Design of a Toolbox of RNA Thermometers

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 e�cient control of large volume chemical reactors, precise spatiotemporal control of gene expression as well as tools to engineer robustness to temperature in biomolecular circuits.

• Conference Paper: http://biorxiv.org/content/early/2015/03/30/017269
• Project(s): NSF MPP