Both Mingdao and NCHU_Taichung have long been acting as great partners for CSMU_Taiwan. This year we collaborated with Mingdao High School for their experience in the iGEM competition while working with NCHU_Taichung for insights and comments from a different biological system. In our partnership with these great teams, we had successfully helped one another in various stages throughout the preparation for iGEM Competition 2022. Following is every section where we assisted each other and kept moving on during our journey. We want to thank the iGEM Team of Mingdao High School and NCHU_Taichung for being such excellent partners along the way, and we hope to continue working together in the future!
Owing to geographic proximity, we decided to cooperate with NCHU_Taichung. NCHU_Taichung was interested in our project and has made a sound plan of cooperation though our projects are respectively based on eukaryotic and prokaryotic systems. Therefore, we determined to carry out long-term cooperation. In addition to the monthly meeting of every department, we held two team-wide meetings. During these two meetings, we discussed the entire projects of both teams and judge each project from different perspectives. Not only did they give us many constructive comments, but they also suggest some solutions to the problems we encounter when operating eukaryotic experiments.
There were several problems emerging after we started implementing our experiment. To overcome these hardships, we decided to consult NCHU_Taichung for help. When we exchanged ideas about construct design, they reminded us that Green Fluorescent Protein (GFP) expression could deteriorate over time. Also, GFP-tagged cells are prone to death owing to the cytotoxicity of GFP, causing obstacles when we conduct experiments and undermining our results. Therefore, we tried to find if there were other reporter genes suitable for our construct. Fortunately, we replaced GFP with Red Fluorescent Protein (RFP) and solved this concern.
During our discussion, NCHU_Taichung also proposed their hardships. We have chosen to use electroporation to transfer the plasmid into the Bacillus subtilis natto. However, the result failed due to some potential problems that had harassed them for a long time. After understanding the circumstances, an idea came to our mind. Since they were using electroporation for transformation, the Bacillus subtilis natto may be burned to death if there was any electrolyte remaining. On this basis, they noticed that the only material that may contain electrolytes was the plasmid extracted from the bacteria. Therefore, they implemented the transformation process again after dialyzing DNA with ethanol and a nitrocellulose filter to wash out the electrolyte. This adjustment finally brought a successful outcome.
At the beginning of the experiments, we did our best to mentor the iGEM team of Mingdao High School in lab skills. During our discussion, we recognized that high school students might not be as familiar as undergraduates with some essential synthetic biology lab skills and the required experiments. Therefore, we held a workshop for about two days at Mingdao High School to teach them several lab skills they need during the execution of their project, such as the usage of pipettes, gel electrophoresis, and western blot. Through this lecture, we may not have them completely master all the lab skills in the field of biology, but we were firm that they could find it easier to move on more smoothly to their experiments.
Western blot was the last step of our project this year, which was also complicated and time-consuming. However, the time was not enough for us to verify the result of the western blot multiple times due to the COVID-19 pandemic. For the sake of saving time and double-checking our final results, they helped us with the second western blot verification under the circumstances that they had spare capacity to do so. Fortunately, the second outcome turned out to be identical to the previous one. Thanks to their assistance, we could confirm that our results were dependable and without error.
The main contributing factors that made us choose to work with NCHU_Taichung in the dry lab were their bio-technical sources, whether in software or hardware. Our teams decided to exchange ideas and collaborate with NCHU to refine our dry lab work. After we proposed the concept of a microfluidic system, they gave us a wide range of insights and optimized some of our designs.
The second version of our blueprint used a Z-shape channel with dead ends to separate CHO cells and antibodies. However, they were concerned that this design cannot do cell retention, meaning the cells would accumulate at the dead end and block the channel. In order to tackle this problem, they suggested that tangential flow filtration may be a proper solution. This advice made it a crucial factor for us to complete the design of our bioreactor.
Through our discussion, NCHU_Taichung tried using GDH enzymatic assay to quantify the concentration of PQQ due to past research. However, the more efficient and sensitive PQQ quantitative method was established in the following experiments. It indicated that the GDH enzymatic assay was not as crucial as before due to the precipitation of electron transfer, and PMS made insufficient sensitivity. Therefore, we suggested the LDH enzymatic assay be an excellent way to go further for dry lab and modeling. It was because one of the main functions of PQQ in vivo was to facilitate the conversion of pyruvate to lactate which makes enormous benefits to the human body.
The Wiki of both sides had multiple interactions to make our websites better. Since Wiki's editing platform was quite different from that of the past, both teams encountered magnificent obstacles. As a peer of Mingdao High School, members in charge of Wiki from both sides decided to have online meetings monthly to learn the rope mutually.
Our cooperation did not stop there. After we were all gradually familiar with the operation of Wiki, we still frequently contacted each other to check if everything was on the right track while also tackling the problems that emerge as soon as possible. For instance, Mingdao once told us about their plight and that they could not seek the method to upload their videos onto their website. Understanding the situation, our members rapidly gave them some tips, solved the problems, and modified their pages better.
Since there was not many professors studying in hybridoma technology in our school, we had to reach other resource and school for the opportunities of interviewing these experts. While discussing and exchanging ideas and plans with NCHU, we found that Prof. Zhao from their school had extensive experience in hybridoma research. Thanks to their introduction, we interviewed Prof. Zhao as a downstream customer of our plan, making our project more complete. （For more information, please see the Integrated Human Practice page）
The project of NCHU_Taichung this year is highly related to diabetes. However, during our discussion, we found that they were undergoing the hardship that there is no medical staff in their university. As a team from a medical university rich in medical and research resources, we spared no effort to assist them with searching for doctors engaged in diabetes research. In addition, since the target subjects were patients with diabetes, they found it hard to find a group of patients who were willing to fill in the form during the process of questionnaire distribution. Therefore, we tried our best to share the Google Form as far as possible. That way, they could successfully conduct interviews and collect feedback for their Human Practice section.
For the Human Practice compartment, we helped each other with thorough judging before we sent out our questionnaires. This year, both CSMU_Taiwan and Mingdao chose topics about animals to dig in. Although our primary goals were different from each other, we could take this advantage and advised questions of both sides from different views. Also, we worked together and tried to come up with methods to obtain more reliable results, such as deleting the middle option and making the target subjects’ opinions clearer.
After adjusting the questionnaires, our team also helped them to promote the link of Google Forms as far as we could, hoping to help them get a better and more extensive group of subjects. Through this cooperation, we were deeply convinced that our team and Mingdao both gained an opportunity to find the precise value of our projects.
In the first meeting of the Human Practice department, we discussed the educational activities we worked on. They mentioned a YouTube project aiming to collect short videos for about one minute from all iGEM teams in Taiwan to introduce every project. They invited us to join in this plan and hoped to help us promote our project. Through this plan, not only did we get an excellent chance of promotion, but we also get to know the goals of other iGEM teams through this collaboration, which profoundly broadened our horizons.
To let the public understand that iGEM is not fighting individually, but through cooperation and mutual help to make the plan more perfect, we cooperated with NCHU_Taichung on two podcast episodes. During the process, we not only shared the team's life but also learned about each other's plans through interviews. In addition, intending to promote iGEM and synthetic biology more widely, they also participated in recording our "Theme Song of Synthetic Biology" and let the melody connect us more intimately. For more information, please see the Collaboration page.
For the section of Education, we helped Mingdao with the design of their board game. Mingdao devised a board game about epidemic prevention this year according to their project. Through this intriguing game, they hope to train elementary school students in Arithmetic while memorizing several vital viruses. In order to make sure that this game was suitable for elementary school students, we played with the members of their team as a test. After one round, we found that the game was somehow complicated for elementary school students. Therefore, we suggested that they extended the target of this game and introduced this game to junior high school students to make their demonstration in elementary school smoother.
Apart from the board game, we also collaborated with Mingdao on the section of the theme song of synthetic biology. We originally wanted to write a song no more than the Mandarin version. However, after they heard about our project, they considered it an innovative method to promote synthetic biology by writing a song and suggested that we translate the lyrics into the English version. Also, after we decided to have other iGEM teams sing the song for us, Mingdao was the first team we reach. Their sweet voice was utterly recorded in our song. For more information, please see the Education and Collaboration page.
