FCB:UANL Synbiofoam


Lab practices

One of the reasons why we decided to formulate a biotechnological firefighting foam, besides the environmental threat current foams represent, is the inaccessibility of foams as a tool for fire combat in our country. Throughout the year, after talking with several stakeholders including potential customers and users (go to our human practices section to get to know their feedback), we started searching for information we needed to know for our foam to be actually offered in the market. For this purpose, we took into consideration the different legislations regarding the biosafety and biosecurity aspects around the use of biotechnology and its products in Mexico. In order to comply with them, we have analyzed Mexico’s laws and statutes, as well as the way our project stands in relation to the regulations that originate from them.


This year, we took a different approach for the analysis of Mexico’s legal framework; we decided to scrutinize not only the legal aspect of GMO products, but we also focused our efforts on how the legislation would be specifically applied to our product Synbiofoam, which led us to analyze the legal requirements for the commercial development of it, to consider the proper steps for our proposed implementation. The following section explains how the different legislations helped us to shape our project.

Objective: According to Article #1 of the CBD, the objectives of this convention are the conservation of biological diversity, the sustainable utilization of genetic resources derived from this diversity, ensuring access to these resources as well as appropriate transportation of pertinent technologies, among other objectives.

How does it involve our project: Our project aims to utilize genetic resources found in the very same biological diversity that is threatened by forest fires. Also, it is a strong example of how synthetic biology can be used to protect biological diversity. Finally, during the development of our project, we have noticed that the Convention’s objective regarding accessibility to these resources is not completely fulfilled since the transportation of the pertinent technologies to utilize genetic resources of our biological diversity is not appropriate because it takes several months for us to receive genetic material from abroad.

Integration of the topics covered in this protocol: Thanks to the support of Youth Biotech, we presented our project, SYNBIOFOAM, at the Virtual Pre-COP-MOP15 during June 2021. In this event, we participated showing the importance of synthetic biology for biological diversity conservation.

Objective: According to Article #1, the Protocol was developed as a result of the Declaration of Rio(1992). This protocol aims to offer a guideline to achieve a satisfactory level of protection when transferring, handling or manipulating living modified organisms, developed using modern biotechnology, that can have adverse effects in the sustainable utilization and conservation of biological diversity and in human health.

How does it involve with our project: Since our project aims to use B. subtilis, we developed a thorough Risk Assessment in which we identified the risks related with the endospores produced by this organism, and based on our observations, we thus decided to generate a Killswitch relying on a CRISPR/Cas9 system. We also noticed that the protocol establishes different methods to allow importation and exportation of products developed with modern biotechnology, nevertheless, we as an iGEM team have experienced first hand how much time it takes for us to receive any piece of genetic material. Our experience with this protocol also helped us to develop a proposal in this topic for the shipping policy hackathon from iGEM, our participation is further explained in this section.

Integration of the topics covered in this protocol: In compliance with Article #15 of the Protocol, we decided to develop a Risk Assessment for our project, in which we identified the risks related to our project. And in compliance with Article # 16, we developed a killswitch based on CRISPR/Cas9 (further explained in our project description section) that could mitigate the potential risks identified in the risk assessment.

Objective: Article #1 of this protocol establishes three different objectives, the conservation and sustainable usage of the biological biodiversity , the utilization of its components, and the fair and equitable use of the advantages that arise from the utilization of genetic resources.

How does it involve our project: This protocol further develops what is established in the Cartagena Protocol by introducing codes of conduct and rules for the collaboration and contact between organizations in charge of transferring technology related to modern biotechnology, this allowed our team to understand the standards that our future product should have in order to export it, it is important to note that the importation/exportation process is something intricately involved in the journey of thousands of iGEM teams worldwide.

Integration of the topics covered in this protocol: As explained in the previous protocol, we developed a risk assessment and a strategy to mitigate the identified risks.

Objective: Article #1 establishes that this law aims to regulate activities related to genetically modified organisms, such activities include confined use, commercial release, experimental release, importation, exportation, among others.

How does it involve our project: Since our project implies the use of genetically modified organisms, we have to comply with this law. This document establishes different ways in which a GMO can be used and the corresponding organizations in charge of overviewing our work. In our case, the Secretary for Natural Resources and Environment, SEMARNAT, for its acronym in Spanish, is the organization to which our project would report its activities.

Integration of the topics covered in this law: Once we identified the organization in charge of our project, we decided to establish contact with them, and we had the chance to talk with Aída Juarez, a former member of this organization. She provided us guidance to fill a document that we would need to deliver in case we wanted to take our project outside our university. This document is called “SEMARNAT-04-025 C Notice”. To get to know her contributions yo our project, please go to our Human Practices section.

Objective: Similar to the Nagoya Protocol in the way it complements the Cartagena Protocol, this regulation aims to further explain details related to the information that a genetically modified product should include in the process of importation and exportation, among other topics that were not clear enough in the LBOGM.

How does it involve our project: The regulation indicates the requirements for the documents we will need for our genetically modified organism, such as the importation and exportation processes to take into account when working with GMOs.

Integration of the topics covered in this law: As explained in the LBOGM, the main product derived from this law was the SEMARNAT-04-025 C Notice.

Objective: CIBIOGEM is an organization focused on overseeing the governance of genetically modified organisms and biotechnology in Mexico. According to article #5 of the Regulation, the CIBIOGEM has the objective to formulate and coordinate the national biosecurity and biosafety policies for GMOs with help of other organizations like SAGARPA, SEMARNAT, SHCP, among others.

How does it involve our project: CIBIOGEM mainly focuses on regulations involving biotechnology and its derivatives. Also, this organization has a consultory group made up of citizen scientists that act as advisors for this organization in certains topics, such as synthetic biology, in which some iGEM alumni have participated as advisors for this organization.

Integration of the topics covered in this law: We could not contact current members of this organization since apparently the contact information published in their webpage is outdated. Nevertheless, we established contact with former members of the advisory group for this organization.

Nowadays, there is a great deficiency in firefighting foams regulation in Mexico, as there are not enough laws related to this matter.

Currently, there is only one law related to foam concentrates, and there are other four laws related to fire extinguishers; however, regulations for our foam are at a dead point due to the nature of our foam, which is made with synthetic biology and is does not strictly enter to the category of fire extinguisher.

The only legislation related to foam concentrates in our country is the NOM-202-SCFI-2017 It addresses the extinguishing capability, performance and construction of firefighting foams, as well as other important specifications for its correct use and test methods.

Some of the most relevant information this law mentions is the temperature at which fire extinguishers need to operate, which can cover a range from -30°C to + 60°C, nevertheless there is still a lot of work to do for the development of laws related to foam concentrates in Mexico.

We were able to understand the role of different legal documents that regulate the use of GMOs in our country thanks to the valuable contributions of MSc Aída Juárez and our advisor Ediner Fuentes. A more detailed explanation of how they helped us and the actions we took after their feedback is provided on our human practices section.

A common thing most of the documents reviewed shared was the requirement of a risk assessment, as a way to demonstrate and ensure that a GMO is safe to take into the market. In consequence, we developed a thorough risk assessment in which we analyzed the potential risks associated with our project. It is mainly composed of three parts: Risk Identification, Risk Management and Risk Communication, that are explained below.


We first identified and analyzed the possible sources of risk, where we concluded that all the bacterial strains used for our project (Escherichia coli K12, Top10, and Bacillus subtilis ATCC 6633) have a history of being safe organisms for its use in the laboratory. Therefore, they are unlikely to cause illness to humans, animals, or plants; and the genetic modifications we designed are not expected to alter these characteristics. In addition, the pUC-derived vectors have also a history of safe use, and do not give the recipient organism characteristics that may cause disease or ill effects on the environment.

In conclusion, the produced GMOs can be classified as Risk Level 1 organisms, which means that the possibility of harmful effects is very low, since they do not cause any type of illness. Based on this, we estimate that the overall risk involved and the probability of an incident are both low, considering as well the constant expert supervision and strict safety measures taken into consideration (which are explained later on in this section). Hence, the possibility of risk is insignificant

The following document shows the complete Risk Assessment made by our team, based on the information presented by team FCB-UANL 2020.


Still, a plan on the management of the project according to correct biosafety and biosecurity measures was done. Some of the strategies adopted are described in the following paragraphs.


  • All of the laboratory activities were performed according to our institution's rules and safety regulations. At all times in the laboratory, the team members were supervised by at least one of the instructors. Only the people who have had previous training in lab equipment and safety protocols were allowed to perform experiments. Regarding our self-education activities, where we had apprentices into our laboratory (for more information, go to our communication section), everyone without experience had several previous theoretical explanations of how things work regarding the lab rules. Every apprentice developed their activities with the supervision of both a wet-lab team member, and at least one of our instructors.
  • Besides the risk assessment carried out, we made the decision to better prepare ourselves. With this purpose, our team members working with the safety aspects of our project enrolled on the Future Learn’s course “Next Generation Biosecurity: Responding to 21st century biorisks”. This allowed us to better understand the definition of biological risk, as well as to establish measures to mitigate them. Topics covered by this course include: biosecurity, biological threat, responsible conduct in science, among others.


Once we identified the risks associated with our project, we came to the conclusion that we had to have a method to ensure that risks related to unintentional GMO release (mainly spores of B. subtilis) would be significantly reduced. Hence, we carried out a predictive model of the competence switch in B. subtilis in order to develop a strategy to avoid sporulation on the molecular level. Nevertheless, the obtained results showed these measures could not stand alone to mitigate the risk of unintentional release by spores; more information of the model and the decision-making process is available in our model section.

Hence, we decided to design a killswitch mechanism based on a CRISPR/Cas9 system that will activate during a specific point in the sporulation process. With the addition of this measure we have a more robust system to control the production of spores, in addition to the ability to degrade the modified genetic material introduced to the bacteria we worked with. We developed this construction along with the UNILA_LatAm team (go to our partnership section to see the things we did together). The complete mechanisms and components of this system are explicitly detailed in our project description section.

The following images briefly explain the mechanisms of the killswitch based on CRISPR/Cas9:


The last step in the process of risk identification, is the communication of our findings, in other words, informing the corresponding authorities about the risks we identified in our project and the measures we took in order to mitigate them, this is a very important step for any biotechnological product produced in Mexico, since this step will decide if your product can keep developing, or on the other hand, not be allowed to reach the market. In the case of our project SEMARNAT is the legal organization in charge of deciding the future steps of our product, additionally, SEMARNAT has laid out a specific way to communicate this information through the , submission of the notice “SEMARNAT-04-025-C”(1) in compliance with LBOGM.

This notice gathers information about the type of GMO we aim to produce, its modifications and the final objective of its production, as well as the kind of facilities in which we plan to produce it and the kind of biosafety and biosecurity measures these facilities have. In this notice we considered the up-scaling in bioreactors for the future production of our product (considered in our model and entrepreneurship sections). Besides communicating the risks identified to the corresponding legal organizations, we decided to also share the path we followed during our risks assessment and the corresponding measures we took in order to mitigate the risks we identified. Next, we present our risk assessment.

In addition, we decided to take the term communication a little bit further into our society, since we know first hand that many people in Mexico have a strong perspective on the production and usage of GMOs, thus we decided to reach our community and share about the processes for the production of GMOs as well as the actual risks that could involve this technology (visit ourcommunication and education sections for more information about this activities).


As part of risk communication, we also decided to generate a “Dual-Use Research of Concern” (DURC) analysis in order to demonstrate that our product has undergone different safety and security analysis to ensure the attainment of the greatest number of benefits with the least amount of risks. With this analysis, we concluded that the benefits of our project outweigh the risk associated with its development and final product. In addition, the possibility of misuse of this technology is low. Thus, we do not consider our project as DURC. Next, we attach the document with the complete analysis.


The Shipping Policy Hackathon 2021 was an event organized by the iGEM organization and After iGEM this year, and it focused on the challenges of shipping biological material or DNA across countries, in order to make teams acquire a better perspective of the issues everyone faces worldwide. During the previous months, we worked on researching the process of shipping DNA to our country, and the requirements to do so. Finally, during the three days long Hackathon, we worked on a document about the Mexican policy for importation and exportation of GMO and biotechnological products, shown below.

With this event, we learned that Mexican regulations directed towards the transit of biotechnological products require an update since they lack the scope necessary to offer reliable methods to protect the environment and society from danger while promoting the development of new technologies.

Basing our analysis in the LBOGM (2) and the NOM-002-SCT-2011(3), and comparing them with the USDA’s Rule of Movement of GMOs we noticed that there is a lack of regulation related to technologies outside agriculture in Mexico, this represents a huge blockade for developing technologies involving biotechnology, this could be related to the importance given to the methods of production for the corresponding GMO, instead of focusing on the characteristics of the resulting GMO, like is done in USDA’s regulation.

Those are the reasons why our proposal is focused on filling up the legal vacuum that the current Mexican regulations have by adding aspects of the USDA's evaluation of the GMOs and other biological materials into the LBOGM, and we expect that by adding specific procedures for the transport of GMO and genetic material, scientists will find fewer legal loopholes to develop their projects while the government maintains the appropriate security measures.


Before carrying out the fire tests for our final formulation (to get to know its composition and how we determined it go to our engineering success section) we developed several experiments with the aim of ensuring [1] that our formulation do not have living bacteria, and [2] that the remnant DNA after the sonication is not able to confer antibiotic resistance. Following the Do Not Release Policy, we sent the protocols to the iGEM Safety and Security Committee, and they accepted our submission. Then, the next step was to experimentally prove our protocol.

For this purpose, our sonicated extract was exposed to UV light for three and ten minutes to determine the best time to avoid bacterial growth; this protocol is based on the work developed by Hernández-Arias, et al. (4). Right after that, we used the same sonicated extract to transform Ca++ competents cells, which were inoculated on petri dishes containing agar and kanamycin, and incubated overnight at 37°C to analyze whether they presented bacterial growth.

On both experiments we obtained positive results, since the UV exposition did work for avoiding bacterial presence on our sample; in the same way, we concluded that the genetic material present in the supernatant does not have the capacity to confer antibiotic resistance. Next, in the following images you can see our experimental outcome.

In the first one you can see the agar plates with the UV irradiated supernatant. The plate on top was the control, on the left is 10 min exposure to 280 nm and the one in the right is 3 minutes exposure to 280nm. In the second one, there are the results of the usage of the supernatant exposed during 10 min to 280 nm to transform E. coli Top10 cells. On the left side is the transformation control (RFP), meanwhile on the right is the test we made. As a result, the genetic material present in the supernatant does not have the capacity to confer antibiotic resistance.

Following the considerations described in the previously mentioned protocol regarding the volume and UV exposure conditions, 128 mL of each one of the Ranaspumins' sonicates were placed in a flat container in order to obtain a wide distribution of the solutions and optimize its exposure to UV radiation for 3 to 10 minutes. Based on the results obtained, we observed that a 10 minute exposure to UV radiation was the most effective in eliminating living bacteria. Once all the samples were irradiated, the presence of living organisms and of DNA confering antibiotic resistance in each one of the samples was determined as previously described. After observing the absence of living organisms or DNA conferring antibiotic resistance in all the samples, 120 mL of the Rsn-2, 40 mL of the Rsn-3 and 40 mL of the Rsn3-5 concentrates were mixed and tested on the fires.


All the fire tests were developed in the facilities of Rescueteam Mx, where they had several fire extinguishers available, and gave us a brief description of how to use them if necessary. The fire and foam manipulation were carried out by the firefighters and experts on the subject, however, all the team members present during the tests wore the safety equipment (safety jacket, pants and gloves) provided by Rescueteam Mx.

All the surfaces that were in contact or around the areas of the fire tests (as well as the safety equipment we wore) were covered by three layers: [1] flame-resistant, [2] water-resistant, and [3] heat-resistant. In addition, for testing the different fuels, they were set on fire over heat-resistant glass surfaces. In addition, the tests were carried out on a wide and well-ventilated space, to avoid breathing the fuel vapors and smoke generated. Overall, those were the recommendations and rules established by the experts. For a detailed explanation of the tests, as well as their results, please go to our proof of concept section.


  1. Secretaría de Medio Ambiente y Recursos Naturales. (2016, 5 de septiembre). Avisos de Utilización Confinada de Organismos Genéticamente Modificados (c) SEMARNAT-04-015-C.
  2. Regulation of the LBOGM. Further development of the terms established on the LBOGM. DOF-30-10-2013.
  3. Secretaria de comunicaciones y transporte (2012, 27 de enero). Para el transporte de materiales peligrosos NOM-002-SCT/2011.
  4. Hernández-Arias, A. N., Jaramillo-Sierra, B., Rodríguez-Méndez, B. G., Peña-Eguiluz, R., López-Callejas, R., Mercado-Cabrera, A. & Alcántara-Díaz, D. (2019). Escherichia coli bacteria inactivation employing ozone and ultraviolet radiation using a reactor with continuously flowing water. Journal of applied research and technology, 17(3), 195-202.
  5. Convention on biological diversity. Related to use and protection of genetic materials and biodiversity. August 1992.
  6. Cartagena protocol on biosafety. Related to risk assessment and transference of technology. 2003.
  7. Nagoya Protocol. Complementation of the process to import and export internationally. 2010.
  8. Law on genetically modified organisms biosafety “Ley de Bioseguridad de Organismos Genéticamente Modificados”. National regulations on genetically modified organisms. DOF-18-03-2005.
  9. Regulation of the CIBIOGEM. Responsibilities of the CIBIOGEM. DOF-30-08-2016.

Our 2020-2021 iGEM project is generously supported by