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

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dc.contributor.authorMcDonnell, Brian
dc.contributor.authorMahony, Jennifer
dc.contributor.authorHanemaaijer, Laurens
dc.contributor.authorKouwen, Thijs R.H.M.
dc.contributor.authorvan Sinderen, Douwe
dc.contributor.funderScience Foundation Irelanden
dc.date.accessioned2018-01-09T10:12:18Z
dc.date.available2018-01-09T10:12:18Z
dc.date.issued2017-11-27
dc.date.updated2018-01-09T09:59:50Z
dc.description.abstractPredation 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.description.statusPeer revieweden
dc.description.versionAccepted Versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.articleide01733-17
dc.identifier.citationMcDonnell, 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.01733-17en
dc.identifier.doi10.1128/aem.01733-17
dc.identifier.endpage14en
dc.identifier.issn0099-2240
dc.identifier.issn1098-5336
dc.identifier.issued4
dc.identifier.journaltitleApplied and Environmental Microbiologyen
dc.identifier.startpage1en
dc.identifier.urihttps://hdl.handle.net/10468/5246
dc.identifier.volume84
dc.language.isoenen
dc.publisherAmerican Society for Microbiologyen
dc.relation.projectinfo: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.projectinfo: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
dc.relation.urihttp://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.01733-17en
dc.subjectBacteriophageen
dc.subjectResistanceen
dc.subjectInsensitiveen
dc.subjectMutanten
dc.subjectmRNAen
dc.subjectAdsorptionen
dc.subjectReceptor,en
dc.subjectSedimentationen
dc.subjectStreptococcus thermophilusen
dc.subjectDairy industryen
dc.subjectPhage infectionen
dc.subjectDairy processingen
dc.titleGeneration of bacteriophage-insensitive mutants of Streptococcus thermophilus using an antisense RNA CRISPR-Cas silencing approachen
dc.typeArticle (peer-reviewed)en
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