Overview

“Partnership with iBowu-China”
This year, there was an online project exchange meeting took place before teams chose the final topics, participants shared about their brain storm to get more inspiration. In this meeting, we noticed that there are something in common between iBowu_China team and our team. We tried to contact the leader of iBowu-China, we all felt we can help each other in many aspects, so here we began our partnership in this year’s iGEM competition with iBowu_China.
Our partner iBowu_China is a well-organized team which had great score in last year’s iGEM competition. We had studied a lot from them. This year iBowu_China major focused on Alzheimer's disease (AD) and tried to use E.coli to produce astragalin, which could provide strategies for AD treatment. Surprisingly, the astragalin production pathway share some enzymes with our anthocyanins production pathway. So we established a pretty close partnership in the whole process of competition.

Communication of Project Design Online

After we established a cooperative relationship, we first convened all the team members to have a project discussion. Due to the pandemic, our meetings can only be completed online. At the meeting, the team members shared the original literature and introduced the project design in detail, which not only included our experimental plan, but also the arrangement of HP. Here is the useful information we exchanged:
① We initially planned to construct the enzymes for anthocyanin synthesis on the same vector for direct transfection, but due to the long pathway for anthocyanin synthesis, iBowu_China's team suggested that we could try to construct multiple plasmids or use two kinds of bacteria to co-culture to achieve the goal.
② The original idea of iBowu_China was to express the proteins in yeast, but we thought that the expression in E.coli was more efficient and convenient. Under our suggestion, iBowu_China decided to carry out codon optimization and try it in E.coli.

Design the Naringenin Detector

We noticed that the iBowu-China team would use naringenin as the substrate to generate astragalin, and naringenin is exactly our intermediate product. In the literature survey before, members in our team have found a naringenin detector, so we jointly constructed and used this detector with iBowu_China to mutually verify the experimental results.

Expermient Training

This year, our team and iBowu-China conducted experimental training together. From PCR of target genes to Gibson cloning, we studied basic molecular biology experiments and the basic ideas of synthetic biology. This experimental training deepened the connection between our two teams. There are excellent students in each team, and we are very lucky to know such excellent partners.

Interview Together to Get Suggestions for Both Team

During the process, both teams had interviewed some experts. We mainly contacted ophthalmologists and some botanists, while iBowu_China contacted with some neurobiologists and doctors for AD treatment. Although we're concentrated on different diseases, we all need to express proteins in microorganisms. So, together with iBowu_China, we successfully found Mrs.Geng who very familiar with the expression of proteins in E.coli, and we asked her to provide some experimental suggestions for us and iBowu_China. We showed some of our experiment result to Mrs.Geng. For our team, she told us to consider whether different anthocyanins can be produced by mixing and how can we purify the specific one. For iBowu_China, she asked them to increase the expression of individual enzymes under 18℃, which truly provided more production.

Hold a Meetup

June 26: First meetup
This year, we jointly held a large-scale meetup with the team of iBowu_China.
After more than a month's preparation and discussion, this meetup was officially held on June 26. A total of 20 teams participated in our discussion online, 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. This was a pretty large-scale meetup held by high school students.
Our Meetup consists of the following steps:
First of all, we made extensive publicity and promotion before the meetup, we also made a good plan and built a good platform.
Secondly, during the meeting, we focused on the team project presentation, and we provided time for audience questions and discussions as well. It was 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. Many teams found partners on our meetup and gained more support and help.
Thirdly, we invited the iGEM Asian Ambassador to conclude.
Finally, we recorded the whole process of the meeting and fed back the video of each team's speech and questions to the team itself. We also uploaded these videos on the open video website Bilibili for people interested in synthetic biology to watch. So far, all the videos have been played more than 1000 times.

[August 26: Second meetup] After the first meeting, we received a lot of feedback. Among them, we were surprised to see that many teams felt very helpful to this meetup, and they hoped to have more opportunities to participate in the discussion. Therefore, we actively coordinated with our partner iBowu_China, and finally the second Meetup was successfully held on August 26, exactly two month later. This meeting was an offline roundtable discussion, and we named it iGEM Beijing Regional Roundtable Meetup.
• A Squad from 10+ Different High Schools
• A Union across China and US Schools
In this meeting, we finally communicated with our partners face to face, which was more efficient. We cheered and applauded each other's progress in the experiment, and provided each other with resources and help. Many students became good friends in this meeting.

Communication to Improve Experimental Conditions

After this joint interview, we went back to the laboratory to push forward the project separately. Soon after, we received the happy news from iBowu_China team. After changing the expression conditions, they significantly increased the protein expression level, which indicated that they had found a more suitable condition for E. coli to express protein. Their experimental results showed that the protein expression level reached the maximum under the condition of (conditions needed here). So, in the subsequent experiment, we took their advice and tried the expression condition.

Promotion Video Preparation

Time comes to an important deadline, we will submit promotion video. Since it was a big job for both teams, we cooperated mainly in the following parts:
First, we exchanged shooting equipment and shooting skills. There was a student in the iBowu_China team who was very good at video production, and he taught us many ways to make a video stunning.
Second, we communicated with each other on the text of the video. Our team found a student who was good at English to polish the text. This student helped the two teams modify the subtitles of the video to make our expression clearer.
Third, our promotion video encountered a big problem in the process of uploading, the students of iBowu_china gave us a hand, and we finally completed the submission before the deadline.

Online communication: Zu Yong with ibowu-china

Zu Yong, male, 2017's doctoral graduate in biochemistry and molecular science from the School of Pharmacy of East China University of Technology, General Manager of Prokaryotic Biotechnology (Shanghai) Co., Ltd., and currently self founded a digital medical technology enterprise.
On September 20, 2022, we, together with iBowu_China, interviewed Zu Yong, General Manager of Prokaryotic Biotechnology (Shanghai) Co., Ltd.
We introduced our project to Professor Zu Yong and asked the following questions:

1.In what possible way can we maximize our productivity, and the expressiveness of the proteins.
2.In real-world production we dyed the contact lenses, is there anything that we need to pay attention to on safety aspects?

Answer:
1.The concentration of coumaric acid oleoresin substrate is not enough.
2.The intermediate product, the outcome quantities and reaction rates of each enzyme need separate testing.
3.For temperature, you can try personalizing in accordance to the condition of each enzyme, for instance the initial temperature, how long is this temperature kept, what temperature will later be set, for how long.
4.The safety of contact lenses indeed requires consideration. We produced contact lenses dyeing only to prove the feasibility of the product, and the craftsmanship in the industrial production in reality, but its toxicity and safety aspects still need consideration.
5.Since the eyepatch can be made, then more storytelling is needed, like what the principle is, how the effects look like.
6.Later on we will do some improvements on the safety descriptions: we are now testing the effect of dyeing on the daily-thrown contact lenses. In the case of the formal production, we need to strictly follow the production safety protocols, which, in China, belongs to Category III of the Regulation on the Supervision and Administration of Medical Devices and the Specification for the Production Quality Management of Medical Devices, we will verify further as a future plan.

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 bio doodle from 2021, and team SHSBNU_China raised a few points for improvements for us. Later, team SHSBNU_China also joined the Biology Art Exhibition and started a project on molecular biology education card game.

Pre-jamboree

To have a better performance in the final jamboree, the two teams held a pre-jamboree activity, we are pleased to invite school teachers and students. Some people were very familiar with the two projects, some of them had never been involved in project, and some junior students even haven’t learned biology, we showed the project and got some advice for them. I would like to thank my partner sincerely for growing up together in the competition and their help in experiments, HP and many other aspects.

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.