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    Next Generation Sequencing Reveals the Expression of a Unique miRNA Profile in Response to a Gram-Positive Bacterial Infection

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    Author
    Lawless, Nathan
    Foroushani, Amir K.
    McCabe, Matthew
    O'Farrelly, Cliona
    Lynn, David J
    Keyword
    MicroRNAs
    Next generation sequencing
    Bovine
    Innate immune system
    Epithelial cells
    RNA sequencing
    Date
    2013-01-25
    
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    URI
    http://hdl.handle.net/11019/714
    Citation
    Lawless N, Foroushani ABK, McCabe MS, O’Farrelly C, Lynn DJ (2013) Next Generation Sequencing Reveals the Expression of a Unique miRNA Profile in Response to a Gram-Positive Bacterial Infection. PLoS ONE 8(3): e57543. doi:10.1371/journal.pone.0057543
    Abstract
    MicroRNAs (miRNAs) are short, non-coding RNAs, which post-transcriptionally regulate gene expression and are proposed to play a key role in the regulation of innate and adaptive immunity. Here, we report a next generation sequencing (NGS) approach profiling the expression of miRNAs in primary bovine mammary epithelial cells (BMEs) at 1, 2, 4 and 6 hours post-infection with Streptococcus uberis, a causative agent of bovine mastitis. Analysing over 450 million sequencing reads, we found that 20% of the approximately 1,300 currently known bovine miRNAs are expressed in unchallenged BMEs. We also identified the expression of more than 20 potentially novel bovine miRNAs. There is, however, a significant dynamic range in the expression of known miRNAs. The top 10 highly expressed miRNAs account for >80% of all aligned reads, with the remaining miRNAs showing much lower expression. Twenty-one miRNAs were identified as significantly differentially expressed post-infection with S. uberis. Several of these miRNAs have characterised roles in the immune systems of other species. This miRNA response to the Gram-positive S. uberis is markedly different, however, to lipopolysaccharide (LPS) induced miRNA expression. Of 145 miRNAs identified in the literature as being LPS responsive, only 9 were also differentially expressed in response to S. uberis. Computational analysis has also revealed that the predicted target genes of miRNAs, which are down-regulated in BMEs following S. uberis infection, are statistically enriched for roles in innate immunity. This suggests that miRNAs, which potentially act as central regulators of gene expression responses to a Gram-positive bacterial infection, may significantly regulate the sentinel capacity of mammary epithelial cells to mobilise the innate immune system.
    Funder
    Teagasc Walsh Fellowship Programme
    ae974a485f413a2113503eed53cd6c53
    http://dx.doi.org/10.1371/journal.pone.0057543
    Scopus Count
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    Animal & Bioscience

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