Team:DTU-Denmark-2/Safety

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Revision as of 08:40, 19 September 2011

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    • Mammalian Cells
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Safety

1. Would the materials used in your project and/or your final product pose

a. Risks to the safety and health of team members or others in the lab

The biological parts and devices that we designed and constructed do not pose any risks to the safety and health of team members or others in the lab. In this project we work with organisms and cells that are widely used in laboratories and generally regarded as safe, namely the E.coli strain DH5α, and the fungus Aspergillus nidulans. One part of the team worked with these two organisms in a biosafety 1 lab. The other part of the team worked with the human osteosarcoma cell line U-2 OS in a biosafety 2 lab. To avoid cross-contamination the part of the team working with the microorganisms were not allowed to enter the mammalian cell lab.

There are potential risks in a number of standard practices performed in the lab, therefore appropriate measures were taken when preparing media and buffers, performing gel extractions, and using molecular biology kits for DNA purification etc. In general nitrile gloves, labcoats, and goggles were worn when required to ensure the safety of our team members in the lab.

b. Risks to the safety and health of the general public if released by design or accident?

Our standard assembly system is intended for research and development only and does therefore not pose a threat to public safety. If the biological parts or devices should be released by accident there would be no apparent risks, since the biological parts we are working with are not hazardous. They would not constitute any danger or other negative effects, if they were released by accidents. Furthermore our working strain E. coli DH5α does not thrive outside the laboratory, because it is an auxotroph and cannot transmit plasmids to other strains (it is a F- strain).

We have constructed resistance marker cassettes expressing ampicillin, hygromycin, or neomycin resistance. The antibiotics mentioned are used in public health settings to treat infections and therefore the spread of antibiotic resistance is very problematic and can have great implications on the public health. If these marker cassettes were accidentally released and a given microorganism was naturally transformed with the DNA there could be a small chance that the organism would have acquired resistance to the respective antibiotic. However, this is unlikely to happen. Should our E. coli DH5α containing a plasmid with antibiotic resistance be released into the environment, it would be unlikely for the strain to transfer the plasmid, as the strain as earlier mentioned is F-. Therefore the spread of antibiotic resistance genes poses a low safety risk. The Aspergillus nidulans strain used in this project is also an auxotroph and to survive outside the lab, it requires high concentrations of arginine, uracil, and uridine. The mammalian cells transfected with plasmids containing antibiotic resistance would not be able to survive outside the lab, because the U-2 OS cell line like many other mammalian cell lines requires very specific growth conditions.

c. Risks to environmental quality if released by design or accident?

We have evaluated that there would be no risks to environmental quality if our biobricks or devices were released by design or accident, because the coding sequences mainly encode fluorescent proteins. As mentioned before the strains we were working with are not able to grow outside the lab and therefore do not pose any risks to environmental quality. It is important to dispose biological waste correctly to prevent accidental release of the biological material from the laboratory. If all safety precautions are followed according to all local and federal regulations our project will not raise any safety issues. The guidelines for disposal of biological waste as defined by the Center for Microbial Biotechnology (CMB) at the Technical University of Denmark (DTU) was followed to prevent possible release of genetically modified organisms (GMO).

d. Risks to security through malicious misuse by individuals, groups or states?

No, the parts we have constructed are in themselves or combined not a threat to security. The aim of our project is to develop an assembly standard that is easy and fast to use, but the assembly system can of course be misused, and thereby pose a security risk. However, this is the case for all technologies within biotechnology, and would require the knowledge and expertise of designing experiments and conducting them.

e. Specifically, are any parts or devices in your project associated with (or known to cause)

- pathogenicity, infectivity, or toxicity? None of our parts or devices are associated with, or known to cause pathogenicity, infectivity or toxicity.
- threats to environmental quality? No parts or devices have been suspected to be a threat to environmental quality.
- security concerns? There are no security concerns associated with our project or the parts and devices constructed.

2. If your response to any of the questions above is yes:

a. Explain how you addressed these issues in project design and while conducting laboratory work.

As non of our parts or devices are associated with or known to cause pathogenicity, infectivity, or toxicity it was not necessary to address such issues in the project design or while conducting laboratory work.

b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry.

There where no safety, security, health and/or environmental issues regarding our submitted parts.

3. Under what biosafety provisions will / do you operate?

a. Does your institution have its own biosafety rules and if so what are they?

At the Center for Microbial Biotechnology at DTU, there are safety rules that all employees and students have to follow. There are some general rules regarding fire, accidents, preventing accidents, and personal safety outfit. The general safety rules and the biological safety rules of CMB can be found here.

b. Does your institution have an Institutional Biosafety Committee or equivalent group?If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review.

At CMB we have a local biosafety group that ensures the laboratory safety. Before initiating our project at CMB we signed a formula concerning the safety rules. Furthermore, we had to write short description of the type of experiments that are expected to be performed during the project, which is standard procedure at the institute. This description included the toxicity of the chemicals used, how to dispose them, and which precautions to make in order to perform the project in a responsible and safe way. The safety group found that the safety aspects of the project were sufficient, and the project was initiated after the approval. The safety group furthermore advised us to work according to standard safety protocols for genetic engineering and molecular biology.

c. Did you receive any biosafety and/or lab training before beginning your project?If so, describe this training.

All the team members had already been through extensive lab training and biosafety rules, because of previous experimental projects at the Center for Microbial Biotechnology, DTU. It was therefore decided that the laboratory work could be initiated after signing the formula concerning CMB’s safety rules and writing the project description without further laboratory training.

d. Does your country have national biosafety regulations or guidelines?If so, provide a link to them online if possible.

In Denmark, specific laws and guidelines exist, that are to be considered when working with GMO. These guidelines are based on “Bekendtgørelse om genteknologi og arbejdsmiljø” (“The Order of Gene Technology and Working Environment) from 2008, which is based on guidelines/rules determined by the European Union (1). Our project has followed the guidelines and laws set by the authority in Denmark and the European Union. In conclusion, the organisms and biobricks used in this project do not represent any hazard to the researchers, when working in the laboratory or to the public safety. If the safety standard procedure is followed there is also no risk for contamination of the environment.

References

1. the GenTSV §5 Abs. 2 i.V.m.Anhang Teil B, Teil A II, and the statement of the ZKBS (Central committee for Biological Safety, Germany).