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Project Summary

Our idea

The aim of our project is the design of a standard device to control the synthesis of any protein of interest. This "general-purpose" device, implemented in E. coli, acts at the translational level to allow silencing of protein expression faster than using regulated promoters. 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, left panel). When the promoter controlling the TRANS coding sequence is active, it drives the transcription of an oligoribonucleotide complementary to the CIS mRNA sequence. The TRANS/CIS RNA duplex prevents ribosomes from binding to RBS on target mRNA, thus silencing protein synthesis. The amount of the TRANS-repressor regulates the rate of translation of the target mRNA (see Fig. 1, right 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 to test CIS and TRANS' mRNA affinity

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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-The project aims to design a new device to control the synthesis of a protein in Escherichia coli regardless of the protein to be controlled. This "general-purpose" standard device acts on translation to allow a faster silencing of protein expression if compared to standard regulated promoters. We named this device <b>T-Rex</b> (<b>T</b>rans <b>R</b>epressor of <b>Ex</b>pression).
+The aim of our project is the design of a standard device to control the synthesis of any protein of interest. This "general-purpose" device, implemented in <i>E. coli</i>, acts at the translational level to allow silencing of protein expression faster than using regulated promoters. We named this device <b>T-REX</b> (<b>T</b>rans <b>R</b>epressor of <b>Ex</b>pression).
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-'''How does T-Rex work?'''
+'''How T-REX works'''
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+<html>
 The device consists of two new BioBricks:
-<br><br>
+<br>
 <ul>
 <li><font color="#000080"><b>CIS-repressing</b></font>, to be assembled upstream of the target coding sequence.
 </ul>
-<br>
 <ul>
 <li><font color="#000080"><b>TRANS-repressor</b></font>, complementary to the CIS-repressing and placed under the control of a different promoter.
-</ul>
+</ul></html>
 <br>
-CIS-repressing and TRANS-repressor were designed using our [[Team:Bologna/Software#1|BASER]] tool.
+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. 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. </font>
+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, left panel</i>). When the promoter controlling the TRANS coding sequence is active, it drives the transcription of an oligoribonucleotide complementary to the CIS mRNA sequence. The TRANS/CIS <b>RNA duplex</b> prevents ribosomes from binding to RBS on target mRNA, thus <b>silencing protein synthesis</b>. The amount of the TRANS-repressor regulates the rate of translation of the target mRNA (<i>see Fig. 1, right panel</i>)
-</html>
-<br><br><br>
-[[Image:project3b.png|center|950px|]]
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+[[Image:project3b.png|center|950px|thumb|<center>Figure 1 - T-REX device</center>]]
+</font>
+<br><br>
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-'''How can we test the device?'''
+'''How we can test the device'''
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-In order to test and characterize our T-REX device, we developed the following genetic circuit:</font>
+In order to test and characterize our T-REX device, we developed the following genetic circuit (Fig 2):
-</html>
+</font></html>
 <br><br>
-[[Image:circuit2.jpg|center|900px]]
+[[Image:circuit2OK.jpg|center|900px|thumb|<center>Figure 2 - Genetic Circuit to test CIS and TRANS' mRNA affinity</center>]]
-<font face="Calibri" font size="4" color="#000000">
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-The T-REX device is proposed as a universal and fast switch in synthetic gene circuits.
+<br>
-<br><br><br>
+<font size="4">
+More details about our work are reported in the [[Team:Bologna/Project|Project]] section.
-More details about our work in the [[Team:Bologna/Project|Project]] section.
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-* '''Cultural Association San Sebastiano'''
+* <font color=#0000cd>'''Cultural Association San Sebastiano'''</font>
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