Project description:We used microarrays to study the global gene expression and identified differentially expressed genes in CLOCK inducibel shRNA knockdown GSC272 cells, aiming to identify genes and pathways that are regulated by CLOCK.
Project description:The circadian clock is intricately connected with metabolism, however the precise details of these connections are incomplete. Here we used high temporal resolution metabolite profiling to determine circadian regulation of mouse liver and cell autonomous metabolism. In mouse liver, we found ~50% of metabolites were circadian, with strong enrichment of the nucleotide, amino acid, and methylation pathways. In U2OS cells, 27% of metabolites were circadian, including amino acids and NAD biosynthesis, also clock controlled in liver. To assess whether cell autonomous metabolite rhythms were clock-dependent, we used RNAi to perturb Bmal1, Cry1, and Cry2. Bmal1 knockdown eliminated most metabolite rhythms, while Cry1 generally shortened and Cry2 lengthened rhythms. Surprisingly, we found Cry1 knockdown induced 8 hr rhythms in amino acid, methylation, and vitamin metabolites, decoupling metabolite and transcriptional rhythms. These results provide the first comprehensive views of circadian liver and cell autonomous metabolism.
Project description:Here, we report that the disruption of circadian clock obtained through shRNA-mediated knockdown of Clock and Bmal1 genes negatively impacted the in vitro growth of a T-cell acute lymphoblastic leukemia (T-ALL) cell line. We performed gene expression profiling analysis of RPMI-8402 cell line upon knock-down of the circadian genes Clock and Bmal1. We found that circadian clock directly infuences the expression of genes, such as IL20RB, involved in JAK/STAT signaling, making the T-ALL cells more responsive to Interleukin 20 (IL20).