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Contents 1 iGEM 2012 Biosafety Questionaire 2 iGEM 2012 Team Bonn Biosafety 2.1 1. Safety issues 2.2 2. New Biobricks and safety issues 2.3 3. Regulations in Germany, Lab-training of team members 2.4 4. Future iGEM competitions and safety issues

iGEM 2012 Biosafety Questionaire

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

Our project does not raise any foreseeable safety issues for researchers or for the public or the environment. We use Escheria coli DH5alpha and XL-1blue strains, which are considered lab-safe in biosafety level 1 labs. They have been well characterised and pose minimal risks for humans or the environment. In nature, our new parts are expressed in organisms such as E.coli (ccdB) or Avena Sativa (LOV) and don't cause any threat to humans. Furthermore, we follow standard lab protocols and are supervised during our work by our instructors (Professors Mayer and Famulok) and our advisors (PhD students in our instructors' labs), who work side-by-side with us in the same lab During our work in the lab we had to take more safety measures concerning the use of ethidium bromide. The use of nitrile gloves is mandatory, as well as only to work under a designated ethidium bromide fume hood. The waste is disposed of separately from any other waste in the lab. Due to our lab work taking place in an area where radioactive markers are employed, we carry a dosimeter with us while in the lab.

To work in a level 1 biosafety laboratory, each team member had to take a course about biosafety and regulations in Germany (as required in §8-12). Our advisors showed us how to correctly use the equipment in the laboratory and informed us about safety measures, e.g. protective clothing.

There aren’t any apparent risks, even if organisms or parts of our project are accidentally released. As we are not using any pathogenic bacteria they alone wouldn’t pose any considerable threat for the health of humans, animals or plants. Furthermore, E.coli DH5 alpha is F- , so it won’t be able to transfer any of its plasmids to other organisms. E.coli XL1-blue doesn’t pose a high risk as it can’t survive outside of the laboratory. In addition the parts we use aren’t hazardous (see Question 2).

2. 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?

Our new BioBricks do not raise any foreseeable safety issues. Our two new component biobricks function either as a light-switchable protein, which is inert and non-toxic without a fused effector protein (LOV Protein, BBa_K820000) or as a bacterial nuclease (mazF, BBa_K820001), which although upon expression will lead to cell death in infected bacteria, it will complete its activity before replication or plasmid transfer can be completed. Our new devices function either as a galactase, which is severs 1,4-glycosidic bonds, but poses no danger to organisms, or as kill switches (using a gyrase inhibition or nuclease mechanism), which - like BBa_K820001 - will lead to cell death before the plasmid can be passed on to other cells in the population.

3. Is 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?

The University of Bonn does not currently have a biosafety committee, however, the federal government has instituted a central committee on biosafety, which issues regular rulings, statements and updates on biosafety issues ranging from safe lab organisms to environmental clearance of GMOs. At a departmental and laboratory level, a safety officer supervises radioactive and biological hazards in the lab and has instructed all team members.

A biosafety laboratory such as where we are working has to fulfill several standards and is subjected to many rules (e.g. no mouth pipetting, no eating, drinking or storage of food in the laboratory). GMOs (genetically modified organisms) are subjected to restrictions in Germany like the GenTSV (German act of genetics). We work with biosafety level 1 (S1) organisms, which are defined as neither human-, phyto- nor animal pathogenic.

4. 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?

iGEM 2012 Team Bonn Biosafety

1. Safety issues

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2. New Biobricks and safety issues

The new BioBricks we designed do not raise any safety issues. The LOV plasmid is a light sensor which is uneffective without effector protein but releases the effector protein under the influence of blue light. The function of LOV-lacZ-alpha is to create a part of split-beta-galactosidase, whereas LOV-ccdb leads to cell death of the bacteria.

3. Regulations in Germany, Lab-training of team members

A biosafety laboratory such as where we are working has to fulfill several [http://www.gesetze-im-internet.de/gentsv/anhang_iii_34.html standards and is subjected to many rules] (e.g. no pipetting with the mouth, no food, drinking or storage of food in the laboratory). GMOs (genetically modified organisms) are subjected to restrictions in Germany. We work with biosecurity stage 1 (S1) organisms. The [http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320 GenTSV] (German act of genetics) defines them as neither human-, phyto- nor animalpathogenic.

To work in a biosecurity laboratory of the stage 1 (S1), each team member had to take a course about biosafety and regulations in Germany (as required in [http://www.gesetze-im-internet.de/gentsv/BJNR023400990.html#BJNR023400990BJNG000301308 §8-12]). Our advisors showed us how to correctly us the appliances in the laboratory. Every person working in the laboratory had to wear protective clothing (laboratory coat, protective gloves).

During our work in the lab we had to take more safety measures concerning the use of ethidium bromid. The use of nitril gloves is mandatory, we are only allowed to use it in a special ethidium bromid fume hood and the waste is disposed of seperately from any other waste in the lab. Due to work in an area where radioactive chemicals are used we had to carry a dosimeter with us

4. Future iGEM competitions and safety issues

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