After defining the scope of the project, we conducted a public questionnaire survey that examined the level of public awareness and acceptance of biosensors and CRISPR technologies to assess the feasibility of our project. The survey involved 500 participants from various backgrounds and occupations, with the majority being enterprise staff and students.

We found that more than 50% of the respondents were familiar with biosensors, and their knowledge of the application of biosensors varied. Some participants had used biosensors to monitor their health indicators, while others only had a conceptual understanding without practical experience.

We also encouraged participants to envision the future possibilities of biosensors. Most participants believed that biosensors would have widespread applications in daily life and expressed their willingness to adopt them to their everyday lives and health care. Therefore, we initially aimed to utilize biosensors as a powerful tool to monitor physical conditions and make informed health decisions.

We then assessed their perception of the key technology employed in our project: CRISPR/Cas9. 60% of the respondents demonstrated an understanding of CRISPR technology. We were pleasantly surprised by the public’s comprehension of scientific advancements. Moreover, nearly three-quarters of the participants agreed with our project and showed optimism about storing biological signals in DNA. They recognized the stability and potential benefits of this new generation of technology.

The questionnaire survey revealed that under the context of the vigorous development of the synthetic biology industry in China, the public has a knowledge and acceptance of biosensors.

Next, we interviewed stakeholders from different fields, and investigated the design of CRISPReporter as well as its application in different fields, aiming to improve our project and promote the development of the biosensor industry.

A. Wet lab

a. Selection of photosensitive promoters

In the initial stage of the project design, we consulted with Professor Fei Gan from Wuhan University and Professor Andreas Möglich from the University of Bayreuth on different signal sensing methods.

Through consultation with the two professors, we gained a detailed understanding of the principles and application of different light induction systems. From a literature review introduced by Professor Gan, we learned that the EL222 system can precisely control the level of gene expression in Escherichia coli Top10 by adjusting the intensity of blue light pulses, with less metabolic pressure. Therefore, we adopted EL222 as the signal input design, which proved to be suitable for our system in subsequent investigations.

What's more, following by professors' advice, we also determined to perform directed evolution on the promoter to reduce leakage and increase sensitivity, to achieve precise recording.

b. CRISPR technique

Our project is designed to adopt the CRISPR technique. However, we are still confused about how to select and flexibly apply the multiple derivative technologies of CRISPR at the molecular level. Therefore, we visited Professor Ying Zhang from Wuhan University for some guidance.

Professor Zhang systematically analyzed various CRISPR types from a global perspective and offered us some suggestions. At the same time, the professor believed that the core technology used by our project is essentially homologous recombination, which has some shortcomings in the diversity of signal storage. In addition, Lambda homologous recombination technology has the risk of abnormal recombination during plasmid replication, potentially leading to signal recording disruption.

During our discussion with Professor Zhang, we realized that CRISPReporter may still have deficiencies in project positioning and technical foundation. To assess the gravity of the issue raised by the professor, we designed confirmatory experiments on the system, which you can find on the Results page.

B. Dry lab

Producing a large number of high-quality sgRNAs is essential for building an efficient signal recording system. In the early stage of the project, we consulted Professor Lihua Zhang of Wuhan University on the gRNA optimization model. Professor Zhang patiently discussed the selection of the screening method with us. The professor also offered guidance on positioning, research significance, and presentation form of our model.

We were in need of a large number of high-quality sgRNAs that are widely applicable to different biological chassis. We learned that most of the existing research results are based on deep learning methods to predict the activity scores of a certain species. Considering the existing research foundation and the overall characteristics of the project, we finally positioned the numerical simulation software part as "providing pre-screening and heuristic suggestions for biological experiments".

At the same time, Professor Zhang also provided us with some suggestions on method design, reminding us to pay attention to the collection and selection of data sets. The professor inspired us that, in the context of cross-fusion, we can report the results of our model by comparing it with public databases, or by conducting contrast experiments with randomly identified sequences in wet experiments, achieving deep learning correction of the model.

C. Application

a. Diagnosis

We noticed that some chronic diseases with recurrent episodes, such as inflammatory bowel disease (IBD), require continuous monitoring after diagnosis. This is consistent with the multi-level recording system in our project design. To further explore the potential applications of CRISPReporter in the detection of IBD and other diseases, we decided to pay a visit to Dr. Fei Liao, a chief physician and an IBD expert at Renmin Hospital of Wuhan University.

Dr. Liao pointed out several challenges that need to be addressed before applying the engineered bacteria to clinical practice. These challenges include assessing the sensitivity of the induction markers, evaluating the colonization ability of probiotics in the gut tracts, and ensuring detection specificity for different types of IBD. He recommended that we concentrate on ulcerative colitis as our target disease, as it is more feasible to test in animal models. Additionally, he emphasized the importance of seriously considering human safety and ethical aspects when contemplating the application of the CRISPReporter system for human use.

Finally, Dr. Liao shared his insights on the future development of the CRISPReporter system. He pointed out that our engineered bacteria could be more than just a diagnostic tool, but also a therapeutic agent that could achieve the integration of diagnosis and treatment. Through the exchange and discussion with Dr. Liao, we gained a deeper understanding of the clinical applications and future prospects of the CRISPReporter system in the diagnosis field.

b. Environmental monitoring

Environmental protection and ecological conservation have become increasingly important issues in the modern world. Among them, the detection of various pollutants in the environment is a key challenge.

To better understand the potential applications of CRISPReporter in the field of environmental monitoring, we consulted Professor Chaoqi Chen in the College of Resource and Environmental Sciences, Wuhan University. Professor Chen showed great interest in our system and pointed out its advantages: it can simultaneously detect several different environmental pollutants with less sample pretreatment compared to conventional methods. However, he also raised some questions that need to be addressed before applying the engineered bacteria to practical scenarios, such as the sensitivity, the cost, and the safety of the system. He also mentioned that certain methods for detecting environmental pollutants are well-established and cost-effective. Evaluating whether our new method can compete with these existing approaches is of utmost importance.

Furthermore, Professor Chen suggested that we design a system targeting a specific emerging pollutant, such as antibiotics, as it might have an easier path to enter the market. Based on his advice, we decided to focus on macrolide antibiotics, a new type of pollutant. Learn more on Application

A. Shenzhen Synthetic Biology Infrastructure

To broaden our horizons, learn about cutting-edge synthetic biology technology, and improve the completion of our project, we visited the Shenzhen Synthetic Biology Infrastructure.

The building contains both laboratories and corporate offices, as the builders of the building said, "The scientists upstairs can visit three companies in one day, and maybe a problem will be solved in the elevator."

The tour to Shenzhen Synthetic Biology Infrastructure made us realize that we are living in the world of bio-engineering and the era of synthetic biology. The biology of the future is highly reproducible and high-throughput. In this building, thousands of biological experimental robots can perform the work in just a few days, which may cost millions of scientists more than a year in the past. These infrastructures tremendously free manpower from heavy and repetitive labor. It inspires us to explore the future of CRISPReporter that we can use robots and artificial intelligence to create biosensors to meet our needs.

What we are operating are not screws or gears, but deoxyribonucleic acid and protein.

B. Center for Inflammatory Bowel Disease Research and Diagnosis

Our initial aim is to develop a novel noninvasive method for diagnosing inflammatory bowel disease (IBD) based on CRISPReporter, which can record multiple episodes of IBD patients. To gain a more tangible understanding of the current developments in IBD diagnosis, we visited the Center for Inflammatory Bowel Disease Research and Diagnosis. During the visit, we learned that although the clinical diagnosis and treatment methods of IBD are constantly updated, there are still many deficiencies:

• The etiology and pathogenesis of IBD are still unknown, leading to a lack of targeted and personalized treatment.
• The new treatment method for IBD is still in the clinical trial stage, and its long-term effect needs to be clarified.
• There is insufficient patient education and psychological support for IBD, which leads to low awareness of IBD and affects the treatment outcome.

This visit made us realize that the improvement of IBD diagnosis needs to be conducted from multiple aspects, including strengthening basic research, improving diagnostic criteria, and enhancing patient education and psychological support. It also inspired us to make CRISPReporter into a recorder that can provide long-term personalized monitoring for patients, which has considerable prospects.

The Sustainable Development Goals (SDGs) are a set of 17 global goals established by the United Nations in 2015. They are designed to address various critical issues facing the world, including poverty, hunger, health, education, equality, climate change, environmental protection, and more, to achieve global sustainable development.

All SDGs are interrelated and mutually supportive, and we decided to focus on two specific ones that our project most contributed to: good health and well-being (SDG 3), and clean water and sanitation (SDG 6). As WHU iGEMers, we are concerned with these worldwide problems and their impacts on human health and the environment. Our project may offer a novel approach to these challenges- we wish to do our bit to make the world better!

Our survey on the application of CRISPReporter revealed that most participants believed CRISPReporter would have widespread applications in daily life. They expressed their willingness to adopt them to improve their lifestyles and health. Inspired by the survey, we focused on Inflammatory Bowel Disease (IBD), which can cause symptoms such as diarrhea, blood in stool, abdominal pain, and fatigue.

We aim to develop an innovative non-invasive diagnostic method for IBD based on engineered bacteria capable of recording multiple episodes of IBD. (see Applications page). We hope to help patients with IBD overcome their illness by providing a more convenient, affordable, and efficient approach to monitoring their condition. Moreover, the specialist we interviewed expressed his interest and appreciation for the scope and positive impact of our project on the diagnosis and management of IBD, meaning that our project contributes to building good health and well-being.

Nowadays, not only traditional pollutants like heavy metals and pesticides but also novel pollutants like antibiotics and prescription medication are threatening people’s health. To improve the quality of life and promote health equity, it is of great significance to monitor water pollutants, improve water quality, and manage water resources.

To address these problems, we intend to apply CRISPReporter to environmental monitoring. Our environmental detection chassis will be encased in a specialized material and deployed into the aquatic environment for real-time monitoring. In this way, we can monitor water efficiently and economically (see Application page).

The experts in environmental pollutant detection we interviewed told us that our project contributes towards better societal acceptance of biocontrol, contributes towards better societal acceptance of biocontrol, as it can long-term monitoring detect several different environmental pollutants with less sample pretreatment compared to conventional methods.

We have gained valuable insights from various stakeholders, who confirmed the demand for CRISPReporter to address these goals. We aspire to use synthetic biology to solve more SDGs and make a positive impact on the world. Our tool is well-documented on our wiki, which enables future teams to improve its performance. Our tool can also be adapted to many different scenarios, offering possibilities for diverse users and aims.

WHU-iGEM launched the Plasmid Marathon, which uses plasmids to introduce synthetic biology to more people while enhancing their physical fitness. The process of running is actually the process of plasmid construction. At each checkpoint, participants obtain information about the corresponding fragments by completing game tasks and then assemble the fragments into a complete plasmid at the endpoint. The Plasmid Marathon allows each participant to gain a deeper understanding of plasmid construction while doing aerobic exercise, truly achieving the goal of combining education and entertainment.

On April 25, 1953, Watson and Crick published a paper titled "Molecular Structure of Nucleic Acids: A Structure for Deoxyribose Nucleic Acid", which was hailed as "a milestone in biology, ushering in a new era". In 1962, they received the Nobel Prize in Physiology or Medicine along with Wilkins for their discovery of the DNA double helix structure. However, Franklin, who also made significant contributions to this discovery, died of ovarian cancer in 1958 and lost the recognition she deserved.

To commemorate the 70th anniversary of the discovery of the DNA double helix structure and to honor Franklin's remarkable achievements in various fields, WHU-iGEM organized the DNA Marathon. We set up a booth to promote Franklin's spirit and to inspire young people to seek truth from facts. We designed a special Franklin metal badge and distributed it to the participants. The badge was also sold online and received an overwhelming response.

Our activity attracted synthetic biology enthusiasts from all over the country, who shared their photos with the Franklin badge and their scientific research life on social media.

The DNA Marathon was not only an opportunity to revisit history with a dialectical and inclusive perspective, but also a Marathon of the spirit of seeking truth from facts in scientific research.

To commemorate the 20th anniversary of iGEM, we organized the WHU-iGEMarathon to explore the development of iGEM and synthetic biology in the past two decades. We invited the senior members of WHU-iGEM and the alumni who are engaged in synthetic biology to give advice on our project and share their valuable experiences and insights with us. The senior WHU-iGEM members pass on their knowledge and passion to the younger generations, inspiring every WHU-iGEM participant with their own stories and fostering the communication and collaboration of brilliant ideas.

Sun said that when he organized his team to participate in iGEM, completing a project that seems rather conventional nowadays was already a very exciting thing. But now, iGEM projects have achieved endless exploration and innovation on the level of components, such as logic gates and oscillators. He pointed out that the basic process of synthetic biology, the DBTL cycle, will have a bigger impact on the future of the synthetic biology industry and the whole biological industry.

Mao mentioned that she thinks the main reason why WHU iGEM won the Best Measurement nomination in 2021 was the meticulous design of the induction gradient in directed evolution, complete data collection, and persuasive demonstration of the improvement of the promoters after evolution. This work also provided us with an excellent example of how to deal with the data related to gradient induction, as well as how to effectively present experimental results.

Liang has extensive experience in applying the CRISPR system to yeast cells. Liang recommended we perform multiplex editing detection from the group. To determine whether the plasmid can be edited and whether our designed system can function properly, Liang suggested that we first select the CRISPR editing site on the genome and then on the plasmid, to distinguish between a system issue or a plasmid editing issue.

Wen has extensive experience in plant developmental molecular biology. In our process of inducing editing, Wen advised us to conduct functional verification of the working feasibility of the CRISPReporter editing system by replacing the Lac constitutive promoter with the T7 before starting induction. This would make our experimental design more refined and more logical.

At the WHU-IGEMarathon, we witnessed the rapid development of synthetic biology over the past 20 years. We saw the innovations and contributions made by our seniors in iGEM, and we also heard their expectations and encouragement for us. We will successfully take over the baton at the iGEMarathon, and contribute WHU's strength to the development of synthetic biology.

At the first stop, we worked with the Nanjing-China iGEM team and went to Nanning No. 3 High School in southwest China. We used a combination of online and offline methods to show our recorded synthetic biology popular science videos and carry out basic biology education.

At the second stop, we worked with LZU-China and went to Lanzhou No. 1 High School in northwest China. We gave lectures on synthetic biology to the students there and demonstrated its various applications.

At the third stop, we went to Zibo No. 7 High School in eastern China and introduced the development of biology to the students. We stimulated their enthusiasm for life sciences and guided more students to pursue scientific research.

At the fourth stop, we went to Wuhan, a central city in China, and introduced the overall framework of contemporary molecular biology to the students of Wuhan No. 2 High School. We demonstrated to them the high level of automation and engineering present in the field of biological sciences in the 21st century.

Finally, WHU-China introduced the content, significance, and charm of the iGEM competition to the freshmen who had just entered Wuhan University. We aroused their interest and curiosity in synthetic biology. We were very happy with their enthusiastic feedback. It was through the continuous influx of new students that this interdisciplinary field could develop and innovate rapidly.

This Education trip was a valuable opportunity for us to communicate with younger students. The exchange with young students made us impressed by their infinite creativity. For us young students, only by making original and pioneering progress can we further promote the rapid development of synthetic biology.

SynBio Hourstory is a synthetic biology history popular science activity jointly initiated by eight iGEM teams. It aims to use comics as a simple and beautiful way to popularize the major events of synthetic biology development to the public.

The main characters are:

Gemi represents human curiosity and the personification of iGEM.
Coli leads Gemi into the world of microorganisms and synthetic biology.

Our popular science content includes:

WHU-China: 1953 - The discovery of DNA helical structure
OUC-China: 1957 - The establishment of the central dogma
LZU-China: 1965 - The artificial synthesis of bovine insulin
HainanU-China: 1970s - The discovery of plasmids
UCAS-China: 1990s - The Human Genome Project
GXU-China: 2002 - The artificial synthesis of poliovirus genome
BNU-China: 2003 - The engineering of artemisinin precursor pathway
Tianjin: 2017 - The artificial synthesis of yeast chromosomes

Download comics PDF

In addition to comics, we invite readers to participate in the "Did you learn it?" section. We will attach simple multiple-choice questions at the end of each post to consolidate the knowledge that this comic brings to you. At the same time, the discussion topics for each issue will be timely published on Quora, inviting readers to exchange views with each other. We have collected many really surprising answers.

To arouse the interest of the general public, especially youngsters interested in Science, iGEM Teams from the Hong Kong University of Science and Technology (HKUST) and Wuhan University (WHU) are launching iGEM SynBio Online, a Podcast Series to share stories of distinguished Synthetic Biology researchers, experienced iGEMers and experts in the industry. The Series would be a great educational program to provide enrichment for budding synthetic biologists like us.