Project description:RNA silencing pathways are conserved gene regulation mechanisms that lead to degradation, translational repression or transcriptional silencing of target-transcripts selected based on complementarity with small RNA molecules. Using firefly luciferase reporters, we previously were able show that the RNA binding protein GW182 plays a role in microRNA-mediated gene silencing (Rehwinkel, J. et al., RNA J. 2005). In this study, we provide further evidence for the role GW182 in the miRNA pathway by determining targets regulated by this protein at the genomic level. We examined expression profiles in Drosophila cells depleted of GW182 and compared these profiles to data obtained from cells depleted of AGO1, a key effector of miRNA-mediated gene silencing.
Project description:Ribo-seq with firefly luciferase optimality reporters to determine whether incomplete translation and/or inhibited translation initiation is responsible for the reduced ribosome occupancy on nonoptimal transcripts.
Project description:The RNA-binding protein hnRNP K was knocked down using siRNA in human SH-SY5Y. As a control, cells were treated with an siRNA against firefly luciferase.
Project description:Gene expression profiles of primary lymphatic endothelial cells (LECs) isolated from human foreskin were analyzed after siRNA-mediated knockdown of control (firefly luciferase), Prox1, NR2F2 or Prox1/NR2F2 for 48 hours.
Project description:The miRNA profile between EXO from different pancreatic adenocarcinoma cells (A818.4, Capan-1) and different colorectal carcinoma cells (SW948, HT-29). The impact of a knockdown (kd) of function-relevant cancer stem cell markers (CD44v6, Tspan8, CD151, claudin7) on the EXO miRNA profile. The EXO kd cell miRNA profiles were compared with the wt cell EXO as well as between the different kd EXO miRNA profiles. These analyses were important to elaborate joint miRNA in the EXO profile from different cell lines as a starting point to elaborate possible functional activities of EXO including therapeutic translation.