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

• University of Bologna

• Ser.In.Ar. Cesena