Biosecurity is a paramount global health and well-being guardian, serving as an unwavering defense. In an increasingly interconnected world, the significance of biosecurity cannot be overemphasized. It serves as a beacon of hope in the face of pandemics, safeguarding our communities, agricultural systems, and ecosystems from the devastating impact of diseases and invasive species.
Biosecurity not only underscores our collective duty to safeguard ourselves but also highlights the intricate interdependence of life forms that sustain us. It implores us to maintain vigilance and consolidate our knowledge, resources, and determination in our ceaseless pursuit of a healthier and safer world. It is a testament to our capacity for adaptation and innovation, as we courageously confront the unknown, securing a brighter future for generations to come.
Let us wholeheartedly embrace the paramount importance of biosecurity, for it signifies our commitment to nature and one another, a demonstration of our resilience, and the key to a world thriving in harmony, thanks to iGEM we have achieved the expected safety, and we are eager to learn more about it.
Objectives
Plants exhibit remarkable and ubstantial sensory capabilities. We have posited the utilization of plants themselves as sensing instruments for the detection of diverse compounds and environmental circumstances. Employing plant promoters selectively triggered in the presence of elevated concentrations of CO2, as judiciously curated by the Navarra_BG research team, we have successfully engineered Arabidopsis thaliana plants to function as biosensors attuned to this particular metabolite. The outcomes thus garnered engender fresh vistas for the potential deployment of plants as biosensors for other soil-based pollutant molecules and specific environmental contexts.
Within the ambit of this project, we have harnessed the tenets of Synthetic Biology to devise innovative gene reporter constructs, thereby streamlining the visual assessment of the biosensor plant's response, obviating the necessity for intricate instrumentation. Furthermore, our inquiry has encompassed the exploration of appended sequences fused to the reporter protein, serving as modulators of its degradation kinetics subsequent to the cessation of the stimulus.
In the field of safety, our project has different aims:
1st aim: To protect our health by detecting toxic substances
2nd aim: To protect the environment by detecting hazardous substances
3rd aim: To keep our spaces clean and safe
4th aim: To give a sustainable alternative to the electrical methods used nowadays.
Design
Throughout our whole project we have used two groups of living beings: plants and bacteria. Two groups that are completely innocuous for human beings.
| Bacteria #1 | Bacteria #2 | Plant | |
|---|---|---|---|
| Name | Escherichia coli | Agrobacterium tumefaciens | Arabidopsis thaliana |
| Specifications | Bacteria Gram negative, facultative anaerobe. | Agrobacterium tumefaciens is a Gram-negative, rod-shaped bacterium with motility and peritrichous flagella. | Flowering plant of the Brassicaceae family. Tall, thin and of fast proliferation. |
| Statue | Widely used in synthetic biology and genetic engineering in cultures and transformations. | Used in genetic engineering to modify plants. | Widely used in synthetic biology. All the genome of this plant is known, so it makes it easier to work with. |
| Advantages | The capacity of rapid proliferation. We have used it to grow our plasmids in a very short period. | The amazing capacity of infecting plants. This capacity allows synbio students to introduce bacterial DNA into plants. | We have considered the very fast period of growth (21 days) and the known genomic sequence; this quality is recognised by researchers all around the world. |
| Role in our project | Vector of transportation and incubation of plasmid growth. | Introduce our constructions into Arabidopsis thaliana. | It is used as a modified plant that contains our constructions and it is the final product. |
| Level of danger | None (BSL1) | None (BSL1) | None (BSL1) |
Both; bacterial and vegetal chassis are completely safe to work with and with very low dangerousness. The first two bacteria cannot infect humans, the first one is not very dangerous, while the second one can only infect plants. Furthermore, Arabidopsis thaliana does not emit any dangerous spores or pollen.
Important note about A. tumefaciens: Normally, the bacteria used is a pathogen for plants, that introduces its DNA into the vegetal cells and provokes tumours inside the plant. The version used has been Agrobacterium tumefaciens EHA105 for Arabidopsis thaliana transformation. The vector Ti is disarmed so it is an apathogenic bacteria. This means that the bacteria do not provoke tumours or the damages that they could provoke as a wild-type bacteria, so it infects the plant by sharing its DNA, but it doesn’t damage it severely.
Regarding standard parts or bio-bricks, our constructions are completely innocuous. The DNA that is included in our plasmids codifies only for safe genes such as GFP, recombination sites or even antibiotic resistance that applies solely for bacteria.
Work
During the whole project, we have followed strictly the security rules according to the current regulation in iGEM, the government of Spain, the Committee Bioethics, and other governmental corporations related to STEM disciplines.
The use of equipment has been exhaustive. Not only we have used the safety protocols but physical equipment that always protect us:
- Laboratory coats: Protection of body and clothing from possible spills. Lab coats are cleaned and washed every two months in a regular form. Is it being any serious spill the cleaning is immediate.
- Safety goggles: Always used when required, they assure the safety of our eyes in various procedures while working with bacteria. Especially used when working while doing the Agroinfiltration of Agrobacterium tumefaciens (this task can squirt bacteria into the air, in this way, we can protect our eyes), likewise, we also use them in the management of liquid nitrogen.
- Gloves: Always used except when working with fire. They are always of paramount importance due to the protection of our hands. With them we protect each other from spills, possible mutagenic substances such as Midori Green in Electrophoresis Gels. As a result, we use them the entire time except when working with fire, as they are made up from Latex.
- Mittens: When working with liquid nitrogen, the temperature of the compound is of − 196 °C [− 320 °F], so good protection is of the utmost importance. That is why we use a pair of mittens that are tolerable to extremely cold temperatures. So, with these gloves, we assure the caution and well-being of us, iGEMers, our PIs and our Instructors.
During our work in the laboratory, we have been always supervised by our instructors, receiving professional training. All the team has passed through safety training. Qualified and trained scientists have taught us about all the safety lab measures, the use of biosafety cabins, and emergency procedures (among very others). Also, we have had a responsible for Biosecurity Policies inside the instructors/Pis and another one inside the Team Members.
As we have worked in a level 1 laboratory, the level of dangerousness has been very low. The material treated can’t affect humans’ health and can’t provoke serious problems.
As we handle plant material, we have used not only laboratories but also the infrastructure for plant growth as Biosafety cabinets and risk-free greenhouses and growth chambers, places in which our plants have been growing safely and without risking human health.
We adhered to the required security precautions while we were inside the lab:
- We have always been under the watchful eye of the lab managers, who oversee the actions taken and directing us towards the necessary information and skills.
- Prior to entering the lab, we were required to attend lectures on health and biosafety, during which the procedures and things to do in an emergency were also covered.
- To operate in the lab, we are provided with the proper attire and tools, including lab coats, safety goggles, respirators, nitrile and/or latex gloves, and more.
- Any waste that we generated was disposed of in the appropriate container and processed according to the law.
We have performed our tasks in different places inside the facilities in the laboratory to assure safety for avoiding release beyond containment:
- Safety cabins: To work in a sterile environment we have performed different tasks in them. Some of them are exclusively for plant material and others for bacterial material.
- Autoclave: A sterilization method inside the heat-humid group. With a temperature of 150º C and 5 bars of pressure. All the material used in the laboratory has been processed at those extreme conditions.
Policies
iGEM Responsibility and iGEM Community have also been considered thoroughly during the whole project.
As we are working on a proof of concept, the specimens used and transformed in the laboratory will not exit the facilities by any means. All plants would be treated with heat and pressure before throwing them in garbage disposal. The release of specimens is restricted. We are very concerned about the Release Beyond Containment policies and under no circumstance a GMO exit the laboratory facilities even to the Grand Jamboree or for showcasing our project to the community outside the lab.
As aforementioned, the living beings used during the project are characteristic of level 1 laboratories and all of them are completely innocuous and all of them are included in the Whitelist.
No animal chassis has been used in any part of the project, whether humans, animals from a lower order or human subject research. We know that human research experimentation is completely forbidden and goes against the culture of bioethics, and we would never encourage non-regulated experiments with humans or with animals from superior orders.
Finally, we considered the information provided by iGEM in the webinar ‘Responsibility Workshop: Designing a BWC Compliance Regime’ of March 2022 given by iGEM Responsibility and the Safety and Security Committee. We understood the importance of Safety in the society and how can our projects change the world in a negative way if not treated safely with the correct
Outside the laboratory
As we all know, the competition iGEM does not only focus on the lab work but also on the outside of the lab work. We perform multiple tasks such as wiki construction, integrated human practices, and internal work of iGEM that is not to be fulfilled in the lab. But it is also important to be safe in those fields because it is inside of the project development. These tasks are done in our other workspace, the Planetarium of Pamplona. Given the nature of this office-like space, we count on the following elements to provide our safety throughout the project.
- Fire extinguisher: Type ABC with dust filling. Useful for fires with solid, liquid, and gaseous comburent. Not suitable for use with voltages greater than 50,000. The last revision of this equipment was in July 2023.
- Emergency exits: Three emergency exits with proper flow of people, and in strategic placement for allowing full evacuation of all the people at the planetarium. It allows the fast evacuation of staff, visitants and our team.
- First aid kit: Full of equipment that ought to be needed in case of an emergency. Allows quick and easy response in case of an emergency. Along with the training received, it is useful in cases in which any of us could be injured.
As we are practicing science with a fellowship from the Government of Navarre, we count on health insurance from NICDO (a branch of the Government) that covers any type of injury if needed, we are protected not only by the equipment and policies but by our own Government.
In summation, the paramount significance of biosecurity cannot be overstated; it stands as a cornerstone of global health and ecological stability. The meticulous preservation of biosecurity measures is not merely a matter of choice but an imperative for the preservation of biodiversity, human health, and the sustenance of our planet's delicate ecosystems. The relentless commitment to biosecurity safeguards the intricate web of life upon which we depend, offering a pathway toward a more resilient and harmonious world.
Our proposal for biosecurity
To improve biosecurity for iGEM teams and for the scientific community, we have developed a guide for all the scientific community that includes definitions, protocols, description of material, iGEM policies given by iGEM Responsibility.
The NavarraBG’s Safety Handbook, is available in PDF format, and you will be able to find it in this section. We are very proud of our work, and we hope that this contribution can make our world safer.
Safety image album
