The collaboration with BUCT-China runs through our project, and the common goal that we both work towards is not only the vision of space but also the future expectations of food production.
At the initial stage of the project, the two teams hit it off and appreciated each other's projects.
When determining the project direction, ShanghaiTech_China learned that BUCT-China uses E. coli to produce edible products-cultured meat, so they changed the chassis bacteria from spirulina to E. coli, which preserves the production of food and has been greatly improved by synthetic biology. At the same time, BUCT-China's cultured meat food can be used as a downstream industry to land the project.
BUCT-China also knows that our two teams can utilize our chassis bacteria as platforms for each other, and ShanghaiTech_China is our vision. Keeping our eyes on the stars while with both feet on the ground. Since then, taking space exploration and interstellar migration as the background of our grand vision has been firmly established. We and ShanghaiTech_China contacted each other and conducted preliminary exchanges and discussions. After introducing our respective inspirations and preliminary plans, we put forward each team’s opinions on the overall idea together.
However, in the early and middle stages of the project, Shanghai suffered from COVID-19 and was therefore subject to lockdown control. ShanghaiTech_China’s projects had been affected a lot which is very difficult to develop the experimental part furtherly. BUCT-China was also in quarantine and lockdown for a long time in the middle and last stages of the project due to the impact of the Beijing epidemic, so it was unable to carry out experiments. However, COVID-19 has not knocked us down but allowed our relationships to get closer, and we can support each other through cooperation.
It is not only the support of concepts but also our mutual help in experiments. When the progress of the experiment was limited by the epidemic, we worked together to tap stakeholders and conduct research, and jointly publicized the knowledge of synthetic biology.
The experiment design of BUCT-China offered inspiration to ShanghaiTech_China who adjusted their project plan and initially established the project track. Due to the impact of the epidemic, we held online meetings to better communicate. The two teams sincerely support each other in concept and discussion on experimental design. We arranged meetings together frequently in order to have a further understanding, which allowed us to provide some valuable advice for both teams.
BUCT-China asked us about related issues that the growth conditions and nutritional supply that E. coli can provide in the process of moving towards a alien planet. We provide them with some related production theory and experimental support.
In addition, we have maintained continuous contact with every team member.
Idea of signal molecule degradable design
The members of the BUCT-China suggested that we use degradable label-related elements in the process of information exchange, which provides another new direction for our subsequent experimental design.
For the BUCT-China project, we have proposed relevant suggestions for designing alternating group sensing systems
Design,Human Practice,Education
We found that mussel protein can increase adhesion.BUCT-China proposed that it may be necessary to improve cell adhesion in current cell meat production. ShanghaiTech_China, based on the experience of past projects, told us that mussel mucins have strong adhesion which is completely harmless and biodegradable, so maybe mfp-5 can be used to promote the adhesion of cell meat. As a result, we decided to cooperate with Synmeta (Shanghai Yiru Technology Co., Ltd.), a synthetic biology company derived from the iGEM competition of ShanghaiTech_China, to develop cell meat and solve the adhesion problem in cell meat synthesis.
ShanghaiTech_China proposed that it is necessary to show that the three chassis bacteria have different fluorescence, and the fluorescence brightness is insufficient
BUCT-China send the sequence of the fluorescent protein to ShanghaiTech_China. The fluorescent protein has high brightness and is red. It can be compared with E.coli (blue) and cyanobacteria (green) to identify.
Meanwhile, the events we hold together benefit us, including joining scientific education and designing the questionnaire.
BUCT attended the Science Lecture (metabolism) held by ShanghaiTech_China
Collaboration questionnaire with ShanghaiTech_China
The questionnaire “Just the right question of the thing we care about” is mainly focused on the public awareness and recognition of cultured meat and the Mini Bioproduction Circle System on Mars base (MBCS). The content includes the food safety problem and Mars immigration plans which offers us the requirement and feedback for our project.
We ShanghaiTech_China and BUCT-China collaborated to design and complete the questionnaire.
ShanghaiTech_China has analyzed the data from the survey.
Through this questionnaire, we hope to understand the public's opinions on food safety in terms of food production by microbial circulation production system and artificial meat production, and to improve the project based on the suggestions of our product users.
We received 88 responses from people of different ages and educational backgrounds. 35.2% of them said they were very concerned about food safety, while only 3.4% of them were not concerned about food safety. And more than half of the respondents wanted more than basic safety. This means that for our projects, it is necessary to pay attention to potential food safety hazards, conduct relevant research and improve the projects.
Buct-china survey, compared with traditional meat, artificial meat can avoid animal feeding safety and animal infectious disease safety issues. But BUCt-China needs to know more about its users' awareness and their concerns about eating artificial meat in more ways. We designed questions about public perceptions of cell-grown meat and the safety of artificial versus conventional meat.
We learned that most of the respondents believe that the safety issues faced by artificial cell meat and traditional meat are not fundamentally different. We also need to manage the production of cell meat from the aspects of raw material freshness, food hygiene, food source and so on. A small number of people who think there is a safety difference between the two also expressed a preference for the safety of artificial meat.
Further, ShanghaiTech_China would like to learn about the public acceptance of artificial meat as a possible food for a Mars base and suggestions on the idea of combining a micro-biological fermentation cycle production system with cellular meat.
Through the survey, we learned that more than half of the respondents believe that there is a high probability that humans will migrate to Mars in the future, and that food production is a very urgent need. Most of the respondents also said that food production through microorganisms is an acceptable way of food production. This has led ShanghaiTech_China to further refine their project concept, making it clear that producing food on Mars via a micro-biological recycling production system is a necessary, achievable and acceptable idea.
At the same time, nearly 90 percent of respondents agreed that the combination of microbiosphere and cellular meat was a good way to increase the diversity and nutrition of food in the early days of migration.
Some respondents also made suggestions to us directly through questionnaires and the contact information we left, saying that they hoped to increase the taste of artificial meat and ensure the safety of the microbial system. They said they would be willing to try artificial meat, provided it was safe to eat, and would try to buy such meat on Earth if costs could be reduced to no more than 30 percent of existing meat.
Thanks to these interviewees, we received many positive comments about the idea and some suggestions for improvements.
Modeling team of ShanghaitechTech_China shared their modeling ideas, that is, to simulate gene oscillation using ODEs , quorum sensing, and some ideas about physical modeling, broadening the modeling selection and thinking of BUCT-China.
In the modeling of the gene oscillation circuit, the modeling members of ShanghaitechTech_China suggested that BUCT-China members write codes to solve the ODEs using ode45 solver, instead of the Simbiology toolbox in MATLAB, so that it is easy to better modify, trial and iteration. Although Simbiology is very convenient, it is inconvenient to correct parameters and check the code because of the black box in it. And according to the experience provided by ShanghaitechTech_China, when the ODE system becomes more complicated and ode45 isn't helpful, it is recommended that BUCT try to switch to the ode15s solver. The latter is a solver based on multi-step method and applicable to the rigid differential equation group. ShanghaitechTech_China helped BUCT to check the equation very patiently and iterate some parameters. They reminded that increasing the initial rate of transcription and translation can make the gene oscillation more stable. BUCT-China members finally ran successfully with the suggestions and help of Shanghaitech_China.
At the same time, the two sides exchanged the modeling problem in the process of modeling:
During the modeling of metabolic pathways, BUCT-China provided Shanghaitech-China with valuable experiences to simultaneous assumptions, and explained the use of Michaelis-Menten equation on multi-substrate enzymatic reactions, helping them complete the metabolic pathway modeling of S.elongatus.
Due to the lockdown control of COVID-19, experimental progress of BUCT has been greatly affected. Therefore, they sent the three synthetic plasmids to ShanghaiTech_China, and we helped them measure the fluorescent expression content, and alternate expression was exhibited using fluorescence intensity.
ShanghaiTech_China required the test of the remaining sucrose content in the culture medium furtherly, so BUCT-China helped them using nuclear magnetic resonance spectroscopy to measure the sucrose content in the sterile supernatant they sent to us, to verify whether sucrose hydrolase transferred into E. Coli is effective.
Meanwhile, ShanghaiTech-China hopes to degrade AHL in order to reduce the leakage expression in nitrogen source response. As a result, we discussed that we can add a degrading enzyme to AHL, and a repressor protein with a promoter can be tried. Therefore, a set of promoters P- λ And CI repressor from BUCT-China's expeBUCT_CHINA has designed a system of gene vibration, but there are some difficulties in the results of the results due to the closed campus of the epidemic. The fluorescent intensity is detected, we perform the corresponding detection, and the result is finally handed over to BUCT_CHINA.riment design were sent to ShanghaiTech_China to practice.
In addition, we exchanged the chassis bacteria. For ShanghaiTech_China, BUCT-China E.coli which can produce edible substances, is the downstream industry of the other project, helping the team integrate with the practice and production of the industrialized food industry. We also sent their blue-green algae to them, hoping to integrate photosynthesis into our project. From photosynthesis to PHFA, this is in line with the current environmental protection issues and reduction of carbon emissions. These are our first steps toward the future, showing bright prospects.
ShanghaiTech_China:
In our project, we choose starvation promoters for the nutrient response. However, after two cycles of optimizations, we only get a maximum fold-change of 8.2, which is still not enough for complex genetic circuit design. Some strategies like amplifier (Figure 1A) or positive feedback (Figure 1B) have been used in eukaryotic systems.[1]These strategies are not well developed in prokaryotic systems because of the lack of transcription activators. After communicating with BUCT_China, we learned that an activator delta from phi R73 phage can bind to a specific sequence on DNA and activate a RpoD-dependent transcription.
Based on the two strategies in Figure 1, we constructed seven plasmids with our starvation promoters, PcstA variants. Unfortunately, none of them seem functional, for we didn't see the expected green fluorescence on single clonings on their plates. To test if the delta activator really can work or not, we further constructed two plasmids. One of them expresses the delta activator, while the other only contains the delta activator-related promoter, pF (Figure 2). The mRFP signal of these two constructs didn't show any difference, which may demonstrate that the delta activator itself does not function.
Jointly improve the grand blueprint
The goal of Beijing University of Chemical Engineering is to overcome the public health challenges of the food industry by using the automated production line of artificial breeding meat and reduce carbon emissions in the process. The polytroxy fatty acid produced by E. coli is designed with a dynamic regulatory sequence control system, which builds a modified porous micro-ball to enable the cells to attach and cross-link to make meat. Instrument.
Shanghaitech_China's goal is to provide the initial sustainable food production solutions for the future Mars exploration of human beings. We constructed a micro-artificial ecosystem with microorganisms, and used polyol algae S. Elongatus to convert CO2 into sucrose; nitrogen fixing bacteria A. Caulinodans convert N2 to NH4+.
[1] Hicks, M., Bachmann, T.T., and Wang, B. (2020). Synthetic Biology Enables Programmable CellBased Biosensors. Chemphyschem 21, 131. 10.1002/cphc.201901191