Development and Characterization of Genetic Sensors and Regulators for the Construction of Environmentally-Responsive Genetic Circuits
Date of Award
Doctor of Philosophy (PhD)
Genetic circuits enable engineers to program complex logical behaviors into living organisms. Organisms can be programmed to optimize the production of fuels and chemicals, diagnose and treat diseases, or remediate environmental pollutants. A well-characterized toolbox of genetic sensors and regulators is needed to construct these circuits. Genetic sensors that respond to environmentally-relevant signals allow circuits to evaluate the cell's conditions, and versatile and designable regulators translate information about the cell's environment into the desired response. In this work, we demonstrate the de novo design of RNA thermosensors in Escherichia coli, and integrate these sensors into complex genetic circuits. Next, we provide a large-scale analysis of antisense RNA regulators, generate design rules for these regulators, and validate these design rules through the construction of genetic circuits with predictable behaviors. Finally AND and NAND gates are developed that respond to temperature and pH, and utilize protein and RNA regulators. The sensors, regulators, and circuits developed and characterized here represent a substantial contribution to the synthetic biology toolbox. Furthermore, this work constitutes an important step forward in enabling genetic circuits to overcome challenges in chemical synthesis, medicine, and environmental remediation.
Tae Seok Moon
Ursula Goodenough, Himadri Pakrasi, Hani Zaher, Fuzhong Zhang
Permanent URL: https://doi.org/10.7936/K7GB22GW