The Use of Gateway® Cloning and Modified BAC Trangenesis to Study Zebrafish Craniofacial Development

Presenter:Wade Sugden, Nathan Johnson, Biology

Poster: C-8

Mentor: Mark Sasaki, Institute of Neuroscience

Transgenesis techniques have revolutionized the study of cellular, developmental, and molecular biology by allowing researchers to visualize the proteins they study and manipulate the expression of genes in vivo. Through the use of genetic regulatory elements, transgenes can be tailored to over-express genes of interest, label tissue-specific cell types, and express genes in atypical locations. When expressed in skeletal elements of the zebrafish (Danio rerio), transgenes can be used to explore cell behavior and the genetic pathways involved in craniofacial morphogenesis. Two methods in particular have streamlined the process of creating transgenic animals: Gateway®-mediated and BAC-mediated transgenesis. Here we discuss the use of both techniques to create zebrafish that express transgenes in craniofacial elements using the runx2b, sp7, col11a2, and sox10 promoters. These promoters were chosen because runx2b and sp7 mark bone at different time points during development, while col11a2 and sox10 serve as cartilage markers. In the future, these fish will be vital tools for conducting cell tracking experiments, distinguishing cell types, and expressing genes of interest in craniofacial structures to determine their function.