Description

The retina is a complex biological structure that has lots of similarities with the brain. Retinal degenerative diseases like Age-related Macular Degeneration (AMD) and Retinitis Pigmentosa (RP) cause permanent loss of photoreceptors which there are no current cures or treatments for. The more we understand about retinal development, the better we can work towards finding treatments and cures for these retinal degenerative diseases and restoring the photoreceptors that were lost to these diseases. For my project, I am creating triple fluorescent reporter cell lines from pluripotent stem cells (PSCs) that will allow me to visualize cells, verify gene expression and track retinal development. These cell lines can be used in 2D and 3D differentiations to assay the development of specific retinal cell types. These cell lines will be validated by creating 3D retinal organoids, followed by FACS sorting and RNA sequencing to verify gene expression. I am also creating plasmid constructs to allow for inducible overexpression of genes like NRL, and LHX4 and direct conversion of PSCs into rod and cone photoreceptors. These constructs can be integrated into PSCs via CRISPR-Cas editing to convert the PSCs to eye field progenitor cells and later into photoreceptor-like cells. The organoids and 2D cultures can then be used for further experiments involving drug screening and help us to better understand retinal biology. The culmination of these experiments will allow us to better understand the retina and retina degenerative diseases. The tools that I am making in these experiments will prove to be useful in studying cures and treatments for these diseases for years to come.