Project Summary

Our idea

The project aims to design a new device to control the synthesis of a protein in Escherichia coli regardless of the protein to be controlled. This "general-purpose" standard device acts on translation to allow a faster silencing of protein expression if compared to standard 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 seeked by BASER software.

Transcription of the target gene produces a mRNA strand, starting with the Cis element, which is translated into proteins by ribosome (see Fig. 1, right panel). Trans’ promoter induction produces a transcript that binds with the Cis part. The RNA duplex prevents ribosome from binding to RBS, repressing protein synthesis. Thus, the TRANS-repressor amount regulates the gene mRNA translation rate (see Fig. 1, left 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

More details about our work are reported in the Project section.

Acknowledgements

• University of Bologna

• Ser.In.Ar. Cesena

• Cultural Association San Sebastiano

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