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Team:Freiburg/Results
From 2014.igem.org
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Latest revision as of 21:56, 17 October 2014
Summary
We, the iGEM Team Freiburg 2014, have the goal to combine the capability of viral vectors for stable gene transfer with the spatio-temporal resolution of optogenetics for gene delivery into mammalian cells. We utilized the Murine Leukemia Virus (MuLV) that specifically uses the murine mCAT-1 receptor to infect target cells. By using a blue light inducible expression system, we brought the receptor to the plasma membrane of illuminated cells to enable targeted gene delivery by the virus.
Our project was quite a large success! We:
- produced MuLV viruses containing different reporter proteins,
- optimized MuLV production and transduction protocols to reach close to 100% efficiency,
- created stable mammalian cell lines using the MuLV virus,
- generated patterns of reporter proteins in cell cultures by light exposure,
- demonstrated that the virus exclusively infects cells expressing the mCAT-1 receptor,
- infected cells with the MuLV virus that expressed the mCAT-1 receptor after light induction.
We provide the virus as a tool for the generation of stable mammalian cell lines under biosafety level 1 regulations. Our viral vector, which we propose as a new iGEM RFC, enables the user to introduce any gene of interest stably into mammalian cell lines. Therefore we provide for the iGEM community a fast, easy to handle and safe way of generating stable cell lines.
During the course of iGEM, we learned many new techniques:
- mammalian cell culture,
- fluorescence activated cell sorting,
- virus production under biosafety level 1 and biosafety level 2 conditions,
- many different cloning techniques,
- widefield and confocal microscopy,
- creating a website,
and, last but not least, we had a lot of fun and learned how to work together as a team!
