The Berlin Charité Team
is working, like us, on toehold switches and using cell-free expression systems. We had several skype
calls where we exchanged our knowledge about toeholds and how they are designed, as well as cell-free
expression systems and how to lyophylise them.
EPFL iGEM provided Berlin with a protocol and advice on how to produce their own cell lysate and how to supply it with energy. Berlin Charité provided us with additional knowledge on how to freeze-dry our lysates and systems including the subsequent rehydration of the lyophylised components.
We also collaborated on implementing our toehold designer. Berlin Charité provided us with their understanding of the in-silico procedures of creating a toehold12, which was crucial for the software correctness.
This year's Oxford iGEM team is also working on a cell-free biosensor. We had the pleasure of meeting them in Cambridge at the OpenPlant forum where we started our collaboration. During a skype meeting we exchanged some feedback on our projects. Afterwards, we provided them with a protocol and advice on how to produce their own cell lysate and how to supply it with energy.
City of London School for Boys, UK
CLSB also worked on toehold switches like us. During a skype meeting and several mail exchanges, we talked about the design and specificity of these RNA switches. We helped them overcome their problem with homologous miRNA. Based on that, they designed a second series of toehold switches which are better suited for clinical use.
University College Cork, Ireland
University College Cork was also working on a biosensor. They wanted to test their sensor in a cell-free expression system but were short of time. As we had tested our biosensor in lysate very often, we had a well established protocol. We were unsure though whether it could be applied to a different biosensor, which is why UCC kindly reviewed our protocol, which we adapted to test fluorescence on a platereader.
1. Pardee, Keith, et al. "Rapid, low-cost detection of Zika virus using programmable biomolecular components." Cell 165.5 (2016): 1255-1266.
2. Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939.