Team:Montreal

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Project Overview: Elucidating an Experimentally Viable Repressilator

Although typically used to describe physical phenomena, oscillations are also observed in a range of biological processes such as the circadian rhythm, neuronal communication and nephron function. Elowitz and Liebler (2000) postulated that a relaxation oscillator system called the Repressilator, composed of three genes tied in negative feedback loops, would generate oscillations in protein expression within a cell. Using one of these genes linked with a Yellow Fluorescent Protein (YFP) gene to create a reporter plasmid, we expect to see the cells blinking over time as the concentration of the repressing proteins fluctuate.

The two-gene intercellular system (Macmillen 2002) examined in past iGEM projects developed irregular, triangular-shaped oscillation curves over extended periods of time. In an effort to improve the consistency and regularity of the oscillations, the Repressilator will be used. The system in theory should have better long-term fidelity in oscillation patterns and develop a more natural sinusoidal oscillatory pattern. Despite the foreseeable advantages of Elowitz's theoretical system, the repressilator has yet to be reproduced empirically; in addition, theoretical models indicate that, in general, these synthetic oscillatory systems in bacteria degrade and become irregular, unlike the natural physiological oscillations of larger lifeforms.

Building on previous years of research, the McGill University iGEM team intends to construct a set of Biobricks that will mimic the repressilator behavior and maintain synchronous oscillations in a large population of cells. If accomplished, our theorists and experimentalists will cooperate to refine this system using various modifications to further our understanding of biological clocks and their functioning. •