Team:Freiburg/Safety/safety forms

Safety questions

At the beginning of our research we wanted to be aware of all the probable hazards concerning our project. This included that we tried to identify safety issues. Therefore we concentrated on pathogenicity of the microorganisms and cell lines of interest, the datasheets of chemicals probably used during the project (e.g. DNA stains) and the engineered devices and systems. Thus, we orientated on the hints of the iGEM 2013 safety page as well as the safety constraints for genetic engineering given by the “Stabsstelle Sicherheit” of the University of Freiburg. At this point we are obliged to Dr. M. Zurbriggen, who gave us safety instructions before starting our investigations.

In the next passages, biosafety questions concernig to our project will be answered. There are also some recommendations to other teams, what they could do to optimize biosafety in their labs.

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

• researcher safety,
• public safety, or
• environmental safety?

Our project deals with targeted multiple gene regulation based on the bacterial and archaeal adaptive immune system CRISPR. This tool shall facilitate the daily lab routine of molecular biologists and might enable great progress in almost all genetically characterized disciplines such as cancer research or tissue engineering. Therefore, single biobricks are transformed into E. coli or transfected into eukaryotes such as HEK 293 cells and HeLa cells under safe laboratory conditions. Here, we work on high safety standards given by the University of Freiburg. Furthermore, we try to avoid the use of any highly toxic, acidic or other unhealthy substances. Thus, safety issues do not raise in idem terms.

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 constructs do not raise any safety issues. They should not be able to increase or give pathogenicity to the applied microorganisms or cell lines. The single parts do not encode for any toxins. The devices as they are used in our project should also represent no hazard for public health or environmental safety. In their current state misuse according to bioterrorism should also be unlikely.

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?

Yes, among other things, the department “Stabsstelle Sicherheit” is responsible for questions and issues concerning biosafety at the University of Freiburg.

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?

In our lab, we were especially concerned with carcinogenity. To protect our team we banned ethidium bromide completely and used next generation DNA stains concerned to be less carcinogenic. Additionally, nitrile gloves were used while cutting agarose gels containing DNA intercalating substances. Another hazard is the UV light, which can lead to mutations in DNA. Eyes and skin were protected due to lab coats and UV shield helmets. Theses safety precautions proved themselves in practice and are recommended to all other iGEM-teams.

Systems can be made safer through genetically switches. Our constructs are induced or silenced by stimuli, e.g. light of special wavelengths or hormones. Without the right stimulus the system is not running. Other possibilities are constructs that regulate themselves or self destruction mechanisms in case of dysfunctions.