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| | <p>When melatonin binds to the G protein-coupled receptor MT1, β-arrestins can be recruited. We are going to use this mechanism by designing two fusion-proteins: on the one hand the MT1 receptor will be fused to a fragment of an enzyme, on the other hand, the complementing enzyme fragment will be fused to the β-arrestin. Thus, receptor activation leads to β-arrestin recruitment and therefore to the formation of an active enzyme.</p> | | <p>When melatonin binds to the G protein-coupled receptor MT1, β-arrestins can be recruited. We are going to use this mechanism by designing two fusion-proteins: on the one hand the MT1 receptor will be fused to a fragment of an enzyme, on the other hand, the complementing enzyme fragment will be fused to the β-arrestin. Thus, receptor activation leads to β-arrestin recruitment and therefore to the formation of an active enzyme.</p> |
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| − | <h1> Welcome to iGEM 2018! </h1>
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| − | <p>Your team has been approved and you are ready to start the iGEM season! </p>
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| − | <h3>Before you start</h3>
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| − | <p> Please read the following pages:</p>
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| − | <li> <a href="https://2018.igem.org/Competition">Competition Hub</a> </li>
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| − | <li> <a href="https://2018.igem.org/Competition/Deliverables/Wiki">Wiki Requirements page</a></li>
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| − | <li> <a href="https://2018.igem.org/Resources/Template_Documentation">Template documentation</a></li>
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| − | <h3> Styling your wiki </h3>
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| − | <p>You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.</p>
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| − | <p>While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.</p>
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| − | <p> You must upload any pictures and files to the iGEM 2018 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. </p>
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| − | <p>When you upload, set the "Destination Filename" to <b> T--YourOfficialTeamName--NameOfFile.jpg</b>. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)</p>
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| − | <p>On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world! </p>
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| − | <p>Use WikiTools - Edit in the black menu bar to edit this page</p>
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| − | <p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p>
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| − | <li>State your accomplishments! Tell people what you have achieved from the start. </li>
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| − | <li>You have a global audience! Consider the different backgrounds that your users come from.</li>
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| − | <li>Make sure information is easy to find; nothing should be more than 3 clicks away. </li>
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| − | <li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="https://2018.igem.org/Calendar">iGEM 2018 calendar</a> </li>
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| − | <p> You can also view other team wikis for inspiration! Here are some examples:</p>
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| − | <li> <a href="https://2014.igem.org/Team:SDU-Denmark/"> 2014 SDU Denmark </a> </li>
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| − | <li> <a href="https://2014.igem.org/Team:Aalto-Helsinki">2014 Aalto-Helsinki</a> </li>
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| − | <li> <a href="https://2014.igem.org/Team:LMU-Munich">2014 LMU-Munich</a> </li>
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| − | <li> <a href="https://2014.igem.org/Team:Michigan"> 2014 Michigan</a></li>
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| − | <li> <a href="https://2014.igem.org/Team:ITESM-Guadalajara">2014 ITESM-Guadalajara </a></li>
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| − | <li> <a href="https://2014.igem.org/Team:SCU-China"> 2014 SCU-China </a></li>
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Who are we?
In the following months we will be developing a yeast-based biosensor for the sleeping hormone melatonin. Melatonin regulates the circadian rhythm and plays an important role in depression, Alzheimer’s and Parkinson’s disease. Therefore, we want to facilitate melatonin measurement by building a less expensive biosensor which can be used without special laboratory equipment.
Scientific approach:
Our approach is to genetically modify Saccharomyces cerevisiae by integrating a highly specific human melatonin receptor into the cells. This will allow us to detect melatonin from a saliva sample without previous purification. To get a signal from the melatonin binding, we will use two different signalling systems:
Using a nuclear receptor and a reporter gene:
The membrane permeability for melatonin is high. This permits us to use the nuclear retinoid z receptor (RZR) which is directly regulating gene expression. As the genes regulated by RZR do not exist in S. cerevisiae, we plan to create a chimeric receptor consisting of the RZR and the recognition sequence of the human estrogen receptor alpha (ERα). When melatonin is bound, the modified receptor can bind to the estrogen receptor responsive element (ERE) and as a consequence regulate expression of firefly luciferase reporter genes. The signal intensity of luciferase is dose-dependent in the presence of ligands.
Enzyme-Fragment Complementation Assay:
When melatonin binds to the G protein-coupled receptor MT1, β-arrestins can be recruited. We are going to use this mechanism by designing two fusion-proteins: on the one hand the MT1 receptor will be fused to a fragment of an enzyme, on the other hand, the complementing enzyme fragment will be fused to the β-arrestin. Thus, receptor activation leads to β-arrestin recruitment and therefore to the formation of an active enzyme.
