1. Lab work safety.
1. Instruments.
2. Chemical risk-assessment.
2. Biological risks, biosafety rules.
1. Microorganisms.
2. Used biobriks Parts.
1. Evaluation methods of the biobricks parts.
2. Pathogenicity, Infectivity and Toxicity.
3. Optional question.

1. Would the materials used in your project and/or your final product pose:
1. Risks to the safety and health of team members or others in the lab?
2. Risks to the safety and health of the general public if released by design or accident?
3. Risks to environmental quality if released by design or accident?
4. Risks to security through malicious misuse by individuals, groups or states?
2. If your response to any of the questions above is yes:
1. Explain how you addressed these issues in project design and while conducting laboratory work.
2. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry.
3. Under what biosafety provisions will / do you operate?
1. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible.
2. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them?
4. Describe any concerns or changes that were made based on this review.
1. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.
2. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.
5. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

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 there are safety rules to follow.

During our project, we have seek much information about products and materiel employed in our experiments and the risks associated with these latter. The litterature, the material safety datasheet and moreover the safety engineers of our labs.

At the CEA there is a special section of people in charge of the security and the safety. It is called FLS: Formation locale de sécurité, we may translate: Local Group of Security and Safety. They ensure the safety of the people who are working in the center and the visitors and also of the goods and material. Six members of our team have assists to safety conferences organised by CEA. All team members have met the safety engineer of the labs where we conduct the experiments. He explained the safety rules to be followed.

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 very toxic chemicals like mercury and also about technical aspects 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, speed and costs.

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”, ...

The experiences of our project did not require the use of sophisticated equipment. We have used basic devices that we find in molecular biological laboratory:

Ultra violet lamp:

Centrifuge:

The main risk is for the material. The centrifuge have to be perfectly balance otherwise the rotor could break. The majority of the centrifuge in our lab have detector that warns the operator in case of bad balance.

Autoclave:

The operation of the autoclave require a specific training

Water bath

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. The other source of important chemical risks is BET. To avoid these latters, experiments are performed under chemical hoods and used contaminated materials are sterilized.

During our project, mercury is conserved in the laboratory and is subjected to special treatment for elimination of heavy metals.

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.

Concerning the reprocessing of toxic chemicals or biological wastes, we follow the standard protocols of our lab. The main toxic wastes we have to deal with in our experiments is BET, the other very dangerous chemical is the mercury. Each one of this product is collected in a special barrel (one for each dangerous chemical) that are recovered by a company specialized in the reprocessing of hazardous wastes. The biological wastes are sterilized in an autoclave by heat and pressure before reprocessing by another company.

We work with a strain of E.Coli designed for the lab works. It is commonly used by the students and the researcher.

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 is extracted from Pseudomonas aeruginosa, a highly similar system exists in E. coli.

The system of P.aeruginosa controls numerous genes including virulence factors, a syringe mechanism to inject toxic compounds to a targeted cell, but the bacteria E.Coli we work with does not have this kind of system. We have chosen the system from P.aeruginosa to avoid interferences between our genetic circuit and the metabolism of E.Coli.

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

Even if in our project we don't plan to take our work out of the lab, so this can not be a cause of safety issue, 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.

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.