Team:Uppsala University/Safety

1. Would any of your project ideas raise safety issues in terms o: researcher saety, public saftey or enviromental safety?

There is no risk to the safety nor the health of team members or others in the lab. Neither our material, nor our constructs produce anything toxic. We use E. coli Top10 strain as cloning strain, which has very low fitness outside the lab. We are also working with the E. coli K12 MG1655 strain which is a wild type, well characterized non-pathogenic lab strain that is would not be competitive outside the lab environment. All strains that we come in contact with belong to WHO risk group 1 (no or low individual and community risk), i.e. microorganisms that are unlikely to cause human or animal disease.

The research that we are making provides us with basic information such as how to target microbial resistance genes. In our project there are no genes present that cause toxicity or increased virulence. Therefore the potential for a malicious misuse is low.

In the beginning of the project we were provided with resistance genes CTX-M-15 (confers beta-lactam resistance), AAC(6´) (confers resistance to aminoglucosides) and TetA (confers resistance to tetracycline) cloned from plasmids derived from clinical isolates. These genes are well known and well characterized, and the handling of the genes in this form poses no bigger risk to the lab workers or general public than the traditional selection markers used in standard cloning plasmids. The genes were isolated in accordance to Swedish safety practice by researchers at the Department of Medical Biochemistry and Microbiology at the medical faculty of Uppsala University. The genes were provided to us as single genes without promoters cloned in standard synthetic biology plasmids in an E. Coli TOP10 strain. Our final constructs will be in both MG1655 and the TOP10 strains. In our project we have been working with these isolated genes, but never with the clinical strains, nor the clinical ESBL plasmid. If we would try our constructs in any clinical isolates in the future, it would be done in a lab with the appropriate safety level under supervision of professional staff.

The phage that we are going to use in our project is M13mp18 and the handling will follow standard protocols. This is considered standard procedure, and poses no risk to people working in the lab or the public safety. The phage M13mp18 is restricted to infecting E.coli containing a fertility factor, an F plasmid.

With respect to this, we can conclude that our project raises no safety issues in terms of researcher safety, public safety or environmental safety.


2. Do any of the 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 the parts that we are working with raises any safety issues. None of the clinical resistance genes we are working with will be included in the shipment to the partsregistry.


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?
We work according to the regulations of Work Environment Authority (AFS). The regulations that concern lab work are listed below:

AFS 2000:5 Contained use of genetically modified micro-organisms http://www.av.se/dokument/inenglish/legislations/eng0005.pdf AFS 2005:1 Microbiological health risks - infection, toxin effect, hypersensitivity http://www.av.se/dokument/inenglish/legislations/eng0501.pdf Link to Work Environment Authority: http://www.av.se

The national biosafety regulations and guidelines in Sweden are generally provided by Work Environment Authority and the Swedish Institute for Communicable Disease Control (Smittskyddsinstitutet). http://www.smittskyddsinstitutet.se/amnesomraden/biosakerhet-och-bioskydd/, http://www.av.se/teman/gmm/ In English: http://www.av.se/dokument/inenglish/themes/gmm.pdf

Every individual in the laboratory has had training in the previous courses. Uppsala University has a policy, which states that every individual working in the laboratory has to have gone through a safety session. All bacterial contaminated waste is thrown in a hazardous waste bin. The waste is later handled according to safety protocols and sent for destruction at an external company according to the standard procedures of Uppsala University. All contaminated media and glassware are disinfected with Jodopax (5% iodide, 15% acetic acid) before it is washed.

Uppsala University does not have a biosafety committee, nor a review board. However there is a group at the Centre for Research Ethics & Bioethics (CBR), this group works daily with ethical questions concerning the field natural science. An appointment was booked with Stefan Eriksson, a senior lecture of Research Ethics at CBR and one of the authors of CODEX (http://www.codex.vr.se/en/index.shtml), to discuss the ethical issue that our iGEM project had raised. Stefan Eriksson came to the conclusion that our iGEM project produced no dilemmas although he did emphasize that the major threat that we should be aware of is dual usage amongst researchers.


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?
We in team Uppsala University believe that the most efficient way to confront a problem is preventing it from happening. Thus we believe that raising awareness to other iGEM teams about threats and dual use among researchers. In order to prevent a problem or a threat, deep knowledge and creativity is of the essence. Henceforth it is important that there is a direct connection between Human Practice and bioethics groups so that both the researchers and the public can be supplied with necessary information.