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Project Summary

Which is our idea?

The project aims to implement a protein synthesis regulation system in Escherichia coli that is independent from regulated protein and acting at translational level, to make the control action faster. This "general-purpose" device was named T-Rex (Trans Repressor of Expression). It consists of two new BioBricks: Trans-repressor and Cis-repressing.

How can we achieve this?

The TRANS-repressor is a non-coding DNA sequence, the transcript of which acts as a silencer of the CIS-repressing RNA target. This target includes a region complementary to the sequence of the TRANS-repressor antisense, ends with a ribosome binding site (RBS), and is assembled upstream of the coding sequence of the gene to be silenced. Upon binding of TRANS-repressor and CIS-repressing RNAs, the access to RBS by ribosomes is hampered, silencing translation from the downstream transcript. Accordingly, the amount of TRANS-repressor controls the translation rate of the regulated gene.
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.

What can we use it for?

We developed a circuit in order to test and characterize our T-Rex device:

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]

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

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@@ Line 8: / Line 8: @@
 <div style="text-align:justify">
-= Project Description =
+= Project Summary =
 '''Which is our idea?'''
-Our project aims the realization of a device for gene expression regulation at translation level, regardless of the target gene.
+The project aims to implement a protein synthesis regulation system in Escherichia coli that is independent from regulated protein and acting at translational level, to make the control action faster. This "general-purpose" device was named '''T-Rex''' ('''T'''rans '''R'''epressor of '''Ex'''pression). It consists of two new BioBricks: '''Trans-repressor''' and '''Cis-repressing'''.
@@ Line 18: / Line 18: @@
 '''How can we achieve this?'''
-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.
+The TRANS-repressor is a non-coding DNA sequence, the transcript of which acts as a silencer of the CIS-repressing RNA target. This target includes a region complementary to the sequence of the TRANS-repressor antisense, ends with a ribosome binding site (RBS), and is assembled upstream of the coding sequence of the gene to be silenced. Upon binding of TRANS-repressor and CIS-repressing RNAs, the access to RBS by ribosomes is hampered, silencing translation from the downstream transcript. Accordingly, the amount of TRANS-repressor controls the translation rate of the regulated gene.
-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.
+<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.
@@ Line 28: / Line 29: @@
 '''What can we use it for?'''
-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.
+We developed a circuit in order to test and characterize our T-Rex device:
-Several are the possible future applications, but at the moment we are aiming to oscillators and flip-flop memories.
-More details about our work in the [[Team:Bologna/Project|Project]] section.
-== Project Abstract ==
-The project aims to realize a device with standard biological parts for the post-transcriptional control of gene expression, regardless of the gene sequence to be silenced.
 <br>
-To achieve this, we designed the T-REX device, composed of two standard parts: the TRANS-repressor and the CIS-repressing elements. The TRANS-repressor is a non-coding DNA sequence, the transcript of which acts as a silencer of the CIS-repressing RNA target. This target includes a region complementary to the sequence of the TRANS-repressor antisense, ends with a ribosome binding site (RBS), and is assembled upstream of the coding sequence of the gene to be silenced. Upon binding of TRANS-repressor and CIS-repressing RNAs, the access to RBS by ribosomes is hampered, silencing translation from the downstream transcript. Accordingly, The amount of TRANS-repressor controls the translation rate of the regulated gene.
+[[Image:circuit2.jpg|center|900px|]]
-<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.
 <br>
 The T-REX device is proposed as a universal and fast switch in synthetic gene circuits.
+More details about our work in the [[Team:Bologna/Project|Project]] section.