Amplicon based single molecule footprinting in TC-1 DNMT TKO mESCs after RMCE and after 24h p300 inhibition

ABSTRACT: To quantify the contribution of specific transcription factor binding versus chromatin context in chromatin opening, we inserted libraries containing hundreds of CREs into a landing pad within a neutral chromatin environment, devoid of activating or repressive chromatin modifications, in mESCs using Recombination-Mediated Cassette Exchange (RMCE). Chromatin accessibility of the inserted fragments was then profiled by targeted SMF using PCR primers that anneal to a synthetic flanking sequence, allowing unambiguous differentiation of the ectopic site from its endogenous counterpart. To investigate whether H3K27Ac contributes to chromatin accessibility at enhancers, we globally reduced H3K27Ac levels by chemically inhibiting the histone acetyltransferase p300 with the small molecule A-485 (24

INSTRUMENT(S): Illumina MiSeq, NextSeq 2000

ORGANISM(S): Mus musculus

SUBMITTER: charles girardot 

PROVIDER: E-MTAB-16659 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress