Deletions and translocation in non-coding DNA regions around Sox9 gene are associated with craniofacial disorders. This project will assess changes in Sox9 gene expression via enhancers in the breakpoint regions around Sox9 during neural crest differentiation. Cranial neural crest cells (cNCC) will be differentiated in-vitro from human embryonic stem cells (hESC). Preliminary data has shown enhancer-promoter interactions, luciferase reporter assays and CRISPR will be used to downregulate enhancers and analyze Sox9 transcriptional activity. These enhancers' activity will also be dampened in Sertoli cancer cells to compare activity of Sox9 to that of cNCC. Extensive training will be provided in Illumina-NGS, ChIP sequencing, CRISPR gene editing and hESC culture by Hannah K. Long.