Expression profiling by high throughput sequencing Genome binding/occupancy profiling by high throughput sequencing Other
Summary
To dissect Cohesin-independent mechanisms that compartmentalize the mammalian genome, we combined super-resolution imaging and high-resolution chromatin interactome, chromatin binding and accessibility and RNA sequencing analysis. We found that the bromodomain and extra-terminal domain (BET) family protein BRD2 is one key regulator to promote compartmental interactions in the absence of Cohesin. We also identified competitive, rather than cooperative relationship between BET family proteins in shaping the 3D genome organization. This study uncovers new mechanisitc insights on the 3D organization of the mammalian genome.
Overall design
We generated inducible protein degradation system for Cohesin subunit RAD21, BET family proteins in mouse embryonic stem cells (ESCs). We performed Micro-C, ChIP-seq, ATAC-seq and RNA-seq experiments and analysis after acute depleting individual protein or in combination.