The chromatin dynamics of the TFAP2A/ MITF genetic interaction in melanocyte development.

We have deleted TFAP2A and its redundantly-acting paralog TFAP2C in human melanoma SkMel28 cell lines and are assessing MITF binding using Cleavage Under Targets and Release Using Nuclease (CUT&RUN). Conversely, we deleted all copies of MITF in SkMEL28 cells and are similarly profiling TFAP2A bound loci. We predict TFAP2A functions as a pioneer factor for MITF during melanocyte development and that in the absence of TFAP2A, MITF binding at loci normally co-bound by MITF/TFAP2A will be disrupted. Overall design: We targeted TFAP2A and MITF by CUT&RUN sequencing in wild-type, MITF knockout and TFAP2A; TFAP2C double knockout (two independent clones) SkMel28 cell lines. All experiments were conducted in duplicate. IgG was used as a background control.