Team:Bologna

From 2009.igem.org

(Difference between revisions)
 
(50 intermediate revisions not shown)
Line 1: Line 1:
{{Template:BolognaTemplate}}
{{Template:BolognaTemplate}}
-
 
+
<br><br><br>
-
 
+
-
 
+
[[Image:Ely9Copia.jpg|center|850px]]
[[Image:Ely9Copia.jpg|center|850px]]
-
 
+
<br>
-
 
+
-
 
+
<div style="text-align:justify">
<div style="text-align:justify">
 +
= Project Summary =
= Project Summary =
-
'''Which is our idea?'''
+
<br>
 +
<font face="Calibri" size="5">
 +
'''Our idea'''
 +
</font>
 +
<br><br>
-
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'''.
+
<html>
-
 
+
<font face="Calibri" font size="4" color="#000000">
-
 
+
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).  
-
 
+
</font></html>
-
'''How can we achieve our goal?'''
+
-
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.
 
<br>
<br>
-
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.
+
<font face="Calibri" size="5">
-
 
+
'''How T-REX works'''
-
 
+
</font>
-
 
+
<br><br>
-
[[Image:project3b.png|center|940px|]]
+
-
We developed a circuit in order to test and characterize our T-Rex device:
+
<font face="Calibri" font size="4" color="#000000">
 +
<html>
 +
The device consists of two new BioBricks:
<br>
<br>
-
[[Image:circuit2.jpg|center|900px|]]
+
<ul>
 +
<li><font color="#000080"><b>CIS-repressing</b></font>, to be assembled upstream of the target coding sequence.
 +
</ul>
 +
<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></html>
<br>
<br>
-
The T-REX device is proposed as a universal and fast switch in synthetic gene circuits.  
+
CIS-repressing and TRANS-repressor sequences were designed by [[Team:Bologna/Software#1|BASER]] software.
<br><br>
<br><br>
-
More details about our work in the [[Team:Bologna/Project|Project]] section.
+
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>)
 +
<br><br>
 +
[[Image:project3b.png|center|950px|thumb|<center>Figure 1 - T-REX device</center>]]
 +
</font>
 +
<br><br>
 +
<font face="Calibri" size="5">
 +
'''How we can test the device'''
 +
</font>
 +
<br><br><br>
 +
<html>
 +
<font face="Calibri" font size="4" color="#000000">
 +
In order to test and characterize our T-REX device, we developed the following genetic circuit (Fig 2):
 +
</font></html>
 +
<br><br>
 +
[[Image:circuit2OK.jpg|center|900px|thumb|<center>Figure 2 - Genetic Circuit to test CIS and TRANS' mRNA affinity</center>]]
 +
<br><br>
 +
<br>
 +
<font size="4">
 +
More details about our work are reported in the [[Team:Bologna/Project|Project]] section.
 +
</font>
 +
<br>
 +
<br>
 +
<br>
= Acknowledgements =
= Acknowledgements =
-
 
+
<br>
 +
<font size="3">
* ''' [http://www.unibo.it/Portale/default.htm University of Bologna] '''
* ''' [http://www.unibo.it/Portale/default.htm University of Bologna] '''
Line 52: Line 80:
[[Image:Ser_In_Ar.jpg|left|500px]]
[[Image:Ser_In_Ar.jpg|left|500px]]
-
<br><br><br><br><br><br>
+
<br><br><br><br><br>
 +
 
 +
* <font color=#0000cd>'''Cultural Association San Sebastiano'''</font>
 +
 
 +
[[Image:SSebastiano.jpg|left|200px]]
 +
 
 +
<br><br><br><br><br><br><br><br>
----
----
Line 66: Line 100:
[https://2009.igem.org/Team:Bologna ''Up'']
[https://2009.igem.org/Team:Bologna ''Up'']
 +
</font>

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










Locations of visitors to this page


Up

Retrieved from "http://2009.igem.org/Team:Bologna"