Type

Journal Article

Authors

Douwe van Sinderen
Thijs R.H.M. Kouwen
Laurens Hanemaaijer
Jennifer Mahony
Brian McDonnell

Subjects

Veterinary

Topics
mutant dairy processing dairy industry mrna adsorption sedimentation streptococcus thermophilus resistance insensitive phage infection bacteriophage receptor

Generation of bacteriophage-insensitive mutants of Streptococcus thermophilus using an antisense RNA CRISPR-Cas silencing approach (2017)

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.
Collections Ireland -> University College Cork -> APC Microbiome Institute
Ireland -> University College Cork -> College of Science, Engineering and Food Science
Ireland -> University College Cork -> APC Microbiome Institute- Journal Articles
Ireland -> University College Cork -> Microbiology
Ireland -> University College Cork -> Research Institutes and Centres
Ireland -> University College Cork -> Microbiology - Journal Articles

Full list of authors on original publication

Douwe van Sinderen, Thijs R.H.M. Kouwen, Laurens Hanemaaijer, Jennifer Mahony, Brian McDonnell

Experts in our system

1
Douwe van Sinderen
University College Cork
Total Publications: 144
 
2
Thijs R. Kouwen
University College Cork
Total Publications: 10
 
3
Laurens Hanemaaijer
University College Cork
Total Publications: 10
 
4
Jennifer Mahony
University College Cork
Total Publications: 87
 
5
Brian McDonnell
University College Cork
Total Publications: 6