Project description:The goal of this experiment was to assess whether transcription happens during mitosis. For this, we isolated mitotic HeLa cells and treated them with a transcription inhibitor, ActD.
Project description:To investigate the potential influence of transcription on mRNA stability in mammalian cells, we tested the global relationship between rates of synthesis and decay of mRNAs. We estimated synthesis rates by flash 4sU labeling followed by RNA-seq, and decay rates by treating cells with Actinomycin D (ActD) for 0, 2, 4 and 6 hours and measuring half-lives of all expressed genes by MARS-seq analysis. As short-term perturbations of transcription elongation dynamics diminished mRNA stability, we next wished to examine the effect of extended transcription slowdown. To this end, we treated cells with CPT for 24 hours and profiled mRNA stability in a genome-wide manner using ActD time-course and MARS-seq.
Project description:Developing a psoralen probe (PP)-based method for RNA tagging and RNA-protein complex (RP-complex) enrichment, isolating of both coding and noncoding RNAs from HeLa cells was achieved by PP enrichment. Exploring the type and relative distribution of the PP isolated RNAs by RNA-sequencing analysis. And dynamic investigation of RNA upon transcription inhibition by ActD treatment.
