Neural crest cells are multipotent and migratory progenitors that arise at the border of neural and non-neural ectoderm during early development of vertebrate embryos. The neural crest population gives rise to most of the structural elements of the heard, such as teeth, bone and cartilage, thus being crucial for craniofacial development. Our research focuses on genes involved in migration and differentiation of the neural crest and we are thus poised to shed light on the origins of craniofacial malformations. In particular, we using a candidate gene approach to identify players in craniofacial formation and examine their epistatic relationship to known factors in neural crest development. First we conduct detailed expression pattern analysis followed by functional perturbation using short hairpin RNA interference as well as overexpression of the candidate gene. The effects of the functional perturbation will be evaluated both quantitatively and qualitatively, with quantitative PCR and in situ hybridization for a number of known neural crest factors. Imaging techniques will be used to examine effects of gene perturbations on neural crest migration. The final goal of this project is to assemble a functional gene regulatory network operating during neural crest migration in craniofacial development to allow for better understanding of the ontogeny of craniofacial malformations.