Team:Sao Carlos-Brazil/Human Practices

Human Practices

International Legislation on Biosafety - Space and Stratosphere

Due to the scope of this project, our team had the ambition of performing experiments to test our engineered circuit in a stratospheric probe. A stratospheric probe, also referred as a high-altitude balloon, consists in a helium balloon attached to a box that contains the material that you want to expose to the stratosphere, which is an environment with very low pressure, low temperature, and a high incidence of UV radiation, making it very similar to Mars and, therefore, the ideal place to test our engineered yeast. With the partnership we have established with Zenith Aerospace, an extracurricular group from São Carlos that works with aerospace engineering, we would have easy access to a probe like this if needed.

Our control samples flying over São Carlos. Photo by Zenith Aerospace

We then decided to review the iGEM rules and came across the competition’s “Do Not Release Policy”, which states that the release of genetically modified organisms (GMOs) into the environment is forbidden in any circumstances. However, there were no clarifications whether a stratospheric probe was considered a contained experiment or a release, therefore we decided to contact iGEM’s biosafety committee in order to clarify our doubts.

Representatives from the committee, in special Dr. Piers Millet, also could not assure us in which kind of regimen our probe experiment would fit, while also arising other questions regarding our decision to perform this kind of experiment, as the acceptance of other people outside of the synthetic biology community. Consequently, they have advised us to conduct a review of the Brazilian legislation about this subject. When reading those laws, we noticed that many questions we had still were not clear, and could be subjects to second interpretations, in particular the exact definition of a “containment regimen”, since this kind of regimen is set apart from a laboratory regimen and also from a programmed release by the Brazilian National Committee of Biosafety (CTNBIO) (CNTBIO Normative Resolution nº 7, in Portuguese), but without any major specifications. For this reason, we have decided to research the biosafety legislation about GMOs of different countries all over the world, to try and find information that might answer our questions and, possibly, contribute to the strength of the Brazilian legislation.

To begin our research, we looked up for international GMO legislation from different countries in the Biosafety Clearing House Laws and Regulations database. We have read laws from many different countries but still could not find any information about stratospheric experiments, so we decided to search for keywords related to this kind of study in the database, but the website still returned no results. It is important to note that this database not only contains national laws but also regional and multilateral agreements between different countries.

Biosafety Clearing House’s Laws and Regulations Database

We also contacted over 40 regulatory agencies of space and stratosphere use, and research centers of genetically modified organisms and astrobiology, of all over the world, and we did not get responses from most of them. Among those who have answered us, many also did not know how to answer our questions. Described below is the list of contacted agencies, and the highlighted items are those who have responded to us:

Air Force Space Command (AFSPC) Astrobiology Society of Britain Australian Centre for Astrobiology (ACA)
Australian Space Agency (ASA) Belgian Science Policy (Belspo) Brazilian Air Forces (FAB)
Brazilian Institute of the Environment and Renewable Natural Resources (IBAMA) Brazilian National Biosecurity Technical Commission (CTNBIO) Brazilian Space Agency (AEB)
Canadian Space Agency (CSA) China National Space Administration (CNSA) Colombian Space Comission (CCE)
Department of Industry, Innovation and Science European Astrobiology Network Association (EANA) European Space Agency (ESA)
European Space Research and Technology Centre (ESTEC) European Union Aviation Safety Agency (EASA) Federal Aviation Administration (FAA)
Geo-Informatics and Space Technology Development Agency (GISTDA) German Aerospace Center (DLR) Indian Astrobiology Research Centre (IARC)
Indian Space Research Organisation (ISRO) Italian Space Agency (ASI) Israel Aerospace Industries (IAI)
Israel Space Agency National Aeronautics and Space Administration (NASA) National Agency of Civil Aviation (ANAC)
National Authority for Remote Sensing and Space Sciences (NARSS) National Center of Space Studies (CNES) National Institute of Space Research (INPE)
National Oceanic and Atmospheric Administration (NOAA) National Space Council (NASC) Netherlands Space Office (NSO)
New Zealand Astrobiology Network (NZAN) Roscosmos State Corporation for Space Activities (Roscosmos) Space Application Center (SAC)
SpaceIL Swedish National Space Agency (SNSA) The Planetary Society
UK Space Agency (UKSA) United Nations (UN)

Of all emails sent, we got few relevant responses on the topic: the German Aerospace Center representative has affirmed that the probe’s flight itself could not be considered a release, but preventing the escape of these organisms and assuring the safety of the tests is essential, since any failure of the probe or its integrity and, subsequently, the escape of the experiment may qualify as a release. This representative has also advised us to look for information about space experimentation in Brazilian organizations, but we did not find anything related to sending GMOs to the stratosphere. However, a member of the Australian Centre for Astrobiology declared that our project would definitely be classified as a release, under their legal regime.

A former student from our university, currently working at the National Institute of Space Research (INPE), was also contacted to help our team with the raised doubts, even though stratospheric experiments were not part of his expertise. With his help during the Engineer and Space Technology Conference (Congresso de Engenharia e Tecnologia Espaciais - ETE), we were able to enter in contact with the organizer of the Brazilian Academic Rocket Competition COBRUF (Competição Brasileira Universitária de Foguetes).

We were instructed by the COBRUF representant to look for the rules of the competition, in which it is described that it is forbidden carrying any biological part that may be pathogenic for humans or animals or that offers risk to the local ecosystem on the rockets. He also advised us to contact the Brazilian Space Agency (AEB), National Agency of Civil Aviation (ANAC), Brazilian Institute of the Environment and Renewable Natural Resources (IBAMA) and the United Nations, some of which we had already contacted.

Collaborations between iGEM teams about biosafety

With the absence of fulfilling answers of our first contact with NASA, we got in touch directly with Dr. Lynn Rothschild, designated PI of the Power Cell experiments at the AMES Research Center, with 2011 iGEM team Brown-Stanford. A representative from her laboratory, and also a member from 2019 iGEM team BrownStanfordPrincetn, has responded to our emails and offered to help with NASA regulations. Following his contact with the responsible sectors at NASA, he has shared with us the following information:

A former NASA Planetary Protection Officer has said she is not aware of any legislation regarding GMOs, and that the United States does not have any laws regarding biological experimentation on stratospheric flights, just on space stations, but none about GMOs. On the other hand, a Lead Technologist for NASA's BioSentinel Mission has declared to have already performed experiments with GMOs in stratospheric probes before, but also did not find any legislation on the topic. He also believes that this matter may be resolved with basic biosafety legislation, but he has not shared with us the process he went through to perform his experiment.

CTNBIO Meeting

After many emails exchanged with a CTNBIO Analyst about Brazilian laws on GMO release and containment, we asked if it was possible to arrange a meeting with a CTNBIO representant to share our findings and try to propose a way to improve their regulations to fit stratospheric experimentation. Then, on September 26th, we finally got a phone meeting with Dr. Rubens José do Nascimento, a Human and Animal Health Advisor for CTNBIO.

The call went on for about 40 minutes, while we told him about our findings on other countries' legislation and more about the functioning of stratospheric probes. Dr. Rubens agreed with us that this is a very grey area in the Brazilian legislation. He affirmed to not be sure whether Brazilian laws even applied to the stratosphere since our country's territory only goes so far vertically, and advised us to try to contact the Brazilian Air Forces (FAB), but they have never responded to us.

Nevertheless, we have found a Ph.D. Thesis from Universidade de São Paulo (USP) in which the author defends the need for the delimitation of a country's airspace to protect its sovereignty. He argues that this delimitation should be 50 km of altitude and since the stratosphere is located between 10 and 50 km of altitude, Brazilian laws should apply to this area of airspace, should his arguments be accepted by our government. As far as we know, to this day there is no demarcation of Brazilian airspace regarding altitude.

By the end of our phone meeting, Dr. Rubens advised us to send a letter to CTNBIO asking for a Public Consultation on the topic, as a way for them to ask for society's opinion on this matter, which we have sent (available here, in Portuguese). So far, we have not got any response to the progress of our Public Consultation, but we plan to continue this work even after this year's edition of iGEM.

In conclusion, our research shows that most regulatory agencies' relationship with the public is still very flawed. It is not easy for a common citizen to find this kind of information and many agency websites did not even have a contact page for us to send our questions. We do believe that these agencies may have resolutions for cases similar to ours, but the difficulty to access them makes it harder for scientists to perform their experiments safely. Therefore, the first step for a better, more biosecure world would be the broader dissemination of this type of information, helping scientists at least know the basic procedures they should follow in order to shorten the time it takes to analyze and then approve projects like ours. Another alternative would be the creation of a platform where you could clarify simple doubts, before and without needing to submit a whole project.

Integrated Human Practices

How does our project affect other people (and can be improved by them?)

An important part of the development of an iGEM project is asking ourselves whether what we are building is responsible and good for the world, and there is no better way of knowing that than asking the people that are directly affected by the project. Since Astroshield has many applications in different areas, we have spoken with people from each one of those areas, them being Space (due to Mars colonization applications), Industry (related to sterilization applications), Synthetic Biology (to check if our project complied with the community’s expectations of a synbio project), and General Population (the people that would eat our Astroshield bread in the future). We have listened to all these people, but most importantly, we have applied their insights to improve our ideas, as we describe below.

1. Space

On August 13th, our team had a meeting with Dr. Ivan Gláucio Paulino Lima, of the Blue Marble Space Institute of Science of NASA Ames Research Center, where he works as a part of the research group of Dr. Lynn Rothschild. Dr. Ivan has participated in iGEM before as an advisor for other teams, such as the 2011 team Brown-Stanford, with whom he has developed the project Power Cell.

Dr. Ivan and our team at ONOVOLAB

Our team shared with him the difficulties we had been finding related to our Human Practices project, such as a low rate of responses from the space agencies we had been trying to contact, and the absence of any kind of law or regulation regarding the use of stratospheric balloons in Brazil (as stated on our “International Biosafety Legislation” topic). Dr. Ivan then told us about his experience with team Brown-Stanford, that also had used balloon testing during their experiments. The team, however, did not send any genetically modified organisms in their balloon flights, due to problems related to the cloning of their circuit into S. pasteurii cells, therefore the team members did not look for information on the required protocols for probe experiments with genetically modified strains.

Nowadays, the engineered bacteria constructed for Power Cell are orbiting Earth on a satellite provided by NASA, having complied with the requirements from NASA and the German Space Agency for space experimentation, but the requirements for sending genetically modified organisms (GMOs) to space cannot be applied to stratospheric probes, due to the structural differences between the devices. Nevertheless, this shows the huge potential for projects like Brown-Stanford’s and ours to be evaluated even deeper, with the possibility of becoming actual products in the future, beyond iGEM.

To that end, Dr. Ivan has offered suggestions to help us get the information we wanted for our project, such as submitting it to CTNBio so we would know which rationalizations would be given to us to authorize the probe flight or not under Brazilian law. He also suggested that we looked for information regarding the Beresheet lunar probe, which crashed while trying to land on the Moon on April 2019, consequently pouring over the Moon surface their load which contained genetic materials and tardigrades, microscopic animals known for their high resistance to unfavorable conditions. The lunar probe was developed by the companies Israel Aerospace Industries and SpaceIL, with the support of the Israel Space Agency, with whom our team has tried to get in touch, to inquire about the biosafety aspects of the accidental release of the tardigrades.

Following our team meeting, Dr. Ivan spoke at an event called “Synthetic Biology in Space”, at the innovation center ONOVOLAB, in which he talked about the results from the Power Cell experiments during its orbit around Earth, with the participation of 83 people in the audience (more information on Education & Public Engagement)

2. Industry

Fermentec, a company that provides solutions to the sugar, alcohol, and distillation industry, was extremely important to the development of our project, not only as a sponsor but also providing support, tools, and pointers to the project's progress. All strains, laboratory and industrial (diploids and haploids), used in the project were provided by Fermentec, from their microorganism database, which is the largest Saccharomyces cerevisiae database of the world, with over 2800 isolated industrial fermentation strains.

The enterprise has also given important consulting to the design of our glucose-based kill switch, suggesting research papers that would be useful to the team's objectives, like Dr. Andrea Balan's research paper, which was essential for our obtaining of the glucose-sensitive promoter and the nuclease used in our kill switch's circuit design.

Team member Lucas Boldrini during a skype meeting with Fermentec

Additionally, Fermentec has offered a technical visit to their facilities when delivering the diploid strain PE-2. On this day, the company's Project Manager Officer (PMO), Marcel Salmeron Lorenzi, staged a presentation to the team, introducing the company and its attributions in the industry. After that, the team visited the company's laboratory and the different areas of the corporative facilities. On a second visit, Fermentec offered reagents for yeast transformation (salmon's sperm DNA, lithium acetate and PEG 4000) and the haploid strains (four PE-2's descendants and a laboratory type), which we have used on our project.

Our team members receiving Fermentec’s yeast strains

On top of that, Fermentec has offered guidance about the analysis of successful transformations without 5-FOA and also the yeast transformation protocol used by them, to assist us in the preparation of some reagents.

Finally, to help us verify the applicability of our project in the fermentative processes industry, our team was invited to exhibit the project on the 41st Annual-Renewal Reunion, that occurred on July 25th and 26th, 2019, and possibly meet potential sponsors (more information on Education & Public Engagement)

3. Synthetic Biology Community

In order to listen to the opinion of people in the synthetic biology field and improve the progress of our project, our team attended Jamboré Brasuca, named in honor of the Giant Jamboree, a national reunion about Synthetic Biology annually organized by different Brazilian SynBio groups. The event occurred on March 29th and 30th, 2019, at the Engineering School of Lorena (USP-EEL) and about ten iGEM teams, between former, currently active and teams aspiring to join the competition in the future.

Some of our team members at the Engineering School of Lorena

Brazilian teams trajectory throughout iGEM

The event featured several talks and discussions about Synthetic Biology, innovation, and technology, as well as poster sessions in which teams had the opportunity to present their projects to the attendees and get extremely important feedback for the competition. Our team exhibited the initial outline of our project, and we received important critics and suggestions, mainly about the balloon flight’s biosafety and general project applicability by Dr. Thiago Mendes from the Federal University of Viçosa. We also had the opportunity to talk about the applications of our project, polishing the justifications we brought to the competition about the use of Astroshield yeast in Mars' colonization and on Earth's sugar and alcohol industries.

Our poster presentation at Jamboré Brasuca

The event also allowed us to make contact with members from other teams, letting us outline possible collaborations, and we met Cibele Zolnier, a former member of USP-EEL's team, who later was designated our mentor by iGEM.

We also had the opportunity to meet with two After iGEM's ambassadors to talk about our project in two different situations. In the first moment, we had a meeting with Guilherme Kundlatsch, former ambassador and current coordinator of After iGEM's ambassador program, during which we discussed the progress of our project, as well as the way we should approach its different usages on presentations, on Mars or at alcohol and sugar plants. Guilherme, as an ex-member of iGEM teams from our University in 2014 and 2015, shared his experiences in past editions of the competition, so we could learn with what had been already done by him and his team.

Some of our team members with Guilherme Kundlatsch

After that, we had the opportunity to meet the current iGEM Latin America ambassador, Manuel Giménez, while he was passing by Brazil, at a mini meetup organized on São Carlos by Guilherme Kundlatsch's startup Peabiru, in which members of our team, USP_SaoCarlos-Brazil team and Unesp Araraquara's team that will attend iGEM next year were gathered to show our project ideas. Manuel offered relevant tips and insights about our project's presentation, like the topics that we should focus on during the final presentation and how to tell our history.

São Carlos' Mini iGEM Meetup

4. General Population

Since Astroshield is a genetically modified yeast with not only energetic but also nutritional applications (space bread!), it needs to be approved by potential consumers. With that said, our team made sure to hear the Brazilian population's opinion on the acceptance of the use of genetically modified foods, in general, and about our project specifically. To do so, we have conducted an online survey through Google forms filling in anonymously and voluntary way, which was considered exempt of approval by UFSCar’s Research Ethics Committee due to the nature of the research (CNS Resolution nº 466/12).

We have got 280 responses on our survey, from people from many different backgrounds, ages, and schooling, as shown below. Since we are university students, most of the answers were from people with higher education, which we have felt like could bias our results, hence why we have decided to present the survey results in two categories: people with and without higher education

The demographics of our survey answers

For our survey, we have asked some pretty simple yes-or-no questions: 1. Do you know what a GMO is? 2. Can you identify GMOs in food? 3. Do you buy GMO foods? 4. Would you rather buy non-GMO foods? 5. Are you aware of the legislation behind the development of new GMOs? and 6. Do you believe the development of GMOs is important for society?

After those questions, we explained more about our project and its applications, and asked people whether they would consume it or not and why and also what was their opinion on GMOs in general. At the end of the questionnaire, we have added a brief overview of the definition of GMO foods, how to identify them and the kind of experimentation a new GMO needs to go through to be approved by our government.

As a result, our team could notice that the knowledge around GMOs is directly connected with a person’s level of schooling since 96.6% of people with higher education, finished or ongoing, have answered that they know what a genetically modified organism is. 63.1% of people in this group also know how to identify foods that contain these organisms in their formula. On the other hand, in the group of people without higher education, only 64.6% of people have affirmed to know what a GMO is, and 37.5 % know how to identify their presence in food.

1. Do you know what a GMO is?

2.Can you identify GMOs in food?

When it comes to whether or not they bought GMO foods regularly, the difference isn’t as big between the groups. 78.5% of people with higher education said they were used to buying GMO foods, and only 14.6% said they did not know. As for the people with lower schooling, 66.7% declared to buy GMOs, and 22.9% did not know.

3. Do you buy GMO foods?

When asked about a preference to buy non-GMO foods, both groups behaved similarly, showing that only 37.4% of people would rather buy non-GMO foods, while 19% did not have an opinion on the topic. With this data, we have noticed that a person’s schooling does not directly influence their GMO consumption since, in general, we have noticed that most people do not seem to be resistant against the consumption of GMO foods.

4.Would you rather buy non-GMO foods?

However, the most important result we were able to acquire from this survey was related to people’s information regarding the laws and regulations behind the development of new GMOs. When we asked people with higher education if they knew anything about the process of release of a new GMO, 64.8% said they had no clue of what kind of laws applied to it, while 88.3% of the group without higher education shared the same thought.

5. Are you aware of the legislation behind the development of new GMOs?

Finally, and interestingly enough, we found out that although many people had little to no information on this matter, 71.5% of them believed that the development of genetically modified products was important to society, and some even shared that this kind of research and product development may be necessary to supply future needs of our ever-growing society.

6. Do you believe the development of GMOs is important for society?

Below, we show some of the answers to the final question of our questionnaire, in which we have explained our project idea (a genetically modified radiation-resistant fermenting yeast to be used in food and ethanol production) and asked them whether or not they would consume it and if they had any particular opinions on GMOs, showing positive and negative opinions on the topic:

  • “I wouldn’t consume it due to the medium and long term damage to my health and the environment”
  • “It’s only relevant for the improvement of the production of biofuels like ethanol”
  • “I would consume it if was safely developed, I don’t see it as a problem to create GMOs and use it to produce beers”
  • “I believe GMOs are a valid idea, but I don’t buy them so as not to support big producers. I would rather buy from family farming.”
  • “Yes, I know every change in an organism is exhaustively tested for safety before the product is put to sale, we need to use science in our favor”
  • “I wouldn’t buy it because it doesn’t seem healthy/GMOs cause cancer”
  • “I don’t believe using radiation makes it safe for human consumption”
  • “Marketing actions need to be implemented for the population to know the benefits of GMO production, and the impacts it brings to society”

  • To clarify misinformation around the development and use of GMOs, especially in Brazil, we have also organized an Awareness Action in São Carlos. On August 2nd, we went to the city downtown to speak with the locals and tell them more about this subject. We repeated some of the questions we had on our survey and, when people said they did not know how to identify GMO foods, we showed them packages which contained the "transgenic" symbol (as seen below), an obligatory symbol in Brazil for any product containing more than 1% of GMOs in its composition since 2003 ( Decree 4680/03).

    The transgenic symbol and a bottle of soy oil containing it

    We also told people more about the entire process of release of a new GMO product, explaining the steps, the biosafety requirements, and telling them how many years, on average, it takes to test a product like this and assure that it is completely safe for human consumption.

    Team member Katiane Tostes explaining the importance of the development of GMOs and showing how to identify them


BALAN, A.; SCHENBERG, A. C. G. A conditional suicide system for Saccharomyces cerevisiae relying on the intracellular production of the Serratia marcescens nuclease. Yeast, v. 22, n. 3, p. 203–212, fev. 2005.

BBC NEWS. Tardigrades: 'Water bears' stuck on the moon after crash, 7 ago. 2019.

BIOSAFETY CLEARING-HOUSE. Search for Laws and Regulations.

BITTENCOURT NETO, Olavo de O.. Limite Vertical à Soberania dos Estados: Fronteira entre Espaço Aéreo e Ultraterrestre. 2011. Tese (Doutorado) - Curso de Direito, Universidade de São Paulo, São Paulo, 2011.

BRASIL-SP. 2014.


FERMENTEC. Banco de Microrganismos | Fermentec – Soluções Tecnológicas Industriais.


RESOLUÇÃO NORMATIVA Nº 7, de 27 de abril de 2009 - Resoluções Normativas - Comissão Técnica Nacional de Biossegurança - CTNBio.

RESOLUÇÃO NORMATIVA Nº 466, de 12 de dezembro de 2012 - Resoluções Normativas - Ministério da Saúde - Conselho Nacional de Saúde.

THE DIGITAL LIBRARY USP. Theses and Dissertations.


UCAR CENTER FOR EDUCATION SCIENCE. The Stratosphere - overview. 2019.