Proof of Concept
In the figure above, we can see a model of what we would see if our bacteria and sensors are working effectively. The bacteria (in yellow) has invaded only the colon cancer cells (in red) and has not invaded the non cancerous fibroblast cells (in purple).
In order to prove that our product is working, we must utilize mammalian cell culture.
If our sensors are working, our bacteria will invade colon cancer cells and noncancerous cells (fibroblast control group). In order to test the bacteria, we must therefore co-culture bacteria with colon cancer and fibroblast cells and observe whether or not the organism is able to invade the right type of cell. This will allow us to create similar conditions to the human gut with and without inflammation as we clearly cannot test the efficacy of our product in an actual human gut. In order to differentiate between which cell is which, we utilize nucleofection and marked each type of cell with a different nuclear marker This will allow us to effectively, and efficiently determine which cell the bacteria actually did invade.
In order for the bacteria to actually invade the cell, we thought of using invasin—a protein that allows bacteria to enter cultured mammalian cells; however, we ran into issues with assembling the various components of invasin together. We hope to troubleshoot this process in the future and determine the specific errors involved in the manufacturing of the invasin part.
If product works, bacteria will only invade the colon cancer cells.
