Dynamic Lineage Priming by ERK is Driven by Transcription Factor-Independent Enhancer Regulation [2h_EU]

Extensive analysis if the means by which ERK affects transcription via enhancer regulation in mouse embryonic stem cells. This study combines analysis of mRNA, and nascent RNA dynamics and correlates this with changes in transcription factor, co-factor, and RNAPII binding and activity. Mouse embryonic stem cells, stably expressing a cRAF-ErT2 expression cassette, were treated with 4OHT for 2hrs to activate ERK signalling via cRAF-ErT2. 15 minutes prior to RNA isolation, cells were treated with 5mM ethynyl uridine to label newly synthesized RNA . Subsequent to total RNA isolation, labelled RNA was purified by standard chemistry, libraries prepared, and sequenced.