Partnership with SHSBNU_China
In April, we took part in an online project exchange meeting to get more inspiration. In this meeting, we shared a primary idea which is using astragalin to treat Alzheimer's disease (AD). What surprised us was that after this meeting, members of SHSBNU_China team contacted us: “The projects between us this year had a lot in common. We thought maybe we can help each other to finish the project better.” After a thorough discussion, we began to establish partnership with SHSBNU_China team.
SHSBNU is a high school in Beijing with a beautiful campus, advanced equipment, excellent teachers, and superior teaching quality. It enjoys a high reputation in Beijing and has a certain influence on the world. Our partner SHSBNU_China is a veteran team in the iGEM competition, they had received great scores in the previous competition. This year they major focused on eyesight protection, they want to produce anthocyanins in E.coli. After discussion, we found the astragalin production pathway shares some enzymes with the anthocyanins production pathway, so we established a close partnership in the whole process of competition.
In April:
Communication of project design online
We had a big online meeting with the team members of SHSBNU_China, which was the first time we met the whole team. Although it was an online meeting, we were still happy to meet so many partners, and we were looking forward to further cooperation in the future.
This meeting focused on the design of the project. We introduced the background papers and the design ideas of the project to each other, and also discussed HP in the process of the project. We have gained a lot.
First of all, in the process of presenting our project, we found that there were many details that we were still not very clear about, such as which species the gene we plan to use comes from. Secondly, members from the SHSBNU_China team raised a lot of questions to us, which we totally ignored. For example, they proposed to preferentially use E.coli for protein expression since E.coli is more efficient and convenient for protein expression compared with yeast. Third, when we heard SHSBNU_China planned to construct all the enzymes for anthocyanin synthesis on the same vector for direct transfection, we propose to separate vector expression or bacterial strain expression. This was a great chance for us to review project ideas before beginning.
In May:
Experiment training
This year, we conducted experimental technical training with students from SHSBNU_China team. On the first day in the lab, our teachers explained the safety rules of the laboratory for us and reminded us to pay attention to safety during the experiment. Next, we studied basic molecular biology experiments and the basic ideas of synthetic biology. Moreover, we also conducted experimental operation training. In the beginning, the teacher led us to complete the experiment and taught us PCR, electrophoresis, transformation, Gibson cloning, and other operations. Later, we were able to design and complete the experiment all by ourselves. The whole process was under our teacher’s supervision.
In June:
Hold a meetup
[June 26: First meetup]
This year, we jointly held a large-scale meetup with the team of SHSBNU_China. At the beginning of June, we came up with an idea to build a platform for discussion between teams, we talked about this idea with SHSBNU_China and our meetup was decided to hold on June 26. Our team members were excited, so we started to advertise before the meetup, we also made good plans and debugged software. Because of the pandemic, we had to discuss online. A total of 20 teams participated in our meetup, including both domestic and foreign participants, as well as high school and university teams. We also invited iGEM Asian ambassador Heng Shi for comments and discussions. During the meeting, each team was given 10 min to give a presentation, followed by audience questions and discussions for 2min. Most of the participants were well-prepared, especially the university teams, we were amazed by their idea and data. It was also worth noting that after the meeting, we designed a dedicated time for concentrated discussion between the participating teams, to conduct experimental design or communication on cooperative relationships. A small meeting could be established between several participants to discuss. Many teams found partners at our meetup and gained more support and help. Finally, we recorded the whole process of the meeting and fed back the video to each team. We hoped the speech and questions would help them make more improvements. We also uploaded these videos on the open video website named Bilibili, the most popular open video platform among young people in China. All uploaded materials are free for people interested in synthetic biology to watch and so popular among young Chinese students. So far, all the videos have been played more than 1000 times.
In July:
Design the naringenin detector
Our project this year has a lot in common with the project of SHSBNU_China. First of all, our basic goal is to use E.coli to express different enzymes, which can catalyze substrate and finally get different products. Second, our team uses naringenin as a substrate to produce astragalin, but naringenin is an intermediate in the SHSBNU_China project, so we may be able to link the two teams through naringenin. As for the naringenin we used together, members of SHSBNU_China team found a scientific paper reporting the naringenin detector. We studied the paper together and decided to build this naringenin detector. We will use this part together with SHSBNU_China in subsequent experiments to verify the results in both teams.
The block FdeR is a sensor protein to naringenin. After binding with naringenin, it can activate the expression of GFP in the other direction.
In July
Partnership on Education Projects
iBowu-China team have a strong track record of producing public education products. We introduced to team SHSBNU_China our Biology Art Exhibition this year, showing them (left panel) a outlandish art work submitted by one of artist. We also showed them the biodoodle from 2021, and team SHSBNU_China raised a few points for improvements for us. Later, team SHSBNU_China also joined the Biology Art Exhibtion and started a project on molecular biology education card game.
In August:
Interview together to get suggestions for both team
Into August, our experiments had been carried out steadily, but we still had some problems: some plasmids were so difficult to build, our laboratory had bacteria pollution sometimes, and many intermediate in astragalin production pathway may inhibit the growth of E.coli. We tried to search papers for solution but nothing effective. At this time, we talked to SHSBNU_China, and their experimental process was not going very well either, so we decided to find a professional to help us solve this problem. So, together with SHSBNU_China, we successfully found Prof. Geng who was very familiar with expressing proteins in microorganisms. We showed our experiment result to Prof. Geng and asked her to provide some experimental suggestions for us and SHUBNU_China. Prof. Geng was so nice to told us these are frequently happened things and told us don’t be so worried about them. For our team, she suggested us to express proteins under the condition of 18℃, which maybe better for the expression of individual enzyme. We tried this temperature later, we are so surprised to see it truly provided more production. For SHSBNU_China team, she told them to consider whether different anthocyanins can be produced by mixing and how can we purify the specific one.
Promotion video preparation
There is a very important deadline on September 2nd to submit promotional video. We attach great importance to the production of this video, which will be the first time for us to show our creativity and thinking to the judges. To make a wonderful video, we teamed up with our partner SHSBNU_China to discuss it, we cooperated mainly in the following parts: Firstly, because SHSBNU_China team was lack of some shooting equipment and shooting skills, they felt so difficult to finish this task. However, there was a student in our team who was interested in making videos, so we taught them many ways to make a beautiful video and helped with the shooting process. Secondly, after we successfully shot the video, we need to write subtitles to match the images and explain our thought. SHSBNU_China team found a student who was good at English to polish the text. This student helped us modify the subtitles of the video to make our expression more clearly and our oral English sounds more native. Third, we helped SHSBNU_China promotion video with uploading. There was something wrong in their uploading process, and we felt pleased to offer our help.
Hold a meetup
August 26: Second meetup
Into August, we have put a lot of effort in our experiment. One day, our partner SHSBNU_China team contacted with us and told us “after the first meeting, a lot of positive feedback were received.” “Many teams felt very helpful to the last meetup, and hoped to have more opportunities to discuss.” We are flattered and therefore, we decided to hold the second meetup on August 26, exactly two months later. This meeting was an offline roundtable discussion, and we named it iGEM Beijing Regional Roundtable Meetup. We listed two major highlights: • A Squad from 10+ Different High Schools • A Union across China and US Schools In this meeting, we communicated with participants face to face. We shared the progress and encouraged each other, we also provided each other with resources and help. We all felt cheerful and helpful.
In September:
Communication to improve experimental conditions
After this joint interview with Prof. Geng in August, we gained a lot of information. Back to the lab, we adjusted the experimental conditions, according to Prof. Geng's advice, cleaned up the lab thoroughly, and attempted to use a longer reaction duration for Gibson cloning, we also expressed proteins of temperature changes, and tried the concentration of IPTG changes. After a period of trying, we finally got the best expression conditions at the end of August, we are very excited to share this news with our partner SHSBNU_China and suggested that they could try to use this condition for protein expression.
In September:
Our teams interviewed with Dr. Yong Zu online together. Both of our teams ran into the problem of low expression level of our enzymes. We had tried to do codon optimization for E.coli but still did not improve. We took our result to visit Dr. Yong Zu, who got his doctor's degree in biochemistry and molecular biology. He recommended us to find specific quantitative data on how much product has been produced under different condition, and how labs have improved the synthetic process in general. According to his advice, we started to try a time-course and a temperature-course experiment to find the best expression condition. Detailed information is displayed on our results page.
In October:
Modeling
SHSBNU_China team had more experience in building models. This year they also received a lot of help for college teams, and they shared this experience with us iBowu-China. They explained how the equations work for the synthesis of substances. We learned a lot about the dynamical process and tried to work on our model.
Pre-jamboree
In October, we entered the final stage of the competition. All the students felt nervous, we felt a little bit timid for the unfamiliar competition environment, and did not know how to answer different kinds of questions. In order to show the project in a better way and ease our nervousness, we jointly held the pre-jamboree with the SHSBNU_China team. We invited school teachers and students to join our activity. Some people were very familiar with our two projects, some of them had never been involved in projects, and there are also some junior students who had even never known about biology, we gave a presentation and asked for some advice. We do think the pre-jamboree is a great way to give us confidence.
