complemented by the use of the wild-type (WT) strain which had been transformed with an antisense mRNA-generating plasmid (targeting a crucial CRISPR-associated [cas] gene], in order to facilitate the generation of non-CRISPR-mediated BIMs. Phage sensitivity assays suggest that non-CRISPR-mediated BIMs exhibit some advantages compared to CRISPR-mediated BIMs derived from the same strain.Importance: The outlined approach reveals the presence of a powerful host-imposed barrier for phage infection in S. thermophilus. Considering the detrimental economic consequences of phage infection in the dairy processing environment, the developed methodology has widespread applications, particularly where other methods may not be practical or effective in obtaining robust, phage-tolerant S. thermophilus starter strains. en
dc.format.mimetype application/pdf en
dc.language.iso en en
dc.publisher American Society for Microbiology en
dc.relation.uri http://aem.asm.org/content/early/2017/11/20/AEM.01733-17.abstract
dc.rights © 2017 American Society for Microbiology. All Rights Reserved. This is the accepted manuscript version of an article published in Applied and Environmental Microbiology. To access the published work see http://dx.doi.org/10.1128/AEM.01733-17 en
dc.subject Bacteriophage en
dc.subject Resistance en
dc.subject Insensitive en
dc.subject Mutant en
dc.subject mRNA en
dc.subject Adsorption en
dc.subject Receptor, en
dc.subject Sedimentation en
dc.subject Streptococcus thermophilus en
dc.subject Dairy industry en
dc.subject Phage infection en
dc.subject Dairy processing en
dc.title Generation of bacteriophage-insensitive mutants of Streptococcus thermophilus using an antisense RNA CRISPR-Cas silencing approach en
dc.type Article (peer-reviewed) en
dc.internal.authorcontactother Douwe Van Sinderen, Microbiology, University College Cork, Cork, Ireland. +353-21-490-3000 Email: d.vansinderen@ucc.ie en
dc.internal.availability Full text available en
dc.check.info Access to this article is restricted until 6 months after publication by request of the publisher. en
dc.check.date 2018-05-27
dc.date.updated 2018-01-09T09:59:50Z
dc.description.version Accepted Version en
dc.internal.rssid 421115452
dc.contributor.funder Science Foundation Ireland en
dc.description.status Peer reviewed en
dc.identifier.journaltitle Applied and Environmental Microbiology en
dc.internal.copyrightchecked No !!CORA!! en
dc.internal.licenseacceptance Yes en
dc.internal.IRISemailaddress d.vansinderen@ucc.ie en
dc.identifier.articleid e01733-17
dc.relation.project info:eu-repo/grantAgreement/SFI/SFI Starting Investigator Research Grant (SIRG)/15/SIRG/3430/IE/Phage-host interactome of the dairy bacterium Streptococcus thermophilus (PHIST)/ en
dc.relation.project info:eu-repo/grantAgreement/SFI/SFI Investigator Programme/13/IA/1953/IE/Functional analysis of the host adsorption and DNA injection processes of a lactococcal bacteriophage/ en


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