We also discussed safety of our team in <a href="Javascript:loadContent('Humanpractice-block','Humanpractice-2')">human practice page</a>.
We have not had any safety training officially, but have been taught by learned people.
b) Please briefly describe the topics that you learned about (or will learn about) in your safety training.
We learned about techniques for preventing diffusion of Escherichia coli or other organisms including ogenetically modified organisms into environment and risks concerning DNA assembly experiments.
c) Please give a link to the laboratory safety training requirements of your institution (college, university, community lab, etc). Or, if you cannot give a link, briefly describe the requirements.
Division for Environment, Health and Safety (http://www.adm.u-tokyo.ac.jp/office/anzeneisei/index.html) in our university is responsible for training laboratory safety.
Taking lab safety training course is not mandatory, but strongly recommended for those who are involved in experiments. However, this course is for the graduate school students and senior stuffs, and not open to undergraduate students. We thus had a training directly from the PI.
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| Species name(including strain) | Risk Group | Risk Group Source</td><th>Disease risk to humans?</td><th>Part number/name | <th>Natural function of part</th><th>How did you acquire it?</th><th>How will you use it?</th><th>Notes</th>
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<tr><td>Escherichia coli JM109</td><td>1</td><td>DSMZ</td><td>no</td><td></td><td></td><td>from our Lab</td><td>DNA asssembly</td><td></td></tr>
<tr><td>Escherichia coli MG1655</td><td>1</td><td>DSMZ</td><td>no</td><td></td><td></td><td>from our Lab</td><td>Assay</td><td></td></tr>
<tr><td>Pseudomonas fluorescens</td><td>2</td><td>http://www.absa.org/riskgroups/bacteriasearch.php?genus=Pseudomonas</td><td>yes</td><td>sigma factor</td><td>polymerize RNA with RNA polymerase</td><td>order the part DNA from a synthesis company</td><td>transcriptional control</td><td>The bacteria causes opportunistic infection and it affects with usually patients with compromised immune systems.</td></tr>
<tr><td>Pseudomonas protegens</td><td>1</td><td>http://www.dsmz.de/catalogues/details/culture/DSM-19095.html</td><td>no</td><td>anti sigma factor</td><td>prevent combination between RNA polymerase and specific sigma factor</td><td>order the part DNA from a synthesis company</td><td>transcriptional control</td><td></td></tr>
<tr><td>Pseudomonas syringae</td><td>1</td><td>DSMZ</td><td>no</td><td>sigma factor</td><td>polymerize RNA with RNA polymerase</td><td>order the part DNA from a synthesis company</td><td>transcriptional control</td><td></td></tr>
<tr><td>Pseudomonas syringae</td><td>1</td><td>DSMZ</td><td>no</td><td>anti sigma factor</td><td>prevent combination between RNA polymerase and specific sigma factor</td><td>order the part DNA from a synthesis company</td><td>transcriptional control</td><td></td></tr>
<tr><td>Chlorocebus aethiops COS-1</td><td>1</td><td>DSMZ</td><td>no</td><td></td><td></td><td>receive the cells from another lab</td><td>Assay</td><td></td></tr>
<tr><td>Chlorocebus aethiops COS-7</td><td>1</td><td>DSMZ</td><td>no</td><td></td><td></td><td>receive the cells from another lab</td><td>Assay</td><td></td></tr>
<tr><td>Homo sapiens HL-60</td><td>1</td><td>DSMZ</td><td>no</td><td></td><td></td><td>receive the cells from another lab</td><td>Assay</td><td></td></tr>
<tr><td>Homo sapiens oral epithelial cell</td><td>1</td><td>http://www.lifescience.mext.go.jp/bioethics/data/anzen/syourei_02.pdf</td><td>no</td><td>EGP2 promoter</td><td>the promoter of EpCAM</td><td>from the member of our team</td><td>transcriptional control</td><td></td>
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Risks of Your Project Now
Please describe risks of working with the biological materials (cells, organisms, DNA, etc.) that you are using in your project. If you are taking any safety precautions (even basic ones, like rubber gloves), that is because your work has some risks, however small. Therefore, please discuss possible risks and what you have done (or might do) to minimize them, instead of simply saying that there are no risks at all.
a) Risks to the safety and health of team members, or other people working in the lab
When we are exposed to E. coli cells, we have a chance to have irritation in our eyes, skin, and respiratory system. Furthermore, ethidium bromide which we use to detect DNA band is carcinogen. We also use mammalian cells in assay. These cells can be infected by viruses which can also infect us. We also use harmful reagent, such as membrane binding solution.
b) Risks to the safety and health of the general public (if any biological materials escaped from your lab):
As maintained above, our lab has harmful organisms and substances. If they diffused outside, there might be health hazard.
c) Risks to the environment (from waste disposal, or from materials escaping from your lab)
We might dispose chips or tubes which contain a bit amount of genetically modified organisms without sterilizing. It offenses the law of our country; they may effect on biodiversity in our country. The bacteria we used also have drug resistance against ampicillin or CP, and if escaped, they could not be killed by those drugs. Mammalian cells are so weak that they can hardly live in the natural world.
d) Risks to security through malicious mis-use by individuals, groups, or countries
There seems to be no risks with this subject. In today's cognition, parts that we use do not seem to lead the expression of harmful materials.
e) What measures are you taking to reduce these risks? (For example: safe lab practices, choices of which organisms to use.)
In order to avoid the risks about bacterial cells shown above, we autoclave all wastes, sterilize all equipments that contain the organisms with detergent or hypochlorous acid. When bacteria exposed outside the tubes or examiners, we immediately seterilize them with ehtanol. Furthermore, in order to keep ethidium bromide away from our skin, we use kimwipes to wrap the chips used to taking it up before throwing away, and we take care that our bare hands do not touch the gel polluted by the substance.
Risks of Your Project in the Future
What would happen if all your dreams came true, and your project grew from a small lab study into a commercial/industrial/medical product that was used by many people? We invite you to speculate broadly and discuss possibilities, rather than providing definite answers. Even if the product is "safe", please discuss possible risks and how they could be addressed, rather than simply saying that there are no risks at all.
a) What new risks might arise from your project's growth? (Consider the categories of risk listed in parts a-d of the previous question: lab workers, the general public, the environment, and malicious mis-uses.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available?
In sigma-Recounter project, the circuit we constructed has potential for the application for defeating pests or bacteria by means of the expression of toxic proteins. In this case, there is a risk that you mistakenly output the toxin.
In CTCD project, the role of the circuits is to detect CTCs by GFP, thus it is thought to be difficult to have a risk of doing humans or environment harm.
b) Does your project currently include any design features to reduce risks? Or, if you did all the future work to make your project grow into a popular product, would you plan to design any new features to minimize risks? (For example: auxotrophic chassis, physical containment, etc.) Such features are not required for an iGEM project, but many teams choose to explore them.
In the future study of sigma-Recounter project, in addition to the reset system, we intend to increase the number of nodes and to enable one state to move to any other states. Therefore, if our project is used to express a toxin to defeat pest or bacteria, you can prepare an anti-toxin node or a reset system for mistakenly expressing the toxin.
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