Team:Freiburg/Results

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Latest revision as of 21:56, 17 October 2014

The AcCELLerator

The AcCELLerator

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!

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