Team:Korea U Seoul/hp/safty Q

From 2014.igem.org

Simple Protocols
Experiments were performed in the lab of the Dept. of Biotechnology at Korea University. The lab is confirmed to the standards for bio-safety 1 which is suitable for working on agents that are not known to cause diseases in immune-competent adult humans. Works are conducted on open bench or clean bench using standard microbiological practices. Special containment equipment or facility design is not required, but may be used as determined by appropriate risk assessment techniques. Laboratory personnel must have specific training in the procedures conducted in the laboratory and must be supervised by a scientist with training in microbiology or a related science.
Standard Microbiological practice
  1. Statement of Purpose (SOP) are kept for easy access which enlists all the safety measures each experiment should be abided for.
  2. Mechanical pipetting is used
  3. Work surfaces are decontaminated regularly. Clean bench is decontaminated everytime after being used.
  4. People wash their hands after handling viable materials, after removing gloves, and before leaving the laboratory.
  5. All procedures are done carefully to minimize the creation of splashes or aerosols.
  6. Eating, drinking, and smoking are banned inside the laboratory.
  7. All cultures, stocks, and other regulated wastes are decontaminated before disposal by an approved decontamination method for example Autoclaving. Materials to be decontaminated outside the immediate laboratory are to be placed in a durable, leakproof container and closed for transport from the laboratory. These materials are packaged in accordance with applicable local, state, and federal regulations before removing from the facility.
Safety equipment
  1. Everyone must use appropriate safety equipment (Lab gowns, gloves, and goggles,etc) required for the experiment.
  2. Gloves are necessary when handling toxic materials or if there is a wound on the hand.
  3. Goggles must be worn while performing experiments with hazardous materials or splashes of microorganisms
Labatory Facilities
  1. Our lab has sturdy door, which cuts off the lab from outside, and entrance of people other than the lab members is strictly regulated.
  2. Our lab is equipped with two sinks with which people can wash hands and lab equipment.
  3. Our lab is spacious enough and can be easily cleaned. The lab is cleaned regularly and is kept tidy.
  4. Bench tops are always kept tidy and clean. They are impervious to water and resistant to heat and chemicals involved in most experiments done in the lab.
  5. Lab furniture is appropriate for withstanding anticipated loads and uses.
  6. We are readily available with safety manual and guideline, which we can use in case of accidents.
Simple Questions
1. Would any of your project ideas raise safety issues in terms of:
  1. Researcher Safety
    2. EtBr: Ethidium bromide is used as a fluorescent tag for gel electrophoresis in our lab. EtBr is known to negatively affect important biological processes, such as DNA replication and transcription, because it intercalates and deforms double-stranded DNA. Though we wash our agarose gels with EtBr solution, we strictly make sure that the solution does not make contact with human body by wearing gloves when handling the solution. The gels are safely discarded according to the safety guideline provided by the university.
  2. Public Safety
    3. Genetically modified bacteria are strictly prevented from being released out of the lab.
  3. Environment Safety
    4. We strictly follow the safety guideline provided by the university to take appropriate means of safely discarding all experiment waste.
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?
  1. Did you document these issues in the Registry?
    2. Yes
  2. How did you manage to handle the safety issue?
    3. Our project includes transforming modified Pilin A gene cluster from Corynebacterium diphtheriae into Corynebacterium glutamicum to express pili with the genetically modified C. glutamicum.
    Toxin-emitting strain of C. diphtheriae can cause diphtheria by adhering to the nasopharyngeal tract with their pili and emitting toxin, which causes symptoms, such as sore throat and fever. Our team is well aware of the pathogenicity of C. diphtheriae, but we have definitely made sure that our experiments are safe and do not cause any threats to biosafety. First of all, we are using only a part of the genomic DNA of C. diphtheriae, NOT the whole organism. C. glutamicum, into which we are transforming DNA parts of C. diphtheriae, is a completely non-pathogenic species and does NOT express virulent toxin that causes diphtheria symptoms. Also, we do not use C. diphtheriae pili as it is but instead substitute the original pilin protein into other proteins, such as GFP. Furthermore, we are by all means following the biosafety guideline provided by Korea University and KCDC about dealing with the C. diphtheriae genomic DNA parts.
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, our project is under the supervision of Korea University Safety & Security Team. The team has raised no safety issue regarding our project.
References
  1. Cupillard, L. et al., 2005. Impact of plasmid supercoiling on the efficacy of a rabies DNA vaccines to protect cats. Vaccine 23: 1910-1916.
  2. JM Tiedje et al., 1989. The planned introduction of genetically engineered organisms: Ecological considerations and recommendations Ecology, 70(2), 1989, pp. 298-315
  3. Efstathia V. Scoulica. et al., Spread of blaVIM-1-producing e. coli in a university hospital in Greece. Genetic analysis of the integron carrying the blaVIM-1 metallo-β-lactamase gene,Diagnostic Microbiology and Infectious Disease Volume 48, Issue 3, March 2004, Pages 167-172

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