Team:Bologna

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= Project Summary =
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!align="center"|[[Team:Bologna|<font color="#ffffff">HOME</font>]]
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'''Our idea'''
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!align="center"|[[Team:Bologna/Team|<font color="#ffffff">TEAM</font>]]
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!align="center"|[[Team:Bologna/Project|<font color="#ffffff">PROJECT</font>]]
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!align="center"|[[Team:Bologna/Modeling|<font color="#ffffff">MODELING</font>]]
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!align="center"|[[Team:Bologna/Lab-Notebook|<font color="#ffffff">WETLAB</font>]]
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!align="center"|[[Team:Bologna/Software|<font color="#ffffff">DRY LAB</font>]]
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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 T-REX works'''
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[[Image:TeamBologna.jpg|frame|center|Our Team]]
 
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= Project Description =
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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 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 sequences were designed by [[Team:Bologna/Software#1|BASER]] software.
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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|thumb|<center>Figure 1 - T-REX device</center>]]
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'''Which is our idea?'''
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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 (Fig 2):
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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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More details about our work are reported in the [[Team:Bologna/Project|Project]] section.
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Our project aims the realization of a device for gene expression regulation at translation level, regardless of the target gene.
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= Acknowledgements =
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* ''' [http://www.unibo.it/Portale/default.htm University of Bologna] '''
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[[Image:LogoUnibo.jpg|left|100 px]]
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'''How can we achieve this?'''
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<br><br><br><br><br><br><br>
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We designed a 50bp non coding sequence (CIS - repressing) to be assembled upstream of the target gene RBS, while its complementary sequence (TRANS - repressor) is to be placed under the control of another promoter. The TRANS - repressing contains also a cover, that is a short sequence  complementary to the RBS.
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* ''' [http://serinar.criad.unibo.it Ser.In.Ar. Cesena] '''
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When the TRANS-repressor element is present, it binds to the CIS-repressing, forming a RNA duplex and producing an obstruction that prevents the ribosome binding to the RBS.
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[[Image:Ser_In_Ar.jpg|left|500px]]
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* <font color=#0000cd>'''Cultural Association San Sebastiano'''</font>
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'''What can we use it for?'''
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[[Image:SSebastiano.jpg|left|200px]]
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After doing some tests to prove that the presence of CIS – repressing doesn't alter the cell normal activities, we are trying to realize some simple circuits in order to characterize our device.
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Several are the possible future applications, but at the moment we are aiming to oscillators and flip-flop memories.
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----
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You can find more details about our work at the section.
 
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Latest revision as of 03:12, 22 October 2009

ProvaBol2.png
HOME TEAM PROJECT SOFTWARE MODELING WET LAB PARTS HUMAN PRACTICE JUDGING CRITERIA




Ely9Copia.jpg


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]


LogoUnibo.jpg








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


Ser In Ar.jpg






  • Cultural Association San Sebastiano
SSebastiano.jpg










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