Team:Freiburg

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<td  colspan="3" align="center" height="150px" ><h1 >iGEM Freiburg 2014</h1>
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The wiki of the iGEM Team Freiburg is currently in development.
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<td  align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7><a href="https://2014.igem.org/Team:Freiburg"style="color:#000000">Home </a></td>
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<td  align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7><a href="https://2014.igem.org/Team:Freiburg/Team"style="color:#000000"> Team </a></td>
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<td  align="center"  height ="45px"  onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7><a href="https://igem.org/Team.cgi?year=2014&team_name=Freiburg"style="color:#000000"> Official Team Profile </a></td>
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<td  align="center" height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7><a href="https://2014.igem.org/Team:Freiburg/Modeling"style="color:#000000"> Modeling</a></td>
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<td  align="center"  height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7><a href="https://2014.igem.org/Team:Freiburg/Safety"style=" color:#000000"> Safety </a></td>
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<td  align="center"  height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7><a href="https://2014.igem.org/Team:Freiburg/Attributions"style="color:#000000"> Attributions </a></td>
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<td valign="top" style="padding-left:20px; text-align: left;"><h2 id="title">Abstract:</h1>
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iGEM Freiburg developed an optogenetical agent gadget which allows to safely transmit secret messages.
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<td colspan="3" align="center"><h3>Abstract</h3></td>
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Optogenetics, a novel technology that allows temporal and spatial induction of gene expression by the
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use of light, is of growing importance for fundamental research and clinical applications. However, its
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biggest limitation is the time consuming introduction of transgenes into organisms or cell lines. In
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contrast, easy but unspecific gene delivery can be achieved by viral vectors. We, the iGEM Team
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Freiburg 2014, combine the advantages of both approaches – the temporal and spatial resolution of
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optogenetics, and the simplicity of gene transfer offered by viruses. To this end we designed a system
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where the entry of a virus is enabled or prevented by exposing the target cells to light of distinct
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wavelengths.
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<br />
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<br />
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The ecotropic murine leukemia virus (MuLV) is a retroviral vector which enters a cell by binding to the
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cationic amino acid transporter (CAT-1). CAT-1 is present in cells of all mammals, but displays a high
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variability between different species in the third extracellular loop, which is the recognition sequence
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for the virus. Therefore, even close relatives of mice, e.g. rats or hamsters, are immune to the MuLV,
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but upon exogenous expression of murine CAT-1, cell lines from these species can also be infected.
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Hence we use the popular Chinese hamster ovary cell line CHO in our experiments.
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<br />
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For optogenetic targeting of specific subsets of cells, we employ three different systems: a red/far-red
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light system and a UVB light system based on proteins from the plant model organism Arabidopsis
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thaliana as well as a light-oxygen-voltage system from the marine bacterium Erythrobacter litoralis. In
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all three light systems gene expression is induced by recruitment of engineered transcription factors to
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DNA.
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<br />
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Without illumination the cells are in a dormant state and cannot be infected by the viral vector. Upon
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exposure to the appropriate wavelength, they start expressing the viral entry receptor CAT-1. Addition
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of the viral vector to the culture medium leads to infection of the activated subset of cells.
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In order to demonstrate the functionality of the specific gene delivery we developed a device for
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secure communication. The device is working in a two-step process: First, the sender has to specify
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the subset of cells by illumination through a patterned mask. In the second step, upon receiving the
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device the reader has to visualize the message by adding the appropriate viral vector containing a
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reporter gene. The unintended opening of the device by an unaware third person results in a complete
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illumination of the cells leading to the destruction of the message. The time gap between sending and
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receiving the device is limited by the half-life of the receptor, thus creating an additional safety level.
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To expand into the field of medicine, we furthermore chose CRISPR/Cas as a gene cargo.
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<br />
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<br />
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CRISPR/Cas has attracted much attention in recent past, as it facilitates specific gene editing, e.g.
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knock-outs/ins, with high specifity in a minimum of time. In combination with delivery by viral vectors
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this could provide a powerful tool for the treatment of diseases and genetic disorders in vivo.</h3>
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<td colspan="3" align="center"><h3>Sponsoren</h3></td>
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Revision as of 20:05, 29 July 2014

iGEM Freiburg 2014

Abstract:

iGEM Freiburg developed an optogenetical agent gadget which allows to safely transmit secret messages.