Question 1:

Question

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

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

Answer

In this project two organisms were used, following are the two organisms listed with biosafety level.

Before starting to work in the lab, a risk assessment (developed by the HSE group) was done of all the equipment and procedures to be used in this project. It had to be approved by the HSE group.

The grading system used ranges from A to E, where A is no/very low risk, B is low risk, C is moderate risk, D is high risk and E is very high risk. The risk assessment for the project is given below. For the probability of unwanted incident, the grading system ranges from 1 to 5, where 1 is very low probability (once every 50 years, or less), 2 is low probability (once every 10 years or less), 3 is moderate probability (once a year, or less), 4 is high probability (once a month or more often) and 5 is very high probability (happens every week).

All of the participants have had previous lab courses. In these courses the participants have learnt about use of safety equipment, first aid, fire extinguishing, proper dispose of waste, and reading and understanding of safety data sheets of the substances in use. Use of different kinds of equipment and methods used have been taught, as well as writing scientific reports. The labs in use in this project are all approved for working with gene modified organisms (GMOs).

Lab coats, safety glasses and disposable gloves are used when necessary and when required by the HSE guidelines and rules. Materials such as restriction enzymes, buffers, PCR and Gel Purification kits are all bought from biotechnology companies. There are none additional safety precautions to be made than the ones already described. When making solutions of antibiotics, gloves and a fume hood are used, in addition to lab coat and glasses.

When the lab’s not in use, it is locked with all of the equipment and materials/organisms safely stored away. A person that doesn’t belong there will not be able to enter. There are also security personel at the school at all times.

Since these bacteria have plasmids with antibiotic resistance genes for selection, these can be harmful if released into the environment. To be sure, never bring GMOs outside of the lab and all of the materials and equipment contaminated by the GMOs, will be kept in special containers to be autoclaved. It any of the organisms were released to the environment, the biggest problem would be if the antibiotics resistance genes were transferred to other organisms. The natural occurring organisms can then influence humans, plants and animals. As long as the rules and regulations mentioned above (and also the rules and regulations give by the government, university and institute) are followed, this will not be an issue. The rules and regulations of the government, university and institute are given in the answer to question 3.

Question 2

Question

Do any of the new BioBrick parts (or devices) that you made this year raise 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?

Answer

In this project the goal is to get E. coli to react to specific environmental factors that are found around cancer cells, such as high lactate and low oxygen concentrations. These qualities are not connected to the toxicity, infectivity or pathogenicity of the organism and are therefore considered not harmful. The strain of E. coli used is also not pathogenic (the DH5ɑ strain).

There is also one gene expressed under a constitutive promoter. The protein expressed named Colicin is a bacteriocin, a toxic for bacteria, and is produced by E. coli. Because it's produced in E. coli, it's a toxin humans are exposed all the time, through their intestines. It is then not considered harmful. Some scientists even consider using it as an anti-Listerial agent in meat products. ([http://books.google.no/books?id=jGB7mT7GDkcC&hl=no], 05.09.12)

Question 3

Question

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?

Answer

The Health, Security and Environment (HSE) group at the university has certain rules and regulations used in all laboratories. These are rules of work environment and safety, information about these can be found here: http://www.ntnu.edu/hse. Lab work regulations can be found in the HSE handbook: http://www.ntnu.edu/hse/labhandbook. All of the rules and regulations are according to the rules set by the Norwegian government and the Norwegian Biotechnology Advisory Board (http://www.bion.no/english/norwegian-regulation). The project is not in conflict with any of the rules and regulations from the government, university and institute.

Recombinant gene technology is also regulated in Norway by [http://www.lovdata.no/all/hl-19930402-038.html Lov om framstilling og bruk av genmodifiserte organismer m.m. (genteknologiloven)] [Law on the production and use of genetically modified organisms, link in norwegian] and [http://www.lovdata.no/for/sf/ho/xo-20011221-1600.html Forskrift om innesluttet bruk av genmodifiserte mikroorganismer] (Regulation on the contained use of genetically modified microorganisms, link in norwegian)

In addition, all work at NTNU is regulated by the norwegian Working Environment Act ([http://www.lovdata.no/all/hl-20050617-062.html arbeidsmiljøloven], link in norwegian) and internal regulations. Of particular importance at the Department of Biotechnology is the section on chemical and biological hazards (§4-5). Other governmental regulations also apply.

Question 4

Question

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?

Answer

In synthetic biology today, antibiotics are used as means for selecting the cells with the right plasmid. But, as mentioned, this can be a risk to other organisms if released into the wild. It it would be possible to select the GMOs with the right plasmid on basis of something else, it could make the risks of GMOs lower.

If it is possible, a good thing would be to make the organisms used in the lab unable to live in the wild. If they need very specific conditions to survive, the probability of them surviving in the wild or doing any other damage, will be lower.