In order to further popularize the knowledge of synthetic biology and let more people understand our project, we carried out popular science activities.
The students from Harbin No. 6 High School and Tianjin Xianshuigu No.1 High School.
We prepared customized gifts and the opportunities to visit our lab for the students who answered our questions.
·Teacher: Dr. Pengchao Wang
·Undergraduate: Danjie Song
Since the teaching plan and organization form of the two lectures with Harbin No. 6 Middle School were similar, we recorded the whole process of the interaction in detail this time. We also had fruitful interaction with Harbin No. 6 Middle School last year, which strengthened our determination of long-term collaboration.
First of all, we got in touch with the director of the teaching affair of Harbin No. 6 Middle School at the end of June. Due to the COVID-19 pandemic this year, we used the Tencent conference app to carry out an online discussion and proposed that we would present our project to the first grade students online after the national college entrance examination. After many times of coordination, the presentation was scheduled at 3:00 p.m. on July 21, 2020. The purpose of this presentation was to publicize the knowledge of synthetic biology, share the information of our project in 2020, and discuss the related contents of our project design with the high school students. Our instructor, two directors and several teachers of Harbin No. 6 Middle School also attended the online meeting.
At the beginning of the meeting, Dr. Pengchao Wang, our team instructor, presented the theme of "Synthetic Biology -- Breaking the Boundary of Imagination" to the students. Dr. Wang shared knowledge related to life science and synthetic biology, such as the time relationship between earth formation and life emergence, the evolution of living organisms, human cognitive process and its application in other living organisms, the origin and definition of synthetic biology, and the applications and prospects of synthetic biology.
In addition, Dr. Wang compared the earth history to the time of a day and discussed the long evolutionary process of living organisms and how human beings know and use other living organisms. Dr. Wang then introduced to the students the world of synthetic biology by pointing out that "the best way to prove that you know something is to make it yourself." Next, Dr. Wang took Lego bricks as an example to let students understand the practice of synthetic biology, so that high school students can understand the concept of biological bricks more vividly. Finally, Dr. Wang concluded with the applications and prospects of synthetic biology to motivate students' interest in biology.
After Dr. Wang’s introduction, Danjie Song, a member of our team, explained the origin of the iGEM contest, and its academic goals and its international reputation. She also encouraged students to learn more about the iGEM Contest in the field of synthetic biology in their high school. After that, we described the design and basic biological principles of our project for 2020. The students were very surprised to the current situation and number of landmines worldwide, and showed strong interests in this topic in the following discussion.
After the description of each section, we would initiate a targeted discussion. During the discussion, we found that the students in the Senior One were highly active in thinking and interactive. After a short discussion, we went on to describe the actual system of the project, from the mechanism of our installation to the final mathematical model. We used relatively concise and understandable description to present our project. We also discussed the function and operation mode of the hardware system and how to apply the mathematical analysis of the hardware device to the project. In order to motivate the students’ enthusiasm of participation, we also designed a rewarding mechanism. For example, we promised to invite the students who actively answered questions to visit our lab after they returned to school, and handed out relevant souvenirs. During the discussion, we interacted with a number of students and got the sparks of thinking. We indeed felt the interest and yearning of the senior high students in synthetic biology.
Dr. Wang: If we compare the history of the earth to 24 hours in a day, when should human beings appearred?
Shuyang Zhao: About the last minute.
Dr. Wang: Humans appeared at about 11:59 p.m., the last minute before the end of the day, just a group of primates who had just learned to stand upright. In the last 0.1 second (4000 BC), the first civilization in human history appeared in Shinarin the Middle East.
Zihan Wang: What is the detection range of this device?
Danjie Song: It can reach the order of 1 microgram, which is about 1.4 µg/L.
After the presentation of the main content was completed, the students were still actively asking different questions about biology and expressed great interest in synthetic biology. Finally, we expressed sincere gratitude to the students and teachers who watched our presentation, and took a group photo.
1.Kezuo Houqi Primary School
Our goals include to provide students with relevant knowledge of biology, inspire students' intellectual thinking, broaden students' scientific horizon, motivate students' research interests, and guide students to engage in further discussions. Our major approach is the combination of the actual local environmental situation and microbial specialties.
We have prepared customized gifts for students who answered our questions. We cooperated with the Paipai education assistance group of Northeast Forestry University, and gained a lot teaching experience during these activities.
In the first part, Yangjinghui Zhang from the NEFU_China team shared the theme of “microorganisms in life”.
At first, Jinghui described some food that related to microbes, especially, the regional snacks of Inner Mongolia, to illustrate that microbes are very close to our life.
The second part is the introduction of "the things about landmines" to the students. Jinghui played the classic puzzle game, Minesweeper, with children, and thus introduced the optical biological landmine detection sensor. Then, he gave a brief introduction of its biology, hardware and working principle.
Next, Yifan Bian described to the students about the rough distribution of landmines and their threat to human beings. To help the students understand the knowledge of landmines. Yifan introduced the history, hazard, utilization and classification of landmines.
At last, Jianing Hu gave a speech on "War and Peace", to illustrate the misery that the wars have brought to the people, inspire the children to study diligently, and motivate them to contribute to the country in future. At the end of the presentation, the students were still actively asking various questions about biology. Finally, we expressed sincere gratitude to the students and teachers who participated in our presentation.
In order to broaden students' horizons, cultivate their interest to science, stimulate their curiosity, and guide them to explore and think more deeply relying on the knowledge of biology, we carried out popular science activities.
We have prepared customized gifts for students who answered our questions.
Danjie Song, Zishuo Xu and Jianing Hu.
At the beginning of science popularization, Danjie Song from our NEFU_China team shared the theme of "biological wonder". First of all, Danjie gave examples of some food related to microorganisms and food additives used to prevent food corruption around us, to illustrate that our microorganisms are closely involved in our daily life and human beings have begun to use microorganisms in various production industries.
After that, Danjie guided the students to think about additional applications of microorganisms in a divergent way, which led to the optical biological sensor based on microorganisms for landmine detection. Danjie briefly introduced its biological system, hardware and mechanistic principle.
Next, Zishuo Xu from our team described the general distribution of landmines worldwide and their hazards to help the students improve their understanding of landmines. Zishuo explained the history, hazards, purpose and classification of landmines.
Question & Discussion:
A student: Are there many landmines left underground ? Are they from the previous war?
Danjie Song: Most of them were buried during the previous wars for the purpose of national territory protection.
A student: What is the explosion range of a landmine?
Danjie Song: The explosion range of a landmine depends on how it is made and what it is made of. For example, a fragment landmine can cause human injuries over 60 meters away. Many landmines have generally high lethality.
A student: Does stepping on a landmine really cause its explosion?
Yifan Bian: The principle of triggering most landmines is through pressure contact. Generally speaking, stepping on a landmine will trigger it to explode even if the person does not move. However, if the landmines are buried for a long time, they may have been naturally disabled and will not explode, but they still have considerable risks.
Finally, Jianing Hu from our NEFU_China delivered a speech on the theme of "war and peace". As a result of the war, mothers and children were separated, families were broken, and young children took on the responsibility of the whole family. There were so much mourning and death during the wars and nobody could comfort the victims. Although children cannot do too much to prevent all unjust wars, they can be filial to their parents, strive to grow up and contribute to the construction of their own countries.
In the initial stage of our project design, we conducted a questionnaire survey. This is to optimize the preliminary design of the project, provide direction for our project promotion and the follow-up human practice, and generate reference materials for future project development.
We have released questionnaires in both Chinese and English using the Microsoft questionnaire platform, and spread them through WeChat, QQ and other apps. So far, we have collected about 700 completed questionnaires.
According to the education levels of the surveyed people, we designed 16 questions, covering many aspects of the project to improve the understanding of landmines and synthetic biology among different groups, and to get public feedback and suggestions on our project design. We would like to collect information from different populations regarding their knowledge of landmine’s hazards and their views on the existing landmine detection methods. At the same time, we would like to know the public's suggestions, focuses and concerns to the core of our project, such as survey objects, signal collection, etc., to further optimize our project. In addition, we hope to investigate the levels of public understanding of synthetic biology and engineered bacteria, as well as popular forms of publicity and education, so that we can carry out follow-up publicity and popularization work accordingly.
Result Collection and Analyses:
We surveyed people of all age groups in a variety of industries, most of whom have at least a bachelor's degree.
Firstly, we investigated the public's understanding of synthetic biology. According to the survey, the public have a certain level of understanding of synthetic biology. However, their knowledge is mostly limited to the applications of engineered bacteria in disease treatment, agricultural production, etc., but rarely on the topics of synthetic biology. Therefore, we expanded our publicity, especially the potential applications of synthetic biology in many emerging fields, such as explosive detection, environmental treatment, information engineering and so on. According to the survey results, we chose to use popular science videos and articles in the propaganda and online platform to let more people understand synthetic biology and its applications in the emerging fields.
Secondly, we investigated the public views on landmines and their current detection methods. The respondents generally believed that landmines had a great adverse impact and became an urgent international problem. Meanwhile, according to the survey, the public is not satisfied with the current approaches of landmine detection. They are also concerned about a series of problems of traditional landmine detection methods, such as interference of non-mine targets, high cost, and risk of personal safety. As for the use of engineered bacteria in landmine detection, public concerns focus on the environmental impact on engineered bacteria, weak signals and poor accuracy. Therefore, our project design should be committed to creating a safe, stable, low-cost, accurate and efficient biological landmine detection technology.
In the hardware design module of our project, we use the unmanned aerial vehicle (UAV) delivery device and make a heat map through the feedback signals to intelligently plan an optimal demining route, which will ensure the accuracy of landmine detection and the safety of demining personnel to the greatest extent. According to the results of the questionnaire, we choose to place the engineered bacteria in a device with a size of a can, which not only provides suitable growth conditions for the engineered bacteria at an acceptable cost, but also facilitates the subsequent signal receival and processing, and the recovery of the engineered bacteria.
In addition, to improve the accuracy and signal feedback, which is the key in bacterial mine detection, we chose to detect chemical explosives, which is the core of landmines, so as to reduce the interference of non-mine targets. Aiming at the problem that the public is most worried about: "the feedback signal from the bacteria is too weak and difficult to be detected." Through the communication with the relevant professors, we chose to reduce the detection threshold of the promoter and improve the detection sensitivity through the phage-assisted directed evolution. At the same time, we designed to transform the fluorescence signal generated by the engineered bacteria into electromagnetic wave signal, similar to the WiFi, which can expand the bacterial luminous signal, identify accurate location and forward to the app for intelligent processing. These methods are generally accepted by the public, which gives us the confidence to move forward in the process of our project.
At the end of the questionnaire, the importance ranking of the stakeholders gave us the direction to advance human practice. Based on the survey data, we decided to interview relevant government agencies, mine clearance officers and soldiers, and received recognition and many pertinent suggestions.
Professional Data Analysis(Click View Details)
① Multiple response analysis:
When analyzing the results of multiple-choice questions, we used "0-1" coding, which is to set a separate variable for each option. In the SPSS analysis module, select multiple responses, define the variable magnitude, re-encode the options, define them as binary values by the dichotomy, and define the selection options as "1". If not, the default is "0". Then, define a new variable order of magnitude, rename and add it, and the system will add a "$" symbol before the multiple response.
After entering the SPSS analysis-multiple response, first carry on the frequency analysis, and then select the multiple response level. The analysis results of copy 1 are given below as examples.
·The landmine survey methods you know are (multiple selections) ：
The cross-analysis of landmine survey methods understood by people studying science and liberal arts.
As far as you know, the current application of synthetic biology (multi-selection):
The cross-table analysis of consideration that whether it is necessary to convert the direct response biofluorescence signal into other signals received by the two groups of people who understand the current application of synthetic biology.
·In which ways do you want to know about us, synthetic biology and landmines? (multiple selection)
The cross-table analysis of What are the ways in which people of different age groups learn about us, synthetic biology and landmines.
② AHP Analytic hierarchy process (empowerment) :
The ranking of importance may not only directly show the respondents' attitude to each question, but also rank the options according to the frequency and importance of their responses. The AHP analytic hierarchy process is used to analyze the data onto the problems existing on adaptive engineered bacterial mine detection and the ranking of stakeholders in biological mine detection. After entering the data, the abnormal value of the data is processed; all the options are selected, "missing number" and "number < 0" in the SPSS are selected, and the exception processing item is set to Null, to determine the processing.
The analysis results of the copy 1 are given below as examples.
Through frequency analysis, we can conclude that, among the problems on the use of engineered bacteria for landmine detection, the mostly concerned option from a large surveyed population (125 people) is that the feedback signal of bacteria is too weak and difficult to be detected, and also, it is considered that the high cost is the least important issue in the problems of using engineered bacteria for landmine detection.
In the issue of considering stakeholders in the field of biological landmine detection (any relevant person affected by organizational decisions and actions in the external environment of the organization), the first option selected by the largest population is the minefield residents. The largest number of people think that the interests of landmine detection device manufacturers are the least relevant.
In the survey of the current impact and severity of landmines on us, the preferred option of the largest number of people is "the chemicals contained in mines enter the water and are at the risk of causing cancer"; meanwhile, respondents generally believe that landmines have a direct or potential impact on most people.
We used the weight assignment method to rank importance. Because the selection order is different, its importance is also different. In the analysis, it is necessary to assign a weighted number to each numerical value. That is, the first numerical weight > the second weight > the third weight > the fourth weight. The most conventional method of assignment is to score the data in reverse: for example, there are four rankings, four points in the first place, three points in the second place, two points in the third place, and one point in the fourth place. Use the SPSS data encoding function, select all the options, set the score, confirm the coding, and describe and analyze the results after the setting is successful.
In the importance ranking, the obtained average value can represent the order in which the items are selected. The selection with the highest average score in these questions is related to in the use of the engineered bacteria in landmine detection. The concern is that "bacterial feedback signal is too weak and difficult to be detected", which has the highest average value and top ranked. The frequency order of these selections is difficult to detect > affected by environmental factors > poor detection accuracy > may lead to problems, such as biological pollution > high cost.
In the investigation of the current impact and severity of landmines on us, the average score of "chemicals contained in mines enter the water body, reduce water quality, and have a risk of causing cancer" is top ranked. The order for importance is that the chemicals contained in the mine enter the water body, reduce the water quality, have carcinogenic risk > accidentally enter the minefield, and are likely to be blown up > the chemicals contained in the landmine destroy the soil. Wasting a large area of land > it has no effect on most people.
In the survey of stakeholders in biological landmine detection (anyone affected by organizational decisions and actions in the external environment of the organization), the option of the highest average score is minefield residents. The order for interest correlation is minefield residents > government and related military institutions > minesweeping officers and soldiers > mine detection related researchers > biologists > mine detection device manufacturers.
In addition to our usual public communications and campus publicity activities, we opened a number of multimedia accounts to record the progress of our project, which will increase our influence and report our events. In this year, in addition to daily posted essays through our WeChat Public account, we also actively use twitter, bilibili, facebook and other public media post our articles.
Among them, the general history of landmines in the WeChat has been recognized by many faculty members of our university. Through literature review, we constantly supplement the professional knowledge of landmines history, which not only helps our team members to better understand landmines, but also promote our readers and students in the publicity school to have a more profound understanding of our project. Reaching out to the public through social media platforms is an important part of our project. Through the cooperations and conference communications with other iGEM teams, we published relevant updates on multiple platforms. A series of tweets also let many of our partners know more about our project development and the relevant progress of HP in our team. In today's fast-paced era, online reading is a convenient way to learn new knowledge. Also, making good use of new media tools is the key to access the international world stage, which is also an effective way to promote synthetic biology and expand the scope of our publicity. We will continue to innovate and expand our reach to bring synthetic biology to the forefront of biological research.
Mines, a comprehensive
As a special promotion of our project, we planned to release a resource for the public to help people understand the background of our project. Landmines were chosen as the main focus of our project. For this purpose, we wrote a book, "Mines, a comprehensive", which describes the history and evolution of landmines from their invention to the current situation in detail. The book is divided into three volumes of the ancient, modern and contemporary. It will serve as the further information of landmines and hopefully change the superficial perception of landmines among the general public.
Since our project was related to landmine clearance and detection, we designed a game named as questionnaire survey, which is associated with the classic minesweeper games. In order to maximize the simulation of our modeling and data analysis, we designed the rules of Minefield Elite differently from the traditional game, Minesweeper.
1. Clicking on a cell to show the total number of landmines in 3 x 3 with 9 cells including the clicked cell.
2. Clicking on a landmine that does not explode, leading to the end of the game.
3. Mines can be marked with red flags by right-clicking on the grids.
4. The game ending is determined by the number of planted red flags (i.e. the number of marked landmines) when the number reaches a preset number of landmines.
5. The game will count the game duration, number of clicks and number of correct red flags, which are used to calculate a final score.
Just like our hardware devices used in landmine detection, a player clicks on each frame just like that a device is dropped into a minefield. For each click, the display is the signal feedback. Since dropping devices is obviously not realistic in the game, we designed the game that counts the clicks and time duration used by the player and incentivized them to find the most landmines with the fewest devices.
We have uploaded the game to the GitHub and propagated it through WeChat, Facebook and other self-media platforms. We have also uploaded the game to our wiki, so everyone can play the Minefield Elite. In addition, the game has an internal statistic function allowing us to count its use. As of October 19, 2020, we had 977 engagements with the game.
Science is endless. To get useful suggestions, we had an online meeting with an officer from the Mine-Clearing Force, and learned some very stringent conditions for landmine detection in real scenarios. The officer indicated that our device would be eventually used in open fields or even in battlefields. Thus, we tested long-term preservation of the engineered bacteria and their responsiveness to DNT induction in simulative open field conditions. We were also connected to the current Conference of China iGEMer Community. Through extensive discussions with the judges and other iGEM team members, we collected many useful ideas, some of which were used to improve our project.
Currently, we created our reporter system with EGFP as a reporter gene. However, the green fluorescence emitted by the EGFP protein needs an excitation light, which has been added in our system, but this increased the complexity of the hardware of our device. Thus, we considered whether we should employ an autofluorescence system to simplify the design of our reporter system. After thorough discussions and literature research, as well as incorporating the expert’s suggestions, we decided to explore the Lux luciferase system. To date, we have tested the Lux bioluminescent system and obtained promising preliminary results.
The Lux luciferase system consists of five LuxABCDE genes. Among them, The LuxAB genes encode the Lux luciferase, while the products of the LuxCDE genes can convert long-chain fatty acids to long-chain fatty aldehydes, which can be used as substrates of the Lux luciferase to generate bioluminescence. Thus, we constructed a reporter plasmid with the third generation (yqjF3rd) of the yqjF promoter driving the expression of the Lux and test its responsiveness to DNT induction in bacteria. As shown in figure, the yqjF3rd-LuxCDABE system showed a DNT detection threshold of 5 mg/L. We are currently conducting experiments to improve the Lux reporter system.
Our team has established active collaborations with the QHFZ_China team who provided us the CAHS protein of the Water Bear, which has been proved by them to improve the resuscitation of bacteria stocks. We tested the effect of the CAHS protein on the resuscitation of our engineered bacteria after they were freeze-dried into powder. At the same time, we tested long-term preservation of the engineered bacteria and their responsiveness to DNT induction in simulative open field conditions.
Finally, we wanted to test the responsiveness of our engineered bacteria to DNT released from soil in an open field scenario. We immobilized the bacteria in the alginate gel and loaded the gel onto the soil mixed with different amounts of DNT. As shown in figure, the immobilized bacteria could steadily detect the DNT released from the soil.
Based on these tests, we conclude that the engineered bacteria can be resuscitated and then used to detect DNT released from the soil in an open field. Thus, our project represents a connection between the synthetic biology in laboratories and its applications in natural environment.
The sensors of our device can determine the location of a landmine by sensing its volatile substance, DNT. We also think that the system employed in our project can detect other substances if the promoter is accordingly revised. Firstly, our device can be used to detect metal ions in the soil. The system can be modified to detect the amounts of metal ions in soil. Secondly, we may replace the current promoter with another promoter(s) that is sensitive to drug metabolites. The generated device can be deployed in drainage systems or even rivers to detect drug metabolites, which may aid in the detection of drug addicts or pollutants. Therefore, we believe that the concept of our biosensors has a promising future in the field of chemical detection. We also expect that our project system can be applied to other fields to promote the development of novel technologies.