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From 2011.igem.org

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

a. Researcher safety

The materials used in our project pose no risks to the safety and health of the members. All of our biobrick parts were put into Escherichia coli DH5α. We did not use any pathogenic strains of E.coli, nor were any of our biobrick constructs of any potentially hazardous pathways. Our lab is a BSL 2 rated lab. In addition to putting biobricks into E.coli, we also put some of our parts into the Cyanobacteria Synechocystis PCC 6803.

b. Public safety

As with any recombinant microbes, significant testing needs to be conducted at the laboratory scale before being released to the public. Materials in our project would not pose any significant risk to the public. Since our project does not focus on pathogenic or infectious pathways, even if released by accident there would be no risk to the general public. Parts are cloned into vectors containing antibiotic resistance and thus the bacteria would not be able to maintain the plasmids in an environment outside of the lab.

c. Environmental safety?

There would be no risk to environmental quality if released by design or accident. As mentioned previously, all parts are cloned into vectors containing antibiotic resistance.

2.Do any of the new 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?

Our lab operates under BSL2 safety provisions. This means that we are not handling anything that is potentially toxic or pathogenic. In addition we also follow our university biosafety provisions.

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?

Yes Utah State University has an Institutional Biosafety Committee: http://www.usu.edu/fsenate/PDF/Utah%20State%20University%20Councils%20and%20Committees%20Handbook%202010-11.pdf

We have discussed our project with the committee and they have approved our work. Since our laboratory is used for other synthetic biological research the approval was straightforward.

Utah State University has its own biosafety rules. A link is provided here: http://ehs.usu.edu/htm/programs-and-services/biological-safety

The University has rules and regulations regarding the correct disposal and clean up of biological waste material.

All undergraduate students are trained by the Environmental Health and Safety department (EH&S) at Utah State University. This training covers basic laboratory safety procedures and practice. It is required that everyone (including graduate students and staff) who works in a laboratory at Utah State University take this day long course. In addition, all undergraduates were trained specifically on biological safety by experienced graduate advisors and faculty before they were allowed to work in the laboratory. For more information about Utah State University laboratory safety training please see: http://ehs.usu.edu/htm/training/health-and-safety-training-courses

The United States has strict regulations and guidelines for biosafety. The National Institute of Health (NIH) has guidelines for working with recombinant DNA. See: http://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.htm The centers for disease control and prevention (CDC) also has guidelines for biosafety. See: http://www.cdc.gov/biosafety/

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?

Biosafety Topic 4.2 Designing and using a safer host organims/chassis

In regards to designing safer host organisms, most of the currently used lab strains of E. coli have such demanding environmental requirements, that they are highly unlike to grow if they leave the lab environment. That being said, public perception of the actual risks from these "escapes" is quite high, and additional safeguards would be useful to prevent both unexpected survival in the environment and the associated media outcry.

Many iGEM teams and other synthetic biology projects have been based around developing "kill-gene" systems, where through various methods, the system detects that the bacteria is outside its designated habitat, and then proceeds to produce a toxic compound or enzyme that kills the cell. While this procedure works well from an engineering standpoint (and is employed in computers when a program begins to take up all available processing power or act in unexpected ways), it is problematic when implemented in biological systems due to evolutionary pressures. There would be immense evolutionary pressure against organisms possessing these systems, and any individual cell that had a deletion or loss of function mutation in this system would quickly represent the entire population. If this system was present in the organism in a plasmid, it would be highly unstable as plasmid loss in a non-selecting environment can occur rapidly, and even if integrated into a genome would eventually be selected against.

A better solution to prevent accidental release of the organisms would be to "hard code" the environment restriction by developing strains of E. coli with multiple (and I stress multiple for redundant protection) amino acid synthesis pathways knocked out. This will provide an easy-to-supply method of keeping the cells alive in the lab, but prevent them from growing in almost all environments outside the lab. As it is an absence of genes, rather than the addition of a kill-gene, this would be incredibly difficult to be circumvented evolutionarily, requiring the acquisition of several entire genes or pathways, rather than a fairly common loss of a single gene or a single mutation of a gene.

However, public perception of these two systems may be counter to the level of safety that each provides, as a "kill-gene" system might appear safer and more absolute than nutritional restrictions. Also, members of the public who pay attention to the science in science fiction movies might recognize the nutrient limitation system as being similar to the concept in the Jurassic Park series. The multiple knockout modification to this system makes it more reliable, in addition to the fact that bacteria are less likely to digest significant organic matter in the environment to acquire the necessary amounts of limited amino acids.