Team:Bologna

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= Project Summary =
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= Project Summary =
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'''Which is our idea?'''
'''Which is our idea?'''
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The project aims to implement a protein synthesis regulation system in ''Escherichia coli'' that acts at translational level regardless of the target gene to be downregulated. This "general-purpose" device could allow a faster control of protein expression. Our device was named '''T-Rex''' ('''T'''rans '''R'''epressor of '''Ex'''pression). It consists of two new BioBricks, i.e. the '''Trans-repressor''' and the '''Cis-repressing'''.
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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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'''How can we achieve our goal?'''
'''How can we achieve our goal?'''
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The TRANS-repressor is a non-coding DNA sequence that acts as a silencer of the CIS-repressing mRNA. In fact, the Cis-repressing sequence includes a TRANS-repressor complementary region ending with a ribosome binding site (RBS). Moreover, it is assembled upstream of the target gene coding sequence. When the TRANS-repressor and the CIS-repressing mRNAs bind together, the RBS recognition by the ribosome is prevented. Thus, the TRANS-repressor amount regulates the gene mRNA translation rate.
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T-Rex 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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The TRANS-repressor sequence was determined by a computational analysis performed to minimize the interference with the genomic mRNAs and to maximize the base-pairing interaction to the CIS-repressing RNA.  
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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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CIS-repressing and TRANS-repressor were designed using our [[Team:Bologna/Software#1|BASER]] tool.
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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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'''How can we test the device?'''
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We developed the following genetic circuit in order to test and characterize our T-Rex 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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The T-REX device is proposed as a universal and fast switch in synthetic gene circuits.  
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 in the [[Team:Bologna/Project|Project]] section.
More details about our work in the [[Team:Bologna/Project|Project]] section.
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= Acknowledgements =
= Acknowledgements =
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* ''' [http://www.unibo.it/Portale/default.htm University of Bologna] '''
* ''' [http://www.unibo.it/Portale/default.htm University of Bologna] '''

Revision as of 14:37, 21 October 2009

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HOME TEAM PROJECT SOFTWARE MODELING WET LAB PARTS HUMAN PRACTICE JUDGING CRITERIA




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Project Summary


Which is our idea?

The project aims to implement a protein synthesis regulation system in Escherichia coli that acts at translational level, regardless of the target gene. This "general-purpose" device allows a faster control of protein expression.
The device was named T-Rex (Trans Repressor of Expression).


How can we achieve our goal?

T-Rex device consists of two new BioBricks:

  • CIS-repressing, to be assembled upstream of the target protein coding sequence; it contains a ribosomal binding site; (RBS)

  • TRANS-repressor, to be placed under the control of a different promoter; it is complementary to the CIS-repressing, and contains a RBS cover in two versions of different length (4 and 7 nucleotides).

CIS-repressing and TRANS-repressor were designed using our [[Team:Bologna/Software#1|BASER]] tool.

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 RNA duplex prevents ribosome from binding to RBS, repressing protein synthesis. Thus, the TRANS-repressor amount regulates the gene mRNA translation rate.

Project3b.png



How can we test the device?


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

Circuit2.jpg



The T-REX device is proposed as a universal and fast switch in synthetic gene circuits.


More details about our work 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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