Characterization of Conserved Non-Coding Elements Essential for wnt1 Expression in Zebrafish



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Brain development has risen as a point of interest due to the lack of information about its complex biological processes. Wnt signaling pathway has been studied due the role it serves in synthesizing important tissue segments that make up the brain. Studies have shown that mutations in the pathway give rise to many developmental diseases yet there is no clear understanding of the mechanisms underlying these errors. We have focused on the role of conserved non-coding elements (CNE) that enhance similar transcriptional patterning with wnt1 and wnt10b. Using a comparative genetics approach, we characterized transcriptional regulation of zebrafish CNE20 and CNE27 with its spotted gar orthologs. With the in-situ hybridization technique, we characterized CNE transcriptional activity of EGFP driven reporter over different developmental stages. Essentially, this would give us insight of where the similarities and differences lay between the organisms CNE activity. Analysis of resulting signal strength show similarities in zebrafish CNE20 and gar CNE27 which could indicate similar enhancer function even if they are not their respective orthologs. Gar CNE20 ortholog has significantly weaker expression than the zebrafish respective enhancer. A network of enhancer activity is also necessary to tissue development due to the overlapping reporter expression. Lastly, expression across all transgenic lines continue to have a similar expression pattern at the observed developmental cycles. This suggests that enhancer regulation could hold highly conserved mechanisms that determine cell fates across many species.