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Team:BGU Israel/safetyQuestions
From 2013.igem.org
In laboratory work, proper safety precautions must be taken: management of experiments, the correct storage of materials, emergency safety measures, removal of unused hazardous materials and safety training. Safety measures must take into account employee safety, public and environmental implications. In our project, we worked with the lowest risk level, defined as Biosafety Level 1 (BL 1). The highest Risk Group in this project, included organisms and materials is Risk Group 1.
1. List of organisms:
- Escherichia coli, BL21
- Escherichia coli, DH5α
2. List of Biobricks:
- Lac/Ara-1 IPTG inducible promoter(BBa_K354000)- the promoter is synthetic and nonhazardous, and therefore is not associated with a risk group level.
- LVA-tagged cI repressor (BBa_K327018)- Risk Group 1, not harmful to humans or the environment.
- Promoter Lambada cI regulated (BBa_R0051)- Risk Group 1, not harmful to humans or the environment.
- T4 Holin (BBa_K112805) - Risk Group 1, not harmful to humans or the environment (toxin only to bacteria).
- P.A.S.E. 1 cassette
- P.A.S.E. 2 cassette
- KanR (promoter+CDS)
- Lambd cI CDS
- cI translational unit
- HisTag + stop codon
- cI translational unit with His-tag
- pUC57-P.A.S.E.2
- pUC57 backbone (REVERSED)
- pUC-57-P.A.S.E.1
- ampR translational unit (ampicilin resistance CDS+promoter)
- pKD46 functional unit
- Pyrolysyl-tRNA synthetase CDS
- tRNA-Pyl (pylT) gene from Methanosarcina barkeri str. Fusaro
3. Terms of safety and risks regarding laboratory workers, the public and the environment
In the Lab
The E.coli we worked with are common laboratory strains, considered Level 1 Biosafety Containment Agents.
Safety training took place prior to the start of lab work. The goal of the training was to maintain safety in the lab, and to maintain personal safety by preventing incidents such as inhaling, ingesting or getting contact and exposure to the skin with some of the chemical materials that were used. Laboratory work was performed with closed shoes, gloves and lab coats to reduce risk. Additionally, proper biological waste disposal was performed, in order to keep the lab sterile and control contamination. Glassware, needles, and other sharp objects were disposed of in appropriate containers to prevent injury to employees. Examples of potential hazards: work with materials such as ethidium bromide, sodium dodecyl sulfate (SDS), and hazardous halogenated materials; work with ultraviolet lamps, employees; work with open-flame Bunsen burners.
Environmental & Public Safety
The first step towards proper environmental and public safety is working according to regulations safety rules from the beginning of lab work until the end. All biological waste was sterilized with an Autoclave before leaving the lab, and all waste was disposed of in appropriate containers and removal procedures (organic chemical waste, glass, biohazard).
The contamination of the surroundings by biological and chemical materials or microorganisms was prevented by keeping lab work sterile, maintaining correct work habits, and washing hands upon leaving the lab.
The main environmental issue concerning synthetic biology and genetic engineering is the unintentional release of genetically modified micro-organisms (GMOs) into the environment. Currently, the possible consequences of such an event cannot be fully predicted.
Bacteria undergo a natural process called horizontal gene transfer, in which DNA can be transferred from one bacteria to another. They are also subject to evolutionary forces, in which the best adapted bacteria survive. Therefore, it is possible that GMOs that escaped the containment of the laboratory could cause unwanted and even dangerous effects in the environment. A few possible scenarios are the transfer of genes to native pathogenic bacteria that would give them a new evolutionary advantage, or released GMOs spreading in an ecosystem or even our bodies, overpowering populations of natural bacteria.
These concerns were our motivation for our project: our self-destruct mechanism could substantially reduce concerns over using genetic engineering and the risk of the unknown that currently shadows the release of GMOs.
The E.coli strains we used in our project are weakened laboratory strains that are not pathogenic and would not survive in the environment. We chose to work with these strains in order to reduce risks for lab workers and for the surroundings, in the case of unintentional release. For example, if an employee forgets to wash his or her hands upon leaving the lab, even if an infection took hold, the bacteria could be easily killed by antibiotic treatment.
Security
Our project could possibly be utilized for in biological weapons against human beings to provide a killing switch, in order to attack a specific target without spreading the pathogenic bacteria further.
4. If the project moved from a small scale lab study to become widely used as commercial/industrial product, what new risks might arise?
No significant risks would arise. Our construct would be independent and would not harm the environment nor public health. The only possible risk would be the slight possibility of the transfer of genes and certain DNA fragments from our construct to other bacterial populations, with unknown results; however, this is a risk of any release of GMOs.
5. Does the project include any design features to address safety risks?
We decided to utilize holin and lysozyme as our toxin system, because both affect only bacteria. Holin forms pores on the inner membranes of bacteria, and lysozyme degrades the peptidoglycan layer. Both toxins are harmless to other organisms and to the environment.
6. Safety in Ben Gurion University
Each student completed the university’s biological and chemical safety course. Our team also collaborated with the university’s Department of Safety, to ensure that we fulfilled safety standards and that the materials and organisms were Biosafety Level 1.
According to the WHO BioSafety Manual, our lab is qualified as category 2, which is higher than the risk group rating of our organisms (Biosafety Level 1). Therefore, our laboratory was more than matched the safety requirements needed for working with these organisms.
Our safety forms were approved on September 29, 2013 by the iGEM Safety Committee.
Continue the journey: meet the Team .
References
[1] Laboratory biosafety manual (World Health Organization, Geneva, Ed. 3, 2004), pp. 1-66.
