Project description:We used microarrays to identified new factor that regulates key factors in siRNA DYRK2 cells. We silenced DYRK2 in MCF-7 cells (siDYRK2 cells) using siRNA. For control, non-silencing siRNA was used. We obtained expression profiles from siDYRK2 cells and sControl cells.
Project description:We used microarrays to identified new factor that regulates cancer stem cell population in shRNA DYRK2 cells using stable DYRK2 knockdown cells and mammosphere.
Project description:We used microarrays to identified new factor that regulates cancer stem cell population in shRNA DYRK2 cells using stable DYRK2 knockdown cells and mammosphere. We stably silenced DYRK2 in MCF-7 cells (shRNA-DYRK2 cells) using pSuper vector. For control, pSuper control cells were created. We obtained expression profiles from shRNA-DYRK2 cells and pSuper control cells. We also obtained RNA from mammosphere which are stably expressing pSuper vector or shRNA DYRK2.
Project description:We generated liver-specific Dyrk2 knockout mice mating Dyrk2 flox mice with Alb-cre mice and co-introduced SB13-transposase-, myrAkt-, Myc- and mutant Hras-expressing plasmids with either HA- or Dyrk2-expressing plasmid into the knockout mice by HTVi. Dyrk2-expressing suppressed tumorigenesis compared with HA-expressing.
Project description:We generated DYRK2-deficient mice using the CRISPR/Cas9 genome editing method and demonstrated that loss of DYRK2 gene causes fetal growth retardation and neonatal lethality at birth. Total RNA from DYRK2-/- whole embryo was compared with those of WT mice by microarray.
Project description:To understand the molecular mechanisms underlying developmental abnormalities in Dyrk2-/- mice, we performed comprehensive whole-genome RNA sequencing of the MEFs from wild-type and Dyrk2-/- embryos
Project description:The dual-specificity tyrosine phosphorylation-regulated kinase DYRK2 has emerged as a key regulator of cellular processes such as proteasome-mediated protein degradation. To gain further insights into its function, we took a chemical biology approach and developed C17, a potent small-molecule DYRK2 inhibitor, through multiple rounds of structure-based optimization guided by a number of co-crystallized structures. C17 displayed an effect on DYRK2 at a single-digit nanomolar IC50 and showed outstanding selectivity for the human kinome containing 467 other human kinases. Using C17 as a chemical probe, we further performed quantitative phosphoproteomic assays and identified several novel DYRK2 targets, including eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) and stromal interaction molecule 1 (STIM1). DYRK2 phosphorylated 4E-BP1 at multiple sites, and the combined treatment of C17 with AKT and MEK inhibitors showed synergistic 4E-BP1 phosphorylation suppression. The phosphorylation of STIM1 by DYRK2 substantially increased the interaction of STIM1 with the ORAI1 channel, and C17 impeded the store-operated calcium entry process. Collectively, these studies further expand our understanding of DYRK2 and provide a valuable tool to further pinpoint its biological function.
Project description:The dual-specificity tyrosine phosphorylation-regulated kinase DYRK2 has emerged as a key regulator of cellular processes such as proteasome-mediated protein degradation. To gain further insights into its function, we took a chemical biology approach and developed C17, a potent small-molecule DYRK2 inhibitor, through multiple rounds of structure-based optimization guided by a number of co-crystallized structures. C17 displayed an effect on DYRK2 at a single-digit nanomolar IC50 and showed outstanding selectivity for the human kinome containing 467 other human kinases. Using C17 as a chemical probe, we further performed quantitative phosphoproteomic assays and identified several novel DYRK2 targets, including eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) and stromal interaction molecule 1 (STIM1). DYRK2 phosphorylated 4E-BP1 at multiple sites, and the combined treatment of C17 with AKT and MEK inhibitors showed synergistic 4E-BP1 phosphorylation suppression. The phosphorylation of STIM1 by DYRK2 substantially increased the interaction of STIM1 with the ORAI1 channel, and C17 impeded the store-operated calcium entry process. Collectively, these studies further expand our understanding of DYRK2 and provide a valuable tool to further pinpoint its biological function.