Team:UCAS-China/Contribution

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• Contribution
• Part contribution
• Gene expression regulation
• Biosafety module
• Cell-free system
• Education Contribution

part contribution

Gene expression regulation

BBa_K3817018 is an outstanding composite part which is composed by BBa_K3817006 and BBa_K3817007. This part is used for efficient production of downstream proteins, even when they are not on the same plasmid, and CRP protein output can regulate promoter expression so that as much downstream protein is produced as possible. Two efficient cascades can lead to a significant increase in protein production. Under the regulation of this part, we can obtain the targeted protein as much as possible, which is an exciting method to express low-expressed proteins. Furthermore, it is not particularly mature so that other teams are able to explore and search their usability in other possible expression systems.

Temperature-sensitive suicide switch (biosafety module)

BBa_K3817015 is a temperature-sensitive suicide switch designed to prevent possible accidents and leaks in the lab. Although following the planned process, our engineered bacteria will not cause safety problems, we have added this insurance policy to guarantee the safety.

This genetic line was inherited from our project last year. In the gene circuit, CI434ts as a transcription factor can repress TEVts and doc toxin expression, while TEVts as a protease can cut CI434ts, thus forming a balanced relationship. And when the temperature decreases, CI434ts activity decreases and loses its inhibitory effect on downstream toxins, thus initiating the engineered bacteria suicide program. To improve the performance of this suicide switch, we also fused a hybrid tevS/ssrA tag to the C-terminal structural domain of Doc toxin to reduce the leakage expression of Doc toxin. This is a mature section so the future teams can use it directly.

Cell-free system

Cell-free system is a new field in synthetic biology. Specifically, in cell-free projects, components within the reaction system are produced and extracted by synthetic or engineered bacteria and later assembled to function in the same system without the need to introduce any genetically modified living organisms into the system. It will allow us to know in advance what substances may be produced in the reaction system and ensure that it does not pose a threat to the human body.

This is particularly important for environmental, medical or food projects, where leakage and contamination of engineered bacteria are most likely to have serious consequences. In fact, a number of previous iGEM projects have taken note of this and used cell-free systems, and some of them, like us, have used enzyme immobilization processes.

According to article, cell-free systems have several advantages over traditional methods: first, they cannot replicate and spread, reducing the risk of biological contamination; second, they do not have the same reduced efficiency as living cells that must allocate energy to metabolic processes; furthermore, their reduced complexity makes it easier to modify, for example, in our project, we could adapt the system to high-temperature coffee through directed evolution environment.

Some views have raised concerns about the efficiency of enzymatic reactions in cell-free reaction systems. We believe that the enzyme immobilization system we use can address this issue, as enzyme immobilization is a relatively mature bioengineering technology and has been used in industry, such as in juice production. Therefore, we believe that the efficiency of the reaction to remove caffeine from our cartridge can be ensured if we pay attention to the immobilization process and ensure that it does not seriously affect the enzyme activity. Our experiments and experiences can be a reference for later teams and hope they would go forward further.

Education Contribution

To better train iGEM newcomers, we are endeavoring to develop an online education platform: iGEM Online, inviting schools to share their training materials and cooperate to achieve high-quality and efficient training for newcomers.

As the initiator of the project, we have made some initial plans for the platform. The platform mainly consists of two parts: the courses and the forum.

So far, we have now completed the structure of the platform and designed the user interface. We have also found dozens of collaborative teams and set up a club for the communication of course resources and web technology.

We plan to have this website online and operational in the near future to provide a convenient, complete, professional and systematic educational platform for the future training of newcomers to iGEM teams. We also extremely welcome all iGEM teams willing to participate in the construction of this online educational platform.

This blueprint of educating iGEM newcomers in a cooperative and informative way makes contributions to iGEM teams’ construction and development. A good intermediary between teams is what we consider to be the most contributing in the field of education.