Revision as of 06:06, 7 October 2018

Heat-inducible RNA-based thermosensor-1

A RNA thermosensor that can be used for temperature sensitive post-transcriptional regulation which is based on the change of RNA sencondary structure.The heat induced RNA thermosensors can initiate translation of downstream genes at high temperature.

Usage and Biology

RNA thermosensors are RNA-based genetic control systems that sense temperature changes. At low temperature, the mRNA adopts a conformation that masks the ribosome binding site [Shine–Dalgarno (SD) sequence] within the 5′-untranslated region (5′-UTR) and, in this way, prevents ribosome binding and translation. At elevated temperature, the RNA secondary structure melts locally, thereby giving the ribosomes access to the ribosome binding site to initiate translation. Our team designed synthetic RNA thermosensors that are considerably simpler than naturally occurring thermosensors and can be exploited as convenient on/off switches of gene expression.

Characterization

The thermosensor is constructed on the pSB1C3 vector by goldengate assembly method. Ii is composed of Anderson promotor (BBa_J23104), thermosensor (BBa_K2541001) and sfGFP_optimism(BBa_K2541400). The following device is used to characterize the thermosensor. Then we tranfform the device to E.coli DH5a. We measured the sfGFP_optimism expression to get the actual sensitive temperature of the heat induced RNA thermosensor.

As shown in the figure, the thermosensor sensitive temperature range is [ ]. Our data show that efficient RNA thermosensors can be built from a single small RNA stem-loop structure masking the ribosome binding site, thus providing useful RNA-only toolkit for the regulation of gene expression.

Sequence and Features