| Popis: |
Interferons (IFNs) are cytokines that function in the innate immune response to viral infection. Type I and type III IFNs canonically signal through the Janus kinase–signal transducer and activator of transcription (JAK-STAT) pathway, although non-canonical signaling pathways have been described. It is unclear how the differences between these canonical and non-canonical signaling pathways and their roles in type I vs. type III IFN signaling influence infection outcomes. Notably, our group has shown that type III IFN signaling at the maternal-fetal interface is associated with adverse fetal outcomes during Zika virus infection in mice. To study the differences between the type I and type III IFN signaling pathways, we constructed monoclonal knockout (KO) cell lines for the type I and type III IFN receptors (IFNAR1 and IFNLR1), as well as a key component of the JAK-STAT pathway (STAT2) in a lung epithelial cell line (A549). Monoclonal KO cell populations were genotypically validated via Sanger sequencing and phenotypically validated by assays assessing ISG induction and viral inhibition. RNA-Seq was performed on IFNAR1 and IFNLR1 KO A549 cells, revealing possible gene targets that are uniquely regulated by expression of the IFNLR1 receptor. Our STAT2 KO cell model generated herein provides a valuable system to interrogate pathways induced via non-canonical mechanisms. Combined with the results of our existing RNA-Seq analysis, we can examine which genes may be preferentially induced by type III IFN signaling and via non-canonical molecular mechanisms. |
