Throughout our project, we have made several contributions to support future iGEM teams by enhancing the registry, developing custom hardware, sharing both standard and custom protocols, and offering educational resources for students interested in synthetic biology.
We contributed two unique parts to the iGEM registry—EPG and an elongated Wsc1—while also improving the information on a previously documented part, TcI. Additionally, we designed and built a hardware system that enables the spatial activation of magnetic fields, with design details made publicly available for future teams to replicate. Along the way, we encountered various challenges that required creative problem-solving, resulting in the development of custom protocols, which we hope will assist other teams in their research. Moreover, we are dedicated to making synthetic biology more accessible to students, whether they have prior knowledge of the field or not. To that end, we organised educational outreach events to promote synthetic biology iGEM, for example we created a mini-project tutorial suitable first-time Benchling users. Additionally, we have collated educational resources from a variety of teams in our outreach bank to increase accessibility and ease.
We hope that the work we have done on our project will inspire and support future iGEM teams as they strive to advance synthetic biology. Additionally, we aim for the impact of our project to extend beyond the iGEM community, benefiting academics who require spatiotemporal control of gene expression and may find value in our innovations.
One of the new parts we have added to the iGEM registry is a variation of the Wsc1 protein. Wsc1 is a cell membrane protein that acts as a sensor for the cell wall integrity pathway by activating the Rom2 GEF, which in turn triggers the downstream MAPK signalling cascade. This cascade regulates the expression of at least 25 genes involved in cell wall biogenesis (Philip and Levin, 2000). An elongated version of Wsc1 was engineered by Dupres et al. (2009), designed to have wsc1-mid2 fused and extended beyond the cell wall with a His tag that could interact with NTA-Ni+ functionalized atomic force microscopy tips. Professor Heinisch kindly provided us with the Y1 strain expressing this form of Wsc1, which allowed it to interact with our NTA-Ni+ magnetic nanoparticles. This elongated Wsc1 variant is one of our new parts!
Future teams can find our characterization and the sequence of this part on the new elongated Wsc1 registry page.
Additionally, we have added electromagnetic perceptive gene (EPG) to the iGEM registry. EPG is a magnetically sensitive protein derived from Kryptopterus bicirrhis (glass catfish). Its expression in mammalian cells suggests that EPG is a cell membrane protein that mediates calcium influx (Krishnan et al., 2018; Hwang et al., 2020; Ricker et al., 2023). While Krishnan et al. (2018) initially proposed that the mechanism did not involve a conformational change, more recent studies indicate that proteins split across the EPG termini are activated, suggesting a structural change (Grady et al., 2024).
In our project, we developed a system using EPG-Split Nanoluc fusion protein and EPG-Split TEV fusion protein, and other reporter proteins that can demonstrate the control over protein activity through external magnetic field.
The characterization and sequence of this part can be found on the new EPG registry page.
A core component of our project was the hardware we developed, which aimed to enable spatiotemporal control of gene expression using magnetic fields. To ensure accessibility for future teams or researchers, we kept costs to a minimum (approximately £330 or $440) and provided a comprehensive guide detailing how to replicate our hardware system.
Learn more, and read the guide on our hardware page.
In this year's project, we frequently encountered challenges in finding relevant protocols for our experiments, particularly those involving magnetic nanoparticles. To address this, we developed and implemented our own protocols to fill the gaps left by existing literature and official sources. We hope that the protocols provided below will assist future iGEM teams and researchers in their own projects or inspire the creation of similar protocols tailored to their needs.
All our custom protocols, alongside standard protocols used, can be found on our protocols page.
For a biomedical engineering summer course outreach event, we designed and developed materials for a hands-on project. This project provided clear, step-by-step instructions and background information on how to create an E. coli biosensor using Benchling. It was designed for beginners, requiring no prior knowledge, and served to teach basic synthetic biology principles in a practical manner while also introducing learners to the Benchling platform.
As a team, we invested significant time in learning how to use Benchling, and we believe this well-documented project could be a valuable resource for future iGEM teams, helping them navigate the software more efficiently.
To find the relevant resource, look under "Biomedical Engineering Summer Course: Project" on our education page.
We hope that our work inspires young minds to explore synthetic biology, potentially encouraging them to pursue a degree in the field or even consider starting their own iGEM project in the future. More broadly, our contributions to events aimed to also give young people opportunities to learn and explore science beyond their school curriculum. Since we invested a considerable amount of time into developing our outreach resources for our events, we wanted to ensure future iGEM teams can easily use and build upon what we have created. In order to create our outreach presentations, we looked through many other wikis for inspiration and a starting point. This gave us an understanding of how best to upload outreach materials so that they can easily be built upon and re-used. Hence, we uploaded not only the presentations, but documents with handover notes. Handover notes contain feedback from the levels of engagement for each part of the session, what we would change if we were to run it again, and who the event is suitable for.
Read more on our education page.
The iGEM Outreach Bank is a collection of outreach activities and resources that previous teams have created, which can be sorted based on variables such as age group and documentation level. The aim of this is to enable future iGEM teams to quickly search for resources to be adapted and reused, to make preparing for quality outreach events more time effective.
Philip, B. and Levin, D. E. (2001) ‘Wsc1 and Mid2 Are Cell Surface Sensors for Cell Wall Integrity Signaling That Act through Rom2, a Guanine Nucleotide Exchange Factor for Rho1’, Molecular and Cellular Biology, 21(1), pp. 271–280. doi: 10.1128/MCB.21.1.271-280.2001.
Dupres, V., Alsteens, D., Wilk, S. et al. The yeast Wsc1 cell surface sensor behaves like a nanospring in vivo. Nat Chem Biol 5, 857–862 (2009). https://doi.org/10.1038/nchembio.220
Krishnan, V., Park, S.A., Shin, S.S. et al. Wireless control of cellular function by activation of a novel protein responsive to electromagnetic fields. Sci Rep 8, 8764 (2018). https://doi.org/10.1038/s41598-018-27087-9
Hwang, J.; Choi, Y.; Lee, K.; Krishnan, V.; Pelled, G.; Gilad, A.A.; Choi, J. Regulation of Electromagnetic Perceptive Gene Using Ferromagnetic Particles for the External Control of Calcium Ion Transport. Biomolecules 2020, 10, 308. https://doi.org/10.3390/biom10020308
Ricker B, Mitra S, Castellanos A.E., Grady C.J., Woldring D, Pelled G, Gilad A.A., 2023 Proposed three phenylalanine-motif involved in magnetoreception signalling of an Actinopterygii protein expressed in mammalian cells. Open Biol. 3: 230019 http://doi.org/10.1098/rsob.230019
Grady C.J., Castellanos F.E.A, Schossau J., Ashbaugh R.C., Pelled G., Gilad A.A., A putative design for the electromagnetic activation of split proteins for molecular and cellular manipulation Frontiers in Bioengineering and Biotechnology 2024 12 https://doi.org/10.3389/fbioe.2024.1355915