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From 2013hs.igem.org

Abstract

The number of melanoma cases has been increasing in populations worldwide. Nearly every hour one American dies from melanoma. The technology currently available for melanoma forces a person to get a biopsy in order to diagnose it, another less invasive tool for diagnosis would be very efficient. Places where health care is not as readily available, a skin detecting cream for melanoma would allow people to ‘self-check’ for cancerous skin spots. The test would be easy to read and would let a person know that they need to visit their doctor immediately for further action. The purpose of our experiment is to eventually produce a topical cream or salve that can be used to detect the early onset of melanoma. In order to do so, the project will start with Saccharomyces cerevisiae [http://bioweb.uwlax.edu/bio203/s2007/nelson_andr/] and then insert a plasmid containing genes that glow in the presence of precancerous cells. The plasmid will consist of a biosensor for melanocytes specifically the mutated EGFR protein. After detection, our plasmid will code for the production of EPIC Firefly Luciferase and LRE, which will cause the yeast to glow and show the patient where the melanocytes are located. The cream that will be created will act as a diagnostic tool for the early detection of melanoma, allowing for earlier and faster treatment to occur. In order for the system to be effective, a protein receptor on the surface membrane of the precancerous cell must be identified. The most applicable protein receptor would be the mutated version of the Epidermal Growth Factor Protein Receptor. We will look for a protein on the surface of the yeast that will bind to the mutated EGFR. If this cannot be found we will find a suitable protein on the yeast surface and alter the DNA to create a match. The protein receptor on the yeast will send a chemical signal from the binding site to promote the production of arabinose which will cause the firefly glow. The EPIC Firefly Luciferase and LRE biobrick is the most compatible part to use because it has a high luminescent output; it is highly efficient and reliable. In order to use this specific part, there will need to be the production of arabinose within the cell. A metabolic pathway for arabinose production will have to be determined or arabinose will need to be included in the cream itself.