Engineering Protein Structure and Interactions to Create Next Generation Optogenetic Tools

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Light-based technologies are revolutionizing our understanding of biological systems at the most fundamental level and steadily guiding researchers towards new diagnostic and therapeutic approaches. As a form of energy, light enables us to perceive shape and colour, and it is the primary means by which most of us make sense of the world around us. By virtue of its very ubiquity and innocuousness, light is a powerful and attractive tool for biological research. If properly harnessed, light can allow us to non-invasively visualize and control biology as it happens in cells and model organisms. ***Over the past two decades, researchers have been figuring out an ever-increasing number of ways to harness light energy to probe cells and tissues in model organisms. One of the first major breakthroughs in this direction dates to 1994, when researchers first reported that a particular protein from jellyfish could be introduced into the cells of other animals, and cause them to glow green when illuminated by blue light. Another major breakthrough came a decade later, when researchers reported a protein from algae that, when introduced into mammalian neurons, would make the neurons fire upon illumination with blue light. These two breakthroughs stand tall as the prototypical examples of “optogenetic” tools proteins that interact with light and enable researchers to either visualize or manipulate cells. ***In this proposal we aim to add entirely new capabilities to the toolbox of optogenetic tools. As optogenetic tools are necessarily proteins, we will use the strategy of protein engineering to change the properties of optogenetic tools and bestow them with properties that do not occur in nature. As one research Theme, we will work to create singular “optogenetic beacon” structures that exist inside the cell of interest and provide a colourful readout of the cell's activity. A second research Theme builds upon our recent report of an engineered protein that can be split into two pieces using light. We now propose to use this protein to build cages inside the cell that can be opened or closed with light. These cages can be used to non-invasively trap or release proteins and will give biologists a powerful new tool for deciphering intracellular signalling pathways. As a third Theme, we will work to develop an improved system that converts changes in protein interactions or function into colour changes that can be easily visualized through a microscope. ***Altogether, we are certain that our work will represent the next step forward for the field of optogenetics and lay the foundation for future breakthroughs in our understanding of cell biology, in health and disease.