In the following, we explain how our project fulfilled the deliverables for the different medal criteria.
After handing in all the deliverables and assessing the judging session at the Grand Jamboree 2023 in Paris, our project has been rewarded a silver medal!
Our team operates with a high degree of self-sufficiency, handling a wide range of responsibilities such as paperwork, financial management, legal matters, and lab operations. The bulk of our project, around 95%, is managed internally by our team members. We did, however, seek external assistance for specific tasks. For instance, we collaborated with an illustrator to create visuals for our project and enlisted the services of a company to synthesize our primers.
This years project tackles nitrate pollution of groundwaters primarily caused by excessive fertiliser usage in Germany but also representing a problem of worldwide significance. Bioremediative measures currently in place are costly and difficult due to the complexity of maintaining the corresponding denitrifying bacteria, and the necessity of anaerobic conditions since oxygen is the favoured terminal electron acceptor and therefore will be used instead of nitrate by the bacteria if present.
To overcome these problems our project focuses on extracting the catalytic sites of the reducing enzymes contributing to denitrification in these bacteria and to connect them to electrodes together with a cytochrome c acting as the electron mediator between electrode and enzyme. This provides the necessary electrons for the reducing steps in a cell free system, ultimately eliminating the previously mentioned difficulties seen in current bioremediation techniques.
Additionally, this approach avoids the risk of releasing GMOs into the environment and hence can be regarded as a method with improved safety comparing to other methods that would for example aim at increasing enzyme expression using synthetic biology.
By this project we aim to establish a cheaper, novel and safer denitrification system that can be set in place when it is already to late for preventative measures.
This year our contributions to the iGEM community are categorized into two parts - Laboratory and Management. Through our project we managed to design 4 pairs of primers selective to genes coding for denitrifying enzymes in the bacterium Paracoccus denitrificans (strain: DMS104981). The primers identified the genes napA, nirS, norB, and nosZ. These primers also included restriction sites (for identification and introduction into plasmids) and some were his-tagged, allowing for later protein purification. The information about the forward and reverse primers can be found on the contribution and basic parts pages and used by any other team with similar interests in the future.
Additionally, we created a simple tool for quick calculations of reagents and buffers. We believe that such a tool would help consistently plan experiments, document and utilize time and resources effectively. This would also cater to students who are new to a laboratory or have difficulties in mathematics or chemistry.
Drawn from personal experiences of working independently and establishing the iGEM team from scratch, we believe it would be very helpful for other teams to have a good starting point. We created a document listing trusted vendors, the product names, catalog numbers and prices. The products include DNA, RNA, Plasmid isolation kits, PCR, His-Tag and Nitrate testing kits. Most of these kits are used for basic experiments and having such a tool saves on valuable time that would have been spent on searching for reliable suppliers. These companies are known to sponsor iGEM teams, and we hope to expand on it for other products as well.
Our team operates autonomously, with limited external guidance. Consequently, many of our endeavors have been characterized by a steep learning curve. In the pursuit of engineering success, we have repeatedly refined our learning process, which plays a pivotal role in shaping our project. Our foremost priorities are to ensure that our project is not only beneficial to the world but also safe. Furthermore, we've also reiterated our engineering cycle of our primer design.
We find it necessary to be fully aware of how our project can benefit its end users. Our team members did thorough literature research on the causes of nitrate pollution, its consequences for the environment, the main strategies for the treatment and prevention, as well as the main groups of the population affected by it the most. Alongside the extensive research we did on our own, we conducted an interview with one of the field experts Dr. Nils Cremer, who gave us further insight into the topic. Having informed ourselves of the relevance of our project, we adjusted it to be accessible to as many end-user groups as possible.
As soon as we knew which enzyme parts, the active parts of four different reductases, and from which bacterium we were going to use in our project, we looked for the exact gene sequences of those parts in Paracoccus denitrificans and found them on KEGG.
Afterwards, we designed our primers based on these sequences and added the gene sequences and the primer sequences as basic parts to the registry.
This year, we wanted to focus on science education that was informative, fun and inclusive to all kinds of students. We believe that science education helps build critical thinking and problem-solving skills among children. Introducing this at a young age by making it creative not only helps mold the child’s mind but also makes them inquisitive to learn more. There may be many barriers to such an education - lack of motivated teachers, socio-economic constraints, etc. Keeping these in mind we created a book “Joy’s Journey”. Here we explain concepts from water cycle to basic synthetic biology. This tool can either be a teaching aid at a school or can be used by the child independently. Its simple language and illustrations enhance visual learning. We intend to accompany the book with puzzles through which students can recall or practice what was learnt from the book. This allows them to better engage with the content of the book.
We hoped this book could cater to a wider audience that includes students with learning disabilities. We understand the importance of colors and images in their learning process and utilize the same in creating vibrant pictures that captures their attention and keeps them motivated to learn.
With the critical role of inclusivity, our team utilized certain methods targeting students with special needs including specific learning disorders, autism, and more. Tied to education, our book, “Joy’s Journey,” incorporated these techniques in order to create a wider audience and convey this information to as many children as we could. The language barrier had been reduced as the book had been translated into many languages. An accompanying puzzle was also created which aimed to provide further simple yet stimulating visual aid as a method of inclusion. With positive feedback from the institution it was implemented at, the book was able to successfully help children with special needs comprehend complex scientific concepts in an easier way. Furthermore, an interview was conducted with a student in the field of natural sciences struggling with memory loss in order to demonstrate the difficulties she faced and how certain accommodations to other learning disorders may not work. The interview allows one to put into perspective how challenging the current standards of education tend to be for anyone with special needs especially in the field of science.