Safety

Safety forms were approved on September 22, 2013 by Evan Appleton.

iGEM Safety Questions

1. Please describe the chassis organism(s) you will be using for this project. If you will be using more than one chassis organism, provide information on each of them:

We used the strain E. coli K12, the MG1655 strain and its derivatives, which all belong to Risk group 1 (http://www.absa.org/riskgroups/bacteriasearch.php?genus=Escherichia&species=coli) and is not associated with disease in healthy adult humans.

2. Highest Risk Group Listed:

The highest risk group listed is 1.

3. List and describe all new or modified coding regions you will be using in your project. (If you use parts from the 2013 iGEM Distribution without modifying them, you do not need to list those parts.)

4. Do the biological materials used in your lab work pose any of the following risks? Please describe.

a. Risks to the safety and health of team members or others working in the lab?

All bacteria (Escherichia coli K12 and Pseudomonas pseudoalcaligenes KF 707) used in our project belong to the risk group 1, so they do not represent any risk for the health of the members of the team.

b. Risks to the safety and health of the general public, if released by design or by accident?

Our final system or any of the intermediate strains constructed do not represent any risk or danger for the general public as they are all derived from Escherichia coli K12 (belonging to the risk group 1), a strain that cannot colonize the human colon and does not produce toxins.

c. Risks to the environment, if released by design or by accident?

The strains used and constructed derive from E. coli K12, a strain whose survival under environmental conditions (soil or water) is limited. Moreover, the bacteria do not produce harmful compounds.

Source: http://epa.gov/biotech_rule/pubs/fra/fra004.htm

d. Risks to security through malicious misuse by individuals, groups, or countries?

The risk of malicious misuses of our project system is limited as this system is built for bioremediation, namely the detection and the degradation of the PCB. The genes used in this system do not code for toxins or for enzymes synthetizing products toxic to humans or harmful for the environment.

5. If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? (Consider the different categories of risks that are listed in parts a-d of the previous question.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? (Note: This is meant to be a somewhat open-ended discussion question.)

If used at the industrial scale, the risk of accidental release might increase. But in such a case, a different chassis could be used (different species, designed features to limit the impact of accidental release).

6. Does your project include any design features to address safety risks? (For example: kill switches, auxotrophic chassis,etc.) Note that including such features is not mandatory to participate in iGEM, but many groups choose to include them.

No, not at this stage. This project was a proof of concept. A different chassis could be used for the large scale implementation of this project. The GEMOTE system, imagined by the Paris-Saclay 2012 team, could for exemple be used to limit the survival of the strains outside a specific temperature range.

7. What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution’s safety training requirements, if available.

We learned about laboratory safety rules with our advisors during the three months of lab work (gas usage, handling of BEt (ethidium bromide), processing of chemical and biological wastes, precautions with UV light, manipulation of hot melted media, use of phenol...).

8. Under what biosafety provisions will / do you work? a. Please provide a link to your institution biosafety guidelines.

Biosafety in CNRS: http://www.dgdr.cnrs.fr/cnps/guides/risques/pdf/GuideIGHS_p27.pdf

b. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you made to your project plan based on their review.

There is in the Institute of Genetics and Microbiology (Université Paris-Sud) where we performed our experiments, a comity in charge of hygiene and security. One of our advisors, J-L Pernodet, is in charge of questions regarding GMO. He made sure that we only manipulated class 1 micro-organisms.

c. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible.

In France:

INRS: http://www.inrs.fr/accueil/risques/biologiques/prevention-risques/cadre-prevention.html

Haut Conseil de Biotechnologies: http://www.hautconseildesbiotechnologies.fr/IMG/pdf/Manuel_HCB_utilisation_confinee_OGM.pdf

In Europe: http://europa.eu/legislation_summaries/employment_and_social_policy/health_hygiene_safety_at_work/

d. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? (Check the summary table on page 3, and the fuller description that starts on page 9.) If your lab does not fit neatly into category 1, 2, 3, or 4, please describe its safety features [see 2013.igem.org/Safety for help].

The BioSafety Level rating of our lab is Class 1.

e. What is the Risk Group of your chassis organism(s), as you stated in question 1? If it does not match the BSL rating of your laboratory, please explain what additional safety measures you are taking.

Our chassis, Escherichia coli K12 MG1655, belongs to the biosafety risk group 1 and matches the BSL rating of the laboratory.

Written by Eric