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Project Summary

Our idea

The aim of our project is the design of a new device to control the synthesis of any protein of interest. This "general-purpose" standard device, implemented in E. coli, acts at the translational level to allow a switch in protein expression faster than transcriptional promoter regulation. We named this device T-Rex (Trans Repressor of Expression).

How T-Rex works

The device consists of two new BioBricks:

CIS-repressing, to be assembled upstream of the target coding sequence.

TRANS-repressor, complementary to the CIS-repressing and placed under the control of a different promoter.

CIS-repressing and TRANS-repressor sequences were designed by BASER software.

Transcription of the target gene yields a mRNA strand - containing the CIS-repressing sequence at its 5' end - available for translation into protein by ribosomes (see Fig. 1, right panel). Induction of the promoter controlling the TRANS coding sequence, releases a transcript complementary to the CIS mRNA sequence. The TRANS/CIS RNA duplex prevents ribosomes from binding to RBS on target mRNA, thus repressing protein synthesis. The amount of the TRANS-repressor regulates the rate of translation of the target mRNA (see Fig. 1, left panel)

Figure 1 - T-Rex device

How we can test the device

In order to test and characterize our T-REX device, we developed the following genetic circuit (Fig 2):

Figure 2 - Genetic Circuit

More details about our work are reported in the Project section.

Acknowledgements

[http://www.unibo.it/Portale/default.htm University of Bologna]

[http://serinar.criad.unibo.it Ser.In.Ar. Cesena]

Cultural Association San Sebastiano

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@@ Line 38: / Line 38: @@
 CIS-repressing and TRANS-repressor sequences were designed by [[Team:Bologna/Software#1|BASER]] software.
 <br><br>
-Transcription of the target gene produces a mRNA strand, starting with the Cis element, which is translated into proteins by ribosome (<i>see Fig. 1, right panel</i>). Trans’ promoter induction produces a transcript that binds with the Cis part. The <b>RNA duplex</b> prevents ribosome from binding to RBS, <b>repressing protein synthesis</b>. Thus, the TRANS-repressor amount regulates the gene mRNA translation rate (<i>see Fig. 1, left panel</i>)
+Transcription of the target gene yields a mRNA strand - containing the CIS-repressing sequence at its 5' end - available for translation into protein by ribosomes (<i>see Fig. 1, right panel</i>). Induction of the promoter controlling the TRANS coding sequence, releases a transcript complementary to the CIS mRNA sequence. The TRANS/CIS <b>RNA duplex</b> prevents ribosomes from binding to RBS on target mRNA, thus <b>repressing protein synthesis</b>. The amount of the TRANS-repressor regulates the rate of translation of the target mRNA (<i>see Fig. 1, left panel</i>)
 <br><br>
 [[Image:project3b.png|center|950px|thumb|<center>Figure 1 - T-Rex device</center>]]