Team:ETH Zurich/Safety
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== Questions I == | == Questions I == | ||
=== Risks to the safety and health of team members or others in the lab? === | === Risks to the safety and health of team members or others in the lab? === | ||
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Working in the lab entails certain risks for lab members. Our project does not raise specific chemical and biological safety issues beyond those risks normally involved in working in a microbiological environment of biosafety level 1. Our experiments are carried out in a S1 lab minding general precautionary measures and safety standards. This implies wearing cloves, potective glasses and lab coats. We are using E.coli strains TOP10 and JT2 (MC4100-derived) for our experiments, both of which are not pathogenic. All material contaminated with bacteria is autoclaved at 121 °C and 1 bar overpressure. | Working in the lab entails certain risks for lab members. Our project does not raise specific chemical and biological safety issues beyond those risks normally involved in working in a microbiological environment of biosafety level 1. Our experiments are carried out in a S1 lab minding general precautionary measures and safety standards. This implies wearing cloves, potective glasses and lab coats. We are using E.coli strains TOP10 and JT2 (MC4100-derived) for our experiments, both of which are not pathogenic. All material contaminated with bacteria is autoclaved at 121 °C and 1 bar overpressure. |
Revision as of 21:32, 26 September 2012
Contents |
Questions I
Risks to the safety and health of team members or others in the lab?
Working in the lab entails certain risks for lab members. Our project does not raise specific chemical and biological safety issues beyond those risks normally involved in working in a microbiological environment of biosafety level 1. Our experiments are carried out in a S1 lab minding general precautionary measures and safety standards. This implies wearing cloves, potective glasses and lab coats. We are using E.coli strains TOP10 and JT2 (MC4100-derived) for our experiments, both of which are not pathogenic. All material contaminated with bacteria is autoclaved at 121 °C and 1 bar overpressure.
Our lab work also includes using UV light to trigger our sensor. UV light is a known carcinogen and can cause skin irritation, therefore special precautions are needed. In general we are using the UV lamp in a darkhood, which poses no hazards as no UV light can escape. While handling a portable UV lamp, we use special glasses and avoid any exposure to the skin.
Risks to the safety and health of the general public and Environmental quality if released by design or accident?
Though we pay attention to keep our system isolated from the external environment, there is always the possibility of modified organisms leaking by accident. Therefore a (horizontal) transfer of genetic material between our bacteria and other bacteria in the wild is possible. Instead of causing a risk to health of the general public, our device aims for protecting the skin and prevent damage from UV radiation. Testing is needed to guarantee the safety of our device before it is used to this purpose.
Risks to security through malicious misuses by individuals, groups or states?
Our system is designed to sense UV light and produce a fluorescent protein or a pigment as output. We do not see a realistic way to use this system to cause harm. With a malicious intent, it is nonetheless possible to switch the reporter to a more dangerous gene.
Question II
Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?
No, our new BioBrick parts do not raise any safety issues apart from the fact that the switch needs UV light to be activated. Warnings will be put in the Registry for people that will use our part. We managed the UV light safety issues as described above.
Questions III
Does your university have a Biosafety Committee or equivalent?
Our lab follows the general- and bio-safety guidelines provided by the Safety, Security, Health and Environmental Protection (SSHE) unit of ETH Zurich (http://www.sicherheit.ethz.ch/index_EN). For an S1 lab two relevant documents are the Safety and Waste Disposal Manual (http://www.sicherheit.ethz.ch/docs/health_docs/safety_manual_en.pdf) and the Biosafety-Concept (http://www.sicherheit.ethz.ch/docs/concepts_safety_at_work_hygiene/Biosicherheitskonzept).
Have you consulted with your Institutional Biosafety Committee regarding your project?
We are not working with any organisms or parts which would require containment arrangements above BSL1.
Will / did you receive any biosafety and/or lab training before beginning your project?
All iGEM team member attended lectures on lab safety. There was no special training for iGEM because all people already attended a special training on chemical and biological safety in the lab and a fire prevention course in previous practical courses.
Question IV
Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions?
One important thing is to keep genetically modified organism (GMOs) isolated form the environment. It is crucial to optimize all steps during the project to avoid the leaking of used organisms. Obeying the general safety standards has been effective in the past handling GMOs.
To augment safety, another approach lies in employing only such modified organisms that can be sustained in a controlled lab environment. The growth of such bacteria depends on a simple chemical supplement to the medium which is not present in the environment
In the case of our project that aims for being used to protect human skin, one needs ample experiments to guarantee environmental safety.
References
- Brown, B. a, Headland, L. R., & Jenkins, G. I. (2009). UV-B action spectrum for UVR8-mediated HY5 transcript accumulation in Arabidopsis. Photochemistry and photobiology, 85(5), 1147–55.
- Christie, J. M., Salomon, M., Nozue, K., Wada, M., & Briggs, W. R. (1999): LOV (light, oxygen, or voltage) domains of the blue-light photoreceptor phototropin (nph1): binding sites for the chromophore flavin mononucleotide. Proceedings of the National Academy of Sciences of the United States of America, 96(15), 8779–83.
- Christie, J. M., Arvai, A. S., Baxter, K. J., Heilmann, M., Pratt, A. J., O’Hara, A., Kelly, S. M., et al. (2012). Plant UVR8 photoreceptor senses UV-B by tryptophan-mediated disruption of cross-dimer salt bridges. Science (New York, N.Y.), 335(6075), 1492–6.
- Cloix, C., & Jenkins, G. I. (2008). Interaction of the Arabidopsis UV-B-specific signaling component UVR8 with chromatin. Molecular plant, 1(1), 118–28.
- Cox, R. S., Surette, M. G., & Elowitz, M. B. (2007). Programming gene expression with combinatorial promoters. Molecular systems biology, 3(145), 145. doi:10.1038/msb4100187
- Drepper, T., Eggert, T., Circolone, F., Heck, A., Krauss, U., Guterl, J.-K., Wendorff, M., et al. (2007). Reporter proteins for in vivo fluorescence without oxygen. Nature biotechnology, 25(4), 443–5
- Drepper, T., Krauss, U., & Berstenhorst, S. M. zu. (2011). Lights on and action! Controlling microbial gene expression by light. Applied microbiology, 23–40.
- EuropeanCommission (2006). SCIENTIFIC COMMITTEE ON CONSUMER PRODUCTS SCCP Opinion on Biological effects of ultraviolet radiation relevant to health with particular reference to sunbeds for cosmetic purposes.
- Elvidge, C. D., Keith, D. M., Tuttle, B. T., & Baugh, K. E. (2010). Spectral identification of lighting type and character. Sensors (Basel, Switzerland), 10(4), 3961–88.
- GarciaOjalvo, J., Elowitz, M. B., & Strogatz, S. H. (2004). Modeling a synthetic multicellular clock: repressilators coupled by quorum sensing. Proceedings of the National Academy of Sciences of the United States of America, 101(30), 10955–60.
- Gao Q, Garcia-Pichel F. (2011). Microbial ultraviolet sunscreens. Nat Rev Microbiol. 9(11):791-802.
- Goosen N, Moolenaar GF. (2008) Repair of UV damage in bacteria. DNA Repair (Amst).7(3):353-79.
- Heijde, M., & Ulm, R. (2012). UV-B photoreceptor-mediated signalling in plants. Trends in plant science, 17(4), 230–7.
- Hirose, Y., Narikawa, R., Katayama, M., & Ikeuchi, M. (2010). Cyanobacteriochrome CcaS regulates phycoerythrin accumulation in Nostoc punctiforme, a group II chromatic adapter. Proceedings of the National Academy of Sciences of the United States of America, 107(19), 8854–9.
- Hirose, Y., Shimada, T., Narikawa, R., Katayama, M., & Ikeuchi, M. (2008). Cyanobacteriochrome CcaS is the green light receptor that induces the expression of phycobilisome linker protein. Proceedings of the National Academy of Sciences of the United States of America, 105(28), 9528–33.
- Kast, Asif-Ullah & Hilvert (1996) Tetrahedron Lett. 37, 2691 - 2694., Kast, Asif-Ullah, Jiang & Hilvert (1996) Proc. Natl. Acad. Sci. USA 93, 5043 - 5048
- Kiefer, J., Ebel, N., Schlücker, E., & Leipertz, A. (2010). Characterization of Escherichia coli suspensions using UV/Vis/NIR absorption spectroscopy. Analytical Methods, 9660. doi:10.1039/b9ay00185a
- Kinkhabwala, A., & Guet, C. C. (2008). Uncovering cis regulatory codes using synthetic promoter shuffling. PloS one, 3(4), e2030.
- Krebs in Deutschland 2005/2006. Häufigkeiten und Trends. 7. Auflage, 2010, Robert Koch-Institut (Hrsg) und die Gesellschaft der epidemiologischen Krebsregister in Deutschland e. V. (Hrsg). Berlin.
- Lamparter, T., Michael, N., Mittmann, F., & Esteban, B. (2002). Phytochrome from Agrobacterium tumefaciens has unusual spectral properties and reveals an N-terminal chromophore attachment site. Proceedings of the National Academy of Sciences of the United States of America, 99(18), 11628–33.
- Levskaya, A. et al (2005). Engineering Escherichia coli to see light. Nature, 438(7067), 442.
- Mancinelli, A. (1986). Comparison of spectral properties of phytochromes from different preparations. Plant physiology, 82(4), 956–61.
- Nakasone, Y., Ono, T., Ishii, A., Masuda, S., & Terazima, M. (2007). Transient dimerization and conformational change of a BLUF protein: YcgF. Journal of the American Chemical Society, 129(22), 7028–35.
- Orth, P., & Schnappinger, D. (2000). Structural basis of gene regulation by the tetracycline inducible Tet repressor-operator system. Nature structural biology, 215–219.
- Parkin, D.M., et al., Global cancer statistics, 2002. CA: a cancer journal for clinicians, 2005. 55(2): p. 74-108.
- Rajagopal, S., Key, J. M., Purcell, E. B., Boerema, D. J., & Moffat, K. (2004). Purification and initial characterization of a putative blue light-regulated phosphodiesterase from Escherichia coli. Photochemistry and photobiology, 80(3), 542–7.
- Rizzini, L., Favory, J.-J., Cloix, C., Faggionato, D., O’Hara, A., Kaiserli, E., Baumeister, R., et al. (2011). Perception of UV-B by the Arabidopsis UVR8 protein. Science (New York, N.Y.), 332(6025), 103–6.
- Roux, B., & Walsh, C. T. (1992). p-aminobenzoate synthesis in Escherichia coli: kinetic and mechanistic characterization of the amidotransferase PabA. Biochemistry, 31(30), 6904–10.
- Strickland, D. (2008). Light-activated DNA binding in a designed allosteric protein. Proceedings of the National Academy of Sciences of the United States of America, 105(31), 10709–10714.
- Sinha RP, Häder DP. UV-induced DNA damage and repair: a review. Photochem Photobiol Sci. (2002). 1(4):225-36
- Sambandan DR, Ratner D. (2011). Sunscreens: an overview and update. J Am Acad Dermatol. 2011 Apr;64(4):748-58.
- Tabor, J. J., Levskaya, A., & Voigt, C. A. (2011). Multichromatic Control of Gene Expression in Escherichia coli. Journal of Molecular Biology, 405(2), 315–324.
- Thibodeaux, G., & Cowmeadow, R. (2009). A tetracycline repressor-based mammalian two-hybrid system to detect protein–protein interactions in vivo. Analytical biochemistry, 386(1), 129–131.
- Tschowri, N., & Busse, S. (2009). The BLUF-EAL protein YcgF acts as a direct anti-repressor in a blue-light response of Escherichia coli. Genes & development, 522–534.
- Tschowri, N., Lindenberg, S., & Hengge, R. (2012). Molecular function and potential evolution of the biofilm-modulating blue light-signalling pathway of Escherichia coli. Molecular microbiology.
- Tyagi, A. (2009). Photodynamics of a flavin based blue-light regulated phosphodiesterase protein and its photoreceptor BLUF domain.
- Vainio, H. & Bianchini, F. (2001). IARC Handbooks of Cancer Prevention: Volume 5: Sunscreens. Oxford University Press, USA
- Quinlivan, Eoin P & Roje, Sanja & Basset, Gilles & Shachar-Hill, Yair & Gregory, Jesse F & Hanson, Andrew D. (2003). The folate precursor p-aminobenzoate is reversibly converted to its glucose ester in the plant cytosol. The Journal of biological chemistry, 278.
- van Thor, J. J., Borucki, B., Crielaard, W., Otto, H., Lamparter, T., Hughes, J., Hellingwerf, K. J., et al. (2001). Light-induced proton release and proton uptake reactions in the cyanobacterial phytochrome Cph1. Biochemistry, 40(38), 11460–71.
- Wegkamp A, van Oorschot W, de Vos WM, Smid EJ. (2007 )Characterization of the role of para-aminobenzoic acid biosynthesis in folate production by Lactococcus lactis. Appl Environ Microbiol. Apr;73(8):2673-81.