Team:Concordia-Montreal/Partnership
Partnership
We partnered with the iGEM UToulouse 2020 team to create vitamin-producing AstroYeasts
iGEM Toulouse 2020 wikiOverview
Our Partnership with iGEM Toulouse was a fun and valuable asset to AstroYeast and AstroBio. Our projects are complementary as iGEM Toulouse is using an engineered yeast to provide flavoured nutrients for astronauts, while we are designing space-compatible yeast for bioproduction in microgravity conditions. Partnering with them was important as it offered us an application for our space-compatible yeast strains (bionutrient production) while iGEM Toulouse gained insight as to how the cell is affected in microgravity conditions. We also shared networking contacts and provided continuous constructive feedback for each other’s work throughout the year to strengthen our projects.
Essential guide for starting a space project
We had numerous and continuous exchanges with iGEM Toulouse throughout the year as we met biweekly to grow our Partnership. As both of our teams would like to bridge the gap between synthetic biology and space exploration, we embarked on iGEM Toulouse’s 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 of this guide which 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.
You can find our essential guide for undertaking a space project in our contribution page
AstroBio and Video Game Testing
iGEM Toulouse tested our AstroBio database and provided feedback as we progressed. We reciprocated with feedback for their video game regarding explaining certain terms that were not clear, english edits and future improvements such as offering more details as to how organisms are engineered.
AstroBio Feedback from iGEM Toulouse
Toulouse team member: Laurène
"Easy to use database."
"For results: more clear information about the informations available in the table should be delivered, for example: FC fold, what does that mean? What can I do with this result?"
"As maybe an extension in the future of your database, you should think about synthetic biology users and correlate the gene with its promoter or doing a second part of your database with only yeast promoters. So the user can directly see if the promoter that he’s using is well/not well expressed in conditions of microgravity."
"The idea of doing a guide beforehand is great. It will allow people to really understand the whole database. I’m quite impressed that microgravity has this huge effect sometimes on gene expression."
Toulouse team member: Arno
"Make the search a little more flexible (allowing mixed cases, gene alternative names or nicknames if they have one)"
"Search by gene function (Ex: alcohol dehydrogenase, ribosome subunit, …)"
"Allow ordering by names, logFC, species, etc. and a “Show only [number] of [max] function ?”
"Display a little “?” after each column name to get helpful advice"
"About the More Info button: a more user-friendly organization, with tables, colors, etc. Would it be better to just unfold the info below the row we are interested in? or even open it on a new page?"
Toulouse team member: Cécilia
"The database is clear and quite easy to use. Nevertheless, I think that you should add the gene function in the results line and in the search options. Maybe, there should be in the search options a key word option instead of the gene function."
"Since the database is made to find the effect of microgravity on gene expression, there should be the information of the “fold change” in the results of the research. People could therefore rapidly see if the gene expression is either up, down or unchanged."
"I think that the study type and the assay type should be in the results also."
"Maybe you could also add an item “help” in the homepage which would explain how to use the information in the database."
Vitamin A production in space
We were most excited to both be working with yeast with a similar end application in mind- bionutrient production in space. As a proof of concept, we have included GAL10, a promoter from iGEM Toulouse’s iGEMINI design, in our promoter selection.
Figure 1: Reporter for iGEM Toulouse’s GAL10 promoter constructed by iGEM Concordia
We did not have lab access this year, whereas iGEM Toulouse did. We shared our protocols and they were able to execute a growth curve for GAL10. This offers us a strong beginning to our research next year. In exchange, we have access to testing in microgravity conditions, while they do not, and next year, we will test the GAL10 promoter in simulated microgravity conditions.
Experimental Results
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 done over 4 hours in a 96-well plate. Figure 1 shows the plate with the fluorescence result for each condition. The first three columns contain the H2O2 stressor and the last three columns contain the NaCl stressor. 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) and then after the fluorescence rises again.
Figure 2: Representation of the 96-well plate for fluorescence monitoring of different stress conditions.
Figure 2 shows the fluorescence of a sample (in red expressed in RFU, Relative Fluorescence Units) and the mean fluorescence of all the samples (in blue) over time. We found similar results for all the samples. They followed the average each time, which suggests that the stress inducers have no impact at the conditions tested. This is confirmed when we look at the negative control which corresponds to the wells of the last line which have exactly the same fluorescence profile.
Figure 3: Representation of the average fluorescence (blue) compared to the fluorescence of A1 sample (red).
We can conclude about this experiment in two ways. 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.
Protocol developed by iGEM Concordia and adapted by iGEM Toulouse
Partnership Notebook
We are grateful for this Partnership and how it has furthered both of our projects as they grow together. We could imagine merging both of our projects in the future to create a space-compatible provitamin A enriched yeast for long duration space travels.
Partnership Notebook
Our 2020-2021 iGEM project is generously supported by
