April
Week 1
After the school competition selections, students from different faculties joined the UESTC-China team, and everyone is full of energy and enthusiasm! So far, we've successfully recruited members responsible for the wet lab section, but we're still looking for more students to handle mathematical modeling, human practices, and design. We held our first team meeting, got to know each other, and elected our student leader and finance manager. In addition, we had an initial discussion with our PIs about the project's direction. Everyone spent quite some time reading papers to dive deeper into the research on PET degradation and detection. Meanwhile, our WIKI team is gearing up to learn web development basics, familiarizing themselves with frameworks to prepare for the website building phase later on.
Week 2
This week was mainly focused on summarizing and discussing a massive amount of literature we've gathered. Reading papers and brainstorming can be exhausting, but thanks to our PIs' guidance, we quickly formulated an initial project idea. Now, we’re thinking about the specific technical roadmap. Our WIKI team continues to dive into web-related knowledge. It’s been a busy week, with everyone working tirelessly towards the same goal, sometimes barely getting any rest.
Week 3
We successfully recruited new members for mathematical modeling, human practices, and design, making our team officially complete! After our routine Wednesday meeting, we took a group photo in front of the main building on campus. This week, our project design finally started taking shape. The team working on degradation is planning to solve the issue of plastic macromolecule breakdown using a surface display system. Meanwhile, the members responsible for the upcycling section have begun constructing a multi-enzyme cascade conversion system inside cells. The detection team has decided to pursue a colorimetric approach, and we've also discussed the feasibility of various techniques like gene editing. Our WIKI members are now gathering materials, and our HP team member has presented her overall plan for team activities—big things are definitely coming!
Week 4
This week, the identification team confirmed the sequences of most expression vectors and conducted a review, steadily advancing.
The fastest-progressing degradation group continues to work on the construction of the surface display system.
The colleague in charge of upgrading and rebuilding the plate has designed a multi-enzyme cascade system and completed the construction of a three-plasmid system.
The colleagues in the gene editing module identified the target genes for knockout - aldosterone reductase and ethanol dehydrogenase
The graphic designer's colleagues have started to design team uniforms for the team.
May
Week 1
The start of this month was incredibly busy! We’re deep in vector construction, which means every day is basically a PCR marathon.
The degradation group successfully verified the construction of the surface display system, and the PET degradation experiments are steadily progressing. Meanwhile, the upcycling team completed the first plasmid transformation and confirmed it using ampicillin resistance and gene sequencing. After successfully preserving the bacterial strains, they began working on the conversion of terephthalic acid into vanillin. The gene-editing team also designed a gene knockout strategy based on the λ-RED system.
The WIKI group’s progress remains steady, as they continue to learn the framework and gather materials.
Week 2
This week, the detection team expressed four types of anchoring peptides fused with sfGFP and conducted affinity experiments with plastics. We also analyzed the preliminary results of fluorescence experiments. The degradation experiments are starting to take shape. The detection techniques are being optimized step by step, with everyone testing various conditions—things are getting more intense in the lab!
The upcycling team began validating the expression of target proteins in E. coli transformed with the first plasmid. Using SDS-PAGE, they explored enzyme protein expression and, after finding poor results, decided to switch bacterial strains to improve protein yield. The gene-editing team attempted three rounds of experiments based on the designed λ-RED knockout route, but all attempts unfortunately failed.
After multiple discussions during our team meetings, we finally nailed down the direction for our dry lab work. The dry lab team started looking for models suited to directed enzyme evolution.
Week 3
The hectic pace continues, but this week was relatively quiet.
The detection team has been exploring the cleavage conditions for TEV protease, which will take more time.
Both the degradation and gene knockout teams are still combing through literature and adjusting experimental conditions.
The upcycling team continued refining experimental methods and, after switching strains, observed good protein expression. They then harvested the cells and began the upcycling process for terephthalic acid. However, liquid chromatography analysis didn’t detect the expected product, protocatechuic acid, prompting the team to further improve the degradation system.
The modeling group started testing models for protein evolution, while the HP team had a discussion with Professor Quan to finalize the framework structure. Meanwhile, the WIKI group made a surprising leap forward this week, officially starting the framework construction for the website.
Week 4
The lab teams continued to adjust the conditions for the degradation experiments. Repeating experiments requires patience, but it feels like everyone is gaining more confidence with each attempt.
The detection team has completed the initial exploration of the TEV protease cleavage system, while the modeling team began testing molecular docking of the mutants.
The upcycling team is still working on refining the transformation system conditions. This week, they made significant adjustments to the degradation system components, but high-performance liquid chromatography still didn’t detect protocatechuic acid, so they’re continuing to tweak the experimental conditions.
The gene-editing team, after consulting protocols from Nature and various forums, tried changing electroporation conditions, cell recovery times, and medium components, but success remains elusive.
The HP and WIKI groups had a relatively smooth week. In particular, the WIKI team has started drafting some basic documentation, marking steady progress.
June
Week 1
As June begins, our experimental progress has really picked up speed. However, the detection team faced some setbacks this week—modifying the gold nanoparticles didn’t go as planned, and we found that the concentration of the anchoring peptide was still too low. To make things more challenging, we ran out of bacterial culture, so we need to re-express the target protein.
On a more positive note, the degradation team made some key adjustments and finally obtained critical data, which has everyone really excited!
The upcycling team has not stopped tweaking the transformation system. Unlike last time, this week they used ultrasound to lyse the cell suspension and successfully detected the target product. This narrowed the problem down to the transmembrane transport of terephthalic acid.
The upcycling team has not stopped tweaking the transformation system. Unlike last time, this week they used ultrasound to lyse the cell suspension and successfully detected the target product. This narrowed the problem down to the transmembrane transport of terephthalic acid.
The WIKI group is progressing steadily with development.
Week 2
The lab was buzzing with activity again this week. The detection team successfully re-expressed and purified their proteins.
The degradation team continued adjusting experimental conditions, testing more optimization schemes.
The upcycling team, after numerous trials, finally nailed down the optimal transformation system. By adding 1% n-butanol, they successfully enabled the transmembrane transport of terephthalic acid and converted it into protocatechuic acid. This marked a major milestone in the success of the first step of the multi-enzyme cascade system.
Meanwhile, the gene-editing team started learning the principles and structure of the pEcCas gene knockout system.
The modeling team continued their experiments on protein mutations.
The WIKI team made solid progress by refining the structure of the website, and it looks like it’s going to be quite impressive when we showcase it later.
On the HP side, plans for upcoming outreach activities are already in the works.
Week 3
This week, the degradation team’s experimental results gave us renewed hope! The data looks incredibly promising, and the focus will now shift to analyzing the results in depth.
The upcycling team successfully transformed the remaining two plasmids separately and used SDS-PAGE to analyze enzyme protein expression. The results showed that both plasmids could express independently.
The detection team also made progress, exploring new TEV protease cleavage conditions, resulting in a significant increase in concentration.
The gene-editing team started designing a technical route for the pEcCas gene knockout system, working on plasmid construction and other tools.
The modeling team continued testing various mutants—it’s a complex process, but the progress is steady.
The HP team began drafting promotional materials, while the WIKI team continued enhancing the website’s functionality.
Week 4
This week was full of energy!
The degradation team’s preliminary data is in, and now they’re diving into a deeper analysis of the results.
The upcycling team completed the co-transformation of three plasmids, and after screening with antibiotics, they’re ready for the next phase of work.
The detection team finished preparing all the anchoring peptides. The modeling team is still focused on studying mutants, and although the process is tedious, the results are looking promising.
The gene-editing team conducted three experimental attempts based on the pEcCas gene knockout system’s technical route, but all resulted in false positives.
The HP team has finalized their promotional plan and is ready to put it into action.
July
Week 1
As July begins, our experiments are progressing exceptionally well.
The degradation team completed the final round of PET degradation experiments this week. Everyone is busy collecting and analyzing data, and the atmosphere in the lab is both tense and fulfilling.
The upcycling team moved on to verifying the second key pathway. Under new conditions, they successfully detected the final product, vanillin, confirming the feasibility of the second step in the multi-enzyme cascade pathway.
The detection team is also in full swing with their colorimetric experiments, continuously exploring optimal conditions for nanoscale modifications.
Unfortunately, the gene-editing team has faced a series of setbacks. They’re going back to square one, diving into literature and forums to find a workable gene-editing system.
The modeling team is applying mutant data to their models, preparing to provide more theoretical support for the experiments.
On the HP front, the team participated in the first Chengdu Metropolitan Ecological and Environmental Volunteer Service Competition and won the first prize in the Creative Group. This recognition of our earlier plans has been a great encouragement, and we’re hopeful for continued progress in our HP work moving forward!
Week 2
The lab remains as busy as ever, with both the degradation and detection experiments reaching critical stages.
The upcycling team successfully induced expression using the final constructed strain. Under the optimized conditions, they once again detected vanillin as the final product, confirming the viability of the entire pathway.
This marks a major success for the upcycling project!
The detection team encountered some difficulties with their colorimetric experiments, but we’re confident that teamwork and collaboration will help overcome these challenges.
The gene-editing team made a breakthrough by discovering the PHCY-25A/PHCY-26D dual plasmid gene-editing system, allowing them to design a new technical route for targeted gene knockout.
The modeling team continued refining their models, and everyone is excited to see how well the experiments and theoretical models can come together.
Meanwhile, the HP team had a busy week as well. They organized a summer community event titled "Green Future: Reducing Plastic, Cutting Pollution" at Xinglin Community in Yongning Street. The activities included educational talks, DIY bottle-recycling crafts, talent shows, eco-protection poetry collages, and even an environmental song performance. The kids were highly enthusiastic, and everyone walked away with a sense of accomplishment!
Week 3
The progress of our experiments is becoming smoother by the day! The degradation team is nearing the final stages of data collection, while the upcycling group has completed their experimental validation phase. We’re gaining more confidence in the stability of our experimental data.
The detection team finally achieved a breakthrough in their colorimetric experiments this week, and the next step will be to replicate the results for consistency.
The gene-editing team decided to abandon the ethanol dehydrogenase gene knockout and instead completed the design of sgRNAs for five aldehyde-ketone reductase genes, successfully introducing the vectors.
The modeling team has reached a point where their model can accurately predict the performance of various mutants, which is highly satisfying for everyone.
Despite the hectic pace in the lab, the WIKI preparations are progressing in parallel.
On the HP front, our team recently engaged with students from Malaysia to explore new solutions to microplastic pollution, enriching the research and learning experience while brainstorming innovative approaches to tackle this global issue.
Week 4
The experimental tasks for July are finally complete! The degradation and upcycling teams have successfully gathered all the necessary data and are now shifting their focus entirely to data analysis. The detection team wrapped up their experiments beautifully, with the colorimetric process now fully optimized and ready.
However, the gene-editing module hit a snag in the design of sgRNA plasmids, which will require some problem-solving moving forward.
The modeling team is in the final stages of fine-tuning their mutant models, and they’ll soon begin cross-verifying them with experimental data. The conclusion of this experimental phase brought a sense of relief to everyone, but we all know the next steps will be just as challenging.
On the WIKI and HP fronts, the teams are collaborating to discuss the design of our IP character and refine the storyline. More communication with the experimental groups will be needed to ensure that the designs align closely with the lab’s work.
August
Week 1
As we enter August, the experiments are entering their final stages. The degradation, upcycling, and detection teams have all shifted their focus to deep analysis of the experimental data. The teams discussed how the data performed under different conditions and are working to find the optimal solutions.
The gene-editing team made significant progress this week—after adjusting the recovery time, they successfully constructed the sgRNA plasmid based on the YahK gene.
The modeling team combined the experimental data with their mutant models, successfully validating the accuracy of their predictions.
At the start of August, the HP team held a collaborative exchange session with students from the University of Science and Technology of China (USTC). We learned about their innovative design ideas and strategies, leading to an exciting exchange of thoughts and inspiration that left both teams with valuable insights.
Week 2
The analysis of experimental data continued this week. Members from the degradation, upcycling, and detection teams began summarizing the key findings of their experiments and started drafting technical documentation. The gene-editing team made attempts to construct sgRNA plasmids for four other aldehyde-ketone reductase genes but encountered repeated failures in obtaining the sgRNA plasmid for the YahK gene.
Meanwhile, the modeling team further refined their model to ensure consistency with experimental results.
We held a few internal meetings to discuss the best ways to present our experimental data, with everyone eagerly contributing ideas.
The HP and WIKI teams collaborated on designing the storyline and finalizing the project name, narrowing down their focus after multiple rounds of discussion and revisions.
Week 3
This week’s main focus was organizing the experimental results and drafting the experimental reports. The degradation, upcycling, and detection teams completed their data analysis and plots, successfully addressing some of the critical issues encountered during the experiments. The gene-editing team successfully extracted the sgRNA plasmid for the YahK gene from the strain and began testing it in gene knockout experiments.
They also continued working on the sgRNA plasmids for the other aldehyde-ketone reductase genes.
The modeling team finished optimizing their model, and the next step will be to integrate these results into our project presentation. We aim to complete all reports and related materials by mid-September.
The HP team has been busy organizing a social media campaign to connect with universities, planning to complete their outreach posts by the end of August. They are also designing an online environmental and innovation competition, with plans for an offline event in September, which involved active discussions with the school.
Week 4
This week, the degradation, upcycling, and detection teams meticulously organized all their experimental data.
The gene-editing team, after multiple setbacks, decided to focus on the YahK gene knockout. The experimental team collaborated closely with the HP and design teams to discuss the presentation of the project, creating detailed flowcharts and project designs. The modeling team incorporated their finalized data into the reports, ensuring the results were clear and convincing.
September
Week 1
The focus for September is the writing and organization of the WIKI documentation. The degradation, upcycling, and detection teams have fully organized their experimental results, and everyone is now focused on systematically integrating these findings into the WIKI.
The modeling team has also started drafting documentation for their mutant model, ensuring that both the experimental and theoretical components are well-represented in the WIKI.
The WIKI team has been instrumental in assisting with the document structure, ensuring that our team’s achievements are presented clearly and accurately.
Meanwhile, the gene-editing team successfully knocked out the YahK gene in E. coli BL21(DE3) and confirmed the knockout using nucleic acid electrophoresis.
Week 2
This week, the WIKI writing process entered a critical phase. The experimental content from the degradation, upcycling, gene-editing, and detection teams has mostly been incorporated into the WIKI, while the modeling team’s documentation is beginning to take shape. The team is now focused on refining the language and structure to ensure that the content is not only scientifically rigorous but also easy to understand.
The WIKI team has been working hard to integrate the various modules, and overall, the progress is smooth.
On the HP front, the team hosted an online environmental design and knowledge competition. We’ve prepared a variety of exciting prizes for the participants, and I must say, I’ve taken a real liking to that adorable pillow!
Week 3
The WIKI documentation is nearing completion. All experimental data, model results, and relevant discussions have been fully integrated into the document. We had several rounds of discussions with our PIs to ensure that the content is both comprehensive and scientifically accurate. The degradation, upcycling, gene-editing, and detection teams have carefully reviewed their sections multiple times to ensure nothing was missed or incorrectly presented. The modeling team also finalized their mutant model documentation, successfully linking the theoretical and experimental aspects of the project. The team has now moved on to the final stages of formatting and layout adjustments.
Week 4
The final version of the WIKI documentation is complete!
All the content from the various experimental modules has been fully integrated, with the modeling team’s theoretical contributions seamlessly included.
The WIKI team has done an excellent job in formatting and proofreading the document, and the overall presentation of the project is highly professional and polished.
Thanks to everyone's hard work, the final document showcases our team’s efforts in a way that is both clear and impressive, highlighting the full scope of our accomplishments.