Toward tunable RNA thermo-switches for temperature dependent gene expression

Details Toward tunable RNA thermo-switches for temperature dependent gene expression Toward tunable RNA thermo-switches for temperature dependent gene expression

2011-09-25

arxiv.org/abs/1109.5402v1

Biology

Quantitative Biology

Subcellular Processes

12 pages, 10 figures

Scientific paper

RNA thermometers are mRNA strands with a temperature dependent secondary structure: depending on the spatial conformation, the mRNA strand can get translated (on-state) or can be inaccessible for ribosomes binding (off-state). These have been found in a number of microorganisms (mainly pathogens), where they are used to adaptively regulate the gene expression, in response to changes in the environmental temperature. Besides naturally occurring RNA thermometers, synthetic RNA thermometers have been recently designed by modifying their natural counterparts (Hofacker et al., 2003). The newly designed RNA thermometers are simpler, and exhibit a sharper switching between off- and on-states. However, the proposed trial-and-error design procedure has not been algorithmically formalized, and the switching temperature is rigidly determined by the natural RNA thermometer used as template for the design. We developed a general algorithmic procedure (consensus distribution) for the design of RNA thermo-switches with a tunable switching temperature that can be decided in advance by the designer. A software tool with a user friendly GUI has been written to automate the design of RNA thermo-switches with a desired threshold temperature. Starting from a natural template, a new RNA thermometer has been designed by our method for a new desired threshold temperature of 32C. The designed RNA thermo-switch has been experimentally validated by using it to control the expression of lucifarase. A 9.2 fold increase of luminescence has been observed between 30C and 37C, whereas between 20C and 30C the luminescence increase is less than 3-fold. This work represents a first step towards the design of flexible and tunable RNA thermometers that can be used for a precise control of gene expression without the need of external chemicals and possibly for temperature measurements at a nano-scale resolution.

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