The chassis organism we are using for this project.
- E. coli (K 12) TOP10
Highest Risk Group ListedRisk Group 1
Description of the biological materials we are using in the lab
Risks to the safety and health of team members or others working in the labSome materials possess risks to team members. For example, Ethidium Bromide is an intercalating agent so it must be used by with personal safety gear. All lab staffs were trained according to safety manual provided by Hokkaido University, to prevent risks. We took on ourselves to compile a shortlist of often used dangerous materials and safety procedures in our project.
- Ethidium Bromide
- intercalating agent : must be used with personal safety gear
- flammable : must not be used near open flame or in large quantities
Procedures and equipment
- Agarose gel production
- heating in sealed container (rupture risk), scalding hot and vicious during preparation (burn injury risk) - remove container lid before heating in microwave, use safety gear, wait for a few moments before removing from microwave
- Benson burner
- fire risk: DO NOT use flammable materials especially ethanol near open fire
- high velocity: balance appropriately, observe the machine till it reaches top velocity
- high pressure: check the water level, DO NOT open when pressurized
- UV radiation
- damage to eyes and skin: use glove and UV box or UV shield
Non-pathogenic bacteria (policy requires treating as pathogenic, as precaution)
- E. coli (K 12) TOP10
- UV Box
- UV shield
- Clean bench
Waste disposal and sterilization
- All equipment and wastes coming in contact with bacterial were sterilized by autoclave or bleach.
- All chemical compounds were disposed according to requirements for their disposal.
- Table surface used for work were sterilized with 70% ethanol before and after a procedure.
Chemical UsageAll chemical compounds were used according to their manuals and respective material safety data sheets.
Genetic materialAll genes used in this project come from non-pathogenic bacterial strains of E. coli. Expressed proteins did not show any toxic effect to their host. Our biobricks do not have any foreseeable selective advantage if released to the environment. After consideration we could not find any usage pausing a security concern.
Risks to the safety and health of the general public, if released by design or by accidentSome materials pose risks to the general public. For example, Ethanol is a flammable solution so it must not be used by open fire. Not to release those materials, all lab staff is trained according to safety manual provided by Hokkaido University.
Risks to the environment, if released by design or by accidentThe E. coli strains we use in our lab, are saved in our lab. As a precaution all materials coming in contact are sterilized before and after. Reference Federal Register, (1986) Vol. V1: 88, 6952-16985.
Risks to security through malicious misuse by individuals, groups, or countriesThere is no foreseeable risk in misuse of our generated genetic material. Our generated genetic material performs basic functions in biology. However, it is impossible to guard against the incorporation of our parts in malicious settings.
Risks which might arise when our project move from a small-scale lab study to become widely used as a commercial/industrial productOur project is about optimizing the expression of genes. Our device does not contain a coding site. Therefore, risk will arise when other users assemble our parts with dangerous coding sites. We have to caution the users when assembling with dangerous coding sites.
Design features to address safety risksOur device only contains sequences that regulate the expression of genes (Promoter, RBS, and terminator). Therefore, our device in itself does not contain any safety risks and does not have design feature to address safety risks.
Safety training we received8 members of our team received a lecture class regarding gene recombination that was held in Hokkaido University, and shared the information with the whole team. It is based on 'Act on the Conservation and Sustainable Use of Biological Diversity through Regulations on the Use of Living Modified Organisms'.
Link to our institution biosafety guidelineshttp://www.hokudai.ac.jp/jimuk/reiki/reiki_honbun/u010RG00000583.html
Our Institutional Biosafety CommitteeWe have a permission to engage in the experiments from the safety officer of genetic recombination of Hokkaido University. All members participating in the experiments are registered with this office. All members are trained according to the safety demands of safety officer of genetic recombination.
Our country's national biosafety regulations and guidelinesJapan is participating in cartagena act.
Biosafety Level rating of our labOur lab's Bio safety level is 2.
The Risk Group of our chassis organismsThe Risk Group of our chassis organisms is 1.
Faculty AdvisorSone Teruo