Team:UCL/Project

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