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Applications

The bistable is a very interesting tool. Its structure allows using any combination of either markers or functional genes which can have many applications in basic and applied sicence.

Here we show some ideas for possible uses for our basic and improved flip-flop system:

Application 1 – Big scale production of killer proteins

An applied use of the flip-flop could be the production of proteins that are highly toxic or even lethal for those bacteria that produce them (called “killer proteins”). Using the bistable we could get bacteria that synthesize those proteins only after a certain signal given in an optimal physiologycal moment or growth state that we want. This way we could be able to have bacteria that grow until reaching a certain density, and then inducing the change in the flip-flop state making our bacteria to synthesize fast an efficiently our favorite toxic protein. This has obvious advantages for antibody production, purification of useful molecules in clinic or any other industry etc.

Application 2 – Basic Research

A very extended way to regulate protein function among all life beings is postranslational modifications that set a protein or a complex in two different states. One of the best known of these modifications is phosphorylation. A protein can have several sites where can be phosphorylated and depending on where the phosphates are, the protein can be active, inactive, or can do different tasks. To study this phenomenon, scientist use either phosphomimetics, which are aminoacids (specifically glutamic acid and aspartic acid) that imitate the shape of a canonical phosphorylation but do not function as such or non-phophorylatable residues. That way it is possible to study the two states but only in separate cells. Thus, cloning the two mutant versions of a desired phohoprotein (constitutive phosphorylation and non-phosphorylatable) it would allow to study how this protein works with any phosphorylation pattern and to have a reversible system inside the same cell.

Application 3 – Gene therapy

It could be used as an expression system for genetic therapy. A usual vector would need the addition of the inducer for the whole life of the patient, which may cause health problems. This new approach allows that only one dose promotes the expression of the desired gene for a long time, leading to a reduction in the amount of inducer injected.

Application 4 – Biosensors

Also biosensors seem like a plausible application, changing the system so it responds to different inducers. Toxics substances or pollutants may be good targets for the development of new regulatory pathways.

Application 5 – Biological memories

In a long-term future, if synthetic biology continues its actual development, it is possible to imaging biologial memories that work like a flash memory (that is why our project was called Flash-bacter). To achieve that goal, the first step is to obtain very efficient flip-flops. After this, we have to think in a way to distinguish among different bacteria, that is, to be able to assign addresses to singular bits (ideally individual bacterium, or a given set of bacteria) and a way to write and erase the information only for a given address (bacteria). With the current development state of synthetic biology this still pertains to the theoretical realm, but we have the hope that some day the idea could become real. We want to make engineering with genes to create complex system and we are sure that is possible because nature does it.