Team:Copenhagen/Safety

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Guidelines for working with GMO


Biosafety Board


Ethics


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Safety

 

 

Synthetic Biology and Safety

By combining biology and engineering, Synthetic Biology is seen as the design and construction of new biological functions and systems not found in nature. The aim of this biological construction holds high promise in the discovery of cures for various diseases, methods for biofuel production, plants of the future, functional foods and molecular bioelectronics. The development of better medicine and sustainable energy are some of the main focuses.

Rapid advances within this new emerging field have the potential of greatly benefitting mankind, though posits tremendous dangers in the wrong hands or if mistakes are made. It is therefore imperative that we as future researchers are completely aware of the consequences we might have on society and the environment. It is therefore imperative that we as a team are completely aware of the safety and security of our labs and our work, and that we recognize the importance of each member’s responsibility to uphold both international and national laws, regulations and guidelines.


Biosafety and Biosecurity

The Danish laws for working with GMO’s are set forth by the Danish Labour Inspectorate, Arbejdstilsynet, and apply to both commercial and academical institutes. Under these laws, The Faculty of Health Sciences at the University of Copenhagen has set forth a list of procedures for working in a GMO level 1 laboratory. These are followed thoroughly by all members of the Copenhagen iGEM team 2011.

The WHO defines biosafety as the prevention of unintentional exposure to pathogens and toxins, or their accidental release, whereas biosecurity is the prevention of loss, theft, misuse, diversion or intentional release of pathogens and toxins.


Biosecurity:

It is important to keep unqualified personnel and trespassers away from the laboratory and the samples. Access to the laboratories during afterhours is locked down automatically, and a magnetic keycard (only approved by the administration at the University of Copenhagen) is necessary for admittance to the lab areas.

Following the guidelines to the WHO Laboratory biosafety manual, the safety of our iGEM team is ensured by having our laboratories equipped with fire extinguishers, emergency showers, eye showers and heart defibrillators. The same laboratories, which are in use for our iGEM project, are classified as BSL level 1 on the WHO terminology of biosafety levels (BSL).


Biosafety:

The Copenhagen iGEM Team 2011 is currently working with the non-pathetogenic Escherichia Coli derivates XL-Blue and BL21 (DE3), classified under BSL-1. These are generally regarded as safe.


iGEM Questionnaire

1. Would the materials used in your project and/or your final products pose:
  • Risks to the safety and health of team members or others in the lab?
We see no such risks to our safety and health in the lab. All research with Genetically Modified Organisms (GMO’s) is contained in a GMO-class 1 laboratory at the Faculty of Life Sciences, at Copenhagen University. All processes and methods carried out have been done so only after training and/or under the supervision of an instructor to ensure all safety protocols are maintained. In addition every team member underwent safety and waste disposal training before performing any experiments in the lab. Furthermore, the team will be supervised throughout the project by more experienced research scientist.
  • Risks to the safety and health of the general public if released by design or accident?
There should not be any risks for the general public as our project is intended to function as a fundamental research project more than a product to be used “as it is” in the environment. Therefore, all experiments with our BioBricks and use of GMO’s will occur in a contained laboratory environment, with established biosafety and biosecurity requirements. Our modified bacteria is at no risk to the safety of the general public. The reasoning behind this is that the bacteria we work with are not dangerous to human beings. Furthermore, the public safety is ensured as no member of the public is permitted access within the GMO-class 1 labs, unless approved by the university.
  • Risks to environmental quality if released by design or accident?

Environmental safety is ensured as all of our experiments are contained and disposed safely following to both international (WHO) and Danish (Arbejdstilsynet) laws. We use the E.coli strains Bl21 and XL blue, two strains which are generally used in the laboratories and considered safe. The antibiotic resistance genes used are only for development of the design. The project is only on a general research level. No GMO will intentionally be released into the environment during the course of this project. In case of an accidental release of our genetically modified E.coli strain, the added antibiotic resistance gene will quickly be lost to natural selection and therefore not be a problem.



Specific questions
  • This project is designed to produce an agent intended for environmental release. This would require a thorough examination of the safety issues surrounding such a release. For example, would a bacteria designed to kill fungi also kill mushrooms? Or beneficial fungi? If so, what measures would be needed to stop this from happening? It is noted that such a release is not intended but could still happen by accident. If that was to happen, what would the implications be for people that come into contact with the organism, or the environment?

The oximes our E. Coli are designed to produce, have a documented toxic effect on a wide range of fungi and mushrooms. It is very likely that all fungi and mushrooms in the area of release will die. The best way to limit the death of any beneficial species would be to limit the expression of oximes to high priority areas, such as the leaves of the plants or the area surrounding the roots ect. One way to accomplish this could be to genetically modify the plants or weeds of interest to only express the CYP in the leaves. Resulting in a limited explosion of the oxime to the soil. Such vegetation will of course have to be grown in a way corresponding to the individual countries safety regulation regarding GMO crops.

Synthetic production of oximes has a perspective to be used e.g. in tree impregnating paint, which it already is an important component of. The use of this paint could therefore only be limited to areas where fungi are unwanted. This should of course be emphasized on the product.

In case of an accidental release of the modified bacteria the antibiotic containing plasmid will rapidly be lost due to natural selection rendering it harmless to the environment. Also the bacteria we use are so weakened that they will die in a natural environment. Furthermore the promoter we use to induce the transcription of the CYP gene is inducible and only activated by IPTG, which is not present in the nature. Which means that if the bacteria is released, it will quickly lose its resistance gene and it will not produce oximes.



Specifically, are any parts or devices in your project associated with (or known to cause)
  • Risks to security through malicious misuse by individuals, groups or states?

There will be no risk for malicious misuse because the oximes that are produced are not dangerous and can therefore not be used as a biological weapon.

  • Pathogenicity, infectivity, or toxicity?

It is only toxic for fungus.
  • Threats to environmental quality?
Until it has been thoroughly established that it is not harmful to the environment and the level of its toxicity is known, no oxime producing bacteria will be released. So far studies show that the toxicity is limited to fungi and mushrooms. During an accidental release the effect would be very limited because of the scale of oxime production needed to impact larger areas.
  • Security concerns?
As long as production of oximes is under control of e.g. an inducible promoter there will not be any security concerns regarding to an unwanted release of oximes.
Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? None of the BioBricks we have created raises any specific safety issues. All GMO’s are set to biosafety level 1 (BSL1), there are no risks of hazard or biological safety issues to mankind. Any safety issues discovered will naturally be fully documented in the Parts Registry. Although the iGEM project timescale is insufficient for BioBricks characteristics such as genetic reliability and performance reliability. Additional work is usually required if the Parts Registry is to be fully updated and reliable.

Under what biosafety provisions will / do you operate?
  • Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible.
Yes, the institution Faculty of Life Sciences, University of Copenhagen has their own biosafety rules (see the link to the left). These rules are lined up to the right.
  • Does your institution have an Institutional Biosafety Committee or equivalent group?
There is a biosafety board in our laboratory at the Faculty of Life Sciences, Copenhagen University (biosafety). All protocols set forth by the biosafety board are being followed by the Copenhagen iGEM Team.
  • If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review.
The committee is familiar with our project and our presence in the laboratory. The only condition for our labwork is that one of our advisers is at the institute at all times.
  • Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.
Yes, in our first week in the lab we were introduced to the different machines and safety procedures by our adviser Peter Naur. When using new methods we are thoroughly introduced by an adviser about the procedure and the safety regarding to this.
  • Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.
Yes Denmark has it own regulations. See link to the left. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?
As the iGEM competition is gradually becoming more and more popular with each year passing – then the number of BioBricks created will also increase. Therefore, to keep the Parts Registry fully documented and reliable, some form of testing of the BioBricks will have to occur before sending them for use in the Parts Registry database. Safety guidelines when working with GMO’s should be set forth by iGEM – making sure to inform the participants that each country/university may have their own deviations in protocol.
Safety Procedures followed thoroughly by the Copenhagen iGEM Team:

1. Needless traffic in and out of the laboratory must be limited. All visitors must wear a lab coat. Visitor lab coats are located at the entrance to the laboratory.


2. The laboratory must be kept clean and tidy and in good order. After work, all surfaces must be cleared and decontaminated by the user. In case of GMO spill, immediate decontamination must be carried out.


3. All biological waste, including non-GMO waste, is discarded in special bins labelled “Biohazard” and “GMO”.  The material is sealed before being transported for autoclaving. Containers with liquid waste must be clearly labelled with autoclave tape labelled “GMO-waste”.


4. Pipetting by mouth is absolutely forbidden.


5. It is absolutely forbidden to bring any kind of food or drink into the laboratory.


6. Personnel must wash the hands after having been in contact with any type of organisms, GMO or non-GMO, before leaving the laboratory.


7. Special caution must be taken to minimise aerosol production.


8. All experiments in the laboratory must be conducted according to this procedure. The procedures are also valid for experiments including recombinant nucleic acids.


9. Transport of GMO-material between classified areas must be performed using a closed container clearly labelled ”Biohazard” and/or ”GMO”.


10. If the laboratory is no longer to be used for GMO work, down-classification is carried out after main cleaning.



Comments or questions to the team? Please mail us at igemcopenhagen@gmail.com

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