Team:Paris Bettencourt/Safety

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Would any of your project ideas raise safety issues in terms of :

Researcher safety

We only use the Bacteria Escherichia coli DH5a NEB Turbo and K12 which are common laboratory strains [http://www.openwetware.org/wiki/E._coli_genotypes|1], also considered as Level 1 Biosafety Containment agent.

However, there is a number of standard lab reagents that we require for our project, that are harmful on contact. These include:

• Ethidium Bromide:

Acute: Hazardous when ingested or inhaled, and is an irritant of the skin and eye. Chronic: In the long term exposure can have carcinogenic, mutagenic, and teratogenic effects, and can cause developmental toxicity.

• Polyethyleneimine:

May be harmful if inhaled. Causes respiratory tract irritation. May be harmful if absorbed through skin. Causes skin irritation. Causes eye irritation. May be harmful if swallowed.

• Glutaraldehyde:

Eye: Causes eye irritation and burns. May cause permanent visual impairment. May cause chemical conjunctivitis and corneal damage. Skin: May cause skin sensitization, an allergic reaction, which becomes evident upon re-exposure to this material. May cause hives. Causes skin irritation and burns. May cause staining of the hands (brownish or tan). Ingestion: Harmful if swallowed. Causes gastrointestinal tract burns. May cause central nervous system depression, characterized by excitement, followed by headache, dizziness, drowsiness, and nausea. Advanced stages may cause collapse, unconsciousness, coma and possible death due to respiratory failure. Possible aspiration hazard. May cause lung damage. Inhalation: Harmful if inhaled. Causes chemical burns to the respiratory tract. May cause asthma and shortness of breath. May cause nausea, dizziness, and headache. Chronic: Effects may be delayed. Repeated or prolonged exposure may cause allergic reactions in sensitive individuals.

Environmental safety

Even though it is it is generally assumed that E. coli would be out-competed by natural strains once it is outside of a lab, we assume the fact that in case of accidental release of any GEB, it would raise safety issues because we don't know the potential effects. So any Genetically Engineered Bacteria (GEB) can be potentially dangerous if released in the environment, either on purpose or by accident. The concern about Horizontal Gene Transfer or spread of GEB lead us to develop this project to protect the environment from synthetic devices. During the work on our Biosafety system, we protected the environment from contamination by waste products: all hazardous waste was placed in the correct container (e.g. biohazard containers for biological waste such as E. coli colonies), autoclaved and disposed of responsibly by the university. Team members were taught proper molecular biology skills and aseptic techniques. Team members followed all necessary procedures like washing their hands with disinfectant before leaving the laboratory to avoid transmitting potentially harmful material to the public/environment.

Public safety

Apart from the general concern about the potential harm of GEB to the Environment and Public upon release outside of the lab, none of our designs have the potential to harm the public if released by design or accident. Our lab is especially equipped for microbial manipulation and everything is done to avoid the release. Public safety is ensured, as no member of the public is permitted access within the labs unless approved by the university, and team members followed all necessary procedures like washing their hands with disinfectant before leaving the laboratory to avoid transmitting potentially harmful material to the public/environment. However, we are conscious that as any bacterium, E.coli can be hazardous. For instance, all traumatic wounds, infected burns and any serious lesions can potentially be contaminated, but it is very rare. Treatment: E.coli can be treated with standard antibiotics, which can be prescribed by a doctor if needed.

Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?

Did you document these issues in the Registry?

Yes, we did. As mentioned before, biosafety is the essence of our project, and we're preoccupied by the risk of dissemination of antibiotic resistance genes, as well as any other bioactive compounds such as toxins and antitoxins. We addressed this issue in the characterization of our system. For instance, we made a semantically contained version of the Kanamycin resistance gene. We are also planning to make semantically contained version of the toxin.

How did you manage to handle the safety issue?

Several mechanisms were proposed beyond the classical laboratory safety measures :

• Semantic containment that avoid the expression of synthetic genes outside the GMOs.
• Meganuclease restriction sites that permit the degradation of the antitoxin.
• DNases that degrade GMO's genome.

How could other teams learn from your experience?

We believe that other teams should keep in mind that not only the bacteria are potentially dangerous, but also the eventual release of their DNA could endanger the environment. We suggest that starting from next years, all iGEM teams should add a biosafety part to their project, where they should eithr use one of the existing mechanisms, or design a new one, especially if their project aims at releasing bacteria into the nature. For this purpose, we created a new [http://partsregistry.org/Biosafety Biosefety page] in the Parts Registry.

Is there a local biosafety group, committee, or review board at your institution?

The work has been carried out in the laboratory of Evolutionary Systems Biology at the Molecular, Evolutive and Medical Genetics Unit (U1001, also know as TaMaRa's lab) of the French National Institute of Medical Research ([INSERM]) within the Paris Descartes University's Medical faculty. More importantly, the Biosafety officer of our unit followed our work. Both institutions have their ethical committees though no specific issue concerning our project needed to be raised.

If yes, what does your local biosafety group think about your project?

We met the chair of the ethic committee of Paris Descartes, and they advice us to keep thinking of the human practice part, while providing us some interesting thoughts.

Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

We created [http://partsregistry.org/Biosafety a Biosafety page] on the part registry that should list all the safety mechanisms and systems available on the part registry, with the links to their description and experimental characterization. We would like to suggest next generations of iGEM teams to consult this page, to find a system that would be useful to increase the safety of their project. We also collaborated with the iGEM Grenoble team, who proposed an additional section to the description of Biobricks™ which would explain their potential danger, and the ways to assess the risk.