Team:TU Munich/lab/safety

law for the regulation of genetic engineering , text in German only). There is a total of four Biosafety levels, with BSL 1 being the lowest and BSL 4 being the highest.

Work inside a BSL 1 lab such as ours involves no devices that are harmful to the researchers if they act corresponding to the general precautionary measures. Especially, no pathogenic organisms are used.

BSL 2 and 3 laboratories are necessary to work with biohazardous material that can cause disease (BSL 3 in case of potential severe disease) for wich an effective cure is available. The security measures of these labs include security work benches and an air filter system, for example.

BSL 4 laboratories are required to work with organisms capable of causing severe disease and for which effective treatment is not possible (such as smallpox or the Ebola virus). A BSL 4 lab has to have a broad range of safety measures. To give some examples there have to be hazmat suits, airlocks to maintain a low air pressure inside the lab, and many methods of decontamination to ensure that no traces of biohazardous material can get outside the lab.

The most harmful substance in the our lab is CyberGreen which is used for staining agarosegels after DNA digestion and separation (used a lot in cloning steps). Here everybody has to be careful, switch gloves everytime he touched something containing CyberGreen and in general be responsible and tidy when working with CyberGreen.

Our e.coli strain (MG1655) is derived from e.coli K12 which is modified so it is not harmful to humans. The strain is resistant to kanamycin (by inserted mutation) and ampicillin (during evolution). Furthermore the strain differs from K12 in 260 more mutations which might lead to its heat resistance. The bacterias are not motil and auxotroph, so they cannot survive in minimal medium (with only glucose as C source)but need additional aminoacids to survive. All in all this leads to a secure strain which cannot survive outside the laboratory. Since nothing from the lab is taken into public and stays inside there should be no safety issues considering public or environmental safety. Used e.coli cultures and waste containing biologic material is autoclaved before throwing away. This ensures that no gentetically modified material can reach the outside of the lab.

Our finished construct itself, the optogenetical AND-Gate, does not pose a threat of any kind. Its only purpose is to controll gene expression in immobilized cells in a spatiotemporal manner. A deliberate missuse of this construct is not possible. This also applies for all intermediate constructs.

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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?

None of our BioBrick parts are harmful to humans or the environment. We are working with the red-light sensor and the blue-light sensor combined in an AND Gate. None of those parts should survive outside the lab. ---- '''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?''' Every department at TU Munich needs a safety delegate, in our case Helene Budjarek: She doesn't have any objections against the project due to safety issues. In general, working with genetically modified organisms in Germany is regulated by the '"Gesetz zur Regelung der Gentechnik (GenTG)"'' (law for the regulation of gentetic engineering, see above). ---- '''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?''' The easiest way to increase safety when working with BioBricks is to prevent the uncontrolled multiplication and spreading of parts. This can be acchieved by... * ... the use of uncommon restriction enzymes * ... not using parts containing infectious DNA in combination with parts that can multiply and spread without the help of a host organism (transposons,…)