Team:BUCT-China/Collaborations

BIT-China
SCAU-China
iBowu-China

Collaborations

I.Collaboration with BIT-China

1.Education

We held two popular science activities with BIT-China. In the first science popularization, we will discuss synthetic biology together with BIT-China and spread some basic knowledge of cultured meat. The main audiences are college students in northern China.

This science popularization activity is mainly divided into four parts, team Promotion Video sharing, popular science lectures, project introduction and audience questioning sessions. When sharing the lecture, we gave a vivid description of the development process, synthesis process, and comparison of artificial meat with traditional animal husbandry meat and plant-based artificial meat. The concise content of the presentation and the lively and relaxed background painting style deeply attracted the audience in the live broadcast room.

Then, the student leaders of the two teams gave a detailed introduction to the team's projects. In the process of mutual exchange of projects, everyone has benefited a lot.

At the last Q&A session, everyone actively asked questions and spoke freely. The collision of words and thoughts burst out fierce sparks, benefiting the two teams and the audience a lot and broadening their horizons. This science popularization is an event jointly held by us and our partner BIT-China, so that both parties have a deeper understanding of each other’s projects; What’s more, the communication with the BIT-China team also started to make us think about how to improve the taste of cultured meat and enrich the relevant product. Therefore, when the project was carried out to the end of the experiment, we conducted the third social survey on consumers’ expectations of cultured meat in the future. Among them, there was a variety of personalized taste customization options (this is the same as the BIT-China project) The survey shows that about 59% of consumers hope that cultured meat can have a variety of personalized tastes in the future. The relevant data are as follow:

In the second science popularization, due to the in-depth experimentation, our understanding of cultured meat was further deepened. At the same time, due to the feedback from the audience during the popularization of science, we also had a deeper thinking on the hot topics of this era. Therefore, with BIT -China, BIT, and iBowu-China jointly organized a seminar on synthetic biology ethics. The preaching format is that BIT and BIT-China will preach offline, and BUCT-China and iBowu-China will preach online together.

The presentation includes four processes: iBowu-China explains what biological ethics is by using examples BUCT-China briefly introduces the project + cultured meat ethics BIT-China briefly introduces the project + artificial life ethics BIT brief introduction of project + biomedical ethics After investigation and literature review, we found that everyone's attention to the ethical issues of artificial meat is mainly focused on four points. First is the aspect of food safety: people who do not know much about biology will think that artificial meat is not "natural" enough, and people who have certain biological knowledge are worry about whether unhealthy factors will be introduced in the cell cultured process; the second is to doubt the value and significance of cultured meat, thinking that China has a relatively developed aquaculture industry, there is no need to invest a lot of time and money to research and develop such a problem. Something that is too reliable; third, it is worried that the development of artificial meat will have certain impacts on society, which may have a negative impact or impact on traditional meat-related industries such as traditional husbandry and affect traditional food culture. At last, from the animal's point of view that cultured meat makes people rethink about the relationship between humans and animals. We found that many concerns and thoughts about the ethical issues of cultured meat stem from the fact that people do not have a comprehensive understanding of the entire production process of cultured meat. In fact, it also shows that in many cases we lack a prerequisite combing and scientific foundation of science to the public. Therefore, we first briefly explained the production process and source of cultured meat, and then discussed more valuable ethical issues of cultured meat with the participants.

2.Artwork

We helped BIT-China draw its wiki flowchart, which was adopted; BIT-China wanted to help us draw our wiki flowchart, which was also adopted by us. The pictures we helped BIT-China draw are as follows:

3.Experiment

BUCT-China->BIT-China

BIT-China plans to use experiments to obtain plasmids expressing fresh, sweet, and bitter human receptors, which will be used in conjunction with the detection plasmids that BIT-China has successfully constructed to measure the taste of food through fluorescence intensity. In the start of experiment, BIT-China took the plasmid PCR to obtain linear fragments, and then through the design of primers, after PCR, the target gene fragments and fluorescent gene fragments were obtained. Then use the OE technology to connect the two fragments. After performing the Gibson ligation of the linear fragment and the OE fragment, introducing them into E. coli for verification. If a colony grows, perform bacterial PCR, and sequencing the DNA after the correct bacterial PCR. In this way, BIT-China successfully constructed a detection plasmid. However, BIT-China encountered difficulties in constructing the other three plasmids. When performing plasmid PCR to obtain linear DNA, the gel recovery concentration was too low due to the long fragment length, or there was no band in PCR. In the communication with us, BIT-China raised this question. Our team members gave BIT-China suggestions. For the case of low gel recovery concentration, consider running the verification gel first. If the strip is single, perform column recovery, which has a much higher recovery concentration of DNA than the glue recovery. If there are no bands, consider replacing the primers. So that, we taught BIT-China how to use Oligo to design primers. The specificity of the designed primers will be better, and clear bands can be obtained after PCR. Since then, BIT-China has tried our methods, and the experiment has made good progress. However, the linear fragment of the plasmid is too long and is not a single band, it can only take the form of gel recovery. BIT-China chose fastfu and phanta two enzyme systems for PCR in the laboratory, and tried to change the PCR temperature at the same time, but the results are still not optimistic. We told BIT-China that primstar enzymes could be used to try and proposed to help BIT-China perform PCR. BIT-China delivered the plasmid and primers to us through logistics. Through our PCR, the concentration successfully reached 213ng/ul.

Using the PCR product BIT-China obtained by our help, BIT-China successfully passed Gibson, the sweet receptor fluorescent plasmid was obtained.

When constructing bitter plasmids, a new problem appeared. BIT-China constructed bitter plasmids in the same way as above, but OE always failed to get the correct product. Even if the fragment length is correct after OE runs, there is no way to get the target plasmid by Gibson. Using Oligo to design primers or changing the PCR temperature did not solve this problem. BIT-China also communicated with our experimental group members and raised the confusion of BIT-China. Our team members checked the plasmid map of BIT-China and found that there are restriction sites on both sides of the receptor. This is because BIT-China needs to replace the restriction site designed by the receptor. We told BIT-China that they can try to construct the plasmid by enzyme digestion. First construct the plasmid linking the promoter and the fluorescent protein, and then insert the receptor. BIT-China listened to our suggestions and learned from the seniors to use the enzyme digestion method to conduct experiments. It was the first time to use this method to construct plasmids. The result was unexpected by BIT-China. After the failure of the second experiment, BIT-China successfully constructed the bitter plasmid in the third enzyme digestion experiment.

BIT-China->BUCT-China

In our project, the most important part is the polyester synthesis part. Whether the cells themselves can synthesize polyester is the key to the structure of synthetic cultured meat fiber. And the use of cells to produce polyester materials with a longer carbon skeleton is also a new breakthrough. In this part of the work, in addition to constructing the intracellular polyester synthesis pathway, how to extract the product and how to detect it is also a headache. At the beginning of the experiment, the yield of the product was very low, and the solubility of the product was particularly poor. We use common organic solvents such as n-hexane, cyclohexane, and ethyl acetate for product extraction, and then gas chromatography for product detection. However, after many attempts, it was found that our products are not highly soluble in these conventional solvents, so we did not get ideal data results in the gas phase at the initial stage. To solve this problem, we had a discussion with the BIT-China team, and we provided products that we speculated that might exist, such as nonanoic acid hydroxylaurate, dipolyhydroxylauric acid, etc. After exchanges and discussions, the BIT-China team believed that the extraction efficiency of conventional organic solvents for target products similar to dipolyhydroxylauric acid was not high. They recommended the use of chloroform for extraction. We accepted the suggestion of BIT-China. In the following experiments, we added chloroform to extract the cytolytic solution in a ratio of 1:5. From the gas phase detection, we detected the target product nonanoic acid hydroxydodecanoic acid. Esters, dipolyhydroxydodecanoic acid, etc. Thanks to BIT-China a lot!

II. Collaboration with SCAU-China

1.Dry lab

In the iGEM meetup of Northeast China at May 2021, we were attracted by the SCAU-China project and made suggestions: we recommended that SCAU-China conduct molecular dynamics simulations on the basis of building the protein model structure to better understand the function, and comparing the results of the wet experiment to the analysis. This proposal was adopted by SCAU-China.

2.Wet lab

Out of our constant concern about environmental issues, we are very interested in SCAU-China’s Chlamydomonas installation. The main theme of SCAU-China this year is to transform algae to absorb heavy metal ions in water bodies, and to use sodium alginate for the fixation of algae. The embedded algae are made into microspheres and put into use. We use the sodium alginate + gelatin system to print the scaffold to embed cells to complete cell fixation. We have great similarities in fixation and have developed close cooperation. In the early days, we had some online discussions, mainly including 3D printing fixed algae, improvement of containment device and so on. For the former, we decided to mix our materials with their algae, and then print out a porous support through a 3D printer to fix the algae. In terms of equipment, we suggest that they can add a power system to the original equipment to improve the flexibility of the equipment in the water body. Our proposal was endorsed by SCAU-China and adopted by SCAU-China.

In the mid-term, after we reached a consensus, we carried out a series of experimental cooperation. First, SCAU-China sent us their algae and the corresponding culture medium. Then we started the 3D printing work. We mix the algae solution (because the laboratory requires that the algae are all inactivated) and the prepared materials here to mix 1:1 volume and ensure that the sodium alginate: gelatin ratio is 3:25, and then mix it. The algae material (used to print the support) and the mixed material without algae (used to print the team name) are respectively loaded into the printing cylinder. At the same time, we set up two team abbreviations and the G code file of the bracket to be played. After that, we adjusted the corresponding air pressure and temperature, installed the printing cylinder on the 3D printer, and started printing. We first completed the printing of the team name to commemorate our cooperation, and then we started to print the bracket. After printing, we put the printed brackets into the six-hole plate, packaged and stored the printed products, and sent them back to SCAU-China, completing the mid-term cooperation.

After receiving the product sent by us, the SCAU-China team members began to use a microscope to characterize the product in the later stage. By observing the distribution of the algae in the material, the result of the algae fixing effect is good.

In the later stage, SCAU-China provided us with the "suicide gene" they developed, and proposed that the "suicide gene" can be used in the strains that produce polymer or hydroxylated collagen to reach a certain concentration of product synthesis after the bacteria. Kind of "suicide" to release the product, so as to reduce the cost of purification and increase the concentration of the product. SCAU-China has given us a lot of inspiration. In the future, we will think about the "suicide" gene better into the practical application of the project. (for more information Team:BUCT-China/Design - 2021.igem.org)

3.Hp

In consideration of the differences in the understanding of food culture between North and South, we are supported by SCAU-China and they distributed our social questionnaires from various periods to Guangzhou, China, to expand the diversity of our survey subjects. In order to spread our project and related synthetic biology and tissue engineering knowledge to farther places, we, together with SCAU-China and FAFU-China, held a joint preaching of science popularization by the three universities called “under the microcosm, above life”.

III.Collaboration with iBowu-China

The exchanges with iBowu-China allowed us to attach importance to children’s science popularization. In order to cultivate children’s interest in synthetic biology, and to convey the joy we have gained during the project process, we have drawn our projects as linear sketches. The form gives us a unique style and is handed over to iBowu-China to complete the writing of "Biodoodle”. We look forward to disseminating knowledge related to synthetic biology and tissue engineering in a more colorful and interesting way. The link of coloring book “Biodoodle” is as follow:
https://shop166436390.v.weidian.com/item.html?itemID=4431158156&wfr=wx_wxh5&source=goods_home&ifr=itemdetail&sfr=app&state=H5WXshareOld&distributorId=1327901843&share_relation=35da30ca2552e32b_1327901843_1

Some of the pictures and final products we participated in are as follows：