Modular Design and Directed Evolution for the Development of Genetically Encodable Fluorescent Sensors

Abstract

The development of genetically encodable sensors for optical imaging has revolutionized the way researchers investigate cellular phenomena. Illuminating the molecular dynamics of the cell has become nearly routine with countless improvements in sensor design and directed evolution efforts to enhance their reporting ability. These optical technologies owe their impressive versatility to their genetically encodable nature. As new luminescent and fluorescent reporter domains are discovered, new sensors will inevitably be developed from them. In this thesis, I cover the basics of sensor design while commenting on methods for construction of prototypes, rational structure guided design, directed evolution methods, and briefly touch on new methods for developing split sensors. The goal of this thesis is to explore the currently available technologies for developing genetically encodable optical sensors and provide a methodology for general sensor design with examples provided within each chapter.