Difference between revisions of "Team:BGU ISRAEL/EthicsAndSafety"

All of our project's lab members received safety training by our advisors, and had to partake in a lab safety online course offered by the Ben Gurion University safety department. In this course, we learned about emergency protocols, safety classification of biological labs, and working with new chemicals. In the lab, we wear protective clothing and gloves at all times (unless working with fire). In necessary cases, we also wore protective glasses and face masks, worked in fume hoods / biological hoods etc. All experiments were done considering all appropriate safety protocols, after consulting our PI and advisors.

In our project, we preformed multiple growth experiments using different intermediates in the PET degradation pathway as carbon sources. Some of the chemicals have specific safety instructions according to their MSDS, which were printed out and stored in the lab for all of the members to read and work according to. (For example, at times, it was necessary to work with a chemical like Catechol, or to handle Ethidium bromide waste, in a fume hood).

Safe Project Design (Safety Risks to the General Public):

Our final goal as the BGU iGEM team is to create an operating biofuel cell, which will break down polyethylene terephthalate (PET) using E. coli and P. putida. We chose to work with these bacteria, as they are known chassis organisms, are easy to manipulate, and are risk group 1 (P. Putida - DSMZ) – they do not cause diseases in human. Moreover, we use the P. putida KT2440 strain, which is the first Gram-negative soil bacterium to be certified as a safety strain by the Recombinant DNA Advisory Committee (Federal Register, 1982(3)) and is the preferred host for cloning and gene expression for Gram-negative soil bacteria (Nelson, K. E., et al. 2002(4)).

With the safety of the environment in mind, we designed a metabolic cascade to degrade the toxic intermediates in the PET degradation pathway, to non-toxic compounds that can be used by the bacterium for energy in the TCA cycle. This way, the degradation process does not release any pollutants or toxins to the environment other than the carbon dioxide and water bacteria produce in their own metabolic processes.

In order to ensure the safety of our product, we integrated into our design the self-destruct mechanism developed by the 2013 Ben Gurion University iGEM team. With this mechanism, not only the expression of our genes will be dependent on an IPTG inducer, but it will also force our E. coli to be fully dependent on an unnatural amino acid (UAA). Without supplying it to the bacteria medium, or in case of release to the environment (which will only happen by accident since we do not intend to release our bacteria), our E. coli bacteria will not survive. Our second bacterium, P. putida, which is design to consume TPA (one of the PET degradation products), would not be able to consume PET as it is dependent on LC-Cutinase (the enzyme which breaks the initial bond in the PET polymer into TPA and EG), since LC-Cutinase is expressed and secreted from E. coli. Therefore, the genetic modification done on this bacterium will be useless, since it is responsible for degrading TPA, which is a commodity chemical that usually does not exist abundantly in nature.

Local Rules and Regulations:

• World Health Organization (WHO) – Laboratory biosafety manual, third edition
• Restrictions on Genetically Modified Organisms: Israel
• IBSA – Israel Biological Safety Association
• Ben Gurion University – Safety Department

1. Synthetic biology: a Jewish view. Glick S. Perspectives in Biology and Medicine, Volume 55 (4), 2012, pp. 571-580
2. http://www.azquotes.com/author/9304-Maimonides?p=2-
3. Register, F. (1982). Certified host-vector systems. Washington, DC, 47, 17197. ‏
4. (4) Nelson, K. E., Weinel, C., Paulsen, I. T., Dodson, R. J., Hilbert, H., Martins dos Santos, V. A. P., ... & Brinkac, L. (2002). Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440. Environmental microbiology, 4(12), 799-808.