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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, 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 are 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.

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).

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 [http://www.gesetze-im-internet.de/gentsv/BJNR023400990.html#BJNR023400990BJNG000301308 §8-12] of the [http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320 GenTSV], the German act of genetics). Our advisors showed us how to correctly and safely use the equipment in the laboratory and informed us about safety measures,such as wearing protective clothing.

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.

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. The first of our two new component biobricks (LOV Protein, BBa_K820000) functions as a light-induced protein switch, which is inert and non-toxic without a fused effector protein. The second of our component biobricks is a bacterial nuclease (mazF, BBa_K820001), which although upon expression will lead to cell death in infected bacteria, will complete its activity before replication or plasmid transfer can be completed.

One of our new devices functions as a galactase, which severs 1,4-glycosidic bonds, but poses no danger to organisms, other new devices function as kill switches (using a gyrase inhibition or nuclease mechanism), which 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 [http://www.bvl.bund.de/DE/06_Gentechnik/02_Verbraucher/05_Institutionen_fuer_biologische_Sicherheit/02_ZKBS/gentechnik_zkbs_node.html 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 college, departmental and laboratory level, a safety officer supervises radioactive and biological hazards in the lab. All team members members have taken an extensive safety instruction course with this safety officer.

A biosafety laboratory in Germany 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 mouth pipetting, no eating, drinking or storage of food in the laboratory). GMOs (genetically modified organisms) are subjected to restrictions in Germany like the [http://www.gesetze-im-internet.de/gentsv/index.html#BJNR023400990BJNE001503320 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?

Close control of gene expression and protein function is very important from a biosafety perspective. Our project adds a light-activated control device for protein function, which can prevent uncontrolled protein activity.