Partnership

    We held several meetings in the aspects of experimental design, HP and Wiki production. We have established a continuous partnership from May to October 2022 and achieved good results. We have conducted a lot of discussion and cooperation in HP design, communnication&education activity design, material preparation, etc. We jointly carried out community education, book city publicity and other activities. Because our team is composed of high school students, we are relatively weak in molecular biology technology and experimental operation. Therefore, we have extensively learned from the NNU-China team in these aspects. In particular, we both choose Yarrowia lipolytica as the chassis cell, so we learn about the culture and genetic operation technology of Yarrowia lipolytica from them. They also helped us carry out important gene knockout operations.

    Here are some details about the progress we made together to each other's project.

    In early May, NJXDF-CHN go to Nanjing Normal University to have a meeting with the NNU-China team. NNU-China 2022 and NJXDF-CHN met for the first time in Tiangong Building. Both teams introduced their projects. We learned that the project theme of NNU-China is the synthesis of DHA and EPA by engineering microorganisms. We found that in the experimental part, they also select Yarrowia lipolytica as the chassis. We believed that we could help each other and make progress together in the follow-up experiment.

    Because our foundation in molecular biology is relatively weak. On May 28,2022, we and members of the NNU-China team conducted a series of partnership activities for guidance and teaching of genetic engineering and molecular cloning. In the course of the activity, elders of the NNU-China team taught us the basic principles of molecular cloning the skills of primer design and Golden Gate method to construct plasmids. We are very grateful for their generous teaching and are determined to learn more from our partners in the experiment.

    On June 10, the NNU-China team arranged a laboratory safety manual. They explained the laboratory safety to our team members, making us familiar with the structure of the laboratory and the use of instruments and equipment.

    On July 1, NNU-China and NJXDF-CHN carried out offline communication activity. This time, we mainly had an in-depth exchange on the production of picture books, peripherals and PV. The members making picture books and peripheral items in both teams showed the production results to each other. NNU-China put forward ideas and suggestions for the project of the our team, as well as details that should be paid attention to during the production process. The two members in charge of PV production in NNU-China team explained in detail to NJXDF-CHN about the use of the software they used in PV production, which provided us with the production plan for PV production. We also showed them our ideas on the follow-up work during this communication, which gave us some new ideas.

    On July 13rd-15, NNU-China organized an activity to train NJXDF-CHN on Yarrowia lipotica genetic operation. Because we both choose Yarrowia lipolytica as the chassis cell, this is a new type of unconventional oleaginous yeast. Our team has already had a certain understanding on Yarrowia lipolytic, but has no experience in the cultivation and genetic operation of Yarrowia lipolytic. So the members of NNU-China spent 3 days to teach our team members about the experimental operation techniques of Yarrowia lipolytic, including inoculation, culture, and transformation. We quickly mastered these skills, and we believed that this activity played a key role in promoting the experiments of our project.

    On July 21, 2022, NNU-China 2022 and NJXDF-CHN carried out a propaganda activity in Nanjing Wangzilou Community. The activity centered on DHA/EPA eggs and phenylethanol. During the event planning period, we jointly made the event planning plan. We also designed interactive games related to the content of the presentations for the children participating in the activity. During the activity, the two teams gave a series of explanations on DHA/EPA eggs and phenylethanol. Later, we led the children to observe Saccharomyces cerevisiae with a microscope and play interactive games together. Together, we will bring the fun of science to children and plant the seeds of synthetic biology in their minds.

    NJXDF-CHN attempted to engineer Yarrowia lipolytica as a competitive platform host to produce 2-PE. To improve 2-PE titer, some competing pathways are needed to be deleted. The major byproduct is analysed to be phenylacetate, which is the oxidized product of phenylacetaldehyde catalyzed by aldehyde dehydrogenases (ALD2 and ALD3). Since we have not mastered the gene knockout technology of Yarrowia lipotica, we asked the NNU-China team for help to knock out these two genes. A marker-free gene knockout method based on Cre-lox recombination system was used as previously reported. And NNU-China sequentially knocked out both ALD2 (YALI0D07942g) and ALD3 (YALI0F04444g) in po1fk. The resulting engineering strain was then handed over to our team, and we obtained a very good phenotype on 2-PE production in subsequent experiments. NJXDF-CHN team believes that the gene knockout work they help us to do is an important step for us to complete the engineering success.

    On July 24, 2022, NNU-China and NJXDF-CHN held a publicity campaign on synthetic biology and Omega-3 eggs in Phoenix Book City. In order to adapt to Phoenix Book City, a place with dense flow of people and a wide range of people, we changed our publicity methods. We used fixed booths to display, distributed leaflets, and designed on-site questionnaires to adapt to different age groups on the site. Through the on-site feedback of different groups, we could get more comprehensive public information feedback.

    In September, we had an offline discussion with the NNU-China team. We discussed the problems encountered in Wiki pages, we learned a lot of material writing and wiki production skills from the NNU-China team.