1. Risk-assessment of our project
1. Instrumentation.
2. Chemical risk-assessment.
3. Biological risk-assessment.
1. Microorganisms.
2. Used biobriks Parts.
1. Evaluation methods of the biobricks parts.
2. Pathogenicity, Infectivity and Toxicity.
4. Environmental impact.
2. Biosafety provisions

Laboratory work requires the using of complex equipment or performing delicate operations, it also involves the use of toxic, flammable or explosive. The execution of this work may cause accidents or serious poisoning; the effects can be immediate or insidious. For this all reasons and during of our project .

Manipulation of living organism allows producing artificial form of life and metabolism. These modifications, although well controlled, require application of the precautionary principle.

Our project is based on the utilization of mercury, which raises questions about security for the researcher but also for the public and the environment. Mercury is an element that has toxic effects on brain and renal function.

Engineered bacteria might be accidentally or on purpose released in the environment. So, caution involves the implementation of different blocking to limit the propagation of these organisms in the nature:

• Nutritional blocking: organisms could survive only with artificial substances. In this way, in case of release into the nature such organisms would die.
• Evolutionary blocking: organisms couldn’t adapt themselves and evolve alone in the nature. This blocking prevents mutations of the organisms that allow them to survive.
• Preprogrammed cellular death: implementation of a suicide gene which is inhibited during wet work. In this way, organisms couldn’t survive outside the laboratory.

Through the iGEM competition, we face to the risks of synthetic biology. So it’s important to deal with safety issues our project could cause.

During our project, mercury is conserved in the laboratory and is subjected to special treatment for elimination of heavy metals. About searchers, protections are simpler: it is necessary to work without contaminating the material. If it is contaminated, it shall not be touched with bare hands. Hence the use of protective equipment.

We are making a device that can quantify a component in water, such as heavy metals pollutants. Two models are being developed. One of them involves the use of the Mer sensor. We therefore need to use mercury to test this system. These raise the environmental issue of the toxic waste management. Liquid having Mercury or tips and dishes that are in contact with this toxic are kept in specials bins. This rubbish bin is then given to a society specialized into toxic waste treatment. A slip monitoring is sign up by every organism that is involved into the production, transportation and treatment of the toxic waste. When the later is cremated, the producer of the waste receive and attestation that must be kept as a proof of the appropriate treatment.

The system we develop needs to be kept off until we want to induce it. In order to achieve that, we develop a post-transcriptional switch mechanism. This system, extracted from Pseudomonas aeruginosa, is highly similar to another one of E. coli. The later involve the management of numerous genes including virulence factors. The bacteria we use don’t have any gene coding for virulence factor so the risk doesn’t exist with our strain.

We have FLS (Formation locale de sécurité) that regulates what is coming in the laboratory where we are working; they ensure the safety of the researchers, public, and the environment. Six member of our team have had safety training, and all team members have met the safety engineer, who explained the safety rules to be followed.

Some specific equipment like autoclave is used only by trained personnel.

The toxic wastes like BET that we use during our experiments are collected in special barrels that are recovered by the reprocessing hazardous wastes company.

Rules are more or less restrictive for the scientist but follow same goals: protection of experimenter, persons who are in the experimental area, the local, and environment preservation. Our project is based on biomolecular experimentations. Each partner laboratories where we work are specialized in molecular biology and so have the needed equipment. What the Facility Security Engineer think about our project? Not particular attentions are needed concerning molecular biology experimentation but we are aware about chemical risks that can exist during manipulations. Most important risks are BET and mercury using. To avoid these latters, experiments are performed under chemical hoods and used contaminated materials are sterilized.

Chemical garbages are stored in specific barrels. Decontamination society retrieves these trashes.

From the environmental point of view, simple and efficiency methods can be used like bacteria not able to survive outside by use Amino Acids which not existing in nature. It’s too possible to use rare carbon source for the bacteria. We can also use a suicide gene repress by a chemical molecule not found out of a laboratory. Another ways is to make bacteria weak face to the micro-organisms natural selection. For the researcher’s safety in lab, the work in sterile middle, overall and gloves wearing and all other standard protections things are evidently recommended.

To increase the safety off iGEM competition, we think about bacteria which have an inducible essential gene for binary division by a chemical not existing or rare in nature, by this way the bacteria can’t be divide itself so it will be not selected and going to disappear nearly.

We will soon discuss with people that have already worked with mercury or others heavy metals. At CEA some researchers worked on Microsystems devices to detect and quantify these pollutants. They will share their experience and knowledge with us about the way to conduct safe experiments with these chemicals and also about technical aspect of existing measurement device. We would like to compare our work, our biosystem to “technological only” system that already exist, in terms of precision, sensitivity, reliability and costs.

So our work for the safety section of our wiki is not over, it will continue all over our project and after the jamborees. Indeed, we plan to present our work and the synthetic biology to a larger public: companies which fund us, school in our villages and town, a conference at “Midi Minatec”, ...