Abstract
Human eye morphogenesis is a complex and conserved process involving tissues from varied embryonic origins and a series of reciprocal inductive events. A critical effector of the Hippo signalling pathway, Yap1, plays a role in organ size control by mediating cell growth, apoptosis, and survival. In human patients, as well as zebrafish and mouse models, perturbed regulation of Yap signalling result in ocular-specific disorders including microphthalmia and coloboma. However, underlying mechanisms of Yap in the development of the human eye remain poorly understood. My project aims to use inducible CRISPR interference (CRISPRi) in addition to using chemical Yap inhibitors to down-regulate expression of wild-type alleles in retinal organoids to perturb normal Yap function. This allows characterisation of cellular outcomes and essentially investigate stem cell proliferation and differentiation in ocular disorders. I will show characterisation of wild-type and Yap inhibited retinal organoids and the ongoing generation of an inducible CRISPRi system in human pluripotent stem cells.
