Communication 2

After the meetup, we agreed to have a second meeting in April. We agreed to communicate online at the Tencent conference because of time and distance, and the main discussion was about the design of the project. We learned from iBowu-China where all of their project designs for this year, including but not limited to mitigating cockroach disasters, developing treatments for depression, and synthesizing active ingredients for herbal medicines. We also learned about the content of their projects that participated in 2020 iGEM last year and the year before. Among other things, they introduced iBowu-China 2020 project Sureloc, which aims to treat cancer cells with targeted drugs. We thought it was a really cool thing to do about treating cancer, and after we learned that iBowu-China had only won the silver award for the 2020 anti-cancer drug development project, we decided to tackle this big problem from a new angle: the recurrence of cancer after surgery. We also learned from iBowu-China how to find relevant information from the iGEM history project and learned how to do literature searches.

Communication 3

In May, we also had a Tencent meeting, in which we discussed mainly HP design, and in particular, how to design HP content around experiments.One of the very important things we learned is that HP must complement the experiment. It cannot be made into an unrelated activity.

Communication 4

In the second half of May, we decided on a topic for the project. We found a new research paper on Lon-protein and we shared this with an online meeting with iBowu-China. iBowu-China helped us to evaluate the feasibility of the project. Also their 2020 veteran team members Lucas and Linda, who were also last year's experimental team leaders, gave high marks to the idea of this project. If we can successfully produce the protein in E. coli, the project can be completed. We found that the Lon protein can be synthesized in E. coli from Abcam, a molecular biology company that produces biomaterials.

Communication 5

In the third exchange in May, the iBowu-China suggested us to try pet28a+ plasmid with BL21(DE3) strain, which is the expression system designed by iBowu-China 2021 project. iBowu-China team experimental leader helped us to construct plasmid vector based on our own bG-pet28a plasmid. Also they suggested that if the expression of Lon protease is successful, we can consider using liposomes for delivery, which is fully studied by the iBowu-China team in the 2020 project.

Also, we provided help to test the performance of the T7 promoter used by iBowu-China this year. iBowu-China team has had some success at this time using the T7 promoter to express β-glucuronidase and then plans to tune the strength of that promoter by mutating some promoter sequences. If the new promoter is successful, we can use the mutated promoter to boost Lon protein production, and we will provide more data on Lon protein expression measurement by the new promoter.

Communication 6

In June, our first communication centered around the safety form. Since it was our first time to fill out this form and we had a lot of questions, and iBowu-China had experience in filling out safety forms, we learned from iBowu-China about lab safety and biosafety issues, how to prepare for lab safety, and how to fill out the excel form section correctly. We also double-checked iBowu-China's safety forms as requested. iBowu-China reminded us that it would be better to declare safety measures regarding mammalian cells, i.e. the possible use of HeLa cells to test whether the product glycyrrhizic acid has the effect of inhibiting the growth of cancer cells.

Communication 7

In mid-June, we completed the video shoot with iBowu-China. The back-to-back submissions of the safety forms and the promotional video were not easy, but we felt very excited and productive. The two teams exchanged ideas for the awareness video and shared resources and techniques for filming. iBowu-China shared a lot of background information and graphic material about cancer and cancer treatment for our project presentation in the video. We also provided iBowu-China with a great filmmaker's workshop and came up with some interesting scene ideas for iBowu-China's video, with much of the footage shot at our school.

Communication 8

The first exchange in July kicked off the offline experimental part of the project.

Because our school's lab could not be used during the summer due to the epidemic, we borrowed iBowu-China's lab to conduct the summer experiment content together. At the same time, our team received laboratory skills training from the iBowu-China team. From preparing LB cultures, pouring LB agar plates, to transferring the right amount of PCR mix and DNA template, our team quickly started our lab journey with guidance. We made about 50 agar plates for iBowu-China, and our two teams helped each other during the experiment.

Communication 9

In July, we interviewed medical experts together. Both of our teams wanted to understand how their products could be used to treat cancer. We had the opportunity to interview medical experts and both teams attended the meeting. We came to the conclusion that it was not feasible to make our product into a trustworthy drug within the iGEM season.

Communication 10

In mid-July, we helped iBowu-China conduct the BioDoodle survey Add WeChat survey data.

Communication 11

Optimization of protein expression.

Our experiment encountered difficulties with low protein expression. We asked iBowu-China to come and help us try the protein expression.However, they also did not succeed in expressing the protein. We sought new help in the HP interview.

Communication 12:

iBowu-China constructed the C4 promoter on the plasmid by Gibson assembly. They also suggested us to try whether the effect of multiple promoters could enhance the expression level of the protein. However, we got other methods to enhance protein expression level from HP at that time, so we did not perform the experiment of replacing promoters.

Communication 13:

Modeling was a difficult part for both teams. iBowu-China introduced the NWU-China-B team for a teleconference on modeling, and we found that none of our students were good at modeling and dropped the modeling part after a discussion.

Communication 14:

Education on Campus
We presented a biology illustration book from iBowu-China co-authored with several teams to our students during school education. We got a lot of feedback from the students, and we sent it back to the iBowu-China team. Many "bugs" were found in the feedback.

Communication 15:

October 1 was the last day to submit the security form. iBowu-China and CKWA-China checked and discussed the security form with each other to make sure nothing was missing. Both teams received approval from iGEM headquarters after the initial submission in June, but we were careful in completing the security forms. In particular, we checked the sections we planned to submit to make sure there were no errors. This deadline was also for title and abstract submissions! We proofread each other's titles and abstracts and suggested changes. And iBowu-China suggested that we should include some digital content in the abstract to be more convincing.

Communication 16:

Throughout October, we worked together to complete the wiki content. Among them, because our students were the first time to contact the wiki part of iGEM, they were quite rusty with HTML and CSS, so we thanked iBowu-China for providing us with a set of templates that we could refer to.

Future arrangements:

We will also do a mock defense together at the end of October and many other sessions, because wiki freeze so this part can not be recorded.