Team:Aix-Marseille/safety
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Safety
Our objective is to reduce the propagation of arboviral diseases by tiger mosquitoes. To do so, we planned to modify a bacterium, which would detect a specific viral infection in a mosquito and kill them. This solution would protect the food-chain, which mosquitoes are part of and would also be an eco-friendly alternative to current treatments. In the lab, we used the bacterium E.coli as a way to prove our concept. However, in the following steps of the project, we aim at using Asaia sp., as this bacterium is part of the mosquito microbiota. We used toxins in order to kill infected mosquitoes (Cry11Aa, Cyt1Aa) and a chaperon which will protect the bacterium from these toxins (p20). Finally, these toxins needed to be released in the mosquito microbiota in sufficient amounts to kill the insect hosting the arbovirus. In order to have a safe engineered machine, we designed a timer system, based on a colicin system.
Risks management
We received a safety and security formation (lab rules, biosafety equipment, biosecurity, waste management system, special protocols) as we arrived in the lab. We were also supervised by mentors, who taught us how to manipulate safely in the lab.
We did have an open bench in order to work with our bacteria to limit the risk of contamination. Toxic chemicals were located in special rooms and we had to wear gloves (in addition to our scientist’s blouse). We wore our scientist’s blouse all the time, and we used to disinfect our bench every time we used them and at the end of each day of work. And, as mentioned before, we were sorting waste : contaminated waste (with bacteria) were going in a decontamination tank (empty every day) or directly in a special bin ; not contaminated waste were going in a sterilization tray or directly in another special bin. Every bin was recovered once a week. If anything bad would have arrived, doctors were available nearby.
Identifying possible risks
As mentioned before, we wanted to work with E.coli and Asaia sp. Yet, E.coli is easier to use in the lab in the first place. Therefore, E.coli DH5α cells were used for plasmid production and E.coli MG1655 cells as expression bacteria. E.coli strains are not pathogen, their utilisation does not cause any risk. The important safety point, when using this strain, is not to throw it in the sink.
We used some dangerous product mentioned here:
- Mitomycin C (inducer of our timer system),
- GelRed (in order to reveal our blot membranes),
- Acrylamide (to perform SDS PAGE gels),
- NBT and BCIP (part of the Western Blot reveal solution).
These products are mutagen and may cause injuries, that’s why we followed a security formation before the start of our manipulations.
We worked with toxins, which can be harmful molecules. We therefore needed to understand them better. We have in particular created toxins parts that we have sent to review in order to be authorized to use them.
Our final product would be released in the environment (as an eco-friendly product) and used by different types of people (associations, cities or private companies). If security and bioconfinement parameters are not respected, our final product may cause injuries. Indeed, as Asaia sp. is a pathogen, it could harm humans and animals, if spread in the environment without being engineered as a full-secure system.
Compliance with iGEM’s rules and policies
Nothing was planned to be released derived from our project. We wanted to work with mosquitoes, and as they are animals, we made sure to have the authorization of iGEM to use them.
Legislation about GMOs
In terms of GMOs and GMMs, France does not have its own regulations. The regulation of GMOs and GMMs in biotechnology is regulated in Europe by numerous directives and regulations on the patentability of biotechnology, the use and marketing of GMOs…
European legislation applicable to GMOs has been set up since the early 1990s. This specific legislation has two main objectives:
- to protect human health and the environment, and
- to ensure the free circulation of harmless genetically modified products in the European Union
All the legislation on GMOs was recently modified to create a new legal framework: Directive 2001/18/EC on the deliberate release of GMOs into the environment (1). This regulation is very strict; it takes into account the evaluation of the risks for the environment, limits the duration of the authorization to 10 years, renewable, and sets up a compulsory monitoring after the authorization of the release in order to ensure the good progress of the release on the market. Finally, this regulation constrains the European Commission to publish a report on marketing experience every three years.
When a company that owns a GMO, having completed its studies in a contained environment, wishes to obtain an authorization to place it on the market in the European Union, it must go through :
- Authorization procedure for deliberate release
- Authorization procedure for placing on the market
For each of these procedures, the company must submit an application for authorization and assess the risks before being authorized for use.
Directive 2009/41/EC relates to the contained use of genetically modified microorganisms. It authorizes the use of GMMs only if the purpose of their use is the protection of health and the environment