Team:UNITS Trieste/Safety

Would any of your project ideas raise safety issues in terms of researcher safety, public safety, or environmental safety?

In the wetlab we follow the general behavior rules of the laboratories we work in, like wearing gloves and labcoat when we manage chemicals, cellular cultures and the equipment we need, never eating or drinking inside the work area and smoking outside the building.
In the SynBiome Project we intend to use E. coli, that are categorized as Risk Group 1 microorganisms, so the danger is absent or low for workers and community. The cells we use for the eukaryotic section are Hela cells. The laboratories we use to work with bacteria and Hela cells are both BSL 2 (biosafety level 2), according to the Italian laws.
We are aware that the manipulation of bacteria and eukaryotic cells has always a range of risk for human and environmental safety, so in order to avoid the involuntary spreading of microorganisms we have been trained about the containment and the waste handling procedures, the use of laminar flow cabinets, biosafety hoods and autoclave for tubes and glassware used for cellular cultures and bacteria.

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

• did you document these issues in the Registry?
• How did you manage to handle the safety issue?
• How could other teams learn from your experience?

We made several meetings to make a risk evaluation based on the guidelines of our country and the European Community. We have considered different aspects: the vectors used, the donor and receiver microorganisms, the BioBricks and genes transferred and transfected are some of them. We finally stabilized that our new BioBricks are not dangerous for human and environmental safety. Our bacterial constructs contain promoters and genes that derive by non-pathogenic organisms and produce molecules for Quorum Sensing communication and its regulators (derived by Agrobacterium tumefaciens) and a β-glucosidase by Cellulomonas fimi. These constructs are introduced in a Escherichia coli DH5α strain that is a Risk Group 1. The resulting organisms are still Group 1 bacteria.

The eukaryotic cells are modified to produce a β-lactamase only in the presence of certain QS molecules produced by bacteria (see Neddermann et Al., 2003). The production of a antibiotic resistance is necessary for the bacterial survival in the Synthetic Biome, but in the absence of the genetically modified bacteria the Hela cells won’t be able to transcribe for the enzyme, so it not represent a sensitive risk.

• there a local biosafety group, committee, or review board at your institution?
• If yes, what does your local biosafety group think about your project?
• If no, which specific biosafety rules or guidelines do you have to consider in your country?

We don’t have a local biosafety group or committee. The legal responsible about safety in the laboratories the team in working in is the ICGEB Director Dr. Mauro Giacca, who is following the development of the project since the beginning. The team asked to the Safety Assistant Marco Vegliach more information concerning the Italian Law: the laboratories that accommodate our team follow the Dlgs 81/08 about health and safety issues at workplace, the Dlgs 206/01 and the DM 25.09.2001 (Italian texts only) about the contained use of genetically modified micro-organisms, as suggest by the 2000/608/EC, the 98/81/EC and the 90/219/EEC directives.

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 think that for our SynBiome the safety rules suggested by the iGem competition are enough.