Team:Jilin China/Result/Version 2
VERSION 2.0
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Results
After the integrated human practice work, we update SynRT Toolkit to version 2.0. In version 2.0, we added 23 heat-repressible RNA-based thermosensors to Toolkit.
Heat-repressible RNA-based thermosensors were based on the RNase E. We measured these thermosensors' activity by using measurement device like before. But this time, we designed a new negative control. In the following, it will be called negative control-2. Negative control-2 has a cleavage site of RNase E. It will always be digest by enzyme. Due to the effiency of RNase E, we decided to use negative control-2 instead of traditional negative control.
We designed 100 heat-repressible RNA-based thermosensors, their sequences are different. After measurement, we finally selected 23 out of 100 heat-repressible RNA-based thermosensor in toolkit.
We measured the activities of these thermosensors at 3 temperatures: 29, 37 and 42℃. Figure 1 shows the measurement result of the 23 different thermosensors. Compared with the positive control, all the heat-repressible RNA-based thermosensors' normalized fluorescence decrease at elevated temperature. They also have different intensity and sensitivity. We have added the charaterize result to the parts registry, users can choose their appropriate thermosensor.
We also computed the fold-change from 29 to 37℃ and 37 to 42℃. As the figure shows. These fold-changes were lower than positive control and the fluorescence per Abs600 were higher than negative control-2.
