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

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. Overall design: Mouse ESCs expressing a cRAF-ERT2 construct allowing ERK activation upon the addition of 4OT to the media. Cells were left untreated (0h), or treated with 250nM 4OHT for 1-2-or 4hrs. Total RNA was collected and purified and rRNA contamination was subsequently removed using the ribogold kit from Illumina prior to library preparation.