Safety

• Would the materials used in your project and/or your final product pose:

  • Risks to the safety and health of team members or others in the lab?

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

• • Risks to environmental quality if released by design or accident?

• • Risks to security through malicious misuse by individuals, groups or states?

Our project doesn't raise any safety issues other than usual lab safety issues– whether for the researcher, the publicity or environment.

To assure this in first place, every participant took part in a general safety meeting in order to join the iGEM-Team. This meeting dealt with the common safety rules concerning good laboratory practice, working with genetically modified organisms (GMOs), including storage and disposal. We only worked with non-hazardous, non-pathogenic and organisms like E. coli lab strains (DH5 alpha or BL21). As general lab procedure, we wear a lab coat and single-use gloves. When working with e.g. liquid N₂, we use goggles as well. When operating with dangerous compounds (as e. g. ethidiumbromide), we use nitrile gloves. Furthermore dangerous substances are stored and handled in designated rooms in order to assure the safety of the researchers.

For the protection of the public and the environment against hazardous substances, all GMO-contaminated waste is inactivated by autoclavation. Before leaving the laboratory, every researcher cleans and disinfects his/her hands. Moreover, we leave the windows closed and do not throw jeopardy stuff into the sink.

Our biobricks contain inducible promotors, regulators and reporter genes. None of them are able to cause illnesses or threaten humans in any other way. Most inserts are also derived from non-pathogenic, non-hazardous organisms. One promotor and one gene are dereived from genomic DNA from Ralstonia eutropha and Neisseria meningitidis. Here, we will only work with the DNA of these organsims. The amplified and cloned fragments again belong to the GMO safety class S1.

Strains or DNA derived from our project will not be used outside the lab. The watersamples to be tested are transported to the lab, so there is no risk of contaminating the environment with GMOs.

• If your response to any of the questions above is yes:

• • Explain how you addressed these issues in project design and while conducting laboratory work.

• • Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry.

None of our planned BioBricks raises any special safety issues as detailed above.

Under what biosafety provisions will / do you operate?

At the faculty of biology we have biosafety groups for each area of research. Since we work in the field of microbiology and synthetic biology PD Dr. Ralf Heermann and Prof. Dr. Heinrich Jung are the responsible commissioners for our team. Both checked the project thoroughly had no concerns about the safety of our project.

To ensure that we don't contaminate the environment with our kit we thought will handle the water samples only in the lab.

Before we started to work in our lab, Prof. Dr. Mascher gave us a biosyfety training. He told us about possible dangers in the lab and how to react in a case of emergency. During the lab inspection we learned, where the fire extinguishers and fire blankets are and how to use the emergency shower and the eye douche. For our own safety we usually do not work alone (or if so, e.g. to inoculate cultures, one of the instructors is informed). Furthermore we were told the general rules while working with GMOs. We keep the windows closed and autoclave contaminated waste. We wear labcoats and sometimes goggles. After leaving the lab, we wash and desinfect our hands. We learned how to handle, store and dispose our material and how to document our work properly.

Germany has signed and ratified the [http://www.cbd.int/doc/legal/cartagena-protocol-en.pdf Cartagena Biosafety Protocol]. This protocol ensures safe handling, use and transfer of genetically modified organisms. Furthermore, we have our own laws and guidelines for biosafety here. For example, all laboratories which are handling GMOs have a designated biosafety level, which is stated in a genetic engineering decree ([http://www.gesetze-im-internet.de/bundesrecht/gentsv/gesamt.pdf Gentechnik Sicherheitsverordnung]) and monitored by university officials.

The general safety rules are listed here (This file is derived from Göttingen University, but the rules are identical.)

• 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?

To improve the safety while working with (hazardous) biobricks a new kind of biobrick backbone should be established. Every potentially pathogenic or hazardous biobrick should be cloned in a special backbone containing the sequences of the present backbones (ORI, resistance cassette,restriction sites ...) AND an inducible killing gene cassette. This killing cassette (e.g. ccdB in E. coli with an exchanged promotor or bak for eukaryotic cells) is induced by a normaly absent reagent that could easily be added in case of contamination. Adding this reagent would result in an expression of the killing gene, which leads to the death of the (pathogenic) cells containing the biobrick plasmid.