Team:Jilin China/Result/Version 1
Demonstrate
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Demonstrate
·RNA-based thermosensors can achieve temperature sensing
We designed four different types of RNA-based thermosensors, including heat-inducible RNA-based thermosensor, heat-repressible RNA-based thermosensor, cold-inducible RNA-based thermosensor and cold-repressible RNA-based thermosensor. We designed more than 200 different thermosensors, and constructed their measurement device.
After the experiments, we got some inspired results. As figure 1 shows, heat-inducible RNA-based thermosensors' activities increase at elevated temperature. Heat-repressible RNA-based thermosensors' activity decrease with increasement of temperature. Besides, cold-inducibe RNA-based thermosensors show lower sensing temperature range than heat-repressible RNA thermosensor, their intensity decrease sharply from 15 to 20℃. And cold-repressible RNA-based thermosensors' activity decrease with decreasing temperature even below 29℃.
Figure 1. Heat map of four different types of RNA-based thermosensors. Rows represent activity levels of different thermosensors. The activity levels are the mean of three replications. These values are normalized using the fluorescence/Abs600 of positive control. (A) includes 48 heat-inducible RNA-based thermosensors' activities at 29, 31, 35, 37, 39 and 42℃. (B) includes 23 heat-repressible RNA-based thermosensors' activities at 29, 37 and 42℃. (C) includes 10 cold-repressible RNA-based thermosensors' activities at 15, 25, 29, 35 and 37℃. (D) includes 8 cold-inducible RNA-based thermosensors' activities at 15, 20, 25℃. ·Features of RNA-based thermosensors can be computed using fitted curve
In order to make users select a thermosensor conveniently by getting the melting temperature, intensity and sensitivity of the thermosensors, we fitted a curve to reflect the relationship between the change of temperature and the expression intensity of thermosensors (Figure 2).
Figure 2. The fitted curve of BBa_K2541029. The blak dash line is the tangent line at melting temperature. The intersection of the upper gray dash line and curve represents the stem-loop structures of thermosensors are all destroyed. The intersection of the medial gray dash line and curve represents a 50% switch in expression occurs, and it is defined as melting temperature. The intersection of lower gray dash line and curcve represents the stem-loop structure of thermosensors all exsit. Table 1 shows the features of all the heat-inducible thermosensors we obtained from the fitting curve.
Table 1. Features of the heat-inducible RNA-based thermosensors 
These features points to the diversity in thermosensor response. These thermosensors' melting temperature ranges 33 to 55℃, with different relative intensity and sensitivity.
·SynRT toolkit is developed and updated to version 3.0
Based on these datas, we classified these thermosensors, and develop a search engine -- SynRT Explorer. You can visit our Search Engine page to use it.