Project description:This experiment captures the poly(A)RNAs co-immunoprecipated with Isw1 in WT and npl3-1 cells. Chromatin remodelers play an essential role in regulating DNA transaction processes. The yeast chromatin remodeling complex ISW1 displays an unexpected function in nuclear mRNP surveillance, retaining premature mRNPs in proximity to their transcription site. This activity of ISW1 requires its recruitment onto chromatin, its interaction with mRNPs but not its nucleosome sliding activity. In npl3-1 cells, mRNA export is compromised resulting in an increased association of mRNAs with Isw1.
Project description:To identify the directly bound transcripts of Flag antibody (the backgroud control for our METTL16 RIP-seq), RNA immunoprecipitation sequencing (RIP-seq) was conducted HEK293T. Briefly, HEK293T cells were infected with pmiRNA1-empty vector. Only the GFP-positive cells were used for study and expanded in DMEM medium.
Project description:Sox2 is a master transcriptional regulator of embryonic development. Having found that Sox2 interacts with RNA-binding proteins, we designed an experiment to discover RNAs associated with Sox2 and other pluripotency factors. Briefly, we used RNA immunoprecipitation followed by high-throughput sequencing (RIP-seq) to sequence transcriptomes enriched for RNAs associated with Klf4, Nanog, Oct4, Sox2 and Suz12. For completeness, this submission includes all RIP-seq data from this study, although some of the data was only used for exploratory analyses. Such analysis indicated that it was important to include input samples for each of the the cell lines that were to be compared, to account for differences in gene expression (e.g. between J1-birA with and without bioSox2), and that overnight IP was more informative than 3 h IP. Therefore, the analysis described in the associated manuscript used samples from experiment batch 3 and 4 only (samples 11-18).