Introduction
First of all -- we would like to express how deeply proud we are of the hard work and accomplishments that went into this project! Over the past few months, every member of Team SHSD, together with the advisors, instructor, and partners who supported us, has shared in a truly meaningful competition cycle.
Our concerns about global warming and our interest in Chlamydomonas reinhardtii as a model organism inspired this research project. Through the design and construction of plasmids, cultivation and genetic modification of algae, as well as PCR/qPCR verification and heat stress assessment, we carried out a series of experiments to generate a transformed C. reinhardtii strain and evaluate its potential resilience to high temperature.
At the same time, we actively engaged in human practices, scientific education, and related events. Throughout the project, we continuously optimized experimental conditions and setups to overcome various technical challenges. With the support of external sponsors and advisors, we gathered feedback, absorbed new insights and discoveries, and integrated them into the project to continuously refine and advance our research.
We believe our efforts have made valuable contributions to the iGEM community and beyond: not only in scientific research, but also in team building, external collaboration, cross-cultural exchange, social impact, and in learning to cope with setbacks and overcome challenges. We are honored to present a detailed summary of these achievements:
PART I. Scientific Contributions: We Contribute a Tiny Drop to the Vast Ocean of Algae Studies
As a high school team, we have approached this year’s iGEM with a mindset of learning and growth. We put our full effort into our research, seeking to apply creativity and practical spirit in the hope that it will carry meaningful value. Specifically, our contributions are as follows:
- We provide iGEM teams interested in future research on Chlamydomonas reinhardtii with comparative cultivation experience for a cell wall–intact strain (CC-124) and a cell wall–deficient strain (UVM-4). This includes not only detailed, experimentally validated cultivation conditions, but also differences in viability between the two strains under electroporation, upregulation of mitochondrial chaperone protein expression, and high light exposure, together with their underlying mechanisms. To our knowledge, direct comparative conclusions on the latter are not numerous in existing research. C. reinhardtii may be common in the wild, but in the laboratory, especially for beginners, it can be a fragile organism. This is particularly true for UVM-4, a commonly used cell wall–deficient strain in transgenic experiments. We hope our experience helps subsequent teams avoid pitfalls in Chlamydomonas cultivation.
- Based on our experimental results, we propose 10 μg/mL as the optimal antibiotic concentration for resistance screening in transgenic C. reinhardtii. This concentration is consistent with existing findings while remaining conservative within the optimal range. This choice helped us avoid unintended growth inhibition of UVM-4–positive transformants, which are especially fragile after electroporation and upon upregulation of mitochondrial chaperone protein expression. Our selection provides a targeted reference for subsequent teams with similar technology, experience, laboratory conditions, and target strains (e.g., cell wall–deficient strains).
- Compared with the chloroplast-acting homologous proteins CPN60α and CPN60β, our target gene, CPN60C, has received less attention in prior C. reinhardtii research. Our study adds to this relatively underexplored area and enhances its innovative character. Our experimental results show that increasing expression of the endogenous CPN60C gene enhances the survival and growth of transgenic algae under heat stress.
- We contributed two CPN60C expression cassettes as new composite parts to the iGEM community. We believe CPN60C has further potential for exploration and would be honored if these parts facilitate research by subsequent iGEM teams.
- Our research direction is highly creative and imaginative. Unlike our initial plan to target carbon-concentrating mechanisms directly, during Project Evolution we considered a practical yet often overlooked scenario for algal carbon-concentrating applications: if carbon concentrating is most needed where the greenhouse effect is most pronounced, then algae must first be able to survive in relatively hot environments. This led us to recognize the connection and potential between heat-stress resilience and algae’s capacity to mitigate the greenhouse effect. We believe our experience evolving this research direction can provide insights and references for other iGEM teams.
- We explored statistical modeling to predict the probability of target gene expression from plasmids. While most iGEM modeling focuses on molecular dynamics, given our technical constraints we investigated statistical approaches instead. Drawing extensively on prior research, we used Monte Carlo simulations of sequential events to develop our modeling framework and conclusions.
PART II. Team Organizational Contribution: We Provide a Unique Organizational Model for Other iGEM Teams
Our team implemented a distinctive and challenging organizational structure. During the project timeframe, our members were based in Shanghai, China; Chongqing, China; the United States, and Australia, with additional external support from the Germany, France, and Vanuatu. Collaborations for background research, conceptualization, dry lab work, visual design, and related tasks were primarily conducted remotely, while Human Practices combined online and offline efforts. Wet lab work was carried out entirely offline, spanning the entire summer.
We held weekly online meetings to ensure team cohesion and information sharing. Even across three time zones, members consistently attended on time despite the challenges. Our leadership structure consisted of one captain and two vice-captains, with a detailed and well-balanced division of responsibilities for team management and scientific execution. Each functional sub-group had a specific lead, and more than half of our members gained leadership experience through voluntary roles. We also established a robust reporting mechanism to ensure that any issues were promptly communicated and resolved. We believe this organizational model offers a strong example for cross-cultural, cross-time-zone, and cross-language collaboration, and we look forward to future iGEM teams developing even more creative approaches.
PART III. External Collaborative Contributions: We Provide a Successful Example of Reaching Out to the Broader Research Community
External support was indispensable to our participation in this year’s competition. While we have expressed gratitude in many places on our Team Wiki, we wish to highlight it again here: the construction of Plasmid A was made possible through the generous donation of plasmid backbones by Dr. Lei Zhao and Dr. Yuyong Hou from the Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences. Dr. Hou also helped optimize the expression cassette design, construct the plasmids, and provided valuable suggestions. The construction of Plasmid B was supported by Dr. Kaiyao Huang and Dr. Lian Ye from the Institute of Hydrobiology, Chinese Academy of Sciences, who likewise donated plasmid backbones. We are deeply grateful for their selfless assistance.
This journey began when we bravely reached out by email to ask for help. We had discovered that the so-called “common” plasmid backbone for Chlamydomonas reinhardtii was not actually common in China, that full-sequence synthesis was time-consuming, costly, with a high risk of failure, and that international ordering times were unpredictable due to the policies. Summoning our courage, we contacted Chinese scholars working on Chlamydomonas reinhardtii.
We still remember the apprehension we felt then: Would this be too intrusive? Would the scholars, busy as they are, even notice us? Or dismiss us as reckless high school students?
To our surprise, we received immediate and encouraging responses. The scholars offered support beyond our expectations, demonstrating remarkable generosity. While we felt deeply grateful, we also thanked ourselves for taking that courageous step -- overcoming hesitation and shyness made this valuable scientific exchange possible.
We believe this story can inspire future iGEM teams: be bold, reach out, and ask questions! Whether you are a professor, a PhD candidate, or a high school student eager to embark on scientific research, we are all bound by our shared passion and respect for science.
PART IV. Contribution of Cross-Cultural Communication: We Demonstrated How to Resonate Globally with the Issues We Care About
As noted earlier, our external supporters also included friends from Germany, France, and Vanuatu. We actively engaged with them to share perspectives on climate change and extreme weather, while envisioning future solutions such as the Chlamydomonas solution we are developing.
These cross-cultural exchanges helped us realize that, despite being located in Asia, Europe, the Americas, and Oceania, and despite differences in city size, population, and industrialization, we are all experiencing the consequences of global warming. The issues we care about are not regional but planetary. This recognition underscores the significance of our work and the essence of our mission. The feedback we received from these friends also became a key driver for ongoing reflection and refinement of our project.
We believe that pursuing cross-cultural communication and integrating project reflection can serve as a source of inspiration for future iGEM teams.
PART V. Contribution for Social Impact: We Helped More People Understand the Facts and Principles Behind Global Warming, the Greenhouse Effect, and Algae
Through our Human Practices events, we carried out surveys, launched social media initiatives, and organized outreach activities on campus and in the community. We explored how people of different ages, gender identities, and educational backgrounds perceive global warming and the greenhouse effect. We also gathered their opinions on our proposal to regulate carbon dioxide levels using genetically modified Chlamydomonas reinhardtii. In doing so, we educated communities about the causes and consequences of global warming and the greenhouse effect, while raising awareness both locally and more broadly.
Notably, our campaign also helped correct public misconceptions about Chlamydomonas reinhardtii. We found that many people were less familiar with algae than expected, and often generalized the harmful effects of certain species to all algae. For example, “the red tide” is widely taught in high school biology as being caused by algae, but few realize that it was primarily caused by diatoms and dinoflagellates -- not green algae such as Chlamydomonas reinhardtii.
We would also like to acknowledge the important support from Zihan (Emma) Lyu. In particular, building on the team’s research progress and ongoing outreach efforts, she helped prepare further social media materials and assisted in coordinating and carrying out the subsequent communication plan. Her involvement has definitely strengthened the design, organization, and writing of our social media engagement and next-step project activities.
We believe that even our small-scale outreach efforts contributed valuable information to the iGEM community and beyond, raising awareness and correcting misconceptions, just as we ourselves have learned from the work of other iGEM teams.
PART VI. Contributions to Frustration Management: Our Courage, Creativity, Growth, and Reflection
During our iGEM journey, our team encountered and overcame numerous challenges. Some were expected, such as limited time, resources, and technical capacity, which led to repeated failures in our initial PCR attempts. Others were wholly unexpected: the “common” plasmid backbone we relied on proved unavailable locally; our laboratory relocation rendered equipment unusable while we really need to conduct PCR verification; and midway through the project, we discovered a database-identified target gene that was not part of our original plan.
How do we view such setbacks? We believe no one in the iGEM community would dismiss them as worthless, just as no one assumes scientific research is always smooth sailing. For us, setbacks were moments to confront, resolve, and even transform into opportunities, sparking new ideas in adversity.
Unable to obtain a plasmid backbone? We reached out to researchers working on Chlamydomonas reinhardtii.
Unable to access our lab? We sought temporary external resources and designed alternative protocols.
Identified a new gene of interest? We asked: could this be even better than our original plan? Let’s investigate -- and it was.
In this way, we repeatedly discovered solutions to problems, encountered supportive researchers and peers, and advanced toward our goals. We hope to share our courage, flexibility, and problem-solving experience with other iGEM teams.
Of course, maintaining optimism and perseverance does not mean mistakes are inherently positive. Setbacks remain setbacks. But they brought us lessons and reflections we would like to share with the iGEM community:
Always scrutinize every assumption of “common sense” or “should-be” during project design, especially when working in unfamiliar research areas or with limited resources.
A robust double-checking process can prevent basic errors. Even the most diligent researchers experience fatigue or distraction, and mistakes can occur. Instead of expecting one person to be flawless, it is wiser to build in multiple checkpoints.
Never underestimate the importance of literature review and background research. Comprehensive groundwork helps projects avoid many pitfalls.
- The above represents Team SHSD’s contribution to the 2025 iGEM competition. We sincerely hope that future iGEMers can gain something from our work: whether it be experience, methods, and conclusions, or courage, resilience, and passion for science.