Foxe3 is a DNA-transcription factor (TF) that is vital to the normal development of the lens. The lens is a structural component of the eye that is responsible for transmitting and focusing light onto the retina. Here we examine the role that Foxe3 plays in regulating networks of genes that are essential for cellular differentiation. Ultimately, this regulation results in the proper functioning of the lens and better vision. We will specifically analyze two mice strains homozygous for mutations of Foxe3. In one of these strains, Foxe3KO, the Foxe3 gene is entirely replaced with the sequences encoding mCherry, a red fluorescent protein. The strain, Foxe3Mlr68, consists of a mutation in the C-terminal domain of the Foxe3 gene that was generated in the Robinson lab. FOXE3 knockouts are known to have severe defects in lens development, resulting in numerous phenotypic symptoms of ocular disease. I proposed to compare these mutations and their impacts on lens morphology at three developmental stages. Both Foxe3Mlr68 mice and Foxe3 KO mice showed signs of abnormal lens development in comparison to wild-type mice. In the future, the Robinson lab aims to learn more about the mechanism by which Foxe3 regulates gene expression and why such severe phenotypes are observed in MLR68 mutants. In addition, since the Foxe3 KO mice have mCherry expression in the cells that should express FOXE3, FACS sorting will allow for the separation of lens cells at stages of development earlier than we can reasonably dissect from embryos. We were successful in our goal to develop a standard procedure for FACS in which we determined the enzymes needed to successfully generate a single-cell suspension. In the future, after FACS sorting, we will perform RNA-seq analysis to determine what genes are expressed in these lens cells that lack FOXE3. This project reflects my journey of intellectual growth and development at Miami University and has been purposeful in gaining research experience in preparation for dental school.
Author(s): Jillian Lentz, Biology and Music Major
Advisor(s): Michael Robinson, Department of Biology
Jacob Weaver, Department of Biology
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