Abstract
Purpose :
Development of the central nervous system is dependent on complex gene expression patterns that are both developmental stage and cell-type specific. Yet, traditional methods of understanding gene function usually require direct knock out of the gene, which confounds our ability to discern temporal and tissue specific function. These gene expressions patterns are often regulated by Super-enhancers, vast regions of the genome comprised of multiple smaller conserved elements. Further, understanding how specific elements of a Super-enhancer are regulating these expression patterns is costly and time consuming given traditional experimental methods.
Methods :
Here, we have created a panel of reporters containing specific super-enhancer elements predicted to regulate several genes critical for retinal development. In vivo and ex vivo use of these reporters allow us to visualize which individual super-enhancer elements are able to drive expression during development and in what cell types in mouse retina.
Results :
Interestingly, while several individual enhancer elements do lack specificity, three of thirteen elements tested so far show cell type specific expression patterns of genes critical for retinal development. This supports our central hypothesis that entire Super-Enhancers have diverse rolls across time and space, which can be further dissected by looking at individual regions using a tool that allows for screening efficiency.
Conclusions :
This method of assessing super-enhancer critical regions provides insight into understanding how these individual regulatory mechanisms affect neurogenesis and how their disruption contributes to phenotypic diversity in disease, while also providing a deeper understanding of gene regulation, and gene function that cannot be achieved with traditional knockout mouse approaches.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.