Determining the spatial and temporal expression of transcription factor glis3 during pancreatic development

Gli-similar 3 (glis3) is important for the normal development of the insulin-producing β cells of the pancreas. In addition, Glis3 has been found to be critical for the transcriptional activation of the insulin gene in the mature organ. The hormone insulin is responsible for signaling the uptake of glucose in the blood by peripheral tissues to be used for energy or stored as fat. Accordingly, a number of pathologies have been linked to mutations within the GLIS3 locus in humans including type 1 and type 2 diabetes mellitus. The zebrafish (Danio rario) has emerged as a valuable model organism to study developmental processes due to its rapid external development and transparent embryos. Zebrafish embryos were fixed at key developmental stages and analyzed via whole mount in situ hybridization (WISH) in order to determine the spatial and temporal expression of glis3 throughout development. In order to visualize glis3 expression at the protein level, a transgenic zebrafish line is being engineered using CRISPR/Cas9 technology to produce fish that express chimeric glis3 fused to green fluorescent protein (EGFP). Understanding the spatio-temporal expression of glis3 throughout development could aid in identifying potential therapeutic targets for the treatment of Glis3-associated diseases, including diabetes. Further, the EGFP-glis3 transgenic zebrafish line will provide a powerful tool to identify glis3 target genes, protein-protein interactions, and molecular mechanisms underlying pancreatic development and disease