Project description:The study investigates CTCF/cohesin binding and chromatin looping by ChIP-seq and Micro-C. By ChIP-seq, we determined the genome-wide binding profiles of CTCF and Smc1a cohesin subunit in a knock-in mouse ES cell line (wt-CTCF; clone C59) with endogenously tagged wild type CTCF (FLAG-Halo-mCTCF) and Rad21 (mRad21-SNAPf-V5), and compared them to the same ES line expressing a mutant CTCF (ΔRBR-CTCF; clone C59D2), where we replaced a previously described RNA binding region with a short linker (GDGAGLINS) followed by a 3xHA tag (N576_D611del::3xHA). By Micro-C, we compared nucleosome-resolution chromosome folding maps of the same ES cell lines C59 and C59D2 described above, to determine the effect of deleting CTCF RNA binding region on chromatin looping.
Project description:Using Chromatin immunoprecipitation combined with Selective Isolation of Chromatin Associated Proteins (ChIP-SICAP), we define the chromatin-bound proteins co-localized with CTCF in mouse embryonic stem cells (ES) and neuronal stem (NS) cells. We uncover the proteins that are differentially co-localized with CTCF in ES and NS cells. We show the link between the underlying role of RNA-binding proteins with consolidation of chromatin structure during mammalian development.
