Following iGEM's rules and policies
We have not done any prohibited activities in iGEM, such as testing products on humans, releasing or deploying a genetically modified organism outside the lab or using organisms from Risk Group 3 or 4 and parts from those organisms. All of our work is covered in the White List.
Also, we have not done any special activity that needs permission from iGEM, like working with animals or samples from animals, bringing a product of a genetically modified organism outside the lab, conducting laboratory experiments using human samples, such as blood, DNA, other bodily specimens, and health or psychological outcomes, using parts or organisms obtained from anywhere other than a trusted commercial or institutional supplier, biasing the inheritance frequency of a genetic marker in an organismâs progeny, increasing risks from antimicrobial resistance, such as by using novel resistance factors⊠etc.
In our research, we are not doing surveys, interviews, or other human subjects research.
About our lab
In our laboratory, we have safety equipment and equipment for working with microorganisms and chemical agents:
As shown in the images, our project requires working in a biosafety level 1 laboratory (Level 1- standard microbiological lab), therefore, we only worked with organisms that are not harmful to humans or the environment. To ensure safe handling of the laboratory, all team members received an introduction to the S1 area of the respective laboratory.
All security measures have been approved by our PI, who has supervised us at all times and whenever we have needed it.
About our project
Our bacteria (E.coli) will be engineered to produce resveratrol. Resveratrol is a natural molecule found in wine and grapes, with important antioxidant effects and benefits against health issues related to the process of ageing. Our main goal is to accomplish the efficient bioproduction of this important molecule, using precursors present in the wine industry waste. We have used DH5-É and C2984H strains of E. coli. Our E.coli will act as a vector to produce resveratrol. We will need to introduce three enzymes from the natural biosynthetic pathway into our bacteria (TAL, STS y 4CL) and the PPOR gene. This gene will provide fluorescence to colonies with the Golden Gate L2 construct (all four genes on the same plasmid), as well as being necessary to seal the construct. As substrate for E.coli growth and the production of resveratrol, waste from the wine industry will be used. The Golden Gate technique will be used to perform the theoretical construction and assembly of the genes. Finally, we will measure protein levels by different techniques: targeted metabolomic analysis, chromatography, Western Blots...
We have created new parts that we will upload to the iGEM registry, which are modifications and adaptations of the cloned genes to E. coli. None of the used parts could be hazardous in our context.
We have ordered the used DNA/RNA from a company, a member of the IGSC.
Identifying project risks
Materials and reagents we used that may pose a hazard are listed below:
- UV Light (Transilluminator): the transilluminator presents a danger of skin burns and eye damage from inappropriate exposure to UV light. However, the Gel Doc device we use stops the UV light when open. To cut out gel pieces to isolate DNA, we use an LED-based lamp.
- Autoclave: presents risks of explosion, thermal contact and electrical contact.
- Hazardous chemicals (flammable liquids such as ethanol): we employ kits for the isolation of DNA, thus avoiding organic solvents, and a safe alternative to ethidium bromide to stain DNA in agarose gels, called Sybr Safe. The main hazardous chemicals used are ethanol and isopropanol, flammable liquids. We have also been trained in the handling of concentrated acids and bases, such as the ones used to calibrate the pH meter, which could damage the skin if not properly used. Using liquid nitrogen also has potential. That is why we always use thermal gloves and safety goggles to handle it.
- GMO E.coli: accidental release of GMO E.coli would mean the release of a bacterium with antibiotic resistance, which may trigger modifications of microbial populations and the acquisition of antibiotic resistance in other strains.
Anticipating future risks
If our project could be developed beyond iGEM to a final product, the main use would be in a factory or industrial context. If we are able to synthesize resveratrol, one option would be to scale it up for industrial production and encapsulate it for commercialization. It could be used as a nutritional supplement, given its beneficial properties; but also used in the cosmetic industry. The versatility of our product allows us to incorporate it into facial creams, serums, lotions, and masks.
This future development would therefore involve the release of a product of synthetic biology, but not any living organisms. We conceived our project as a way to improve the availability and access of resveratrol to people. Resveratrol has many properties, so it could be used in multiple ways: in the cosmetic industry, in the food industry... In any case, the extension of our project would require numerous tests and experiments to determine different parameters: maximum efficiency, analysis of safety and toxicity, production in other organismsâŠ
Bioethics is very important, and in our team, we have discussed breaking norms about engineering biology. This project could lead to bad outcomes, like harm to human health and safety or harm to agricultural animals, crops, or domesticated animals. Accidental exposure to a hazardous organism or chemical, or an accidental release of an engineered organism or part into the environment might be the way those bad outcomes come to pass.
Also, this is a very unlikely situation but it would happen that some of the bacteria we are using could accidentally enter the environment through cloth contamination, for instance. However, since our parts correspond to enzymatic processes in the biosynthesis of resveratrol, using gene sequences that are already available in nature, the risk of some adverse effects in the environment is negligible.
Managing risks
We are in contact with the Work Health department in our university as well as professors experts in biosafety and bioethical issues with whom we are going to discuss the risks and implement security plans. We have also received safety and security training, including:
- Lab access and rules
- Responsible individuals
- Differences between biosafety levels
- Biosafety equipment (e.g. biosafety cabinets)
- Good microbial technique
- Disinfection and sterilization
- Emergency procedures
- Rules for transporting samples between labs or shipping between institutions
- Physical biosecurity
- Personnel biosecurity
- Chemical, fire and electrical safety
Furthermore, our research laboratory follows the biosafety and biosecurity standards set by current legislation, including several regulations that apply at the European level and have local counterparts. Some of them are described in the following web pages:
- Classification, labelling and packaging of substances and mixtures.[1]
- Protection of the health and safety of workers against risks related to chemical agents at work. [2]
- Qualitative and simplified methodology for assessing the risk of accidents involving chemical agents. [3]
In addition, while doing our work in the laboratory, we use personal protective equipment, such as lab coats and gloves. It is also equipped with a waste management system, and we have access to medical surveillance and an accident reporting system. We have followed the advice and received training regarding the main safety issues discussed above. We have been closely supervised by our PIs, particularly Lucas Castellani, who is a well-trained postdoctoral fellow who has been primarily with us in the lab.
Other actions we have taken to receive biosafety training have included:
- Consulting with other experts about managing risks
- Crafting a responsible communication plan
Finally, If our project were to be developed further for a real-world use after the iGEM project stage, our project would need support or review to manage the risk we wrote above. Following the recommended instructions and taking the biosafety measures as explained, we are prepared to minimize the risks of our work and respond quickly in case hazards occur.
| [1] | |
| [2] | |
| [3] |