Starting from care and concern for our aging parents, SR-Shenzhen seeks to contribute a scientific solution to societal aging. Throughout the project, our Human Practices (HP) work ensured that from day one of technical design we considered safety, compliance, and inclusivity, aiming for a solution that is safe, accessible, and inclusive.

In our Human Practices process, we consistently emphasized the importance of “responsible research.” Research is not merely about gathering information, but also about demonstrating accountability to interviewees and society. To avoid one-sided, superficial, or even misleading conclusions, we must uphold rigor and respect in both methodology and attitude.

Figure 1. Concept Map of Responsible Research

Why conduct responsible research?

1、Ensure authenticity and validity of information: Different groups have significantly varied needs and perspectives. Only through multi-layered and standardized research can we avoid distortions caused by sampling bias or leading questions.

2、Respect interviewees: An interview is not just about acquiring information; it is also a process of communication and trust-building. Maintaining neutrality and respect encourages interviewees to share their genuine thoughts.

3、Establish a traceable evidence chain: By preparing background documents, structured question outlines, and follow-up mechanisms, we make the entire process open and transparent, ensuring the reliability of our conclusions.

Stakeholder Interview Handbook

Figure 2. Stakeholder Interview Handbook

Before formally engaging stakeholders, smooth, truthful, and effective communication is key to mutual trust and meaningful feedback. We compiled a Stakeholder Interview Handbook to provide a unified communication framework and SOPs, improving standardization of interviews and efficiency in stakeholder interactions.

Initial Inspiration

Figure 3. Public Service Advertisement: “Family”

The public service advertisement “Family” made us acutely aware that our parents are quietly aging. Aging is universal—it concerns everyone, including our future selves. Although we are still in high school and not personally focused on anti-aging, we observed our parents paying attention to wellness and trying anti-aging approaches. More broadly, many middle-aged and older adults are actively seeking ways to delay aging and stay healthy, indicating a real and widespread demand.

After recognizing this need, we began reviewing the landscape of anti-aging products and methods.

Background Research

We realized our parents are aging faster than we had noticed, reminding us that aging is not abstract but present in daily life. We hope to find a scientific, healthy, and sustainable path for anti-aging that can help our parents and others.

In everyday conversations, we learned our parents already try various wellness routines and anti-aging products. Talking with people around us, we also found many middle-aged and elderly individuals want to slow aging, yet have limited awareness of synthetic biology.

Figure 4. Health-Supplement Shop Owner

To better understand the current market, we reviewed literature, interviewed a health-supplement shop owner, and conducted online research. We found the market is crowded with products, but two problems are common: (1) high prices, which limit access; (2) insufficient scientific evidence for some products, raising concerns about efficacy and safety. This reinforced that existing solutions fall short.

Based on this, we decided to explore synthetic biology: by redesigning metabolic pathways, we aim to engineer microbial cell factories that safely, sustainably, and at low cost produce anti-aging actives, overcoming the high-price/low-accessibility barrier.

Our project also takes on a science-communication mission. We plan inclusive outreach for older adults—plain-language talks, Q&A, and everyday examples—so they can understand basic synbio principles and potential applications, and give us feedback. This is not only research, but a family-centered and socially responsive effort.

Consultations with School Teachers

Figure 5. Consultations with Biology Teachers (Multiple Teachers)

We consulted our school biology teachers, who confirmed that aging interventions are a frontier of synthetic biology. However, given how fast the field is moving, they could offer limited specific experimental guidance. This pushed us to seek experts with deeper domain expertise to sharpen our research direction.

Consultation with a Nutritionist

At the beginning of our project, we aimed to identify a substance that could intervene in the aging process while also being scientifically validated. Although the market is filled with various anti-aging products, their complex ingredients and uncertain efficacy quickly made us realize that online information alone cannot ensure scientific reliability. While our school teachers could point us in a general direction, the cutting-edge nature of this field meant they could not provide detailed research guidance. Therefore, we decided to seek more specialized advice from experts with clinical and practical experience.

Figure 6. Communication with Nutritionist Danqing

We reached out to and interviewed a nutritionist. During the discussion, we raised questions regarding aging, nutritional interventions, and the current use of dietary supplements. Drawing on clinical experience, the nutritionist introduced several commonly used anti-aging bioactive compounds and shared insights into consumer concerns and misconceptions when choosing supplements. Through this face-to-face exchange, we gained a deeper understanding of the anti-aging product market and user needs.

In the interview, the nutritionist particularly emphasized the potential of nicotinamide mononucleotide (NMN). As a precursor of NAD⁺, NMN has been supported by numerous academic studies for its effects in delaying cellular senescence and improving energy metabolism. Following this advice, we conducted an extensive literature review and found that NMN indeed showed positive outcomes in animal experiments and early-stage clinical studies, while also being highly sought after in the supplement market. This finding helped us define our research focus.

Based on the survey results and the nutritionist’s feedback, we decided to make NMN the core target molecule of our project. Accordingly, we designed a synthetic biology approach that modifies the metabolic pathway of engineered E. coli to efficiently produce NMN. This method not only reduces production costs but also avoids environmental pressures caused by traditional chemical synthesis.

The nutritionist’s feedback helped us not only determine our research molecule but also recognize the close link between synthetic biology and nutritional health. In our subsequent inclusion activities, we plan to design dietary guides for middle-aged and elderly people to help the public better understand how scientific research connects with daily health needs.

For more details, please refer to Inclusivity.

Power–Interest Matrix Analysis

Figure 7. Power–Interest Matrix of Project Stakeholders

We conducted an analysis of the stakeholders involved in our project, defining them as individuals or groups who could influence the process of product research and development. By constructing a Power–Interest Matrix, we were able to more precisely identify our potential target customer groups—namely, the end users of our product and the decision-makers in purchase processes. According to the results of this analysis, we categorized stakeholders by their level of importance into four key groups: anti-aging research centers, nutrition and health supplement companies, middle-aged corporate executives, and private medical aesthetics institutions.

Figure 8. Lecture and Discussion with Mr. Zhao, Planning Director at a Listed Research Institute

During our communication with Mr. Zhao, the Planning Director of a large listed research institute, we further analyzed the attitudes and perspectives of these target stakeholder groups. As Mr. Zhao emphasized, different stakeholders may hold sharply contrasting views of our project: some might see us as competitors, unwilling to offer constructive feedback or even intentionally providing misleading opinions, while others might regard our project as a socially beneficial initiative and thus be eager to provide help and support. For example, health-supplement companies are potential competitors and therefore are less likely to give impartial feedback. From this, we realized that it is essential to develop differentiated communication strategies for different target groups to ensure that the feedback we obtain is authentic, effective, and representative.

At the same time, we recognized the need to carefully consider the attitudes of our parents when conducting interviews. As the starting point of our project inspiration, our parents naturally became the first people we wanted to interview. They can provide unique user perspectives and lived experiences related to aging and wellness. However, because parents tend to be naturally supportive and encouraging, their feedback is often overly positive and less critical. This reminded us that, when designing interviews and collecting feedback, we must strike a balance between “familiar goodwill” and “objective evaluation”, in order to obtain comprehensive, accurate, and truthful data.

Interviews with Parents

Figure 9. Team Members Communicating with Their Parents

Our parents were the first group of interviewees, as we sought to explore the real-life needs behind the issue of aging.

We actively engaged in conversations with our parents, who shared their personal experiences and opinions regarding wellness routines, health management, and the use of anti-aging products. Through these discussions, we found that our parents showed curiosity and interest toward synthetic biology, though their feedback was mostly from a user-oriented and intuitive perspective.

Overall, our parents provided positive and encouraging feedback, but offered few critical comments. Their responses showed a degree of subjectivity and bias, primarily due to the “closeness effect” inherent in family relationships. From their input, we identified that aging in their lives mainly manifests as declining memory, reduced physical stamina, and skin loosening.

In terms of gender differences, we observed that:

• Men tend to focus more on nutritional supplementation, particularly related to obesity and its associated metabolic health issues (the “three highs”: high blood pressure, high blood sugar, and high cholesterol), often preferring vitamins and functional supplements.
• Women, on the other hand, are more concerned with aesthetic and skin health management, favoring skin care, beauty, and anti-aging treatments.

These insights not only helped us develop a more comprehensive understanding of the physical and emotional impacts of aging but also provided valuable references for our subsequent product positioning and user segmentation.

The feedback from our parents offered us a firsthand user perspective and marked the first essential step in our Human Practices process, laying a foundation for broader societal research in the next stages.

Questionnaire Survey

After completing the initial interviews with our parents, we realized that relying solely on their feedback provided a sample size too small to reflect the real needs of the broader public regarding aging and anti-aging products. Therefore, we decided to conduct a large-scale questionnaire survey to obtain broader and more representative data, helping us better understand differences in needs among various demographic groups.

To encourage greater participation, we distributed the questionnaire online and through local community networks, gathering responses from 383 participants of different genders and age groups.

Figure 10. Key Results of the Questionnaire Survey

Survey Results:

• Gender differences: Women were more likely to engage in diverse forms of health and beauty management, such as skincare and nutritional supplementation, while men showed higher participation in healthy eating and regular exercise.
• Age differences: Each age group showed distinct perceptions of aging. Younger participants focused on healthy diets and basic skincare; middle-aged groups prioritized aesthetic medicine and supplements; older adults emphasized nutrition and senior-friendly exercise products.
• Manifestations of aging:Middle-aged and elderly participants mainly reported skin laxity and memory decline as their key concerns.
• Consumer considerations: Most respondents held positive attitudes toward product efficacy, pricing, and brand reputation, yet had significant concerns about safety. Many also chose “uncertain,” revealing limited access to reliable information.
• Behavioral differences: Across different wellness and beauty habits, efficacy and safety were core priorities, whereas brand and price played larger roles in more commercialized purchasing decisions.

However, we also noted that some questionnaires were incompletely or carelessly filled, limiting the representativeness and reliability of the data.

Recognizing these limitations, we decided to place greater emphasis on face-to-face, in-depth interviews in the next stage of our Human Practices work. Compared with numerical survey data, direct communication allows us to better capture authentic attitudes, underlying concerns, and practical needs. Thus, we gradually shifted our research approach from quantitative questionnaires to qualitative interviews.

The survey provided us with an initial market and user profile, helping us identify how gender, age, and consumption habits shape health and beauty behaviors. These insights formed the foundation for our later experimental design and product positioning. At the same time, we learned that questionnaires alone are insufficient to reach target audiences accurately; a more effective approach involves direct dialogue and multi-channel engagement to gather meaningful feedback.

Street Interviews

After completing the questionnaire survey, we realized that data alone could not fully capture people’s real attitudes toward anti-aging. To better understand public perception and bring our research closer to real-life contexts, we conducted a series of random street interviews. Our goal was to collect more diverse and vivid feedback—but this process also made us aware of the complexity and challenges inherent in field research.

Figure 11. Summary of Street and Community Interviews

After completing the questionnaire survey, we realized that data alone could not fully capture people’s real attitudes toward anti-aging. To better understand public perception and bring our research closer to real-life contexts, we conducted a series of random street interviews. Our goal was to collect more diverse and vivid feedback—but this process also made us aware of the complexity and challenges inherent in field research.

During the random interviews, we encountered several difficulties. Many people were pressed for time and could not provide in-depth or detailed responses, resulting in a limited amount of effective data. Nevertheless, we gathered several valuable insights:

• A doctor suggested that we could explore producing NMN through synthetic biology, noting that this approach was both innovative and feasible.
• A fitness enthusiast mentioned that NMN might significantly enhance muscle strength.
• We also discovered the emergence of “pet NMN” products on the market, which inspired us to reflect on the diversity of the anti-aging market’s demands.

From these scattered yet genuine responses, we derived several key findings:

1、Income differences: People from different income levels exhibited distinct attitudes toward anti-aging. High-income groups were more willing to try novel health products, whereas middle- and lower-income groups tended to be more cautious and found it harder to understand or accept our project. This insight emphasized that our product must aim for lower production costs to achieve broader accessibility.

2、Cognitive differences: Some respondents showed limited understanding of synthetic biology, making it difficult for them to accurately grasp our research. This highlighted the importance of scientific communication and the need for simpler, more inclusive explanations in public engagement.

3、Need differences:Occupations and lifestyles also shaped anti-aging needs. For example, fitness groups focused more on physical performance, while middle-aged and elderly people cared more about memory and skin-related aging issues.

Based on these findings, we decided to make two major adjustments to our subsequent Human Practices work:

• Targeted interviews: Instead of continuing random street interviews, we plan to proactively engage participants from different income levels and life backgrounds. This approach will help us collect more structured, representative, and stratified feedback.
• Inclusion Program: We plan to design a series of science communication and engagement activities for groups less familiar with synthetic biology. By using clear, relatable explanations and inclusive dialogue, we aim to help more people understand and participate in discussions about our project.

Although the random street interviews yielded limited data, they helped us recognize the balance between quantity and quality in social research. More importantly, they revealed how income, cognition, and lifestyle differences create a complex landscape in the anti-aging market. These lessons motivated us to pursue greater precision and inclusivity in future studies—making our research not only more socially grounded but also more valuable in real-world contexts.

Interviews Across Different Demographic Segments

From our earlier random street interviews and questionnaire surveys, we found that different social groups show highly distinct needs and attitudes toward anti-aging. A single sample or population segment is insufficient to represent the full picture. To better understand the real demands of society, we adopted a more neutral and realistic classification of interviewees:

1. Basic livelihood–oriented group — those focused on meeting essential living needs;
2. Balanced lifestyle and health-investment group — those who seek a balance between life quality and moderate health spending;
3. Quality- and brand-driven group — those who prioritize high-end products, innovation, and brand reputation.

This stratified approach allowed us to gain a more precise and multidimensional understanding of societal needs.

Basic livelihood–oriented group:

This group is primarily concerned with essential daily consumption and is highly sensitive to price. In health management, they tend to rely on balanced diets and moderate exercise, but show low acceptance of new anti-aging products. Their focus is on safety and cost-effectiveness.

Balanced lifestyle and health-investment group:

With stable income sources, this group is willing to make moderate investments in health and anti-aging while maintaining quality of life. They are more likely to experience memory decline or reduced stamina due to work pressure, and thus show strong interest in nutritional supplements and functional foods.

Quality- and brand-driven group:

These consumers have high purchasing power and are eager to try new or premium products. They are deeply concerned about brand reputation and user experience, and are more likely to choose aesthetic medicine or personalized nutrition programs. Their decisions are strongly influenced by a product’s scientific credibility and innovation.

Figure 12. Interviews with Different Demographic and Consumer Groups

We later confirmed the validity of this classification. Different consumption and health-investment philosophies directly shape people’s attitudes and choices regarding anti-aging. We also found that beyond economic level, gender and occupation type (manual vs. intellectual work) are equally important influencing factors that must be incorporated into our research.

By interviewing individuals from diverse professions and integrating insights from our previous street and community interviews, we arrived at the following conclusions:

• Basic livelihood–oriented group: Prioritize maintaining daily routines through diet and exercise; weak demand for anti-aging; most concerned about price and safety.
• Balanced lifestyle and health-investment group: Show the strongest demand for anti-aging, especially those in mental or office work. They are sensitive to fatigue and memory decline and are highly interested in scientifically validated products.
• Quality- and brand-driven group: Tend to favor aesthetic medicine and premium supplements, paying close attention to brand image, reputation, and experience quality.

Gender differences were also notable:

• Women focused more on appearance and skin health, preferring aesthetic treatments, beauty care, and skincare.
• Men tended to focus on nutrition and fitness as ways to maintain overall health.

The core challenge we identified is that anti-aging is a long-term, time-consuming, and costly pursuit. Different groups—shaped by disparities in resources and values—naturally follow different decision-making paths.

Based on this stratification, we realized that group differences extend beyond demand intensity and spending power; they also determine preferences for product formats. To make our project more socially relevant, we must move beyond the laboratory and design a diversified product matrix:

• For the basic livelihood–oriented group, develop safe, affordable, and cost-effective solutions.
• For the balanced group, design nutritional supplements that balance efficacy and convenience.
• For the premium group, explore innovative formats combining aesthetic medicine and personalized health care.

At the same time, we recognized that questionnaires and secondary data alone cannot reveal the market’s true pain points. To ensure our product framework truly aligns with user needs, we plan to conduct further interviews with frontline professionals. By directly listening to their practical experience and insights, we can refine our product positioning and application scenarios, making our designs more targeted, realistic, and aligned with future implementation pathways.

Aesthetic Institutions & Online Health Supplement Influencers

During our early research, we discovered significant gender differences in anti-aging needs. Women are generally more concerned with appearance and skin condition, preferring aesthetic medicine, beauty treatments, and skincare routines. In contrast, men tend to focus on nutrition and fitness to maintain health. This observation inspired us to realize that, if we aim to develop an anti-aging product, we must fully account for the distinct preferences of each target demographic to achieve precise product design and positioning.

Figure 13. Online meeting with Ms. Zhu, manager of an aesthetic medicine institution

Figure 14. Interview with a health influencer with 800K followers — sensitive account details hidden

To verify this hypothesis, we conducted interviews with aesthetic medicine institutions and online influencers in the health supplement industry.

From our discussion with the aesthetic institution, we learned that the clientele was overwhelmingly female, with very few male consumers — reinforcing the fact that female demand for aesthetic anti-aging treatments is highly concentrated.

Meanwhile, in the health supplement e-commerce space, we found that male consumers showed a stronger preference for nutritional and functional supplement products.

These first-hand insights helped us confirm that gender-based differentiation in anti-aging preferences is a genuine and pronounced market trend.

Through comparative analysis, we concluded:

• Female consumers: Their anti-aging needs focus on appearance management and skin improvement, favoring aesthetic and beauty-related products.
• Male consumers: They are more interested in nutrition and physical performance, showing higher acceptance of capsule or supplement-based products.

This divergence in needs made us realize that our project must include both functional products as an entry point and aesthetic-integrated innovations for future development. Based on these findings, we decided to position our initial product in capsule form, as it can cater to a broader audience, and is easier to promote and distribute.

Accordingly, we began focusing on synthetic biological production of NMN, aiming to reduce costs sustainably through eco-friendly biosynthesis, ensuring affordability and accessibility. In the long term, we also plan to explore aesthetic applications of NMN, such as developing NMN-infused skincare products to meet the higher-end demands of female consumers.

By continuously refining our product design, we aim to make NMN better serve diverse groups, addressing both health and beauty needs, and ultimately enhancing the social impact of our project.

Generation 1 Strain

At the initial design stage, we conducted an extensive review of academic literature on NMN biosynthesis. Combined with feedback from our parents about anti-aging needs, we realized that if NMN could be efficiently synthesized within a probiotic chassis, it could potentially offer a more affordable and accessible anti-aging solution. This motivation led us to attempt the construction of our first-generation engineered strain.

Figure 15. Discussion with Laboratory Instructor

During the design process, we actively communicated with our laboratory instructor, who reminded us to carefully consider both the feasibility of the metabolic pathway and the operability of the engineered strain. Through literature research, we found that:

• VpNadV, a nicotinamide phosphoribosyltransferase from Vibrio phage KVP40, possesses high catalytic efficiency, enabling the conversion of NAM (nicotinamide) and PRPP (phosphoribosyl pyrophosphate) into NMN.
• NiaP, a nicotinamide transporter, can actively transport NAM into the cell, ensuring a sufficient substrate supply for NadV.

Based on these findings, we decided to construct a Generation 1 strain to verify the feasibility of NMN biosynthesis in E. coli.

We co-expressed NadV and NiaP in E. coli BL21(DE3) and carried out systematic experiments:

• Established a reliable NMN standard curve to ensure the accuracy of subsequent yield measurements.
• Found that high concentrations of NAM inhibited bacterial growth.
• After fermentation, the Generation 1 strain successfully synthesized NMN, confirming the viability of the designed pathway.
• However, we also observed that additional NAM supplementation did not increase NMN yield and even caused inhibitory effects.

Generation 2 Strain

Although the first-generation strain successfully achieved NMN synthesis, we found that even after supplementing additional NAM, the yield could not be further increased—and in some cases, inhibitory effects appeared. This observation made us realize that NMN biosynthesis was limited by deeper metabolic bottlenecks, rather than by the availability of external substrates alone.

Figure 16. Discussion with Laboratory Instructor

We held in-depth discussions with our laboratory instructor and reviewed relevant literature together. The instructor pointed out that NMN synthesis depends not only on NAM but also on PRPP (phosphoribosyl pyrophosphate). The endogenous PRPP synthesis within the engineered E. coli might be insufficient, thereby constraining the pathway. This insight made us recognize that we needed to enhance PRPP supply through metabolic engineering to overcome the rate-limiting step.

Through literature research, we identified the PRPP synthetase gene (BaPRS, EC 2.7.6.1) from Bacillus amyloliquefaciens. This enzyme catalyzes the reaction R5P + ATP → PRPP, serving as a key node in nucleotide metabolism. Previous studies showed that overexpression of BaPRS can significantly increase intracellular PRPP levels, thereby promoting NMN biosynthesis. Based on this finding, we decided to introduce BaPRS into E. coli and construct our second-generation strain.

By introducing BaPRS into the Generation 1 strain, we conducted fermentation experiments and obtained the following results:

• The NMN concentration doubled compared with that of the first-generation strain.
• However, the high intracellular accumulation of NMN imposed metabolic stress on the strain, leading to slower growth rates in some experiments.

By enhancing PRPP supply, we successfully boosted NMN biosynthetic capacity. Nevertheless, a new challenge emerged: excess intracellular NMN not only negatively affected cell vitality but was also difficult to secrete efficiently. This insight provided a clear direction for the design of our third-generation strain.

Generation 3 Strain

In the second-generation experiments, although we successfully increased NMN yield, we also identified a new bottleneck — the excessive intracellular accumulation of NMN. This accumulation not only led to partial conversion of NMN into NAD⁺, consuming the target product, but also imposed metabolic stress on the cells, reducing their vitality.

This observation inspired a new idea: if we could transport NMN out of the cell in time, it would help maintain intracellular metabolic balance while promoting overall NMN accumulation.

We discussed this issue again with our laboratory instructor and conducted an in-depth literature review. Studies showed that the PnuC transporter from Bacillus mycoides can export NMN from the cytoplasm to the extracellular environment. Our instructor suggested introducing this gene in combination with NadV and BaPRS.

According to the literature:

• The PnuC transporter is a membrane protein capable of recognizing and transporting intracellular NMN to the extracellular space, thereby reducing its internal concentration.
• This not only relieves intracellular metabolic pressure but also prevents NMN from being excessively converted into NAD⁺.
• Furthermore, extracellular accumulation facilitates direct product recovery from the fermentation broth, aligning with industrial-scale production needs.

Building on the second-generation strain, we introduced the PnuC transporter and conducted validation experiments:

• The NMN concentration increased to nearly 1000 μM, far exceeding that of the previous two generations.
• Extracellular secretion of NMN was significantly enhanced, reducing intracellular stress and maintaining cell vitality.
• Although glucose consumption increased slightly, it had no adverse impact on the overall metabolic performance of the strain.

This optimization achieved the breakthrough we had envisioned: the third-generation strain not only demonstrated high biosynthetic efficiency but also gained the advantage of extracellular secretion, laying a solid foundation for future scale-up fermentation and industrial application.

GLP-1 Module

After identifying NMN as the core molecule of our project, we decided not to limit our research to a single active compound, but to explore additional anti-aging molecules with clinical potential. Through literature review and user demand analysis, we noticed that male participants expressed strong concerns about obesity and metabolic health, as well as a higher acceptance of oral drugs and supplements. This insight inspired us to consider introducing a compound that could regulate metabolism, improve weight management, and enhance insulin sensitivity — in addition to NMN.

Figure 17. Online discussion with a nutritionist

We first shared our initial experimental progress with a nutritionist through an online discussion. The nutritionist advised us to focus on synergistic effects rather than relying on a single molecule, which led us to identify GLP-1 (Glucagon-Like Peptide-1) as a promising target.

Figure 18. Group photo with Li-Ying Bio

Figure 19. Discussion with Dr. Ying Zhang, founder of Li-Ying Bio

Subsequently, we collaborated with Li-Ying Bio, where we received professional guidance in protein design.

Figure 20. Team discussion with Dr. Zhou, Shenzhen University

Figure 21. Team discussion with Dr. Shuhua Yin and Dr. Wei Li, CUHK

Since the design complexity exceeded our own capabilities, we further sought support from doctoral researchers at the Chinese University of Hong Kong (CUHK) and Shenzhen University. With their help, we worked on the sequence optimization and construction strategy for GLP-1.

Through literature research, we confirmed the anti-aging and metabolic intervention potential of GLP-1. It has been shown to promote insulin secretion, improve energy metabolism, and alleviate metabolic disorders in animal studies. Market research also revealed that GLP-1 analogs are already applied in obesity and diabetes treatment, indicating strong translational potential.With guidance from the doctoral mentors, we designed a strategy involving codon optimization and cloning of the GLP-1 coding gene, followed by expression of active GLP-1 in E. coli.

We successfully constructed a GLP-1 expression system in E. coli and experimentally verified the expression of the target protein. The results confirmed that GLP-1 was successfully synthesized in the engineered strain, demonstrating the feasibility of our design. Although further optimization of secretion efficiency and protein stability is still required, this achievement has laid the experimental foundation for our next step — developing a male-oriented NMN + GLP-1 capsule, combining metabolic regulation with anti-aging benefits.

Glutathione (GSH) Module

During our research on gender differences in user needs, we found that female participants were more concerned about facial skin condition and antioxidant effects. However, they showed limited interest in oral products, preferring beauty and skincare formats instead. This finding prompted us to consider introducing, alongside NMN, a molecule that possesses both cellular protection and skincare value.

Figure 22. Discussion with a nutritionist

We discussed these gender-specific differences with a nutritionist, who suggested that combining antioxidant effects with skin management could be more appealing to women. Following that, we also reviewed feedback from aesthetic medicine institutions and e-commerce platforms, which further confirmed that the female market strongly anticipates “dual-effect” products combining beauty and health benefits.

Figure 23. Group photo with Alpha Bio — biotech face mask company

Figure 23-B. Technical briefing and discussion with Alpha Bio researchers

Through communication with Alpha Bio, a biotech company specializing in face masks, we learned about the market potential of applying active biological molecules to skincare products. During the discussion, we discovered that Glutathione (GSH) — a classic antioxidant — perfectly met this demand.

By consulting academic literature, we found that the GshF enzyme from Streptococcus agalactiae serotype V possesses dual catalytic functions, allowing it to synthesize GSH in a single step, thereby simplifying the metabolic pathway. Compared to traditional two-step GSH synthesis systems, GshF exhibits higher expression efficiency and is more suitable for metabolic engineering applications.

Based on this, we decided to introduce the GshF gene into E. coli to establish a high-efficiency GSH biosynthesis module.

After introducing and overexpressing GshF in E. coli, we successfully achieved a significant increase in GSH yield. Experimental results showed that:

• Overexpression of GshF did not significantly affect strain growth;
• Enzymatic activity assays confirmed its high catalytic efficiency;
• Supplementing precursor amino acids and improving oxygen supply further increased production;
• Functional assays demonstrated that the engineered strain had a strong free-radical scavenging ability.

These results confirmed that engineered E. coli can serve as an effective microbial factory for GSH production, providing a solid experimental foundation for the future development of NMN + GSH oral formulations and skincare masks.

Project Safety Considerations

Safety is the fundamental prerequisite of our entire project. Without safety assurance, all research and product development would lose their meaning. By reviewing previous iGEM team cases and relevant literature, we realized that any anti-aging–related engineered strain must achieve clinical safety and environmental controllability. Therefore, from the very beginning — the chassis selection stage — we established biosafety as our top design criterion.

During the process of determining our chassis, we consulted our laboratory instructor and analyzed experiences from previous iGEM teams through multiple discussions. Our instructor reminded us that although conventional E. coli strains such as K-12 or BL21 are easy to manipulate in the lab, they have significant limitations for clinical or product applications. Ultimately, we focused on E. coli Nissle 1917 (EcN) — a well-validated probiotic chassis with a long record of human use.

Advantages of EcN:

• Long-term safety verification: As a human probiotic, EcN has been clinically used for over a century, showing reliable intestinal colonization and immune-regulatory effects.
• Lack of typical endotoxins: Unlike regular E. coli, EcN’s lipopolysaccharide (LPS) structure lacks the highly immunogenic lipid A component, significantly reducing toxicity risks.
• Engineering flexibility: The synthetic biology community has already developed mature genetic modification tools for EcN, facilitating the efficient construction of heterologous metabolic pathways.

Based on this evidence, we ultimately selected Nissle 1917 as the core chassis for our project.

In project implementation, we also designed a future-oriented factory production workflow:

Most importantly, we clearly specified in our workflow that only purified products will be extracted, and all engineered strains must be completely inactivated before leaving the production facility. This procedure ensures both environmental and public safety, aligning with strict biosafety regulations.

Through this design, we achieved a balance between technical feasibility and application safety, establishing a solid foundation for the sustainable development of future anti-aging products.

Final User Validation

After completing multiple rounds of product iteration, our team decided to return to our original inspiration — our families and parents. We wanted to revisit the people who first inspired this project and test whether our solution truly addressed the real needs of users.

Figure 24. Team members communicating with their parents

Figure 25. Follow-up interviews with different groups — family caregiver (left); university research administrator (right)

We conducted second-round interviews with our parents and follow-up conversations with previous interviewees. The results showed that:

• Parents remained highly supportive and positive, affirming that our project was practical and meaningful;
• However, some respondents found our technical design more difficult to understand due to the project’s increasing scientific complexity.

Through this process, we discovered several key insights:

• People without a scientific background were often unfamiliar or uncertain about concepts such as “synthetic biology” and “engineered bacteria.”
• Middle-aged and elderly participants preferred to understand our project through everyday analogies, such as healthy eating or skincare routines.

These findings revealed the importance of adopting differentiated communication strategies for different audiences. Without such tailored approaches, our project could be misunderstood or fail to gain broader public acceptance.

In response to these challenges, we designed and implemented inclusive science communication activities to make our project more approachable and relatable. We believe that the true value of our work lies not only in laboratory data or product prototypes, but also in whether it can be understood, accepted, and integrated into everyday life by diverse groups.

For more details, please refer to Inclusivity.

Clinical or Application Validation — Interview with Physician

Anti-aging has long been a topic of great public interest — it reflects both the desire to improve quality of life and the vast potential of a growing market. To verify whether our research carries practical clinical value, we decided to interview a physician, seeking a professional medical perspective to evaluate our project and ensure that it not only holds scientific significance but also aligns with real clinical needs.

Figure 26. Online consultation with a physician via WeChat video call

During the interview, the physician first affirmed our research direction. He pointed out that if we could develop an effective oral anti-aging product, it would possess a strong competitive advantage. In particular, if the product could prevent vascular plaque formation and reduce blood lipid levels, it would be especially beneficial for people with “three highs” — high blood pressure, high cholesterol, and high blood sugar. Such outcomes would directly address major social and medical needs in the field of aging intervention.

At the same time, the physician also provided constructive feedback: the use of E. coli as a chassis organism and the involvement of genetic modification could potentially trigger public concern or misunderstanding. He suggested that, for external communication, we might consider adopting a more inclusive and science-friendly terminology, or using language that is easier for the general public to accept and understand.

The physician further outlined the standard preclinical development pathway our project should follow:

1、Animal testing: Future studies should verify safety and efficacy using multiple animal models such as mice, dogs, cats, and pigs.

2、Gradual transition to human trials: Only after collecting comprehensive animal data should the project proceed to human clinical trials.

3、Ethical and regulatory compliance: While our current experimental design poses no ethical or health-related issues, the physician reminded us to remain attentive to public perception and legal frameworks.

From this consultation, we gained three key insights:

1、Clinical value confirmation: The physician emphasized that oral anti-aging formulations have significant potential, particularly for the “three highs” population, where clear clinical applications exist.

2、Experimental roadmap: We clarified the proper development sequence — beginning with animal studies, followed by human clinical trials, in accordance with standard biomedical R&D protocols.

3、Public communication awareness: We recognized the need to use approachable and socially sensitive terminology in science communication to avoid misunderstandings surrounding terms like “E. coli” or “genetic modification.” This also reminded us to strengthen public education and inclusivity efforts, ensuring our project is both scientifically sound and socially accepted.

Legal Consultation — Interview with Lawyer

During the progress of our project, we realized that scientific feasibility is not the only barrier determining whether a product can successfully enter the market. Regulatory compliance and legal risks are equally crucial for ensuring the successful translation of research outcomes into real-world applications. In particular, our core molecule NMN is subject to significantly different regulatory policies across countries and regions. To prevent potential policy or legal obstacles in later stages, we decided to consult a lawyer to obtain professional advice on legal compliance and intellectual property protection.

Figure 27. Discussion with a lawyer from Shanghai Landi Law Firm

During our conversation with the lawyer, we raised several key questions:

1、Regulation of synthetic biology research: If we produce NMN through engineered E. coli and plan to sell it in the future, what safety or approval requirements must we consider?

2、Intellectual property protection: As a student team without legal entity status, how can we legitimately apply for and protect our technological achievements?

Through the discussion, we obtained the following key insights:

• Regulatory compliance: Research involving synthetic biology must comply with the Biosecurity Law of the People’s Republic of China, and certain experiments or public demonstrations may require prior filing or approval.
• Intellectual property: Student teams can apply for invention patents as individual applicants (natural persons). Additionally, internal agreements can be signed to clarify rights and profit distribution among team members. The team may also file a provisional patent, with the option to convert it into a formal application within one year.

From this consultation, we gained two major takeaways:

1、Proactive regulatory communication: The lawyer suggested that we actively engage with regulatory authorities, such as the Food and Drug Administration (FDA) or related oversight departments, to seek early guidance. This approach not only helps us avoid potential risks, but also demonstrates our commitment to safety and legal compliance.

2、Clear intellectual property pathway: We now have a defined strategy for protecting our innovations, including filing patents and formalizing internal agreements to ensure transparent and fair distribution of rights and benefits among team members.

Government Regulation and Public Communication Management

We consulted legal experts from the Market Supervision and Regulation Department to gain a deeper understanding of the current regulatory environment and compliance requirements relevant to our project.

Figure 28. Online consultation with Lawyer Zhang Shihua

During our online interview with Lawyer Zhang, he provided insights from a legal and regulatory perspective, addressing our questions regarding the legality of NMN, restrictions on health product advertising, and approval procedures. Throughout the discussion, Lawyer Zhang not only patiently answered our questions but also explained the regulatory logic behind policy enforcement, using real-world case examples. This helped us develop a comprehensive understanding of the compliance process within the health and biotechnology industry.

From this consultation, we obtained the following key takeaways:

1、Approval Process: To legally enter the market, any new active ingredient must undergo a strict approval procedure and be included in the national regulatory directory. Additionally, all production stages must comply with hygiene and quality standards.

2、Advertising Compliance:Health product promotions must be strictly differentiated from pharmaceuticals. Advertisements cannot exaggerate efficacy or claim therapeutic effects. Only approved functional descriptions may be used, and all materials must include the statement: “This product cannot replace medication.”

3、Regulatory Focus Areas: In recent years, short videos, live streams, and e-commerce platforms have become major channels for health supplement sales. Regulatory authorities are increasingly focused on identifying and penalizing false or exaggerated claims in these online promotions.

After absorbing these insights, we decided to make strategic adjustments to both our project design and communication approach:

• In the R&D phase, we will clearly distinguish between “research-oriented exploration” and “commercial application”, emphasizing our scientific objectives rather than making premature claims about market-ready efficacy.
• In our business plan and promotional materials, we will strictly adhere to legal terminology, avoiding any expressions that could be misinterpreted as therapeutic claims.
• We also plan to continue seeking guidance from legal and regulatory experts to ensure that our future product development and commercialization pathways remain fully compliant with national approval procedures and industry standards.

Establishing Quality Standards — Interview with Mr. Zhang

For any product to truly enter the market, innovative synthetic biology design alone is not enough. The establishment of quality standards is the key to ensuring safety, efficacy, and public credibility. This realization prompted us to reflect on a crucial question: How can we transform scientific research results from “laboratory innovation” into real-world applications?

Figure 29. Discussion with Mr. Zhang Zhenkun

We had an in-depth discussion with Mr. Zhang, an industry expert with extensive experience in biotechnology and product development. He emphasized that quality standards are not only essential for scientific reliability, but also a prerequisite for public acceptance. Through real-world examples from the biotechnology and healthcare industries, he illustrated that even the most cutting-edge scientific innovations cannot achieve large-scale application without standardization and quality assurance.

Inspired by this discussion, we systematically reviewed national and international standards related to health supplements, functional foods, and biological products. Our findings revealed that:

• Different fields have varying quality requirements, particularly in purity analysis, stability testing, and safety evaluation, all of which are subject to strict regulations.
• In the anti-aging product market, consumers are highly sensitive to safety, which is directly tied to the presence of transparent and reliable quality standards.
• As an emerging compound, NMN still lacks a fully developed quality system. The market urgently needs a scientific and transparent quality control framework.

Based on the literature review and Mr. Zhang’s guidance, we established a preliminary quality control process tailored to our project:

1、Production stage: Ensure that engineered bacteria are cultivated under strict and controlled conditions to maintain product consistency.

2、Testing stage: Use validated analytical methods from the literature to confirm NMN purity and concentration.

3、Safety evaluation: Incorporate risk prevention mechanisms during the design stage to avoid potential biosafety issues.

4、Standardized documentation: Create traceable operational protocols and quality records, forming a clear “Synthesis–Detection–Verification” closed-loop system.

Through our conversation with Mr. Zhang and our exploration of quality standards, we developed a deeper understanding that standardization is the bridge between scientific research and societal value. This not only strengthened the scientific rigor and credibility of our project but also encouraged us to think about long-term applications and industrial translation.

For our team, this process marked a transformation — from being pure scientific explorers to becoming practical innovators who understand the relationship between research and society, and between science and industry.

Project Feasibility

Figure 30. Online discussion with graduate students from the Southern University of Science and Technology

The synthesis of NMN can be achieved through two primary approaches: traditional chemical synthesis and biosynthesis using engineered microorganisms. To better understand the advantages and practical potential of our proposed biological route, we conducted a detailed discussion with two chemistry graduate students from the Southern University of Science and Technology — Mr. Xu and Mr. Liang. Both have extensive experience in organic synthesis research and offered valuable, firsthand insights into the strengths and limitations of the two methods.

During the discussion, they patiently answered our questions regarding synthetic pathways, experimental costs, and industrialization challenges.

From this interview, we obtained several key insights:

• Challenges in chemical synthesis: The main difficulties lie in multi-step reactions, which often generate by-products and impurities. The purification process consumes large amounts of solvents and chromatographic materials, leading to higher costs and lower yield and purity.
• Cost structure: The use of complex precursors, rare reagents, and precious metal catalysts (such as palladium and platinum) significantly increases experimental expenses.
• Advantages of biosynthesis: In contrast, enzyme-catalyzed reactions and microbial fermentation operate under milder conditions and simplified steps, reducing raw material consumption and energy use. This approach demonstrates clear advantages in terms of green chemistry and economic feasibility.
• Research-to-application transition:Organic synthesis often remains within the realm of fundamental research and is difficult to scale up for industrial applications in the short term. Biotechnology, however, offers greater engineering potential and practical applicability, particularly in the fields of environmental protection and metabolic engineering.

This exchange helped us gain a clearer understanding of the superior cost-efficiency, sustainability, and application potential of the biological synthesis route, further strengthening our confidence in advancing the synthetic biology–based approach for NMN production.

Project Commercialization

As our scientific research progressed, we gradually realized that science and business are not opposing forces — they complement and strengthen each other. Scientific achievements that remain confined to the laboratory cannot truly benefit society; commercialization, on the other hand, enables scalable application of research outcomes and serves as a way to validate their real social value.

With this understanding, we decided to move from research toward commercialization, aiming to transform our synthetic biology innovations into market-ready, socially impactful products.

During the commercialization exploration phase, we engaged in close communication with stakeholders from various fields:

• Entrepreneurs — shared their experience in capital operations and business models, helping us understand that a sustainable business model is the foundation for translating research into real-world impact.
• Synthetic biology and process engineering experts — analyzed potential bottlenecks in strain optimization and production processes, emphasizing that high yield and low production cost are essential for market competitiveness.
• Investors and industry mentors — introduced us to strategies for market channels, strategic partnerships, and product pricing, enabling us to develop a systematic understanding of the industrial value chain.

To identify the right market entry point, we carried out the following initiatives:

• Business model learning: Studied successful brands to understand the complete process from patent layout to market promotion.
• Differentiated product portfolio design: Based on our survey data and gender-specific needs, we developed concepts for a multi-product matrix targeting different consumer segments.
• Market and pricing research: Examined how income levels affect price sensitivity and brand trust among consumers.
• Patent and compliance research: Learned how to file invention patents as individual applicants (natural persons) to legally protect our team’s technological achievements.

Through these efforts, we began shaping a clearer commercialization roadmap, integrating scientific innovation with practical business strategy, and paving the way for our synthetic biology project to achieve real-world impact and sustainability.

For more details, please refer to Entrepreneurship

Through engagement with parents, the public, doctors, lawyers, and industry mentors, we discovered that the demand for anti-aging solutions is real, yet there are significant differences across user segments, social perceptions, and regulatory thresholds. Human Practices (HP) has continuously helped us calibrate the alignment between science and society: from parents’ lived experiences to doctors’ clinical pathways, from regulatory advice to quality standards, and further to business models and investor feedback, we have established a complete value loop. HP not only helped us refine the technical roadmap (NMN, GLP-1, GSH) and ensure safety and compliance, but also enabled our project to be understood, accepted, and applied by society. In this sense, HP serves as the bridge linking science—society—industry, ensuring that our solution carries true scientific value, social value, and commercial potential.

Value Pathway

Figure 31：Cycle of Responsible Research

Every step along this path is a recalibration of value: aligning not only with user needs, but also with social expectations, regulatory requirements, and industrial implementation.

Inspiration (family/society) → Research (segmentation/doctors/lawyers/regulators/industry) → Design (NMN/GLP-1/GSH & EcN safety chassis) → Iteration (yield/secretion/quality control) → Compliance (registration/naming/communication boundaries) → Translation (product matrix/channels/partnerships) → Inclusivity (science popularization/education/feedback).

Future Outlook

Our project will not stop at the laboratory stage. Moving forward, we will continue to optimize the engineered strains and production processes, advancing from small-scale experiments to pilot and industrial-scale applications. On the clinical side, we will steadily progress along the path of “animal experiments → human validation” to ensure both scientific rigor and safety. At the societal level, we will keep promoting inclusive activities to help middle-aged and elderly groups better understand and participate in synthetic biology. At the industrial level, we will actively explore collaborations with enterprises, investors, and application scenarios, driving our product from research output toward an affordable, sustainable, and widely accessible anti-aging solution. Ultimately, we hope ChronoCure will not remain merely a competition project, but grow into an innovative pathway that truly improves the quality of life for families, society, and future generations.