Difference between revisions of "Team:UCC Ireland/Collaborations"

 
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<p>We collaborated with École Polytechnique Fédérale de Lausanne in Switzerland to test our Erythromycin biosensor to test its viability in a cell-free system. In-vitro protein expression is the production of recombinant proteins in cell lysate using biomolecular translation machinery. Two basic components of are needed to accomplish cell-free synthesis: (1) the genetic template, the pJKR-H-mphR plasmid (our erythromycin biosensor) and (2) reaction solution containing transcriptional and translational machinery and enzymes. After establishing that the presence of a dose-response relationship exists in the inducible gene control system in pJKR-H-mphR plasmid through in-vivo testing in DH5-alpha cells in the lab, we sought to test out system in a cell-free environment. Given that, in general, extracts for cell-free systems are engineered from systems that are known to support high level protein synthesis, they might support a system in which the level of readout protein, either sfGFP or AmilCP, production is  proportional to the concentration of inducer that the sensor is exposed to. In contrast, Dh5-Alpha cells may not be sufficiently protease deficient, resulting in progressive degradation of sfGFP or AmilCP over the course of an experiment, producing inconsistent results.</p>
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<h1 style='text-align: center;'>École Polytechnique Fédérale de Lausanne</h2>
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We collaborated with École Polytechnique Fédérale de Lausanne in Switzerland whose project focuses heavily on using cell-free systems. During the course of our collaboration, we discussed how cell-free systems could be beneficial to both our projects, enabling our systems to be used more widely and with more ease than if they were implemented in live bacterial chassis’. We also discussed cell-free protocols and reporter systems that could be used.
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Specifically, the EPFL iGEM team asked us to review their cell-free protocol and compare it to what we would use with our cell-free collaborator Dr. Thomas Meaney, who aims to bring cell-free technology to a wider market. This comparison of cell-free protocols will help them to optimise their cell-free extraction and build out their project.
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Through the course of our discussions, we learned that the EPFL team had tried to use chromoproteins in their cell free system with limited success. This has informed the future design of our biosensor strategy and has made us aware that we need to be careful in using amilCP as a reporter system and has made us aware that optimisation of the cell-free chassis may be required.
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Overall, our discussions and sharing of ideas were mutually beneficial in the future design and implementation of both our projects and we are very thankful to EPFL for their time.
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Latest revision as of 00:52, 2 November 2017

UCC iGEM 2017

École Polytechnique Fédérale de Lausanne

We collaborated with École Polytechnique Fédérale de Lausanne in Switzerland whose project focuses heavily on using cell-free systems. During the course of our collaboration, we discussed how cell-free systems could be beneficial to both our projects, enabling our systems to be used more widely and with more ease than if they were implemented in live bacterial chassis’. We also discussed cell-free protocols and reporter systems that could be used.

Specifically, the EPFL iGEM team asked us to review their cell-free protocol and compare it to what we would use with our cell-free collaborator Dr. Thomas Meaney, who aims to bring cell-free technology to a wider market. This comparison of cell-free protocols will help them to optimise their cell-free extraction and build out their project.

Through the course of our discussions, we learned that the EPFL team had tried to use chromoproteins in their cell free system with limited success. This has informed the future design of our biosensor strategy and has made us aware that we need to be careful in using amilCP as a reporter system and has made us aware that optimisation of the cell-free chassis may be required.

Overall, our discussions and sharing of ideas were mutually beneficial in the future design and implementation of both our projects and we are very thankful to EPFL for their time.