Show simple item record

dc.contributor.authorMallikarjunappa, Sanjay
dc.contributor.authorShandilya, Umesh K
dc.contributor.authorSharma, Ankita
dc.contributor.authorLamers, Kristen
dc.contributor.authorBissonnette, Nathalie
dc.contributor.authorKarrow, Niel A
dc.contributor.authorMeade, Kieran G
dc.date.accessioned2021-12-22T12:25:52Z
dc.date.available2021-12-22T12:25:52Z
dc.date.issued2020-11-02
dc.identifier.citationMallikarjunappa, S., Shandilya, U.K., Sharma, A. et al. Functional analysis of bovine interleukin-10 receptor alpha in response to Mycobacterium avium subsp. paratuberculosis lysate using CRISPR/Cas9. BMC Genet 21, 121 (2020). https://doi.org/10.1186/s12863-020-00925-4en_US
dc.identifier.urihttps://doi.org/10.1186/s12863-020-00925-4
dc.identifier.urihttp://hdl.handle.net/11019/2731
dc.descriptionpeer-revieweden_US
dc.description.abstractBackground The interleukin-10 receptor alpha (IL10RA) gene codes for the alpha chain of the IL-10 receptor which binds the cytokine IL-10. IL-10 is an anti-inflammatory cytokine with immunoregulatory function during the pathogenesis of many inflammatory disorders in livestock, including Johne’s disease (JD). JD is a chronic enteritis in cattle caused by Mycobacterium avium subsp. paratuberculosis (MAP) and is responsible for significant economic losses to the dairy industry. Several candidate genes including IL10RA have been found to be associated with JD. The aim of this study was to better understand the functional significance of IL10RA in the context of immune stimulation with MAP cell wall lysate. Results An IL10RA knock out (KO) bovine mammary epithelial cell (MAC-T) line was generated using the CRISPR/cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9) gene editing system. These IL10RA KO cells were stimulated with the immune stimulant MAP lysate +/− IL-10, or with LPS as a positive control. In comparison to unedited cells, relative quantification of immune-related genes after stimulation revealed that knocking out IL10RA resulted in upregulation of pro-inflammatory cytokine gene expression (TNFA, IL1A, IL1B and IL6) and downregulation of suppressor of cytokine signaling 3 (SOCS3), a negative regulator of pro-inflammatory cytokine signaling. At the protein level knocking out IL10RA also resulted in upregulation of inflammatory cytokines - TNF-α and IL-6 and chemokines - IL-8, CCL2 and CCL4, relative to unedited cells. Conclusions The findings of this study illustrate the broad and significant effects of knocking out the IL10RA gene in enhancing pro-inflammatory cytokine expression and further support the immunoregulatory role of IL10RA in eliciting an anti-inflammatory response as well as its potential functional involvement during the immune response associated with JD.en_US
dc.language.isoenen_US
dc.relation.ispartofseriesBMC Genetics;
dc.subjectJohne’s disease (JD)en_US
dc.subjectInterleukin-10 receptor alpha gene (IL10RA)en_US
dc.subjectCRISPR/cas9en_US
dc.subjectGene knockouten_US
dc.subjectCandidate geneen_US
dc.titleFunctional analysis of bovine interleukin-10 receptor alpha in response to Mycobacterium avium subsp. paratuberculosis lysate using CRISPR/Cas9en_US
dc.typeArticleen_US
dc.date.updated2020-11-08T07:33:06Z
dc.language.rfc3066en
dc.rights.holderThe Author(s)
dc.identifier.doihttps://doi.org/10.1186/s12863-020-00925-4
dc.contributor.sponsorTeagasc Walsh Fellowship Programmeen_US
dc.contributor.sponsorNatural Sciences and Engineering Research Council of Canada (NSERC)en_US
dc.contributor.sponsorSemex Alliance (Canada)en_US
refterms.dateFOA2021-12-22T12:25:52Z


Files in this item

Thumbnail
Name:
12863_2020_Article_925.pdf
Size:
1.704Mb
Format:
PDF

This item appears in the following Collection(s)

Show simple item record