DYRK1A Inhibition via Conditional Knockout

Chemical inhibition of DYRK1A (e.g. using harmine) inhibits Th17 and promotes Treg differentiation. To better understand the mechanisms by which DYRK1A regulates Th17 differentiation, we are comparing expression profiles of naïve CD4+ T cells during the course of Th17 differentiation in the presence or absence of harmine. Overall design: We MACS-isolated naïve CD4+ T cells from 4 mice on C57/Bl6 background. These were plated in pro-Th17 conditions in the presence or absence of 10 uM harmine. Cells were harvested at 0 and 5 hours and RNA isolated by RNeasy Micro & Mini kits. pro-Th17 conditions: 2ug/mL Anti-Il-4 2ug/mL Anti-Il-12 2ug/mL Anti-IFN? 20ng/mL Il-6 20ng/mL Il1ß 0.1ng/mL TGFB