Team:UCL/Project

From 2013.igem.org

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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.
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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<p class="abstract_title">Chassis</p>
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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.
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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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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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<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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<a href="https://static.igem.org/mediawiki/2013/b/b4/ExperimentsUCLigem2013.gif" data-lightbox="image-1" title="Protocls UCL iGEM 2013">
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<p class="abstract_title">Lab Protocols</p>
<p class="abstract_title">Lab Protocols</p>
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Fill in later.
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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.

Circuit Overview

Our genetic circuit aims primarily to remove amyloid plaques in the brain, which are associated with Alzheimer’s disease, prevent neuroinflammation and support neurons.

Detection

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.

Insertion

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.

Degradation

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.

Selectable Marker

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. 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.

Chassis

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!

Parts

We submitted two parts to the registry; a new eukaryotic and prokaryotic selectable marker, and the protease, MMP-9.

Future Developments

Our circuit design is far from finished; given sufficient time in the lab, we would have included several other important components, including the de-activating agent VIP and the support factor BDNF.

Experiments

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. This section includes experiments for both bacterial and mammalian lab that have been performed before the Lyon jamboree!

Lab Protocols

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. This section includes protocols for both bacterial and mammalian lab experiments that have been performed before the Lyon jamboree!

Safety

It is essential that both personnel and experimental products are as safe as possible to avoid harm. Risk may be minimised by following safety procedures for any plausible dangerous situation that may occur in the laboratory.

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