Week 9 HW: Cell Free Systems

Homework question from Peter Nguyen

1.Write a one-sentence summary pitch sentence describing your concept. I could use cell free systems adapted to producing inaK in order to directly inoculate glaciers with the aim to preserve and boost ice formation, it would help glaciers rebuild and be more resistant to increasing temperatures caused by climate change. However, I would be interested in pushing the idea of geotextiles already helping preserve glaciers and design a living material, with inaK with a boosted ice nucleation function to create proactive glacier covering actively working to rebuild and preserve glacier ice.

2.How will the idea work, in more detail? Write 3-4 sentences or more. The inaK would be synthesized through a cell free model, using an alternative to E. coli which could resist and be active in sub-zero conditions (to stay active in glaciers), such as Oleispira antarctica a psychrophiles bacteria which has evolved to have specialized ribosomes and enzymes able to remain flexible and functional in a frozen environment. Oleispira antarctica contains unique chaperone (Cpn60 and Cpn10) preventing protein misfolds in frozen temperatures. This cell design would include pores in the membrane so it can stay alive in the textile by having an ATP source of input. These then freeze dried cells would be put into a textile (inoculated during the making of the textile), the textile can be brought to location and installed on the glacier and then be rehydrated to allow ice nucleation of the glacier to begin. This would permit me to create a live material that would be dormant in production and transportation and control its freezing function (preventing the textile from accidentally freezing its surrounding). Note that because I am working with ice nucleation there might be challenges in freeze drying these cells.

Reference List Ferrer, M. et al. (2003) ‘Low temperature-induced systems failure in Escherichia coli: Insights from rescue by cold-adapted chaperones’, Journal of Biological Chemistry. Cui, Y. et al. (2022) ‘Cell-free PURE system: evolution and achievements’, Biodesign Research. D’Amico, S. et al. (2006) ‘Psychrophilic microorganisms: challenges for life’, EMBO reports.

3.What societal challenge or market need will this address? This addresses the environmental, social and political issue of melting glaciers caused by climate change, which only increases the power of climate change as glaciers are key factors in slowing climate change. We are actively losing biodiversities and ecosystems and doing very little about it. It is not seen as a profitable income so little motivation is inputted. However, in the longterm, this irreversible damage done to our nature will actively make climate change worse, and there will be many destructive environmental, social and economic consequences driven by this overlooked issue.

4.How do you envision addressing the limitation of cell-free reactions (e.g., activation with water, stability, one-time use)? Working at very large scale, scale of the glacier, it can be a challenge to efficiently rehydrate the living material as it would be very energy consuming and costly to do it manually, but, if the living geotextile is strategically implemented at the right time of the year (early spring, already when they glacier coverings are usually installed) then nature itself through rain could activate the material naturally. The aim is to limit the human labor impact and simply give nature a tool to reinforce what it already knows how to do. Considering the one time use issue, geotextile coverings which are already used to protect glaciers are removed and installed yearly according to their natural ice melting and forming cycles. The next step of my research would be to find a way to keep the textile created and reabsorb it with new inaK cell free protein systems when it is needed next. The goal is to create a regenerative textile and closed loop system to avoid waste through one time solutions.

Homework question from Ally Huang

1.Provide background information that describes the space biology question or challenge you propose to address. Explain why this topic is significant for humanity, relevant for space exploration, and scientifically interesting. (Maximum 100 words) I am interested in exploring the purpose of ice nucleation in cell free design, freeze dried or not, taking the shape of a multipurpose textile which can be used as an alternative to current voluminous refrigeration tools or a freezing textile to activate. In space stations like the ISS a lot of research relies on lab samples being preserved in sub zero temperatures, from human research samples to organisms or protein crystals. Within research some experiments need cold induced phase changes to be activated or triggered. This technology could also be used for food or medical supplies.

2.Name the molecular or genetic target that you propose to study. Examples of molecular targets include individual genes and proteins, DNA and RNA sequences, or broader -omics approaches. (Maximum 30 words) The genetic target of this project would be the inaK ice nucleating protein, commonly found in Pseudomonas syringae, with a wide potential of freezing functions.

3.Describe how your molecular or genetic target relates to the space biology question or challenge your proposal addresses. (Maximum 100 words) The challenge is optimizing cold packaging and storage systems, a material which would require less space or a material able to be activated once in space again having less constraints in terms of space while travelling. The inaK offers a variety of possibilities in a cell free system whether freeze dried or not as it has a focused and controlled function to freeze. According to the development of the product it can be chosen at what temperature it freezes or activates and how resistant it can be to external temperatures. Creating a highly controlled and bespoke design for certain use in space allows for better control on the research done in space, every aspect of the research can be tailored in hopes to improve success rates of experiments. InaK is a relatively easy INP to work with.

4.Clearly state your hypothesis or research goal and explain the reasoning behind it. (Maximum 150 words) I am interested in creating polyvalent designs with multiple usages and applications, this project aims to find an optimal alternative refrigerating system which can have bespoke qualities specific to in space research. As small of a detail it might seem every aspect and tool of experiments impacts the result of research and can lead to better efficiency, results or unexpected breakthroughs. During a space mission all equipment has to be optimized due to lack of space and need for many items and a polyvalent tool that can respond to a wide range of uses can help with the space optimization.

5.Outline your experimental plan - identify the sample(s) you will test in your experiment, including any necessary controls, the type of data or measurements that will be collected, etc. (Maximum 100 words) I would design a cell free system for the inaK ice nucleating protein, freeze dry some and then create living textiles, some active and some dormant. The practicality of a textile is that it can be molded, cut, sewn, layered to adapt to any existing object which would then need a freezing function. I can control the amount of inaK for the freezing rate needed, experiment with the different temperatures it can freeze at and the different temperatures it can stay frozen at, I can explore the threshold of the inaK. I would then test the reactivation rates, how much water is needed and how long it would take.