We contacted Prof. Dr. Sven Panke, Head of Biosystems Science and Engineering Department at ETH, to evaluate our project and its alignment with iGEM format. At this time, we were finalizing our idea and Prof. Panke could give us valuable insights for our design. Finally, we wanted to get feedback regarding feasibility of our aim and thus validate our approach.
Since our project does not involve genetically engineering bacteria or other organisms, we initially questioned whether it fit well within the iGEM framework. Prof. Panke reassured us that our idea is perfectly suitable for the competition, emphasizing that our approach exemplifies the use of synthetic biology at its best in diagnostics.
Prof. Panke recommended starting with a single pathogen. Demonstrating a complete system for one pathogen would make it easier to argue that multiplexing could be achieved in the future. This advice was particularly important given the project’s time constraints.
Our discussion also highlighted potential challenges for a paper-based system. Lysis can be difficult, so pathogen selection is crucial. For example, Gram-negative bacteria like N. gonorrhoeae are very hard to lyse, whereas Gram-positive bacteria, viruses, or parasitic protozoa are more manageable.
We aim to use a colorimetric readout to make the diagnostic user-friendly and the results easy to interpret. Prof. Panke emphasized the importance of calculating the precise amount of dye required for a visible signal, and consequently, how much Cas protein would be necessary for the reaction.
Freeze-drying was initially a concern because we were unsure where to access the required equipment or how to use it. Prof. Panke clarified that any reputable biochemistry lab typically has a freeze-drying machine, making this step feasible.
We incorporated these insights into our project design by prioritizing a single pathogen, selecting it based on lysis feasibility and planning dye and Cas protein quantities for clear colorimetric readouts.