Team:Paris Bettencourt

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Using synthetic biology to investigate newly discovered biological phenomena

The interest: A recent ground-breaking paper [1] described a new cell-to-cell bacterial communication system: nanotubes. Through excellent electronic microscopy images, antibiotic resistance transfer and faint fluorescence transfer , and cross-antibiotic resistance, previously unknown exchange channels were revealed between B.subtilis cells and even between completely different species. Results suggest that protein and/or RNA can travel through these tubes. This discovery may lead to a redefinition of individuality in bacteria. Given the many applications of known communication systems (e.g., quorum sensing, conjugation) in synthetic biology, harnessing the capacity of the nanotubes will open endless possibilities for new applications as amorph computing....

The challenge: Using synthetic biology to characterize the nanotubes, the structure, composition and control of which are still unknown. We decided to work on this challenging problem by providing other proofs to support this discovery.

See our work!

	The project: We bet-hedged our chances using different reporter systems by creating several devices relying on protein or RNA diffusion. Our devices rely on an emitter cell and a receiver cell that amplifies the signal. We chose to work with signaling molecules of different sizes and natures to test thoroughly the diffusion possibilities. Experiments were coupled with a thorough modeling approach tackling both the detection devices as well as diffusion, assisted transfer within the tubes and the membrane fusion within.
	The Team: We are fifteen students from parisian universities coming from many different disciplines who came together to participate in the iGEM competition. Come and meet the Team.
	The values: Ethics and safety are two main concerns when building genetically engineered organisms. You can visit our Human practice work and our safety page.

Achievements

We have:

• Used rational design in combination with modeling to create 6 new couple of emitter and receiver constructs (1) that could help characterize nanotubes' properties, using synthetic biology approach.
• Proofs of principle of 5 working emitter and receiver devices.(2) (3) (4)
• Sucessfully reproduced the GFP experiment of the Ben-Yehuda's team paper (5), indirectly proving the existence of the nanotubes under our microscopes.
• Developped two original models (6) (7) that could explain the observed transport parameters through the nanotubes.
• Builded a new original microfluidic chemostat chip, to monitor single bacterial layers under controled conditions.(8)
• Reproduced the original antibiotic experiment (9), design completary controls and proposed alternative explanations for the results observed.
• Created new original 38 parts (10) and devices for B. subtilis, and send them to the registry.
• Created and characterized an efficient amplifier with positive feedback loop of T7 RNA polymerase on T7 promoter.(11)
• Created devices aiming at hijacking two endogenic bistable switches in B. subtilis. (12)
• Characterized a fully functionning fluorescent concentrator device.(13)
• Collaborated with a nice iGEM team for the human practice(14), building a scientific and citizen proposal on synthetic biology.
• Most importantly, we have learned a lot during this iGEM competition.

References

1. Intercellular Nanotubes Mediate Bacterial Communication, Dubey and Ben-Yehuda, Cell, available here

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