Our project and story present several innovations as well as novel results that benefit the field of Synthetic Biology research. Since the initial development of the POAP pathway[1], we have been the first group to test a minimized carbon fixation cycle in-vivo.

Additionally, throughout our project we developed a pipeline (Fig. 1) that can be implemented to easily engineer a full metabolic pathway into a viable organism from the ground up. This technique could be used by scientists for various other applications, in addition to the implementation of a carbon sequestration cycle.

Adaptive Laboratory Evolution (ALE) has been implemented by synthetic biologists and bioengineers to create novel strains of commonly used single-celled organisms, such as E. Coli, that can tolerate severely hostile environments. For example, this strategy is commonly used to develop biosensors that retain functions while surviving high salinity or extreme pH environments. Our work presents a novel application of the ALE technique (Fig. 2), to develop a thermotolerant strain of the JCVI Minimal Cell. This strategy could be used in future research to develop thermotolerant strains of other single-celled organisms, such as E. Coli, cyanobacteria, or even yeast species. Furthermore, this technique could be used to evolve the JCVI Minimal Cell and serve as a model organism for research to learn how certain genetic evolutionary characteristics developed in more complex organisms.

By using the JCVI Minimal Cell (Syn-3B) as our biological chassis, we have successfully demonstrated that this organism can be used as a platform to screen and analyze the effects of engineered proteins on basic biological functions. The minimal cell genome and proteome is so efficient and simple[2] that it serves as a perfect model organism for observing how engineered proteins might affect the metabolic pathways of other, more complex organisms. Our methods can be used to effectively test the influence of other engineered metabolic pathways (particularly other minimized carbon sequestration pathways) on a more complex organism’s biological functions.

Our project lays the foundation for the development of a novel organism that will efficiently remove carbon from the atmosphere and fix it into a neutral source. Through this, we are fighting climate change.

We hope that our project will contribute to efforts that will work in conjunction with movements such as nationwide movement to end fossil fuels in the US. The climate crisis is an intersectional issue which needs to be addressed via comprehensive sets of solutions including sustainable sustainability research. From marching alongside our cousins nationwide to advocating for climate-friendly housing solutions to investing our energy into climate research, our team has inserted itself into various aspects of the climate justice movement that both include and transcend our research into carbon fixation.

Here we also provide tips and tricks for future iGEM teams!

• Start the website and all videos at least three months in advance of the deadline!
• Look for sponsorships early! It’s much easier to get reliable sponsorships if you make relationships with potential sponsors by emailing them early, meeting them at conferences, and building rapport.
• Create a Gantt chart or some other method of keeping track of which tasks you have to do and the timelines each task is expected to take.
• Document all your work very methodically.
• Reach out to as many possible collaborators as early as you can. Even if you don’t end up collaborating in a big way, they can help you solve a smaller roadblock you may be facing, or you can help them!
• Don’t be afraid to think outside the box.
• Keep an eye out – solutions may present themselves in completely unexpected ways.
• Attend other conferences that are relevant to the work that you are doing. It’s a great opportunity to learn about the innovative work happening in your field!
• Spend some time with each other outside of work!
• Leverage the experience and skills of everyone on your team, especially your mentor! Everyone’s unique point of view can lead them to finding incredibly innovative solutions.

We are the first iGEM team to be officially recognized as a UCSD organization. Our work will lay the foundation for future UCSD iGEM teams to flourish.

We have presented this work at the UCSD Bioengineering Day 2023 and will be presenting once more at Bioengineering Day in 2024. This event is the biggest Bioengineering event at UCSD and serves as an essential component for the education all UCSD Bioengineering students (seniors and juniors) receive.