Medals

Requirements for a Bronze Medal

Attributions

The experimental group spent nearly three and a half months on the experiment, the module group spent one month just solving the problem of an iron-sulfur center in molecular dynamics, and the wiki group started learning about website construction a month before wikifreeze. Since the project was confirmed, human practice students have been in constant communication with others... Each member of our team has done his or her part in this year's project, from initial brainstorming to mid-stage experimental validation to late-stage promotion and outreach. The results of today's project would never have been possible without the combined efforts of everyone involved. In addition, the frequency of the weekly group meetings allowed us to get a lot of help from the instructors. At the same time, as the project progressed, we encountered many problems. In addition to our mentors, we have obtained a large number of experts and faculty to help us with experimental conditions and experimental research guidance, so we believe that we meet this criterion.

Project Description

We present a detailed introduction on the description page. On that page, we give a clear and concise description of our treatment protocol for cardiovascular disease caused by poor diet, as well as a detailed explanation of why our team chose to work on this particular project and how we were able to infiltrate it with synthetic biology ideas. At the same time, we recorded the whole process of project thinking in the form of references to ensure the reliability of our project. Of course, we also use some illustrations and other visual resources to explain our projects. The contents meet the criteria for this award.

Contribution

We noted that the requirement of a contribution profile is in the hope that we can make useful contributions to the iGEM team in the future, so the core of this work revolves around this principle. In this part of the work we supplement some new experimental data such as BBa_K3838613 and BBa_K3733043. We used some convenient verification methods on top of the original and supplement some literature. Second, during the experiment, some experiments, such as the construction of the cre/loxp recombination system, took various detours at an early stage due to our inexperience, which we will very much like to document in this section as a warning to subsequent teams. The iGEM competition should be a large group, a huge family, and helping each other should be our eternal principle.

For more details, please click the following link BBa_K3838613 and BBa_K3733043.

Additional Requirements for a Silver Medal

Engineering Success

This part requires us to demonstrate engineering success in a part of your project by going through at least one iteration of the engineering design cycle. We take this as the core principle of our work in this section. During each design phase of the project, our experimental and mathematical modeling work has been designed and revised several times in the course of communication and discussion with instructors. As stated in the guidance, a good engineering design cycle should go from design to construction, from construction to testing, from testing to learning, and from learning to design again. Whether it is the optimization design of enzyme kinetics selected by the mathematical model for the experiment, or the multiple selection and discussion of the degradation pathway of TMA in the experiment, we have learned a lot in this process, and we are genuinely glad when we see the successful results. Perhaps that's the appeal of synthetic biology versus iGEM.

Collaboration

Collaboration is important in the whole iGEM competition. By collaborating with other teams, we can improve efficiency and reduce the occurrence of the same error. This year, we have cooperated with seven teams. We learned a lot from each other. More specifically, we had an in-depth communication with YiYe-China on modeling and human practice. We have constructed an intestinal information platform with UCAS-China, WHU-China, NWU-China-A and NJMU-China. We have compiled a probiotic manual with LZU-HS-PRO-A team and with NWU-China-A. We have searched the relevant provisions of laws and regulations on probiotics in China, which is the basis for us to continue to further marketization. Further more, we have had in-depth communication with ZJUT-China and WHU-China about some project problems and we proposed solutions for each other. At the same time, we also attend the Central China Exchange Conference. We all have a pleasant cooperation and improved our projects efficiently.

Human Practices

In order to make people realize the importance of healthy and understand our project, we have held activities to encourage people exercise and focus on thrombus. Based on data from questionnaire, we improved our project design and ask for experts to give us some advice. Further more, we have had deep communication with other teams and propose solutions to each other which has greatly improved efficiency. At the same time, we have made popular science app and a probiotics handbook and so on. We always put stakeholders’ feeling at the first consideration. We think Human Practice is an integral part in the whole competition. It is a bridge between public and science and our team has constructed a firm bridge which is also full of love. We believe our project can bring a lot of benefits to people around the world.

Proposed Implementation

Our target group is mainly cardiovascular disease patients, and we have designed the hardware and software to make our project widely available in the market. We hope that the hardware will allow the user to know the condition in real time by drawing blood, and the software will monitor and control the hardware to give the treatment diagnosis and the recommended dose of our product. And our engineered bacteria will be processed and encapsulated to form pills for daily use with theophylline, and users will be able to adjust the amount of theophylline taken according to the software feedback to achieve more accurate medical treatment.

Additional Requirements for a Gold Medal

Integrated Human Practices

Science is at the service of people. Connection is a powerful thing. Seeking truth from facts. When we merge these three sentences together and bring it to reality. We find it easy to change the world. Interact with the outside frequently and effectively, base on reality at key points of key issues, and create connections from reality to solve problems. IGEM is not only a place of inspiration in biology and engineering, but also a place where worldview inspiration sparks. We are willing to make the world a better place.

Improvement of an Existing Part

When we were designing this year's project, we decided to use nattokinase for thrombolytic therapy for thrombus which was already in the blood vessel. Through project query in previous years, we found that NYCU-Taipei had uploaded pelB leader sequence with aprN ( BBa_K4060021 ) in 2021. But for unknown reasons, only proteins were detected in SDS-PAGE, and the activity of nattokinase was not detected in the fibrin plate. Following the literature review, we improve the uploaded part sequence. Finally, we obtained nattokinase that was correctly expressed and active at 37 °C.

Project Modeling

Based on the concept that experiment and Model forming a closed circuit, we developed two models to describe the degradation of TMA in our project under the guidance of experiment, and we proposed an optimization method for gene circuit design and simulated the theophylline induction threshold by point mutations, which will greatly reduce the complexity of experiment and improve our project design. On the other hand, we tried to generalize the project, we propose to reduce the production of TMA by designing enzyme peptide inhibitors thus improving the therapeutic efficiency, which will provide guidance for future iGEMers in the therapeutic works.

Proof of Concept

In proof of concept requires clear guidance, we have detailed records used to verify the experiment thought the whole circuit design and realization of our proof-of-concept experiments in proof of concept page, for example, analog simulation to get ahead of TMD mutations is refer to the corresponding locus mutation to experiment, the results achieved mathematical model and the experimental verification, in addition to this, We also used clever red fluorescent protein to verify whether the riboswitch can function, and then used the successful results for subsequent validation. Finally, we clearly demonstrate the successful verification results of our circuit design using a large number of data graphs and experimental phenomenological graphs. In addition, throughout the validation process, we paid great attention to the safety of the experimental methods, reducing the use of contaminants and carefully documenting toxic substances.

Partnership

This year, we have reached partnership with YiYe-China and ZJUT-China. Affected by the epidemic, we were still on the project under the anti-virus policies formulated by Chinese government. We contributed a lot on YiYe-China's digital module (include the experimental scale and the research principle of references), and ZJUT-China also played an important role in our experiment (about Cre-LoxP system). HZAU-China also conducted in-depth communication on Human Practice with the two teams (include a live broadcast with 9,000 page views with YiYe-China and a discussion on probiotics with ZJUT-China).

Education

and

Communication

For this year's education, we tried a different way to motivate people into participating our activity. Movement is always more useful than just words. In this way, we spread the idea about our project, and achieve a goal that more people acknowledging synthetic biology. But being acknowledged to something is not just knowing it, but to act and to participate in it. We hope that more people would act synthetic biology and the future and pay more attention to vessel disease.

Awards

Education

Hardware

Our "i-Box" consists of a blood collection module and a TMAO detection module, for which we use electrochemical sensing to quantify the concentration of TMAO and give a therapeutic diagnosis and recommended dosage of our products. The user can be informed of the disease in real time through blood sampling, and can intelligently regulate the ratio of theophylline and probiotics by connecting to the "i-box" using a smartphone, which makes our hardware close to life.

Integrated Human Practices

Model

Based on the concept that experiment and Model forming a closed loop, we developed two models to describe the degradation of TMA in our project under the guidance of experiment, and we proposed an optimization method for gene circuit design and simulated the optimized working performance of degradation enzymes by point mutations, which will greatly reduce the complexity of experiment and improve our project design.On the other hand, we tried to generalize the project, we propose to reduce the production of TMA by designing enzyme peptide inhibitors thus improving the therapeutic efficiency, which will provide guidance for future iGEMers in the therapeutic works.

Safety and Security

In our project, we used DH5α and BL21 for gene cloning and expression, and our chassis chose to use a widely recognized probiotics, Escherichia coli Nissile 1917. At the same time, the tmd and dmd genes we used are from Mesorhizobium and Hyphomicrobium, which are not pathogenic. Meanwhile, our genes such as Tes4, aprN, frmA and frmB are not harmful to organisms. Moreover, we design a temperature suicide switch. As the engineered bacterium leaves the body, it switches on the suicide module due to the reduced ambient temperature, killing the engineered bacterium. In addition, the laboratory only authorized teachers and students can enter and exit, and our lab has a special security officer, Chunjiao Xia teacher. At the same time, before the official start of the experiment in the iGEM project, each team member must participate in a safety education conference, and through the corresponding laboratory safety tests online, when operating in the laboratory, strictly follow the laboratory safety operation.

Software Tool

To complete our project, we developed two software tools for different purposes. First, we built Wechat applet to add functional modules such as "about us", "science about thrombosis", "project introduction", "message board",these functions can lie in the communication between patients and let more people know about iGEM in China.this will achieve psychological care and help cardiovascular patients recover as soon as possible. In addition, for our hardware, an application was developed that can control the hardware and interact with the user in real time to adjust the treatment dosage.