Team:Korea U Seoul/hp/safty Q

1. Issues to the researcher safety
1. E.coli(BW25113, DH5a, BL21(DE3)
Moreover, E. coli strain we are using are BW25113, DH5a and BL21(DE3). Those E. colis are used for produce proteins(nacrein) and they use different promoters. Refer to former research they studied widely about safety and by the result founded to be very safe. They also weakened for laboratory use. Therefore would not pose a threat to environment. Thus, this E. coli strain is less likely to harm researchers.

In conclusion, we find our project harmless. Although no significant safety issues were raised during the experiment, it is advised that experimenter should follow safety protocols listed on this website. This includes wearing gloves and goggles as bacteria may cause irritation and skin rash. Please refer to lab safety protocol: http://kusafe.korea.ac.kr/main.asp (university guideline, Korean) http://bric.postech.ac.kr/myboard/read.php?Board=safety&id=1481(national guideline, Korean)
2. Vectors
Vectors We used is pATLIC and pBL plasmid. They used to produce protein which we want to made by using other promoters. pATLIC and pBL vector have same role but protein which made by pBL system is soluble but protein made by pATLIC is insoluble.

This plasmid have resist genes against antibiotics. So it can be danger to nature and human. however to transcript DNA and make protein, they need some key materials. So in nature they can be activated. Also we follow safety protocols about definite by our university. Its include biohazard treatment so we can isolate our vector from nature and human.


2. Risks to the safety and health of the general public
There are no significant risks to the safety and health of the general public. First, we do not use pathogenic bacteria. Second, gene products do not induce immune response. Third, our lab follows rules of bio-safety level limiting reagents and material we use. Here are reasons why our project is less likely to cause any health risks.

BioBrick we made is Carbonic Anhydrase from nacreous layer in oyster pearls. It can hold CO2 by attach that by CaCO3 form. So CO2 fixed by nacrein have special crystal structure because of nacrein between the CaCO3. Nacrein only action as a Carbonic Anhydrase and it is one of the most universal enzyme. One difference is using Ca to fix CO2. But CaCO3 also popular chemical, So there's no risk for nature.

Also it is hard to express in other species because of the vector which we use need special material to active.

1. Regarding local biosafety group
There is no biosafety group, committee, or review board at our institution. The iGEM project has not been reviewed by any local bio-safety committee. However, we follow the bio-safety guidelines for any procedure in the laboratory. Please refer to our safety guideline (includes what biosafety rules we have to consider in our country): http://kusafe.korea.ac.kr/main.asp (university guideline, Korean) http://bric.postech.ac.kr/myboard/read.php?Board=safety&id=1481(national guideline, Korean)


2. Consideration about biosafety rules or guidelines
Based on our university guideline and national guideline, we made 3 major biosafety principles and their detailed guideline.
1. Biosafety level of laboratory
This principle defines boundary of experiment which labs can conducted and limits the type of experiments based on their capability. Each lab should be evaluated for their biosafety capability and assigned any level of biosafety to conduct an experiment. Based on their capability and assigned biosafety level, types of experiments that they can do should be limited. For example, a lab assigned to lowest biosafety level cannot handle pathogenic bacteria.
2. Assessment and evaluation of project and pathogenicity of bacteria
This principle assess and evaluate biosafety of project and pathogenicity of bacteria. When assessing a project, one should evaluate potential threat for researcher, public, and ecosystem. Like medicinal project undergoing IRB examination before the experiment, biological experiment should go though series of assessment and evaluation. Biosafety committees and scientists should communicate each other and give them thorough feedback.
3. Bio-security
This principle covers securing biological organism from lost, theft, and misuse. It also deals with regulation after lost, theft, and misuse is happened. To achieve this, bio-security system should be established.

1. Suggestions for safety issue
Safety is the most important part of iGEM competition. Every member of team should know and follow the safety rules. Here are recommendations we give to the future iGEM teams
1. Every member should know about safety rules in the lab. This includes bio-security level, pathogen, gene product, protocol, and etc.
• Each member should know bio-security level of lab. Can lab deal with pathogen? Does the lab have proper equipments for experiment? This kind of consideration is recommended before designing and conducting experiments. Conducting a survey is recommended.
• Each member should be careful if they are dealing with pathogen. How can we prevent disease outbreak? What are safety requirements? Please study about pathogen and related diseases beforehand.
• Each member should consider their gene product. Gene product may be chemically dangerous and may damage ecosystem.
• Each member should be aware of protocols of instruments to avoid any accidents. This is also related to lab results.
2. Communication matters.
Miscommunication leads to accidents and poor results. Results of the experiment should be reviewed and shared by every member of the group. Weekly group discussion is recommended. Each member should know everything about the experiment before one actually does it.


2. How could parts, devices, and systems be made even safer though bio-safety engineering?
• Changing promoter or other regulatory mechanism to shorten the duration of mRNA Changing promoter sequence effectively controls survival time of mRNA and overall production of protein. If mRNA degrades easily, we can control it.
• Adding ‘time bomb’ to kill themselves as time goes. Internal clock bomb will ensure death of our product. So it will be much safer.