Overview ​

We believe that Human Practices work serves as an important bridge connecting laboratory research with the human world. Therefore, throughout the overall development of our project, we have closely integrated Human Practices with experimental work and applied our understanding of the 3R principles within the Human Practices framework.

Milestone ​

To clearly demonstrate how our HP activities were integrated with the project design, we highlight the key milestones of our HP work.

The Key Milestone of our HP

Our logic in Human practices ​

About our stakeholders ​

During each key milestone, we were guided not only by scientific principles but also by reflections derived from linking our work to the real world through Human Practices. Engaging with various stakeholders who provided valuable insights into their needs helps develop an effective and responsible design to the real world. Therefore, to effectively respond to different types of stakeholders, we categorized them into four groups:

Interviewing with Marine Aquarium Landscape Store Owner

Following these four categories, we successfully identified the key needs of each stakeholder and responded to them through careful consideration of balance, both in our project design and future plans. Additionally, to ensure effective interaction with stakeholders, we refined the classic AREA framework, using it to define the evaluation dimensions of our practices.

About our framework ​

AREA framework is classic and universally used in iGEM HP design. However, we found that the initial AREA framework focused more on the entire project instead of specific events, which encouraged us to do modification according to our need of closing the loop and defining evaluation aspects for every HP activity. Therefore, we modify AREA framework.

Integrated Human Practices ​

Background Investigation ​

In the initial phase of our project, we conducted extensive literature reviews on corals and bleaching, gaining insights into endangered coral systems and their vital connection to humans. To gain a deeper understanding of the realistic impact of coral bleaching on the lives of specific stakeholders, we plan to visit various coral-related industries in Guangzhou for in-depth research.

Name: Interview of Marine Aquarium Landscape Store Owner

Key words: industrial profitability, education need

Expectation: The interview aimed to assess how environmental changes affect the coral supply chain, the owner's understanding of coral ecosystems, and their views on conservation efforts and bleaching relief. We also explored more opportunities for collaboration in education activities.

Description: The owner noted that most corals are now sourced from Indonesia due to lower bleaching impacts, while Australian corals, especially certain species, are harder to obtain. He also emphasized the importance of blue light for coral growth in aquarium systems and shared experiences from diving, highlighting the negative impact of inexperienced divers on coral health, which inspired him to focus more on coral education and conservation, reinforced through social media outreach.

Reflection: The interview highlighted how coral bleaching is impacting the industry's product availability and profitability. And there was the need for more education on coral conservation to prevent damage by inexperienced handlers.

Adjustment: The discussion emphasized the industry's reliance on coral health and the broader significance of bleaching relief, which indicated the importance of coral conservation and our project. We also plan to deepen our research and explore collaboration to raise awareness through social media.

Interviewing with Marine Aquarium Landscape Store Owner

Name: Interview of Coral Craft Shop Owners

Key words: "Not the same! "

Expectation: Our interview aimed to understand the impact of coral bleaching on the coral craft market and assess customer awareness of environmental issues, exploring how our project might address industry needs.

Description: Shop owners, each with over a decade of experience, shared insights on market shifts. Despite legal certification, many customers mistakenly associate coral crafts with bleaching, even though crafted corals aren't affected. This misconception has reduced demand for coral jewelry and crafts.

Reflection: Initially, we overlooked the distinction between crafted corals and bleaching-affected corals, highlighting the need for more thorough research before interviews.

Adjustment: The shop owners emphasized the importance of distinguishing between deep-sea corals used in crafts and those affected by bleaching, which we would like to integrate into our education activities.

Interviewing the Coral Craft Shop Owner

Name: First round of questionnaire survey for the public

**Key words: **necessity for education

Expectation: We aimed to understand the public knowledge of corals and their preferred methods of receiving education, particularly regarding coral bleaching and synthetic biology.

Description: This questionnaire focused on what people knew about coral and coral bleaching, and briefly asked the public about their views on synthetic biology.

In our first survey, we asked about public awareness of coral bleaching and the marine environment. Over 60% recognized coral as an animal, and more than 75% correctly identified healthy coral in images. However, while 90% agreed corals need protection, less than a quarter knew about coral bleaching. This highlighted the need for public education on coral conservation.

Therefore, we add a question at the back of the questionnaire, to understand their preferred way of being educated, with the result shown as follows.

Firgure 5. The way you prefer to be educated

**Reflection: **The survey revealed a general lack of understanding about coral bleaching despite awareness of coral protection. Additionally, we also realized that traditional education method, such as regular stalls and science talks, may not be very effective.

Adjustment: Inspired by this questionnaire, we plan to create more innovative and impactful coral-related educational activities. This led to a series of age-specific programs designed to raise awareness about corals and their protection. You can find more details on our Education page. (hyperlink to education page)

Our Human Practices Goals ​

Through extensive background investigation, we preliminarily came up with a sets of goals for our future Human Practices activities following our interpretation of 3R principle within the context of our project.

• Prioritizing biosafety to prevent unintended environmental impacts in the marine system
• Developing an innovative and effective synthetic biology approach to address coral bleaching
• Ensuring the designs of our project and activities taking in diverse needs of various coral-related stakeholders
• Increasing public awareness of coral bleaching and conservation through various outreach activities. -- Education (hyperlink to education page)

We prioritize biosafety to stress the importance of "Responsibility", "reflect" on the real world context to ensure effectiveness and be "responsive" to various stakeholders. Guided by goals integrated with the 3R principle, we set up to step into our engineering cycle.

cycle 0 ​

After investigating coral bleaching mechanisms, we identified Reactive Oxygen Species (ROS) as a key factor causing oxidative stress. To counter this, we designed a genetic circuit in Zooxanthellae to express antioxidants and catalase with a hydrogen peroxide sensor, aiming to prevent oxidative stress in corals and their algae. We then presented this idea to experienced professors for feedback

Name: Interview with Prof. Peiyuan QIAN on chassis selection

Stakeholder Context：

Professor Peiyuan Qian: Professor Peiyuan QIAN is Chair Professor of the Department of Life Sciences at the Hong Kong University of Science and Technology, Director of the Pacific Institute of Marine Science, Director of the Asia-Pacific Institute of Marine Science, and an internationally renowned marine biologist and ecologist. His research mainly focuses on the health and sustainability of marine ecosystems, with a particular focus on the conservation and restoration of coral reefs. Professor QIAN's research on coral reef ecosystems has had a significant global impact, particularly in the study of climate change, ocean acidification, and other impacts on coral reefs.

Key words: Zooxanthellae × => Symbiotic Bacteria √

Expectation: The interview aimed to refine our experimental design by consulting marine bioscience expert Prof. Peiyuan QIAN, who has extensive experience in marine invertebrates and microorganisms.

**Description: **During our discussion, Prof. QIAN emphasized the complexity of the Zooxanthellae genome and the challenges this poses for gene editing. He suggested alternative approaches using symbiotic bacteria, which may offer a more feasible solution. He also expressed interest in further discussing the dry lab aspects of our project in future meetings.

Reflection: Prof. QIAN's expertise helped us realize the difficulties of working with Zooxanthellae and the need for a more practical approach. His insights into the genome complexity were particularly valuable and highlighted the importance of exploring alternative strategies.

Adjustment: Based on his advice, we decided to shift our focus from Zooxanthellae to symbiotic bacteria, as this offers a more achievable path forward. Following his recommendation, we will conduct preliminary experiments by testing the expression of antioxidants and catalase in E. coli to assess the functionality of our circuit. We look forward to further discussions with Prof. QIAN to continue refining our project direction.

Communicating with Prof. Peiyuan QIAN

Name: Interview with Prof. Xiaomeng LI on Zooxanthellae Preliminary Experiment

Stakeholder Context:

Professor Xiaomeng LI: Prof. Xiaomeng LI is an Assistant Professor in Bioscience and Biomedical Engineering Thrust at the Hong Kong University of Science and Technology (Guangzhou). He developed an approach to identify histone modification-involved interactions under physiological conditions and investigated the functions of histone crotonylation. He designed and established a novel chemical proteomic approach to study bacteria-host interactions during infection. Dr. Xiaomeng LI's research interests focus on answering the epigenetic questions involving histone PTMs and protein-protein interactions, by combining diverse technologies including photo-crosslinking, bio-orthogonal chemistry, quantitative proteomics and cellular and molecular biology tenchniques, as well as developing novel chemical biology approaches to tackle the current challenges and further the investiation on such problems.

Key words: simulation experiments, Zooxanthellae

Expectation: After shifting our chassis to symbiotic bacteria, we consulted Prof. Xiaomeng LI to refine our circuit design and experiment setup.

Description: Prof. LI stressed the importance of simulating environmental factors that cause coral bleaching, especially after changing the bacterial chassis. He advised acquiring Zooxanthellae and setting up experiments to determine optimal culture parameters, such as light intensity, wavelength, and temperature, to replicate stress conditions affecting Zooxanthellae in the wild.

Reflection: The discussion clarified the role of environmental factors in coral bleaching and the need to simulate both optimal and stress-induced scenarios for Zooxanthellae. This is critical for testing our genetic circuit's ability to mitigate bleaching effects.

Adjustment: Following Prof. LI's advice, we will focus on replicating the environmental pressures Zooxanthellae face in their natural habitat. Our experiments will assess our system's protective response under stress, providing a foundation for testing the genetic circuit's effectiveness.

Prof. Xiaomeng LI

Name: Interview with Professor Jun XIA

Stakeholder Context:

Professor Jun XIA: Professor Jun XIA is an expert in the field of neuroscience and is currently a Professor of Biology at the Hong Kong University of Science and Technology. His research focuses on synaptic plasticity and the regulatory mechanisms of neural circuits, especially how neuroplasticity can be used to understand learning, memory, and the pathogenesis of neurological diseases. He has published a series of influential papers in the areas of synaptic plasticity and regulation of neural circuits, involving the molecular mechanisms of learning and memory; molecular and cellular mechanisms of neurological and psychiatric disorders, especially Autism, and their relationship to synaptic formation. He has rich experience in molecular biology, cell biology, transgenic animal models and so on.

Key words: mCherry, threshold, re-design circuit

Expectation: To obtain guidance on the logical progression and validation of our biology project, we asked Professor Jun XIA for help.

Description: Although not an expert on our system's specific parts, Prof. XIA advised us to use reporters to monitor protein and RNA expression, such as OxyR's response to hydrogen peroxide, ensuring adequate levels for downstream reactions.

Reflection: Prof. XIA's advice emphasized the need for empirical data on expression levels. We researched hydrogen peroxide concentrations in coral cells to compare with our sensor's threshold, but couldn't find a suitable sensor for bacterial use that met the requirements, prompting us to re-design our circuit.

Adjustment: Following his guidance, we will integrate mCherry as a reporter in our plasmid and measure fluorescence to track protein expression. We're also exploring another target protein to alleviate coral bleaching, ensuring the component's threshold matches real-world conditions.

Prof. Jun XIA

cycle 1-2 ​

Through our literature review, we found that corals use fluorescent and chromoproteins (FPs and CPs) to defend against stress from excessive light. However, their limited capacity reduces effectiveness. Inspired by this, we propose expressing FPs and CPs in symbiotic bacteria to enhance the coral microenvironment and provide extra protection against bleaching.

Name: The 8th Southern China iGEM Regional Meeting

Key words: ''CPs beat FPs''

Expectation: As novices in the iGEM community, we joined the 8th Southern China iGEM Regional Meeting to connect with other teams, gain insights, and improve our project design.

Description: On May 19, we attended the meeting at Shenzhen University, where 16 teams shared ideas under the theme “Get Ideas Ripple.” We presented our project focused on using FPs or CPs to slow coral bleaching. We also designed an interactive game simulating the impact of fishing nets on coral reefs, inspired by a real-world issue observed by one of our advisors.

Reflection: This in-person exchange provided valuable feedback and highlighted the importance of interactive elements in Human Practices (HP). Discussions with fellow iGEMers revealed that CPs are more efficient at absorbing light than FPs, steering us toward a better solution..

Adjustment: We plan to incorporate the interactive game into more HP activities to raise awareness about coral reef threats. After careful consideration, we chose CPs as our target protein.

Group Photo with All iGEMers Participating in the 8th Southern China iGEM Regional Meeting

Name: Visit to the South China Sea Institute, Chinese Academy of Sciences

Stakeholder Context:

Dr. Sheng LIU: Sheng LIU is a researcher and doctoral supervisor in marine biology. He is mainly engaged in the research of plankton physiology and molecular ecology. Pioneer the use of molecular biology techniques to study biologically based production processes in natural marine areas, including: The study on the circulation route and efficiency of key species, the link between microfood rings and classic food chains, and the construction of plankton food web structure has a significant role in promoting the understanding of the structure, function and operation of ecosystems, and also has important guiding significance for the safe management, prediction and development of regional marine fisheries.

Key words: Zooxanthellae cultivation

Expectation: After selecting chromoprotein as our target protein, we sought guidance from Prof. Sheng LIU at the Institute of Oceanology, Chinese Academy of Sciences, to ensure the efficient cultivation of Zooxanthellae for our preliminary experiment.

Description: We consulted Prof. LIU, Prof. YANG, and Dr. JIANG, discussing coral bleaching dynamics, symbiotic algae cultivation, and managing the algae-coral relationship under stress. Prof. LIU shared insights on the effects of temperature, CP expression, and coral-algae balance. Prof. YANG provided a detailed guide on culturing Zooxanthellae, covering media selection, temperature, light intensity, and surface adherence for growth. Dr. JIANG gave us a lab tour, offering practical insight into the experimental setup for Zooxanthellae cultivation.

Reflection: Prof. LIU's advice highlighted the complexity of coral biology and its symbiotic relationship with Zooxanthellae. Prof. YANG's guidance on attachment and growth was crucial, especially regarding the importance of surface adherence and light gradients. The lab tour gave us hands-on experience in creating the optimal environment for our experiments.

Adjustment: The visit helped refine our experimental design. We are ready to start our preliminary experiments, focusing on environmental factors affecting Zooxanthellae growth. This data will be key in evaluating our approach to mitigating coral bleaching and advancing our research. The collaboration with the Institute’s experts and access to samples will support the development of practical solutions for our project.

cycle 3 ​

Our experiments showed that constant CP expression in E. coli, aimed at protecting Zooxanthellae from light stress, may harm their growth. After reviewing literature and consulting experts, we decided to switch to a light-inducible system using the elighton sensor for controlled CP expression, which started our third cycle.

Name: Consultation at the South China Sea Institute, Chinese Academy of Sciences

Stakeholder Context:

Professor Meixia ZHAO: Professor Meixia ZHAO is a professor and master tutor at the School of Oceanology, University of Chinese Academy of Sciences. Her research interests include coral reef appearance and dynamics, coral reef engineering and ecological restoration, coral reef biogeomorphic change and evolution, and coral reef ecological diversity and ecological functions. She also focuses on how global change and human activity are regulating coral reefs. Professor ZHAO's research interests include marine geology and ecology, coral reefs, global change, ecological restoration and conservation. Since 2008, she has presided over and participated in a number of national and provincial scientific research projects, whose research results are of great significance to the ecological protection and restoration of coral reefs.

Professor Qingsong YANG: Qingsong YANG holds a PhD in marine biology and is currently an assistant researcher at the South China Sea Institute of Oceanology, Chinese Academy of Sciences. His research interest is tropical coral reef ecology. His research focuses on the common-epiphytic microbial community structure of reef-building corals and the biogeochemical cycles of nitrogen in coral reef ecosystems. Dr. YANG has published more than 10 academic papers in several internationally renowned academic journals, providing important scientific basis for understanding the ecological processes of coral reefs and protecting coral reef ecosystems.

Key words: ideal chassis selection, biosafety discussion

Expectation: With limited research on symbiotic bacteria in coral cells and coral synthetic biology, we face challenges in selecting a bacterial chassis and meeting biosafety standards. We organized an exchange at the South China Sea Institute to refine our strategy and gather expert input.

Description: In discussions with researchers Meixia ZHAO and Qingsong YANG, Dr. ZHAO emphasized focusing on light sensors due to the photoprotective role of our chromoprotein and the importance of timing in coral bleaching. Dr. YANG suggested exploring Endozoicomonas elysicola as a potential chassis and using bacterial strain banks for options with better colonization potential. We also discussed the possibility of modified symbiotic bacteria losing survival advantages and the need for evaluation metrics to assess circuit performance.

Reflection: The discussion reinforced the importance of testing light sensor properties, especially since our chromoprotein may affect light absorption. Dr. YANG’s advice on chassis selection clarified our approach, and his question about the unpredictability of bacterial colonization made us reconsider the need for a suicide switch in terms of biosafety.

Adjustment: We will conduct further research on Endozoicomonas elysicola and, if suitable, use it as our chassis. While Dr. YANG downplayed the need for safety circuits, we believe it's still essential to further explore biosafety and ensure our design is safe for marine ecosystems when facing the unknown.

Group Photo with Prof. Meixia Zhao and Prof. Qingsong YANG

Name: Visiting the Hong Kong branch of the Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou)

Key words:biomass, limitation

Expectation: Having received invaluable advice from Prof. QIAN during the early stages of our project, we aimed to visit the Hong Kong branch of the Southern Marine Science and Engineering Guangdong Laboratory (Guangzhou) in HKUST, under Prof. QIAN's leadership, to seek further practical guidance and insights.

Description: We toured the lab and reconnected with a researcher, who, after reviewing our circuit design, raised concerns about the biomass of our expressed proteins. He pointed out that the biomass might be insufficient to effectively alter the light environment or reduce coral bleaching, emphasizing the need to consider this limitation in real-world applications.

Reflection: The researcher’s feedback highlighted the critical role of biomass in our project's success, a factor we had overlooked. It reminded us that even well-designed systems face practical limitations in real-world applications. This insight refocused our efforts on investigation of feasibility.

Adjustment: This feedback prompted us to prioritize the biomass issue in our project. We plan to conduct further literature reviews and, if feasible, simulate the problem in the dry lab. However, we understand that validation experiments will be crucial, and addressing this limitation may be constrained by time and resources.

Visiting the Lab in HKUST

Name: Interview with Professor Liuqian YU

Stakeholder Context:

Prof. Liuqian YU: Prof. YU is a biogeochemical oceanographer interested in using models and statistical methods to advance marine ecosystem understanding, prediction, and management. She is passionate about developing and applying coupled physical-biogeochemical models to improve quantitative understanding of human impacts on marine ecosystems (e.g., eutrophication, hypoxia, aquaculture, and oil spill) and to explore eco-engineering approaches to mitigate human-caused environmental problems. She is also keen to improve the model estimation and prediction of marine ecosystem processes by applying data assimilation methods that can optimally combine observations and dynamical models.

Key words: temperature, daily cycle of light, light attenuation

Expectation: In our interview with Professor Liuqian YU, we aimed to better understand how ocean deterioration impacts coral reefs and clarify the relationship between surface light intensity, wavelength, and underwater light. This is key for evaluating our light sensor.

Description: Prof. YU discussed the effects of ocean warming, acidification, and deoxygenation on reefs. She noted that while surface light helps shallow corals, we should also consider seawater turbidity and light attenuation for accuracy. She emphasized the importance of accounting for the daily light cycle in our design.

Reflection: This conversation made us realize the need to address daily light fluctuations and include a stable mechanism in our circuit. We also saw the importance of researching underwater light and temperature’s role in coral health to refine our approach.

Adjustment: We will conduct hydrological experiments to validate our sensors. To prevent overexpression, we plan to integrate a temperature sensor and implement an "AND Gate" to balance light and temperature inputs for more stable operation in coral environments.

Communicating with Prof. Liuqian YU

cycle 4 ​

After discussions and revisions, we've finalized a stable circuit design with integrated light and heat sensors using an "AND Gate." The chromoprotein is expressed only when both conditions exceed thresholds, absorbing excess light to protect corals from bleaching.

Name: Interview with Professor Zhen LIU

Stakeholder Context:

Dr. Zhen LIU: Dr. Liu is an atmospheric scientist working to advance the understanding of the mechanism and physical processes governing both observed and future climate changes across a wide range of spatiotemporal scales. One of the most important objectives is to reduce uncertainties in future climate projections caused by anthropogenic forcing, such as changes in land use and greenhouse gas concentrations. He has devoted most of his efforts to investigating anthropogenic-induced changes (particularly anthropogenic aerosols) in monsoon circulation and precipitation as well as climate extremes by making use of observations and general circulation models.

Key words: "Why And Gate?''

Expectation: To validate the use of an "And Gate" in our design, we consulted Professor Zhen LIU.

Description: Prof. LIU emphasized that heatwaves, linked to coral bleaching, result from increased sea temperatures. However, he noted that a 1.2°C rise during heatwaves might not cause bleaching alone. He agreed with our "And Gate" design, explaining that heat is primarily driven by light, making the circuit less sensitive to daily fluctuations. He also introduced the concept of the "mixed layer," supporting the use of sea surface temperature as a proxy for coral conditions.

Reflection: Since heatwaves may not reach bleaching thresholds on their own, integrating light and temperature sensors through an "And Gate" is ideal for real-world conditions. This design reflects the natural relationship between heat and light, improving system stability against environmental changes.

Adjustment: With Prof. LIU's validation, we will finalize the inclusion of the "And Gate" in our circuit. Next, we'll conduct modeling to evaluate expression levels and ensure the system effectively prevents coral bleaching under various conditions

Interviewing with Prof. Zhen LIU

Name: Visiting Southern Marine Science and Engineering Guangdong Labolatory

Stakeholder Context:

Prof. Wenliang ZHOU: Professor Wenliang Zhou is a professor and doctoral supervisor of the School of Life Sciences at Sun Yat-sen University, focusing on the research of epithelial cell physiology. His research interests include epithelial cell ion transport mechanisms and their regulation, and the role of these mechanisms in the development of disease, particularly in pathogen-induced immune defense mechanisms.

Professor Zhou has made outstanding contributions in the academic field, published a number of academic papers on epithelial cell regulation and immune defense mechanisms, and is committed to cultivating a new generation of biological research talents.

Key words: complexity in marine synthetic biosafety

Expectation: We sought expert feedback on sensor thresholds, natural conditions, and safety issues related to our coral bleaching relief project, aiming for further refinement.

Description: At the Guangdong Marine Lab, we discussed our project with researchers, focusing on light spectrum at coral depth, genetic circuit expression in symbiotic bacteria, and safety circuit design challenges. We also toured their coral cultivation facilities and received guidance on selecting bacterial strains and working with zooxanthellae or corals.

Reflection: The experts emphasized the importance of light intensity and temperature data from coral habitats and the complexity of designing safety circuits to prevent the release of genetically modified organisms. We recognized the need for thorough testing and validation before practical implementation.

Adjustment: Based on their feedback, we will refine our circuit design with a deeper focus on biosafety. Our Human Practices and Wet Lab teams will collaborate closely to address these issues. The lab visit and expert insights have been invaluable, guiding us toward a more robust and practical approach to coral conservation.

Visit the Southern Marine Science and Engineering Guangdong Labolatory

Name: Lecture at Shimen Middle School

Key words: dialogue,quorum sensing

Expectation: We aimed to introduce synthetic biology and iGEM projects to high school competition students with a strong interest in biology, hoping to inspire innovative thinking.

Description: On August 30, 2024, we gave a talk at Shimen Middle School in Foshan, presenting our project’s background, design, methods, and outcomes. We also included a Q&A session to engage students and encourage their input. Their questions and feedback, particularly on gene circuit challenges, sparked valuable discussions.

Reflection: The students suggested using quorum sensing to design a microbial suicide mechanism that triggers cell death based on chemical concentrations. This idea provided new inspiration for improving our project.

Adjustment: Inspired by the students’ suggestion and reminded of the importance of safety design by experts at Southern Marine Science and Engineering Guangdong Lab, we will explore incorporating quorum sensing into our safety circuit to enhance our project’s reliability in oceanic environments.

Communicating in Shimen Middle School

Name: Interview with Professor Muzhi ZHOU

Stakeholder Context:

Prof. Wenliang ZHOU: Prof. Muzhi ZHOU is an currently Assistant Professor in the Urban Governance and Design Thrust at the Hong Kong University of Science and Technology (Guangzhou). She has expertise in the field of gender and family, life course and digital inequality. She studies how critical life events, such as marriage and childbirth, reshape people’s lives in East Asia and Western Societies, where the culture, social policies and welfare regimes differ. Her recent work focuses on how social inequalities are reflected in the digital/virtual society and their interconnectedness with the physical world. Her work has been published in top journals such as Gender & Society, Population and Development Review, Demographic Research, and Chinese Sociological Review.

Key words: questionnaire revision, interview preparation, HP ethics

Expectation: To acquire expert guidance on the art of conducting interviews for our upcoming field trip to Weizhou Island, Guangxi and crafting questionnaires, we have a communication with Prof. ZHOU with the goal of efficiently collecting insightful data.

Description: Before meeting Prof. ZHOU, we drafted interview frameworks and a public questionnaire. She advised us to create concise and clear questions that align with our goals. She stressed the importance of engaging local fishermen, who may be less educated and cautious, and emphasized the need for thorough preparation when interviewing government officials. Prof. ZHOU also highlighted the ethical considerations, such as obtaining consent and keeping detailed records.

Reflection: Prof. ZHOU pointed out that our initial questionnaire was too narrow, limiting data collection, and noted our lack of experience in interviewing, especially with fishermen. Her walkthrough of the interview process emphasized the importance of preparation, scheduling, and ensuring ethical practices.

Adjustment: We will revise our questionnaire to make it more concise and goal-oriented, and we'll tailor our approach based on the community. Recognizing our inexperience, we plan to conduct a pilot exercise with local fishermen to improve our communication skills before the field trip. We'll also ensure proper consent forms are prepared for ethical approval.

Group Photo with Prof. Muzhi ZHOU

Name：The second round questionnaire

Keywords: synthetic biology, our project's future implementation

Expectation: In the second round of questionnaires, we aimed to understand public views on synthetic biology—its safety and applications—especially related to environmental challenges like coral bleaching, which is crucial for our project's future implementation.

Figure 6. How much do you know about transgene

Figure 7. Select three words that represent your attitude toward transgene biotechnology

We found that while many respondents had heard of synthetic biology, few fully understood it, highlighting a significant knowledge gap. Most were cautiously optimistic about using it for coral protection but raised concerns about safety and long-term impacts.

Figure 8. Attitudes towards applying synethetic biology in coral protection

Nearly 90% stressed the importance of strict safety protocols and government oversight before deploying genetically modified organisms in the ocean.

Figure 9. Whether you prefer GMO or non-GMO food

Figure 10. Why do you prefer non-GMO food

Additionally, a significant number preferred non-GM food, citing health and environmental concerns, which indicated a general skepticism about GM technology that could extend to synthetic biology.

Reflection: The public is open to innovative solutions but has significant safety and ethical concerns. This emphasizes the need for better education to build understanding and trust in synthetic biology.

Adjustment: We will focus on addressing public concerns through Human Practices by educating the public on the safety and ethics of synthetic biology. This will be key to gaining support for coral conservation and synthetic biology applications.

In conclusion, the two rounds of questionnaires provided valuable feedback that guided both our Integrated Human Practices (iHP) and education initiatives. By tailoring our future outreach efforts to these insights, we are confident that we can foster a more informed and supportive public, willing to engage in and contribute to the conservation of coral reefs and the responsible use of synthetic biology.

Name: Biosafety & Bioethics in Synthetic Biology Whitepaper

Key words: further investigation and efforts in biosafety

Expectation: To deepen our understanding of biosafety and bioethics in synthetic biology, we collaborated with JLU-NBBMS to contribute to the Biosafety & Bioethics in Synthetic Biology Whitepaper.

Description: We thoroughly researched domestic and international biosafety guidelines, including the Regulations on the Safety Management of Genetic Engineering, the Convention on Biological Diversity, and the Nagoya Protocol. We explored safety circuits and conservation projects, presenting our project’s design, safety strategies, and unique advantages. This collaboration prompted us to critically reevaluate our approach to safety and ethics, offering valuable insights for future iGEM teams.

Reflection: Collaborating on the whitepaper helped us deepen our understanding of biosafety and ethics in synthetic biology. Reviewing regulations and safety designs revealed gaps in our current approach, pushing us to reconsider our strategies for better alignment with international standards.

Adjustment: Moving forward, we will incorporate more structured biosafety assessments and ethical evaluations throughout our project. We plan to create a detailed biosafety framework that integrates these considerations from the design phase, ensuring a solid foundation for future teams.

JLU-NBBMS's Biosafety & Bioethics in Synthetic Biology Whitepaper

Name: Field Trip to Weizhou Island and Beihai in Guangxi.

Pilot Exercise (Preparation): To enhance our communication and interviewing skills within the fishing community, we visited Nansha Shisan Chong, a locality near our campus known for its fishermen. Initially, we encountered several challenges, such as dialect communication issues and a reluctance among some to be interviewed, which are also potential obstacles we might face in Guangxi. Based on these experiences, we reorganized our team to address the dialect issue and learned to select interviewees based on context and observation. At the beginning, it was quite embarrassing and unusual for us to conduct interviews that were awkward and strange due to our halting conversational pace. However, as we gained experience, our interactions became smoother and more fluent.

Feild Trip in Nansha Shisan Chong

1. Diving Survey and Hydrological Data Measurement

   Key words: wavelength, impressive bleaching

   Expectation**:** To measure the temperature and light intensity in the coral habitat to determine if the sensor thresholds are correctly set, and to measure the absorbance of water samples for the wavelengths detected by the sensor and absorbed by aeBlue (target protein) to assess if our circuit functions as intended. Observations of coral conditions in the diving area are also made.

   Description: Led by professionals, we conducted measurements around 13:25 at a location approximately 150-200m off the northwest coast of Weizhou Island, at a depth of 4.9-5.1m. We used instruments to measure underwater temperature and surface light intensity, collected water samples at different positions next to the coral, and returned to the lab for absorbance measurements. The water temperature was around 30°C, surface luminance was approximately 56-62 lux, and the average absorbance values for water samples were 0.004Abs for 450nm wavelength light and 0.005Abs for 600nm wavelength light. During the underwater observation, a considerable amount of live coral was present, but there was also a significant amount of dead coral covered with algae, approximately 30%.

   Reflection: The measured water temperature and light intensity generally align with the threshold design range of our two sensor groups, and the absorbance values met our expectations, indicating the theoretical feasibility of the light sensor function. The direct observation of coral death due to bleaching was impressive.

2. Interview Records

   1. Expectation: To understand the perspectives of stakeholders closely related to coral (fishermen, coral farming entrepreneurs, coral protection scientists, and government officials) on our project. We aim to optimize the project by considering their thoughts and needs, explore the feasibility and application potential of our project in communication with them, and as a synthetic biology project, discuss with them the use of genetically modified technology for coral to aid in our biosafety investigation and consideration.

(1) Interview with Fishermen on Weizhou Island

Key words: GMOs, tangible influence, awareness of coral conservation

Description: We approached a group of fishermen who had just returned from sea and were mending their nets by the seafood market. They kindly agreed to an interview. The fishermen of Weizhou Island have been fishing for a long time and showed a high awareness and enthusiasm for coral protection. They often mentioned the past abundance of coral on Weizhou Island and the significant reduction in coral quantity now. They also spoke about the difficulty in catching fish that coexist with coral and the high prices due to scarcity. Therefore, they strongly support various coral protection projects, believing they will lead to an increase in fish catch. They generally lack understanding of genetically modified concepts and exhibit a strong aversion to them.

Reflection: Through communication with the fishermen, we found their awareness of coral protection to be very strong, indicating that coral protection has been well-publicized among local groups directly related to coral. However, their lack of understanding and strong aversion to genetically modified technology show significant resistance to the application of synthetic biology in coral protection. This implies that we need to conduct simple popular science and assurance of genetically modified concepts through official channels before the project is implemented, and we must conduct thorough research and design in the aspect of biosafety to ensure the project is carried out without affecting marine life and the environment.

(2) Interview with Xu Zhonghua, Chairman of Beihai Kebei Marine Science and Technology Park Company

Key words: ''Blue is rare'', market, stability

Description: We interviewed Xu Zhonghua, Chairman of Beihai Kebei Marine Science and Technology Park Co., Ltd., focusing on their current coral variety sources, breeding methods, coral cultivation conditions, and market demand. We introduced our project in layman's terms and asked for his insights on the project's needs and whether the use of genetically modified technology in new variety cultivation would cause concerns for merchants and customers. He provided insightful responses, emphasizing that the coral industry, being non-food related, faces less customer concern about genetically modified organisms. He also mentioned that merchants are more concerned about the stability of traits when introducing new varieties, and the color blue is rare in corals, which has sufficient appeal and market potential.

Reflection: We realized the coral industry's inclusiveness towards the application of genetically modified technology, which was unexpected and shows the market's inclusiveness for our project. The rarity of blue coral gives it a certain customer market, and our aeBlue, which is blue, makes us aware of the broader application scenarios of our project's results. We can use the new species to help the market generate greater profits. The stability issues raised by Mr. Xu guide us to consider the degradation rate of aeBlue, the expression rate of the circuit, and the symbiotic bacteria's colonization of coral, and the achievable biomass in more depth, in line with stakeholder needs.

Interviewing with Xu Zhonghua

(3) Interview with the Person in Charge of Beihai Zhulin Seawater Aquaculture Experimental Base

Key words: cultivation condition, demand for new varieties

Description: We interviewed the person in charge of the Beihai Zhulin Seawater Aquaculture Experimental Base and visited their coral, clownfish, and grouper breeding areas. The person in charge mentioned that the large factory building uses natural light, and the temperature cannot be controlled too low, which can only meet the growth of corals with more single colors. Vibrant colored varieties generally have light requirements and cannot grow to their best state under these conditions. Therefore, their customers are mostly large wholesalers such as aquariums, not individual enthusiasts. However, the market for buying corals and playing with marine tanks is considerable, and some companies that modify corals to produce more vibrant colors can sell very high-priced varieties due to the large market demand. Coral farmers and players are very willing to try new varieties and welcome genetically modified varieties.

Reflection: At the breeding base, we realized that the sensitivity of corals to the environment affects the types of corals that breeders can cultivate, which in turn affects their target customer groups and profits. Our project, which enhances the tolerance of corals to the environment, can have a certain impact on them. At the same time, the coral market is considerable, and the demand for new varieties exceeds our expectations, showing us the commercial value of our project's application. Observing that the breeding base also cultivates a large number of symbiotic organisms of corals, such as clownfish and groupers, it is clear that the degradation of the coral reef system has a significant impact on the survival of these symbiotic fish.

Visiting the Beihai Zhulin Seawater Aquaculture Experimental Base

(4) Interview with Dr. WANG Xin from the Coral Research Team of Guangxi Marine Science Institute

Stakeholder Context:

Dr. Xin WANG: Dr. Xin WANG is an associate researcher at the Guangxi Academy of Marine Sciences, and currently serves as the academic leader of the coral Reef ecological protection and restoration Innovation Team, dedicated to the research and practice of coral reef ecological restoration. Dr. Xin WANG and his team have long been involved in the protection and restoration of coral reefs in the Beibu Gulf region, focusing on the potential of coral reef ecosystem recovery under the impact of global climate change and human activities.

In an important cross-regional coral rescue operation, Dr. WANG led a team to successfully rescue a group of live corals in crisis and moved them to temporary ponds for processing and recovery. Some of the corals were released back into the wild after recovery, helping to maintain the local ecological balance.

His research has not only promoted the ecological protection of coral reefs in Guangxi, but also provided scientific basis and technical support for future ecological restoration.

Key words: lack in biosafety, increase resilience

Description: We had a conversation with Dr. Xin WANG from the coral research team about coral protection projects, focusing on the current status of coral bleaching, existing protection methods, and biosafety. Dr. WANG mentioned that the main coral protection methods currently include educational propaganda, legal establishment, protected area creation, and artificial intervention for restoration, affirming the innovation of using synthetic biology for coral protection. Regarding biosafety, he mentioned that there are few relevant regulations restricting the application of synthetic biology in the coral field. Dr. WANG also provided advice and guidance for our upcoming hydrological data testing and offered laboratory equipment for water sample testing.

Reflection: Compared to existing coral protection methods, our approach can directly protect corals by enhancing their resilience, rather than indirectly controlling human activities, which has a higher fault tolerance. Compared to the current manual intervention for restoration by scientific research teams, such as breeding and nurseries, which require advance planning and continuous investment in labor and maintenance costs, our method has advantages in this regard. Dr. WANG pointed out the lack in synthetic biology safety in the coral field, and we will further investigate and clarify this area and do as comprehensive a job as possible in biosafety to ensure biosafety.

(5) Visit and Interview with Dr. Qin at the Weizhou Island Coral Museum of Guangxi University

Key words: microenvironment, necessity, education

Description: We visited the Weizhou Island Coral Museum of Guangxi University and interviewed Dr. Qin, who manages the museum. We first learned about the museum's visitor flow and found out that almost all tourists coming to Weizhou Island visit the coral museum. The museum has a lot of coral-related scientific popularization displays and often cooperates with coral protection organizations to hold coral popular science activities. Regarding circuit design, he affirmed our approach of only changing coral's microenvironment, believing that our use of CP protein does not change the coral cell's metabolic pathways, which is very conservative and safe, and has a higher success rate. He mentioned that some teams in the South China Sea are doing research on inducing corals to recover from bleaching, but the current success rate is very low, and the research is very difficult, while it takes a long time for corals to recover from bleaching under natural conditions.

Reflection: In the current situation where the reverse experiment of bleaching is not optimistic, it is very necessary to optimize the research on methods of coral bleaching, which reflects the importance of our project's future application. The considerable visitor flow to the coral museum shows that the public has a certain interest in the topic of corals, and coral popular science work is continuously progressing. We will also refer to similar forms of popular science exhibitions for educational activities in our education.

(6) Interview with Staff at the Guangxi Weizhou Island Coral Reef National Marine Park Management Station of Beihai Forestry Bureau

Key words: benefits and safety

Description: We interviewed staff at the Guangxi Weizhou Island Coral Reef National Marine Park Management Station of Beihai Forestry Bureau, who are mainly responsible for the daily patrol and maintenance of the coral nature reserve near Weizhou Island, submit annual reports, and cooperate with the Guangxi University of Marine Science to evaluate the protection results. To enhance the public's awareness, they would release several items with the slogan of coral protection, which will be delivered to citizens.

Reflection: The core requirements from the government side are the project's benefits and safety. In terms of benefits, our method has a clear advantage over traditional methods. However, safety poses a challenge for us. We will consider as many safety issues as possible in the project to provide greater possibilities for future implementation.

Communicating with Staff at the Guangxi Weizhou Island Coral Reef National Marine Park Management Station of Beihai Forestry Bureau

**Adjustment：**After the field trip to Guangxi, we first prove the validity of our light sensor design by different wavelength light absorbance of seawater samples, which is significant to our proof of concept. Through on-site data collection, we also have a deeper understanding of environmental factors for threshold of sensors. More importantly, we communicate with various stakeholders about our project, gaining invaluable opinioins which will help us develop our HP goals into next stage, which forms our final HP value and finishes a complete cycle for our iHP activities. The discovery of the potential market of our project product and the attention to biosafety direct us to the key working aspects in future.

Our Human Practices Value ​

Since we have completed the final version of our circuit design, developed a comprehensive understanding of the real-world context, and gathered insights from various stakeholders, we have gained a clearer sense of our responsibilities and objectives. Consequently, we updated our preliminary Human Practices Goals to reflect our final Human Practices Value. This update not only encapsulates the key themes of our previous Human Practices activities but also provides guidance for our future consideration and efforts.

1. Investigating the knowledge gap in coral synthetic biosafety and adopting a comprehensive approach to biosafety considerations
2. Developing an innovative and effective synthetic biology approach to address coral bleaching
3. Introducing a novel coral strain to expand the coral farming industry and boost economic benefits
4. Ensuring our design considering balance between diverse needs of various coral-related stakeholders
5. Enhance public understanding of corals, coral bleaching and conservation through various outreach activities

Recognizing a gap in the application of synthetic biosafety to coral, we now place greater emphasis on investigating this knowledge gap and integrating comprehensive safety considerations into our project design. After our field trip to Beihai, Guangxi, we identified the business potential of our outcome, which has become the third statement in our HP values for further discussion. Values 2 and 3 now guide two main implementation directions, each with distinct end users and concerns. Regarding values 4 and 5, we made slight adjustments as our understanding of HP evolved. Instead of trying to satisfy all stakeholders, we now aim to balance and make trade-offs after full consideration, acknowledging the inevitable conflicts among different viewpoints. For value 5, we added coral-related knowledge to our promotion and education designs, as we realized that a well-informed public can better navigate the coral-related market for their own benefit. For more details on the application of our HP values, please refer to our Future Implementation page.

Conclusion ​

Reflecting on our past human practices activities, we are pleased with how seamlessly they integrated with the engineering cycles of our project design. By engaging with various stakeholders and following our HP logic, we closely adhered to the principles of being "Responsible," "Reflective," and "Responsive." Through continuous two-way dialogue, we received valuable feedback, were affected by the world, and, in turn, affecting it. We firmly believe that, with the insights gained from our integrated human practices efforts, our project will ultimately contribute to alleviating coral bleaching, making a real difference and helping save thousands of corals from the edge of death.

Reference ​

https://responsibility.igem.org/human-practices/what-is-human-practices

https://www.ukri.org/who-we-are/epsrc/our-policies-and-standards/framework-for-responsible-innovation/

https://impact.wp.st-andrews.ac.uk/responsible-research-and-innovation-area/