Team:UCL/Project

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Genetic Engineering

Ethical Questions

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.

Click the abstracts below to read more.

Circuit Overview

Our genetic circuit aims primarily to remove amyloid plaques in the brain, which are strongly associated with Alzheimer’s disease. Our circuit focuses on three core parts; firstly, a promoter which responds to oxidative stress - this can be used as a proxy to detect plaques. Secondly, a plaque-degrading protease, which will be secreted in response to oxidative stress, and thirdly, a chemoattractant, also secreted in response to oxidative stress, which will draw more microglia towards the plaque. We have also developed our own selectable marker, which we can use to select for transformations of mammalian cells.

Detection

Senile plaques increase the rate of production of reactive oxygen species which is damaging to brain cells. We capitalise on this, by creating a promoter sensitive to oxidative stress, in order to initiate the production of other circuit parts.

Chemotaxis

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.

Degradation

We expressed and characterised MMP-9 in several chassis. MMP-9 is 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.

Selectable Marker

We used resistance to zeocin, a cell killing glycoprotein, to act as a selectable marker transformation/transfection in all our chassis, E.coli, HeLa and microglia.

Chassis

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.

Parts

We submitted X parts to the registry, LIST GENE NAMES and improved X, LIST GENE NAMES. To be filled in later

Experiments

Fill in later.

Lab Protocols

Fill in later.

Safety

For experimental procedures, it is essential that both the personnel and the experimental products are as safe as possible to avoid harm. Risk may be minimised by following safety procedures for any likely situations that may occur in the laboratory environment.

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