Engineering of a light-activated Caspase-3 in vitro and in Drosophila neurons

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" Engineering of a light-activated Caspase-3 in vitro and in Drosophila neurons "
Smart, Ashley Danielle Davis, Graeme W; Wells, James A

Document Type	:	Latin Dissertation
Language of Document	:	English
Record Number	:	904732
Doc. No	:	TL4zg5b6mg
Main Entry	:	Smart, Ashley Danielle
Title & Author	:	Engineering of a light-activated Caspase-3 in vitro and in Drosophila neurons\ Smart, Ashley DanielleDavis, Graeme W; Wells, James A
College	:	UCSF
Date	:	2017
student score	:	2017
Abstract	:	A cell is a complex structure filled with proteins that are undertaking hundreds of tasks. Some proteins have multiple different roles within a cell. These proteins must be differentially controlled temporally and/or spatially so the cell can use them to achieve different functional outcomes. Many mechanisms could be used to control these proteins, but we currently do not know how different cell regulation mechanisms lead to different protein roles. To this end, we have engineered one protein with temporal and spatial control to have a tool to begin to understand how different protein regulations lead to different outcomes. The protein we engineered is Caspase-3 which has a variety of roles from apoptosis to synaptic remodeling. This dissertation provides the strategy used to create a light-activated caspase and the testing and optimization of it in vitro and in vivo. Chapter 1 discusses the background of caspases as well as their different roles in cells. Chapter 2 provides the design of the light-activated caspase including in silico modelling and testing in vitro. Chapter 3 discusses the testing of the light-activated caspase in vivo and our findings that different neuronal cell types respond differently to caspase activation. Chapter 4 discusses potential modifications and improvements to the light-activated caspase and how a light-activated caspase can be used to further understand what type of regulation of proteins can lead to different cellular outcomes.
Added Entry	:	Davis, Graeme W; Wells, James A
Added Entry	:	UCSF