Design of a Toolbox of RNA Thermometers

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Rohit Satija, Shaunak Sen, Dan Siegal-Gaskins, Richard M. Murray
Presented at Synthetic Biology: Engineering, Evolution and Design (SEED), 10-13 June 2015

RNA thermometers mediate responses to temperature changes in various natural circuits, and have been developed in a synthetic context as well. However, a toolbox of RNA thermometers with diâµerent sensitivities to temperature is lacking. Here, we address this issue using a combination of computational and experimental methodologies. We analysed a set of available synthetic RNA thermometers through a quantification of their activity as a function of temperatures in a cell- free expression molecular breadboard system as well as through computation of their melt profiles. Based on this, we computed melt profiles of a library of RNA thermometers and found that the library contained RNA thermometers with a range of sensitivities and thresholds in their response to temperature. We constructed this library and found, through preliminary measurements, a wide range of responses to temperature, which in some cases matched the computational predictions. The constructed library represents a toolbox of RNA thermometers with different sensitivities and is foun- dational work towards synthetic biology applications such as efficient control of large volume chemical reactors, precise spatiotemporal control of gene expression as well as tools to engineer robustness to temperature in biomolecular circuits.