Thesis committee members:
Advisor: Robert Gross M.D., Ph.D (BME)
Shawn Hochman, Ph.D (BME)
Nicholas Boulis, M.D. (BME)
Shan Ping Yu, M.D., Ph.D (Emory)
Ute Hochgeschwender, M.D. (Duke)

Title:
Development of genetically encoded bioluminescent light sources for optogenetic applications.

Abstract
The use of optogenetic techniques in vivo currently faces many important challenges that prevent the translation of optogenetic approaches into a clinically meaningful context. Several of these challenges relate to the light source, which currently relies on using lasers or LEDs coupled to optical fibers implanted into the brain. These light sources are both impractical to use in long term in vivo settings (e.g. hardware dependency, limited tissue penetrance), and also pose a significant safety risk (e.g. tissue damage, heat-induced injury). The overall goal of this project is to address the current technical limitations of using external light sources for optogenetic applications in vivo.

We propose to couple luciferase proteins with light-sensitive opsins to demonstrate that bioluminescence can be used as an alternative light source for optogenetic applications. The first two aims of this thesis will consist of developing these biological systems and validating them in vitro. The first aim will consist of demonstrating a valid proof-of-concept showing that luciferases can be used for optogenetic inhibition. The second aim will consist of utilizing responsive luciferases for developing novel optogenetic feedback systems. The third aim will consist of validating these tools in vivo and using them to modulate epileptic activity.

This thesis aims to demonstrate an alternative means for activating light-sensitive opsins in a purely hardware-independent manner, which would add to the versatility and translatability of the optogenetic approach for neuromodulatory applications.