Team:Freiburg

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<title>The AcCELLerator</title>
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  background: -webkit-linear-gradient(0deg, #181617 0%, #181617 10%, rgba(24, 22, 23, 0) 30%), url(https://static.igem.org/mediawiki/2014/5/54/Freiburg2014-10-06_NIH3T3_with_mulv010_header_Thumbnail.jpg);
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          <div class="right" style="background-color: #D94B2B;">
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            <div class="description">
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              <h2>Welcome!</h2>
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              <p class="subtitle">We, the iGEM Team <a href="/Team:Freiburg/Team/Members">Freiburg 2014</a>, are pleased to welcome you on our homepage!</p></br>
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<p class="subtitle">Take the time to explore our project:</p>
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<p> <strong>The Ac<span>CELL</span>erator</strong> </p></br>
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<p> Scroll through the slider or click <a href="https://2014.igem.org/Team:Freiburg/Project" class="read-more">HERE </a>to get directly to our Projectpage.</p>
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              <h2>Our Project</h2>
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              <p class="subtitle">The AcCELLerator facilitates one central process in mammalian cell culture: Gene delivery.</p>
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<p class="subtitle">It provides you with the possibility to stably insert and express multiple genes with high spatio-temporal resolution. The two component system combines the advantages of optogentics and viral vector based gene delivery.</p>
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              <a href="https://2014.igem.org/Team:Freiburg/Project" class="read-more">Read More</a>
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              <h2>Results</h2>
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              <p class="subtitle">To validate our system we used different experimental methods including FACS-analysis, immunoblotting and confocal microscopy.</p>
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              <a href="https://2014.igem.org/Team:Freiburg/Results" class="read-more">Read More</a>
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              <h2>Policy and Safety</h2>
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              <p class="subtitle">The work with viruses and viral vectors provoked safety fears in many people we talked to about our project. Therefore we decided to closely link safety efforts with policy and practice and developed the innovative <a href="https://2014.igem.org/Team:Freiburg/PolicyAndPractices/Link-It-thestory">Link-It</a> approach.</p>
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              <a href="https://2014.igem.org/Team:Freiburg/PolicyAndPractices" class="read-more">Read More</a>
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            <div class="description">
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              <h2>Notebook</h2>
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<p class="subtitle">What we did - and how we did it:</p>
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<p class="subtitle">You can check it here.</p>
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              <a href="https://2014.igem.org/Team:Freiburg/Notebook" class="read-more">Read More</a>
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                <h2>Home</h2>
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                <h2>Our Project</h2>
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                <h2>Results</h2>
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                <h2>Policy and Safety</h2>
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                <h2>Notebook</h2>
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<td  colspan="3" align="center" height="150px" ><h1 >iGEM Freiburg 2014</h1>
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<p style="color:#000000" >
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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://igem.org/Team.cgi?year=2014&team_name=Freiburg"style="color:#000000"> Official Team Profile </a></td>
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<td style="border:1px solid black" align="center"  height ="45px" onMouseOver="this.bgColor='#d3d3d3'" onMouseOut="this.bgColor='#e7e7e7'" bgColor=#e7e7e7><a href="https://2014.igem.org/Team:Freiburg/Project"style="color:#000000"> Project</a></td>
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<td align ="center"><a href="https://2014.igem.org/Main_Page"> <img src="https://static.igem.org/mediawiki/igem.org/6/60/Igemlogo_300px.png" width="55px"></a></td>
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<td colspan="3" align="center"><h3>Abstract</h3></td>
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<p>
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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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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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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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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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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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Latest revision as of 20:58, 17 October 2014

The AcCELLerator
The AcCELLerator

Welcome!

We, the iGEM Team Freiburg 2014, are pleased to welcome you on our homepage!


Take the time to explore our project:

The AcCELLerator


Scroll through the slider or click HERE to get directly to our Projectpage.

Our Project

The AcCELLerator facilitates one central process in mammalian cell culture: Gene delivery.

It provides you with the possibility to stably insert and express multiple genes with high spatio-temporal resolution. The two component system combines the advantages of optogentics and viral vector based gene delivery.

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Results

To validate our system we used different experimental methods including FACS-analysis, immunoblotting and confocal microscopy.

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Policy and Safety

The work with viruses and viral vectors provoked safety fears in many people we talked to about our project. Therefore we decided to closely link safety efforts with policy and practice and developed the innovative Link-It approach.

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Notebook

What we did - and how we did it:

You can check it here.

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Home

Our Project

Results

Policy and Safety

Notebook
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