Identification of mir199 Targets Involved in Xenopus Laevis Eye Development

According to the CDC, between the years 2004-2006 congenital eye defects were present in 1 of every 5,349 births. Our research proposes to uncover the underlying regulation of eye development through identifying proteins that, at high levels, block normal eye development. The network of proteins responsible for eye development (the EFTF) is well understood and conserved across vertebrates. Previous research from our lab has identified post transcriptional regulation of this network through microRNA regulation, in particular through miR199. We found that in addition to its regulation of the EFTFs, miR199 also regulates proteins blocking correct eye development. This allowed us to conclude that balanced expression of miR199 is necessary for normal eye development. Our lab identified several potential targets that could block normal eye development and are predicted to be regulated by miR199. My role in this project has been to validate miR199 regulation of the targets hace1, ptk7.L, and prox1. To do this, I have cloned fragments of their 3’ untranslated regions (3’ UTR) and performed luciferase assays on them. Greater understanding of the underlying systems regulating eye development will pave the way for future research regarding how these proteins lead to congenital eye defects.