Team:Toulouse INSA-UPS/Partnership
Partnerships
Introduction
We decided to engage with two iGEM teams as partners because we shared mutual objectives related to our respective projects and had numerous meetings during several months throughout the season with both teams.
The collaboration with Team AshesiGhana was a valuable asset for our Human Practices part of our iGEMINI project. As for the collaboration with team Concordia, it was central to our project to create new ways of bridging the gap between biotechnology and space and meet our mutual goals for this edition.
Team AshesiGhana
New techniques developed for space could, like many others in the past, be used on Earth and have major repercussions in a world where the population is growing and where resources are diminishing. This is why we thought about the implementation of our β-carotene production device on Earth. We looked for areas in the world where vitamin A deficiency occurs. Ghana, in particular, is part of these areas and by chance, since iGEM is a competition with teams from all over the world, we found the team AshesiGhana.
What does Team AshesiGhana brings to IGEMINI team
Implementation of a vitamin A production device in Ghana
We were thinking about implementing our system on Earth, in areas affected by vitamin A deficiency. Since Ghana is one of them, we needed Ganhaian population feedback in order to make our system evolve towards real life issues. To do so, we created a survey about the usefulness and acceptability of our system.
In this surveyn we asked for insight on how to improve the system according to the needs of the local population. Team AshesiGhana collected and analyzed the data of this survey.
Analysis of AshesiGhana team:
The iGEM Toulouse team wanted to have an idea of basic knowledge of people on microorganisms and GMOs and their opinion about using them in daily life. More than 70% of the 23 people we surveyed somewhat understand what was GMOs. 75% of the people probably eat or definitely would eat GMOs which mean that, without knowing our proposed solution (engineered vitamin-enriched yeast), they would accept to eat engineered microorganisms.
Before the survey, one of the things that iGEMINI thought about was to use alternate forms of energy, that would allow to power the production system. This concern arises since the iGEM Toulouse team thought that it might be difficult to have access to electricity in some areas in Ghana. However, only 25% of respondents answered that they knew people with no access to electricity. Therefore, it might not be necessary to find an alternative to electricity after all.
From this analysis, we see that about 70% of respondents would eat the solution provided by iGEMINI which is in correlation with the first answer but we do not know if people have changed their mind depending on the solution the iGEM Toulouse team provided (e.g. providing a solution for vitamin A deficiency). It allows us to infer that this solution would be relatively popular in Ghana. However, for those who said no, the reasons given were with respect to safety, personal values, and price.
The final question was to evaluate if the iGEMINI solution to prevent vitamin A deficiency would be accepted and needed and 90% of the people thought that this device could respond to this issue.
The general sentiments from the survey are that people are excited about the project! There were even some suggestions from respondents that we hope iGEMINI might potentially take into consideration.
Our conclusion: Even if only 23 people answered our survey, we had a small overview on how our vitamin A production device could be well accepted by the Ghanaian population. Their suggestions implied that using electricity may push some people away but we also considered using solar panels as power generators. After an ethical and practical reflection, we understood that our system is adapted to function in a spacecraft and not to function on Earth to complement deficient populations. The price of our device will probably be a first barrier. Sterilization methods and the use of resources such as electricity will be yet another difficulty, as some people do not have access to them. However, many aspects of our project remain interesting for an application on Earth. The use of nutritive yeast or coculture using minimal resources to produce a compound of interest are avenues that can be explored separately. These are the many possibilities for the evolution of our project that we will deepen if we continue working on it, and that we strongly advise other iGEM teams who would like to take over our project to reflect upon.
NGO contact
Team AshesiGhana also gave us NGO contacts. We contacted "Nutrition connect", people from "Global Programme Lead for Food Fortification" at the "Global Alliance for Improved Nutrition" and the Director of "Nutrition for Africa" at HKI (Helen Keller International). However, we did not receive any response for further interviews.
Broader view for our game-based educational tool
Additionally, the Ghana iGEM team gave us the contact of Eric Fosu, the Head of Biology Department at GCIHS (the Ghana Christian International High School). We sent him the link for the first chapter of our video game: iGEMINI Coculture. Our aim was to have a different point of view from a teacher in Ghana, likely very different from a French vision. He gave us some tips to have better navigation through the video game and advised us to add a "voice-over" to read the texts (in case someone playing has challenges with sight). As a conclusion, this game could be used as a pedagogical tool for highschool students in Ghana “as it explores crucial topics like microbiology and genetic engineering in an interesting way” according to Eric Fosu.
What does Team iGEMINI brings to team AshesiGhana?
Their project
This year, AshesiGhana iGEM team engineered a series of bacteria in order to create a self-repairing, bioconcrete, bioluminescent tetrapod. These engineered bacteria would have the ability to degrade shredded plastic as well as induce biocementation to keep the sand particles of the bioconcrete tetrapod together. Their aim is to prevent coastal erosion and plastic pollution.
Design process advices
The iGEM AshesiGhana team gave us access to their design lab book so we could help them through this crucial step. We gave them feedback from the choice of their chassis to what they determined as new properties for their engineered bacterium.
We advised them to find out what was the temperature range the bacterium could grow with, since it would have to be able to grow at low temperature like in the ocean where the tetrapods are supposed to be.
We encouraged them to ask questions such as: how the bacteria will find nutrients in the tetrapod. From the literature we provided for them, they learned the importance of codon optimization to increase the protein expression from gene transcription. In response, they decided to try and run codon optimization in Benchling for their plasmids. However, since they were not able to go to the lab this year, next year’s team will receive these results and determine what to do with them.
Human practices support
For their Human Practices, we suggested them companies in France that could have an interest for them: Carbios and Glowee. Carbios gave them a great starting point for next year’s team since this company developed an enzyme that is extremely efficient at degrading PET. This company will be contacted next year. We also thought of Glowee, a company that currently uses bioluminescent microorganisms. AshesiGhana will use this contact for Human Practices to understand the possible industrial uses of bioluminescence. In addition, they will try to implement their project in the real world by conceptualizing a bioluminescence start-up in Ghana.
Entrepreneurship support
In the entrepreneurial part, we explained tools such as the RACI matrix or the SWOT analysis, but we also emphasized the need of an approach to define the missions and objectives of the future company. We informed them on how to organize the different tasks to deduce the associated risks and analyze them in order to build a viable company. During our exchanges we also talked about using a life-cycle and Issues table to identify the consequences and impact that our iGEMINI project can have on the world (as an example).
Here is the Team AshesiGhana team feedback of our intervention
“On September 11th, Rosemond and Tamisha had a call with Pierre from the iGEM Toulouse team concerning the entrepreneurship aspect of iGEMINI’s project. Through this meeting, we had a refresher of the basics of design and entrepreneurship we did in first year, and we learned some tips and tricks as to how to potentially implement our project outside of iGEM. One major aspect of this call that was useful was the explanation Pierre made with respect to conducting a risk studies analysis. The risk studies analysis is a table that summarizes the potential risks a project could face, the probability of that risk occurring, the impact of the risk, and the severity of that risk. From this aspect of the meeting, we learned that it is important to sit back and look subjectively at our project as we try to determine the potential risks we face and also what we can do to mitigate such risks. While we did not conduct this at the beginning of the iGEM season, we would definitely consider doing this next year so that we also work out risk mitigation measures. Another way this call helped us is that it made us think critically about the various aspects of our project and not just the synthetic biology aspect. We realized that we had been so focused on the biology and modelling part of our project that we forgot to fully consider the implementation aspect of the project. Thus, this meeting was a great refresher to allow us to refocus on the implementation aspect. It was a great meeting and we are grateful for the help provided”.
Conclusion
Throughout this partnership, we were able to help each other teams aiming to develop both of our projects. We benefit from this collaboration by gaining knowledge about the acceptability of our project in a country that is being affected by vitamin A deficiency. In return, we provided AshesiGhana team with tools and advices for their design and implementation process.
Team Concordia
We started to collaborate with iGEM Concordia through their collaboration request on the iGEM 2020 website. Following our interview with Natalie Leys, head of the Microbiology Research Unit at the Space Life Science, we wondered about microgravity effects on cells and agreed that collaborating with an iGEM team working on yeast in microgravity would be beneficial for our two projects related to space. Since their project will spread over 2-year, their first goal is to create AstroBio, a database for differential gene expression analysis in microgravity studies.
Complementarities between our respective projects
Our projects are complementary since we are using an engineered yeast to provide nutrients for astronauts while iGEM Concordia is trying to focus on the behavior of yeast in microgravity conditions. Next year, iGEM Concordia will bioengineer a space-compatible microorganism yeast that is tolerant to these microgravity effects.
Building together a guide to start a space project for future iGEM teams
We had numerous and continuous exchanges with iGEM Toulouse throughout the year as we met biweekly to grow our Partnership (Notebook).
As both of our teams would like to bridge the gap between synthetic biology and astro exploration, we invited iGEM Concordia to embark on our guide for future iGEM teams who are undertaking space projects. This guide is a thoughtful reader-friendly starting point for future iGEM teams and student space associations as they embark on a space-related project. We collaborated on the content and design and are proud to offer it in English and French. It includes information which we both sought out at the beginning of our projects such as what bionutrient production is, an overview of microgravity conditions, past iGEM space projects, astro resources and more! We are also distributing the document via email to student space associations in North America and Europe.Here is our guide for future iGEM teams undertaking space projects (in English then in French):
Mutual feedbacks to shape our projects
We also shared networking contacts and provided continuous constructive feedback for each other’s work throughout the year to strengthen our projects. iGEM Concordia reviewed the design of our video game, while we reciprocated with feedback for their AstroBio database. AstroBio is a database that hosts a collection of transcriptomics data associated with microgravity effects on organisms. They implemented our feedback in order to shape their database and correspond to real demand.
Starting experiments for iGEM Concordia
We were so excited to both be working with yeast with a similar end application in mind. While we had lab access this year, iGEM Concordia did not yet have access to a microgravity simulator (they will next year). We agreed to add a proof of concept as part of our Partnership. iGEM Concordia has included our GAL10 promoter in their strain design and will perform experiments to make it tolerant to microgravity-induced stress next year. The successful expression of GAL10 in microgravity conditions would validate both of our projects. We have begun these experiments, with the help of a protocol from iGEM Concordia. We have performed fluorescence monitoring for mCherry expression under GAL10 promoter in our lab, as they do not have lab access. This fluorescence monitoring will give them a head start next year when they begin their experiments with our promoter GAL10 as a proof of concept for nutrient-producing microgravity-resistant strains.
For the experiment, we wanted to measure the activity of the GAL10 promoter by using a mCherry fluorescent marker in response to different stress inducers such as NaCl and H2O2. Fluorescence monitoring was conducted over several hours (4 hours) in a 96-well plate. The image below shows the plate with the fluorescence result for each condition. The first three columns contain the H2O2 stressor experiment and the last three columns contain the NaCl stressor experiment for exact well distribution).
We distinctly notice that there is no significant difference in fluorescence between the different experimental conditions. We always observe a decrease in fluorescence at the beginning, which corresponds to the change of environment of the cells (transition from the Erlenmeyer flask to the well), before the fluorescence rises again.
Figure 1: Representation of the 96-well plate with fluorescence monitoring of different stress conditions.
The graph below shows the fluorescence of a sample (in red expressed in RFU, Relative Fluorescence Unit) and the mean fluorescence of all the samples (in blue) over time. We find the same pattern for all samples. They followed the average value each time, which means that the stress inducers have no impact. This is confirmed when we look at the negative control that corresponds to the wells of the last line and which have exactly the same fluorescence profile.
Figure 2: Representation of the average fluorescence (blue) compared to the fluorescence of A1 sample (red).
The amount of stress factors we used does not induce any change in the activity of the promoter GAL10 at the concentrations that we tested.
Conclusion
We are grateful for this Partnership and how it has furthered both of our projects as they grow together. We could imagine merging them in the future to create a space-compatible provitamin A enriched yeast for long duration space missions.
Overall Conclusion
In order to establish a partnership, it is important to have common interests and to split up tasks. For both our partnerships and in each team, one person was in charge of organizing the bimonthly meetings for updates before distributing the tasks to the rest of the team. The assiduity of both teams is essential. Overall, following two teams (in our case) was really rewarding since we were able to discuss and exchange ideas, and develop two important parts of iGEMINI: implementation on Earth and in space.
