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: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: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: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 present NGST, TMT and Q-TOF MS platform should provide unprecedented resources to address such questions as to how hypoxic condition affects gene, miRNA, protein, and metabolite expression and changes the molecular pathways, and whether miRNAs participate in this process. For this purpose, we characterise transcriptomic, miRNAomic, proteomic and metabonomic sequencing of control- and hypoxia-treated P. vachelli muscles to elucidate the molecular mechanisms of hypoxia adaptation. We were able to find the predicted miRNA-mRNA-protein-metabolite regulatory network using bioinformatics analysis and miRNA prediction algorithms (Fig. 1). This is the first report on integrated analysis of transcriptome, miRNAome, and proteome, and metabolome in fishes and as such offers deeper insight into the hypoxia molecular mechanisms. We provide a good case study with which to analyse mRNA, miRNA, protein and metabolite expression and profile non-model fish species.