Team:Edinburgh/Bacterial/Core repressilator

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The overall oscillating design of our system is composed of two parallel networks working in the same direction as an associated repressilator, with the addition of red, blue, and green light as a signaling mechanism. The inner network works in the same manner as the original repressilator designed by Garcia-Ojalvo and M. B. Elowitz in 2004, containing three genes in an ordered circular fashion (λcI, lacI, tetR). The outer network, on the other hand, consists of the three light sensors and their associated light producers.

The integrated network can thus be considered in terms of its three separate components:

• Component 1: red luciferase, lacI, red sensor.
• Component 2: blue luciferase, tetR, blue sensor.
• Component 3: green luciferase, λcI, green sensor.

Within each component, the light (luciferase) and the repressor are both active or inactive at the same time. For example, in Component 1, high levels of λcI repress the activity of both the red luciferase and lacI; as soon as the levels of λcI begin to fall, the concentration of red luciferase and lacI in the system increases.

The expression of the light luciferase, which produces light of the associated colour, inhibits the following promoter through the light sensor; the promoter, on the other hand, represses both the following light luciferase and the following promoter.

As for the light sensors, these work in two different ways. The red light sensor absorbs red photons and represses the phosphorylation of OmpR, thus inhibiting the fusion between the OmpR-dependent ompC promoter and tetR. The blue and green light sensors work in a similar manner, by modifying the α-helical domain linker of LovTAP as a conduit for an allosteric signal.

The significance of our design is in the addition of the outer network (the light repressilator), thus introducing the light signal into the cell community and realising multicellular communication.