Team:Bologna

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'''Which is our idea?'''
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'''Our idea'''
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The project aims to implement a <b>protein synthesis regulation system</b> in <i>Escherichia coli</i> that acts at translational level, regardless of the target gene. This <b>"general-purpose"</b> device allows a faster control of protein expression.<br>The device was named <b>T-Rex</b> (<b>T</b>rans <b>R</b>epressor of <b>Ex</b>pression).
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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 can we achieve our goal?'''
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'''How T-REX works'''
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T-Rex device consists of two new BioBricks:
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The device consists of two new BioBricks:
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<li><font color="#000080"><b>CIS-repressing</b></font>, to be assembled upstream of the target protein coding sequence; it contains a ribosomal binding site; <font color="#228b22"><b>(RBS)</b></font>
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<li><font color="#000080"><b>TRANS-repressor</b></font>, to be placed under the control of a different promoter; it is complementary to the CIS-repressing, and contains a <font color="#228b22"><b>RBS cover</b></font> in two versions of different length (4 and 7 nucleotides).
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<li><font color="#000080"><b>CIS-repressing</b></font>, to be assembled upstream of the target coding sequence.
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<li><font color="#000080"><b>TRANS-repressor</b></font>, complementary to the CIS-repressing and placed under the control of a different promoter.
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CIS-repressing and TRANS-repressor were designed using our [[Team:Bologna/Software#1|BASER]] tool.
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CIS-repressing and TRANS-repressor sequences were designed by [[Team:Bologna/Software#1|BASER]] software.
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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.  
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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>)
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[[Image:project3b.png|center|950px|]]
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[[Image:project3b.png|center|950px|thumb|<center>Figure 1 - T-REX device</center>]]
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'''How can we test the device?'''
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'''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>
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In order to test and characterize our T-REX device, we developed the following genetic circuit (Fig 2):
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[[Image:circuit2.jpg|center|900px]]
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[[Image:circuit2OK.jpg|center|900px|thumb|<center>Figure 2 - Genetic Circuit to test CIS and TRANS' mRNA affinity</center>]]
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The T-REX device is proposed as a universal and fast switch in synthetic gene circuits.
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More details about our work are reported in the [[Team:Bologna/Project|Project]] section.
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More details about our work in the [[Team:Bologna/Project|Project]] section.
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= Acknowledgements =
= Acknowledgements =
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* '''Cultural Association San Sebastiano'''  
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* <font color=#0000cd>'''Cultural Association San Sebastiano'''</font>
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Latest revision as of 03:12, 22 October 2009

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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]


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


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  • Cultural Association San Sebastiano
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