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Team:TU Munich/Project/Safety
From 2012.igem.org
Safety
iGEM Questioniare
Would any of your project ideas raise safety issues in terms of:
- researcher safety,
- public safety, or
- environmental safety?
There are dangerous substances used in our lab. The following list features a few important examples:
- In every laboratory of molecular biology, specific chemicals are required for staining of DNA, in order to make it visible on Agarose gels. Most of them directly intercalate into the double strand of DNA, which makes them cancerogenic. A commonly used, but rather dangerous substance is ethidium bromide. In our lab, we use SYBR® Gold. It is less hazardous than ethidium bromide, but can still be cancerogenic if it comes into direct contact with human skin. Protective gloves should be made from nitrile rubber and changed frequently to prevent contamination with SYBR® Gold. All gels and materials that came into contact with SYBR® Gold need to be disposed of seperately. This is done in order to prevent their unintended leakage into the environment, with subsequent harm to humans, animals and plants.
- Methods of molecular biology often require strong acids or bases, like sulfuric acid, or toxic substances such as methanol. They need to be handled with extreme caution and also need to be seperately disposed.
- Many devices in the lab can be potentially dangerous towards researchers, if they are used carelessly or in the wrong way. There are lamps emitting ultraviolet radiation, which can be cancerogenic.
There are several different bacterial strains used in our lab for transformation of BioBrick parts, which have been manipulated in some way to make sure that they are harmless. The strains used in our lab, like the heat-resistent Escherichia coli strain BH28, are derived from E. coli K-12. K-12 is a safety strain, as the bacteria carry several auxotrophies. This means that they are dependent on certain sources of carbon, amino acids and other nutritients. Without them, growth is not or only in a restricted way possible. BH28 is derived from K-12, but has over 260 mutations, which lead to its heat resistance. The other E. coli strains DH5 alpha, BL21 and CP919 that were used in the lab, are also derived from K-12.
If despite all precautions humans are infected with bacteria, the first weapon of choice are always antibiotics. They specifically kill bacteria, while keeping side effects for the patient to a minimum. Bacteria can be resistant against antibiotics. The genes responsible for this are often used as selection markers during cloning. With respect to killing them however, antibiotic resistences can make bacteria more dangerous. The number of supplied resistences should therefore be reduced to no more than necessary.
The bacteria are not motil and auxotroph, so they cannot survive in minimal medium (with only glucose as C source), but need additional aminoacids to survive. Since nothing from the lab is taken into public and stays inside there should be no safety issues considering public or environmental safety. Furthermore, used E. coli cultures and waste containing biologic material is autoclaved at 121 °C before being thrown away. This ensures that no genetically modified material can reach the outside of the lab. Our final construct itself, the optogenetical AND-Gate, is not associated with pathogenicity, infectivity or toxicity. Furthermore, it has no impact on environmental quality, as it is not able to compete to its natural competitors, due to its auxotrophies. All in all this leads to secure strains which cannot survive outside the laboratory.
A deliberate misuse of this construct is unplausible, as it does not increase pathogenicity of the E. coli strain. Scenarios, in which our construct is used to specifically trigger the production of toxins or other substances to harm humans, animals, or the environment in general are unlikely, as the strains used are not able to survive outside from controlled conditions. Even the use of efficient strains would not be able to promote misuse, e.g. as a biological weapon, because the optogenetical AND-Gate does not feature necessary factors of pathogenicity. This also applies for all intermediate constructs.
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?
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?
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?
