Team:UCL/Project

From 2013.igem.org

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<p class="major_title">Genetic Engineering</p>
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<p class="major_title">IGEM: INTELLIGENTLY GENETICALLY ENGINEERED MICROGLIA</p>
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<p class="minor_title">Ethical Questions</p>
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<p class="minor_title">Synthetic Biology Fights Alzheimer's Disease</p>
<p class="abstract_text" style="color:#404040;">
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This year, the UCL iGEM team is taking a radical new step with synthetic biology. We intend to explore the potential application genetic engineering techniques on the brain, because it is the site of some of the most subtle, and many of the most devastating diseases known to medicine. We have devised a genetic circuit for transfecting into a novel chassis for iGEM that is rarely engineered in research - microglial cells, the resident immune cells of the brain. The circuit aims to boost the ability of the microglial cells to break down senile plaques, which are associated with the onset and progression of Alzheimer’s Disease, as well as to protect neurons under threat from these plaques and from inflammation. Alzheimer’s Disease is a neurodegenerative disease that is characterised by the loss of recent memory and intellectual functions. Late stages of the disease often see patients bedridden, mute and incontinent. It is a horrific condition for which a genetic engineering response is both pertinent and somewhat disconcerting. Therefore, we also delve into the neuroethics of the potential progression of synthetic biology in neuroscience.
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This year, the UCL iGEM team is taking a radical new step with synthetic biology. We intend to explore the potential application genetic engineering techniques on the brain, because it is the site of some of the most subtle, and many of the most devastating medical conditions. Alzheimer’s Disease is a neurodegenerative disease characterised by the loss of recent memory and intellectual functions. We have devised a genetic circuit for transfection into microglia, a novel chassis in which standard assembly has never been used, to boost their ability to break down senile plaques, which are associated with Alzheimer’s disease, as well as to support and protect endangered neurons from microglia-mediated neuroinflammation.
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<a href="http://2013.igem.org/wiki/images/1/1a/CircuitOverviewMicroglia.gif" data-lightbox="image-1" title="Genetic Circuit Overview in Microglia UCL iGEM 2013">
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<a href="http://2013.igem.org/Team:UCL/Project/Circuit">
<p class="abstract_title">Circuit Overview</p>
<p class="abstract_title">Circuit Overview</p>
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Our genetic circuit aims primarily to remove amyloid plaques in the brain, which are associated with Alzheimer’s disease. To target amyloid protease expression to plaque sites, we developed a eukaryotic oxidative stress promoter.  
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Our genetic circuit aims primarily to remove amyloid plaques in the brain, which are associated with Alzheimer’s disease, prevent neuroinflammation and support neurons.
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<a href="http://2013.igem.org/wiki/images/1/18/Detectionucligem.gif" data-lightbox="image-1" title="Oxidative Stress Promoter for Plaque Detection UCL iGEM 2013">
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<a href="http://2013.igem.org/Team:UCL/Practice/Creative">
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<a href="http://2013.igem.org/Team:UCL/Project/Detection">
<p class="abstract_title">Detection</p>
<p class="abstract_title">Detection</p>
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Senile plaques increase the rate of production of reactive oxygen species that are damaging to brain cells. We capitalise on this, by creating an oxidative stress promoter, in order to initiate the production of other circuit parts.
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Senile plaques increase the rate of production of reactive oxygen species that are damaging to brain cells. We developed an oxidative stress promoter, in order to initiate the production of other circuit parts.
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<a href="http://2013.igem.org/wiki/images/5/57/Chemotaxisucligem.gif" data-lightbox="image-1" title="Chemotaxis in Microglia UCL iGEM 2013">
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<a href="http://2013.igem.org/Team:UCL/Practice/Neuroethics">
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<a href="http://2013.igem.org/Team:UCL/Project/Chemotaxis">
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<p class="abstract_title">Chemotaxis</p>
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<p class="abstract_title">Insertion</p>
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Microglia readily migrate towards plaques in vivo, but to see if we could increase migration to plaques, we produced a chemoattractant to help them converge.
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The brain is an immune privileged organ and the security of the blood brain barrier makes it difficult to get all but the smallest molecules, such as glucose, from the rest of the body into the brain. This makes inserting our genetic circuit into the brain a trickier task than in most synthetic biomedical projects. Here we examine some plausible methods.
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<a href="http://2013.igem.org/wiki/images/4/46/Degradationucligem.gif" data-lightbox="image-1" title="Beta-Amyloid degradation by MMP-9 UCL iGEM 2013">
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<a href="http://2013.igem.org/Team:UCL/Practice/Documentary">
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<a href="http://2013.igem.org/Team:UCL/Project/Degradation">
<p class="abstract_title">Degradation</p>
<p class="abstract_title">Degradation</p>
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We expressed and characterised MMP-9, a matrix metalloproteinase that is capable of degrading amyloid. By increasing its expression in de-activated microglia, we hope to be able to reduce amyloid burden in Alzheimer’s disease.
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We expressed and characterised a matrix metalloproteinase that is capable of degrading amyloid. By increasing its expression in de-activated microglia, we hope to reduce amyloid burden in Alzheimer’s disease.
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<a href="http://2013.igem.org/wiki/images/8/8a/Selectablemarkerzeocin.gif" data-lightbox="image-1" title="Selectable Marker, Zeocin UCL iGEM 2013">
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<a href="http://2013.igem.org/Team:UCL/Project/Marker">
<p class="abstract_title">Selectable Marker</p>
<p class="abstract_title">Selectable Marker</p>
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We used resistance to zeocin, a cell killing glycoprotein, to act as a selectable marker for transformation/transfection in all our chassis, E.coli, HeLa and microglia.
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We used resistance to zeocin, a cell killing glycoprotein, to act as a selectable marker for transformation/transfection in our chassis; E.coli, HeLa and microglia.
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The biobrick that we made encoding zeocin resistance is a step forward for selectable markers in iGEM - the first one of its kind tailored for mammalian expression systems.  
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<a href="http://2013.igem.org/wiki/images/e/e1/ChassisUCLigem2013.gif" data-lightbox="image-1" title="Chassis: E.coli, HeLa and Microglia UCL iGEM 2013">
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<a href="http://2013.igem.org/Team:UCL/Practice/Report">
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<a href="http://2013.igem.org/Team:UCL/Project/Chassis">
<p class="abstract_title">Chassis</p>
<p class="abstract_title">Chassis</p>
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We conducted our experiments in three chassis, creating recombinant plasmids in E.coli and expressing them in HeLa, and finally, the much harder to transfect primary and immortalised human microglia lines.
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We conducted our experiments in three chassis, creating recombinant plasmids in E.coli before expressing them in HeLa and finally primary and immortalised human microglia lines. Human brain cells have not been seen before in an iGEM project, with experiments in microglia coming as son as they arrive!
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<a href="http://2013.igem.org/wiki/images/d/da/PartsUCligem2013.gif" data-lightbox="image-1" title="BioBrick Parts Submitted to the Registry UCL iGEM 2013">
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<a href="http://2013.igem.org/Team:UCL/Project/Parts">
<p class="abstract_title">Parts</p>
<p class="abstract_title">Parts</p>
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We submitted X parts to the registry, LIST GENE NAMES and improved X, LIST GENE NAMES. To be filled in later
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We submitted two parts to the registry; a new eukaryotic and prokaryotic selectable marker, and the protease, MMP-9.
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<a href="http://2013.igem.org/Team:UCL/Project/Developments">
<p class="abstract_title">Future Developments</p>
<p class="abstract_title">Future Developments</p>
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<p class="abstract_title">Experiments</p>
<p class="abstract_title">Experiments</p>
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Here we explain the wet-lab experiments that built our project. All of these experiments built the foundation of our project, allowing us to generate, test and subsequently submit biobricks to the registry.
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This section includes experiments for both bacterial and mammalian lab that have been performed before the Lyon jamboree!
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You cannot plan an experiment without the procedure. Here we have stored all of the protocols that were used in order to make out wet lab work possible.
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This section includes protocols for both bacterial and mammalian lab experiments that have been performed before the Lyon jamboree!
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Latest revision as of 03:22, 5 October 2013

IGEM: INTELLIGENTLY GENETICALLY ENGINEERED MICROGLIA

Synthetic Biology Fights Alzheimer's Disease

This year, the UCL iGEM team is taking a radical new step with synthetic biology. We intend to explore the potential application genetic engineering techniques on the brain, because it is the site of some of the most subtle, and many of the most devastating medical conditions. Alzheimer’s Disease is a neurodegenerative disease characterised by the loss of recent memory and intellectual functions. We have devised a genetic circuit for transfection into microglia, a novel chassis in which standard assembly has never been used, to boost their ability to break down senile plaques, which are associated with Alzheimer’s disease, as well as to support and protect endangered neurons from microglia-mediated neuroinflammation.

Click the abstracts below to read more.