GSE130251
TWIST1 direct targets during embryonic stem cell differentiation [ChIP-seq]
To understand TWIST1 function in early neural development, we analyzed genome-wide binding sites of TWIST1 in mouse embryonic stem cells (mESC) induced for neurogenic differentiation by chromatin immunoprecipitation, followed by high-throughput sequencing Overall design: A2loxcre mESC cell line with doxycycline-inducible Flag-Twist1 under Twist1-/- background was generated (Twist1-/-; Twist1 O/E). mESC with genotype Twist1-/-; Flag-Twist1 O/E and Twist1-/- were differentiated into neuroepithelium cells for 3 days following established protocol. Neurogenic differentiation was initiated by plating mESC in AggreWells (1x 10^6 per well) using feeder independent mESC. Colonies were then lifted from AggreWells and grown in suspension in Neurogenic Differentiation Media supplemented with 15% FBS with gentle shaking. Doxycycline was added 16 hours to induce FLAG-TWIST1 induction before cells were collected for ChIP-seq analysis. Control cell line without inducible cassette (Twist1-/-) were treated under the same condition. Four biological replicate experiments were performed.