Integrated Human Practices Timeline
See how our interactions influenced project direction and self-reflection.
Human Practices Timeline
February 2023
Our team had settled on the idea of developing a rapid test kit and mitigation system against Clubroot by designing proteins that would bind via electrostatic interactions to protein targets Pro1 and PBEL04 on Clubroot. The detection system was decided to be implemented as a strip that would be coated with our chimeric protein, upon which soil samples could be placed. However, for the mitigation system a delivery mechanism and means for entrance into the canola plant needed to be researched.
March 2023
After getting a better understanding of plant anatomy our team realized that fully developing both our detection and mitigation systems in one year would be a challenging endeavor. Thus, we decided to expand the span of our project to two years with the focus of this year being our detection system and that of our second year being our mitigation system.
April 2023
Despite shifting the focus of our project to our detection system, our team was still wanting to conduct research related to our mitigation system. At the Ag Expo we learned that bacteria that live in symbiotic relationships with agricultural crops are already being engineered to perform functions that enhance plant growth. We realized that engineering symbiotic bacteria could potentially be a viable delivery mechanism for our mitigation system, and will be considering this method further in the second-year of our project.
May 2023
Concerns regarding the sensitivity of our detection system arose at the MindFuel Prototype Challenge. We knew this was an area that would need to be addressed and decided to seek help from those with expertise in improving test kit sensitivity at FredSENSE Technologies.
June 2023
Although our interview with Emily Hicks did not completely address our questions regarding sensitivity, she did suggest looking into other types of ELISAs to use as a proof of principle. We reflected on what she said, and instead of using an indirect ELISA, our team decided to switch to a direct ELISA as this method would be cheaper and easier as it only involves one antibody.
July 2023
Following the conclusion that it would be most effective for our test to be utilized immediately post-harvest, we realized that we would need more specific information about PbEL04 expression. Specifically, we would need to gather more information on if it is expressed beyond the infection stage. This became a future direction of ours.
August 2023
Optimizing our detection system will be a crucial component of our project. However, as our team recently obtained promising results from one ELISA experiment, we decided to make these optimizations future directions. Experiments which will include serial dilutions to obtain the minimum protein target concentration to receive a positive result will be conducted in the second year of our project.
Summary & Future Directions
At the beginning of this season, our team had the idea of developing two systems directed at targeting two aspects of the issue of Clubroot. Gradually with input from experts our project evolved into what it is now: a two-year project with the first year yielding a detection system and the second hopefully leading to the further development of this detection system into rapid strip tests and potentially a mitigation system.
Early on we quickly learned that our project was ambitious and that more time than we had originally thought would be required, inspiring us to pivot from a one year to a two year project. This was a well-thought out step in our project, as the research and results gathered this year would be instrumental to the further development of our detection system into a rapid one, that could have the potential to develop into a mitigation system against Clubroot.
Our team’s presentations and participation in competitions prior to the final iGEM Jamboree not only helped prepare us for our final presentation, but led to connections that played significant roles in our wet lab testing and overall results. With the encouragement of the MindFuel Judges our team partook in conversations with the founders of FREDSense Technologies. These discussions led to our implementation of a direct ELISA rather than an indirect ELISA as we had originally planned. Our wet lab spent much time troubleshooting and it was not until closer to the end of the season that we obtained successful ELISA results. Had our team not decided to switch from an indirect to direct ELISA, we could have not obtained any results at all.
The trip our team took to Edmonton was key to our plan for furthering our wet lab results in the second year of our project. At our University we do not have access to a biosafety level 2 lab, which is required to work directly with P. brassicae. Our team reached out to Corteva Agriscience, an agriculture biotechnology company, with the hope of gathering more knowledge on Clubroot and of potential collaboration between our team and their company, as a primary research area of Corteva’s is Clubroot. Their research team was very interested in our project and established that they would be open testing some of our samples in their own labs, as they have the resources to do so. During our time in Edmonton we also thought critically about the specifics of our detection system, which lead to the establishment of several future directions including determining the exact time points at which PbEL04 is expressed in the P. brassicae life cycle and how much PbEL04 is expressed at these times.
Our team organized a second consultation with FREDSense where more future directions were decided upon. This meeting took place prior to our team performing a successful direct ELISA, so much of the discussion centered around possible means to increase our wet lab success. Once some of these suggestions were implemented our team did indeed get the results we were hoping for. Additionally, we discussed the next steps to take once successful ELISA results had been obtained. Thus, next year as we further develop our detection system, we plan to optimize it in the wet lab and will implement some of the methods (described above) suggested by Dr. Mayall from FREDSense.