Team:ASU/Collaborations

Collaboration

phototroph community

While working on our project, we had a chance to collaborate and talk with many teams working on synthetic biology projects involving phototrophic chassis. We were able to discuss our project ideas with other teams throughout the entirety of the competition. It was incredibly helpful to have intermittent meetups with these teams and share our successes and our problems, while also learning from theirs. We had meaningful conversations with the teams and some of their mentors, and were able to help improve and strengthen the phototrophic community in iGEM. Despite the barriers we faced due to COVID-19, we had a chance to make great connections and meet teams across the world and be inspired through all their goals.

meeting 1

We began our initial partnership by joining the IGEM-Phototrophs community which was organized by iGEM Marburg for all phototrophic teams in the 2021 competition. In this slack channel, we communicated with 11 teams from places like Oceania, Europe, and the United States. Our first meetup occurred on June 26th and we had the chance to listen to the previous iGEM Kaiserslautern 2019 team (who also worked with Chlamy!) about their experiences and challenges. Here we had a chance to question previous members on possible difficulties we may encounter while working with Chlamy and what tips and tricks they may have for us. A major advantage we gained from this encounter was the advice to be patient, and give Chlamy lots of time to grow. We shaped our initial experimental plans around this advice, and owe our completed timeline to them.

We also spoke with René Inckemann, a PhD student and mentor to the Marburg team, and the other algae team involved in the community. While the other algae team was not working specifically with Chlamy, René worked with C. reinhardtii in his own research and offered ample amounts of feedback for our laboratory plans. We were able to express our ideas regarding future lab protocols like integrating DNA into the mitochondria and our gene gun protocols.

meeting 2

At our next meeting with all of the teams, on Saturday July 24th, we heard from Dr. Osorio and Dr. Andreou, both professors with experience working with phototrophs and plant synthetic biology. By this point, we had been attempting the transformation of Chlamy for a while, with no success. We used this meeting as an opportunity to speak again with René Inckemann during an algae breakout session. He suggested that we use a “dark growth protocol” he had been using with success. We did incorporate it into our experiments and compared it to our “light growth” protocol. There was no significant difference in the success between the two, but it taught us an important lesson. In the field of molecular biology, oftentimes our experiments will not go according to plan. It was useful to have another mentor available with different experiences to troubleshoot our experiments with and share our own experiences.

meeting 3

Our final meeting with the Phototrophs community was on September 18th, where we discussed “How to make the World a Better Place”. During this meeting, we concluded our partnership with a final presentation on the impact of our project both on a practical and broader level for the advancement of science. We prepared and presented a talk that discussed our original motivation behind our project, the current issues of arsenic contamination, health impacts, and our achievements thus far. The image below is a screenshot from this presentation.

phototroph handbook

Throughout the partnership, we also collaborated on the creation of a handbook for working with phototrophic organisms in synthetic biology. We are excited to share our experiences - both successes and struggles - with future iGEM teams, and encourage more work on model organisms outside of E. coli. The final product of the handbook that was compiled by all of the teams on the iGEM 2021 Phototroph community can be found below. Our work can be seen on Section 3.3.3 labeled as Direct Methods. Here we give a deep explanation into our method and experience with transforming the chloroplast in Chlamydomonas reinhardtii. We also include some discussion regarding shortcomings and what to avoid when working with Chlamydomonas reinhardtii as a chassis. Protocols for using a gene gun and performing the glass bead protocol for integrating the plasmid in the C. reinhardti are also included.