A Dynamic Interplay of Klf4 Enhancer Elements Drives Naïve-pluripotency. Mus musculus

We characterized a long-range enhancer cluster driving Klf4 expression in naïve pluripotent stem cells. CRISPR/Cas9 mediated screening and genome editing identified DNA motifs corresponding to key pluripotency TFs on these enhancer regions. We employed ChIP-exo genome-wide sequencing to Esrrb and Stat3 in mESCs and validated their functional engagement on the Klf4 enhancer cluster. Moreover, we performed ATAC-seq on wild type and Oct4/Sox2 composite motif biallelic deletion ESCs on Klf4 enhancer E2 to examine the accessible chromatin structure upon loss of lead TFs Oct4 and Sox2 in a single enhancer region. Overall design: Examine high resolution genome wide TF binding in naïve pluripotent stem cells and the accessible chromatin upon loss of key TF binding.