Reprogramming of Müller Glial Cells by Adeno-Associated Viral mediated delivery of split dCAS9-VP64-MS2-p65

Summer 2017

Annika Anderson : Molecular and Cellular Biology (Neurobiology), Gender and Women's Studies

Donor: Pergo Fund
Mentor: David Schaffer

Retinitis Pigmentosa (RP) is a set of genetically inherited neurodegenerative disorders that leads to degeneration of photoreceptor neurons and loss of vision. While onset of RP can be detected early, there are currently no treatments to prevent disease progression. Cellular reprogramming is a promising approach to replenishing photoreceptors after retinal degeneration. In zebrafish, Müller glial cells can dedifferentiate and reprogram into photoreceptor cells upon retinal damage. However, this restorative activity of Müller glial cells is limited in mammals. A CAS9 mediated genome wide activation screen offers the potential to identify genes important in reprogramming Müller glial cells into photoreceptor neurons.
My project aims to test whether the delivery of a split dCAS9-VP64-MS2-p65 by adeno-associated virus (AAV) can be used to activate and identify genes which promote reprogramming of mammalian Müller glial cells to photoreceptor-like cells. I plan to test whether splitting the CRISPR-CAS9 system for viral packaging can activate these genes of interest in vitro and design a protocol for characterizing mouse genotypes for future in vivo analysis of the split CAS9 system. The goal of this work is to explore the significant potential of gene therapy mediated reprogramming of retinal cells to replace depleted photoreceptor cell density for treating RP.

I am incredibly thankful to the Pergo Foundation for providing me with the opportunity to dedicate this summer to my research project. I have learned an incredible amount through this experience and am excited for the possibilities and doors it has opened for me.