Team Attributions

Throughout the year we have had the great opportunity to work with a range of people who had a significant impact on our project. We would like to acknowledge everyone who contributed to our project and essentially made it all possible!

Core Group Attributions

For our core group, here is what everyone contributed to throughout the project:

Project Support and Supervision

Dr Andrew Hitchcock and Dr Tuck Seng Wong aided us throughout our entire project, supplying insights into the many issues we encountered and spotted them before they even occurred on numerous occasions!

Lab Support and Advice

At the beginning of the summer we were hosted by the Molecular Biology Perak 1 Team who offered lab space, general reagents, and aided us in ordering DNA and lab components, helping us find our feet as we prepared for our project. Coming towards the end of the summer they required their laboratory space back, ready for new students starting in September. This was when Dr Hitchcock stepped in to help out and offered lab space for two of our team members to finalise our wet-lab work right up until the wiki deadline.

Project Advice and Feedback

Coming into our project only a select few of our wet-lab team had history researching directed evolution approaches and the use of the MutaT7 system, and none of us had come across Vibrio natriegens before! Talking to a range of academics in the field prepared us for the difficulties we would face throughout our project.

In particular, we spoke with the supervisor and team lead of the Marburg 2018 team, Daniel Stukenberg and René Inckemann, who offered us help to grow and transform V. natriegens, which appeared to be a difficult task according to literature. René also mentioned that to improve the growth rate of V. natriegens in our growth rate experiments we should use baffled flasks to allow greater aeration of the culture, something V. natriegens favours when growing, and showed greater growth rates in following experiments!

Jan Kalkowski, from GASB Education was invaluable in highlighting a potential problem with identifying, isolating and sequencing an evolved gene of interest. His feedback led us to move our GOI to a low copy number plasmid and to consider genomic integration in the future. He also provided us with many valuable papers regarding bacterial genome copy numbers, which greatly informed our work.

Further, talking to Michael Magaraci, a research scientist at Protein Evolution and past iGEMer, gave us many insights into how to be successful as an iGEM team, particularly with how we should present ourselves in relation to our chosen research area, and how to get the most out of the Grand Jamboree.

Douglas Kell, Chief Scientific Officer of Epoch Biodesign, gave us considerable insight into some of the hidden pitfalls of directed evolution. He emphasised how crucial media optimization is, and that perfecting the right media can pay dividends for all future work. This was what inspired us to perform our media optimization experiments for V. natriegens.

Joe Price, from Evolutor, offered us insights into the commercial side of protein evolution projects. Joe gave us various industries that would be interested in a project such as ours and also gave feedback on our deliverables. Adam Bowie of the University of Sheffield hosted an ‘Introduction to AlphaFold’ session where he helped introduce us to AlphaFold to use in research and how it can be applied to iGEM teams. Lastly, Geno Esposito, a mathematician graduate from the University of Sheffield, helped us estimate natural gene diversity from BLAST data which informed much of our directed evolution modelling.

Team Sponsors

Much of our media optimisation process was guided by the sage advice and generous sponsorship of Owen Jonathan and Volker Kraft from JMP. The conversations with Owen and the JMP software licences they sent over is what made using Design of Experiments feasible for our team!

Further Acknowledgements

We'd like to thank all the iGEM teams this year who took the time to collaborate with us on our projects, modelling, and outreach. You can find more of them on our collaborations page! We would also like to acknowledge the works of previous iGEM teams, Paris Saclay 2021 and Marburg 2018, of which we took much inspiration from and helped monumentally in our initial understanding of directed evolution and working with V. natriegens, respectively.