- Human Pratices
In 2021, we achieved a partnership with HZAU-China team to promote each other in project conception, experimentation, modeling, human practice and other aspects. Throughout the competition, we maintained sincere communications and exchanges with each other.
Both teams faced similar problems during the process of project advancement. HZAU-China has been working hard to effectively diagnose IBD diseases. Our team paid more attention to personalized treatment for IBD patients. Different project footholds make it possible for us two teams to carry out complement for each other.
1. Acquaintance on China iGEM Online Meetup
In May, we met with HZAU-China team for the first time on China iGEM Online Meetup (click meetup for details), which we hosted jointly. During the team project sharing session, we were delighted to find out that both teams' projects pointed to the same disease problem——IBD(Inflammatory Bowel Disease). Lately, we learned that HZAU-China project's target group was dogs and other pets, mainly for IBD detection. Although the two teams chose different tracks and different target objects, both teams faced many common problems.
During the discussion section, we exchanged ideas of the current status of IBD, prevalence, treatment methods, diagnosis methods and shared the opinions of the selection of biomarkers. These discussions have played an essential role in promoting follow-up background writing and human practice activities. After the meeting, we exchanged WeChat and QQ contacts for follow-up cooperation.
Fig.1 China iGEM Online Meetup
2.Promtion on iGEM Southern China
In mid-July, in response to the epidemic prevention and control requirements, iGEM Southern China Regional Meeting (click Meetup for details) was held online as scheduled, and we invited the HZAU-China team to participate. After nearly two months of project advancement, both teams have made some progress and encountered new problems.
In the meeting, we discussed the problems encountered during the experimental verification. Since both teams use the E.coli Nissle 1917 strain chassis, all faced the problem of plasmid transformation. We learned that HZAU-China was trying to use electrotransformation to improve efficiency, while we were using chemical transformation, and the effect was not ideal. At the same time, in the face of more stringent epidemic prevention and control regulations, we also discussed the adjustment of HP researches and interviews. During the exchange, HZAU-China team mentioned that they planned to hold a public benefit activity-pet clinic in July, but the event need to be postponed indefinitely due to the epidemic. We suggest postponing the activity to Oct.4-"World Animal Day", which could better stimulate the public to reflect and attract people to participate in the event. At the end of the exchange meeting, the two teams signed the INTESTINAL Program Reference Manual, which meant that the manual was officially open to the public. (see Contribution for details).
Fig.2 INTESTINAL Program Reference Manual(2021)
3.Experimental Group Exchange Meeting
In August, both teams were conducting experimental verification, so we chose to conduct an online special exchange meeting for the experimental group during the summer vacation.
HZAU-China team talked about their entanglement in the choice of substances to relieve inflammation, and we suggest that they could produce LL-37 antimicrobial peptide. Because the molecular weight of this antimicrobial peptide was relatively small, the circuit's impact could be limited to a low level. HZAU-China accepted our suggestions and chose LL-37 as an anti-inflammatory substance to complete an anti-inflammatory module for their project.
Fig.3 Verification of LL-37 antimicrobial peptide
Fig.4 Expression of LL-37(slide from SZU-China)
In the subsequent communications, LL-37 has always been the focus of our cooperation. Since we did not have Tricine-Tris-SDS, we could not verify LL37 on protein level(cause it has minimal molecular weight). At the suggestion of HZAU-China, we sent some protein samples to them to accomplish verification through their Tricine-Tris-SDS. Meanwhile, we also shared part of the results of our cell experiments with HZAU-China to verify the effects of LL37 mutually.
Fig.5 Experimental Group Exchange Meeting screenshot
4.Modeling Group Exchange Meeting
In September, the experiment was nearing completion, and we conducted a modelling group exchange meeting.
In the communication, we mentioned that in the modelling process to characterize the efficiency of SOD removal of ROS, the relationship between the substrate concentration and the indirect characterization of the substrate was difficult to determine. HZAU-China suggested that we could try to endow different parameters to simulate the situations. And it was also recommended that we use the MCMC algorithm to simulate the parameters.
We found that both teams were interested in the intestinal flora competition model during the discussion, but it was challenging to implement due to the high complexity. We also showed the abundance analysis model of the intestinal flora, which gave HZAU-China some inspirations.
Fig.6 Modelling Group Exchange Meeting screenshot
As our neighbor, SUSTech_Shenzhen has kept close contact with us since the team was established, and we often appeared in the same meeting to exchange opinions and make progress together. From the Guangzhou-Shenzhen District Symposium in April to the online meetup and CCiC in August, we have gained a deeper understanding of each other with the continuous progress of our respective projects. SUSTech_Shenzhen focuses on preventing and treating diarrhea with colonized intestinal engineered bacteria in this year's program, which is similar to the core of our program, namely, the engineered intestinal bacteria for 'cocktail' adjuvant treatment. As a result, we have had a long and deep collaboration on various aspects of our projects, experiments, and Human Practice activities.
1.Discussion and trouble-shooting
at the initial stage of the project
At the beginning of the team establishment, we were invited to meet with the person in charge of Human Practice of SUSTech_Shenzhen and learned about each other's projects. Due to epidemic prevention and control, we can only meet online. We have learned that this year's SUSTech_Shenzhen project mainly focuses on treating diarrhea with engineered intestinal colonized bacteria, which has a lot in common with our project design. After an in-depth understanding of the project details, they raised a series of questions about our project application scenarios, which provided us with the idea of focusing our project design on the production and treatment of adjuvant drugs. In exchange, we also trouble-shoot with SUSTech_Shenzhen in terms of the concept and feasibility of some projects and put forward more localized problems such as specific diarrhea to lactose intolerance. Finally, our two teams reached a consensus on further cooperation.
Fig.7 Online meeting with SUSTech_Shenzhen
2.Wet lab cooperation
SUSTech_Shenzhen communicated with us a lot in the experiment, especially in the wet lab, and they provided us with many original ideas for the investigation. To validate our strains, we exchanged RFP (Red fluorescent protein) and mazF plasmids for mutual validation. At the same time, we also provided the strain of E.coli BL21 for SUSTech_Shenzhen to test. These mutual experimental cooperation and verification are of great help to the progress of our experiments.
Fig.8 The experimental materials we exchanged
At the initial stage of the project, we went to the department of gastroenterology of Shenzhen Second People's Hospital for clinical investigation on intestinal problems. In the process of communication with clinicians and nurses, we conducted preliminary verification of the feasibility of our project. We conducted field research on the clinical needs and pain points related to our project. Fortunately, with the consent of both the patient and the clinician, we were able to interview a patient with inflammatory bowel disease (IBD) and question the patient about our respective programs. We received a great deal of constructive information.
Fig.9 Picture with SUSTech_Shenzhen when conducting human practice in hospital
At the same time, to jointly promote synthetic biology and participate in public welfare undertakings, we planned and participated in the Deep Blue Charity Sale held by Shenzhen University and Southern University of Science and Technology. In this activity, we sold materials related to synthetic biology designed by us and gained great attention. In the end, we donated all the proceeds from the charity sale to public welfare organizations, hoping that this public welfare activity could contribute to the spread of synthetic biology in a more meaningful way.
Fig.10 Picture with SUSTech_Shenzhen
and hardware verification in the later stage of the project
In the end of our project, aiming to verify the feasibility of our project again, demonstrate the robustness of hardware design and apply our brochure into practice, we returned to the attention of patients together with SUSTech_Shenzhen. We went to the Department of Gastroenterology of Southern University of Science and Technology Hospital for an interview. In the interview, we demonstrated the project's feasibility again, collected the latest opinions from professional doctors, and showed our hardware design to doctors and patients. Finally, together with the members of SUSTech_Shenzhen, we put the propaganda manual we made into the hospital for use.