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 to be downregulated. This "general-purpose" device could allow a faster control of protein expression. Our device was named T-Rex (Trans Repressor of Expression). It consists of two new BioBricks, i.e. the Trans-repressor and the Cis-repressing.

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.
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.

We developed the following genetic 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]

• Cultural Association San Sebastiano

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