Team:Michigan/Safety

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Safety

1. Would the materials used in your project and/or your final product pose:

• a. Risks to the safety and health of team members or others in the lab?
• b. Risks to the safety and health of the general public if released by design or accident?
• c. Risks to environmental quality if released by design or accident?
• d. Risks to security through malicious misuse by individuals, groups or states?

a. Ethidum bromide is used as an intercalating agent and a dye when testing double stranded DNA. While used in highly dilute solutions, this compound possesses some acute toxicity and is handled in its stock form with nitrile gloves and stored under a fume hood. All liquid waste contacting this compound is disposed of in dedicated jugs, and solid waste collected in marked containers as required by the University of Michigan.

Hydrochloric acid (HCl) is used sparingly in 1M and 6M concentrations to clean DNA from cuvettes and purification columns. Concentrated solutions of HCl are generally handled with long sleeves, gloves and stored in either a fume hood with diaper pads or a locking, latching acid storage cabinet. All acid waste is collected in a separate, labeled disposal jug.

Sodium Hypochlorite is used in household concentrations as a means of destroying organisms and organic material before disposal. This compound is handled with gloves on a dedicated bench in small amounts.

Solutions and plates containing rich media such as LB, SOB, and SOC are frequently used for the growth of E.Coli for purification and DNA extraction. While the strains of E.Coli used are not pathogenic and listed as biosafety level 1 laboratory environment, leftover stocks and plates are still autoclaved and disposed of separately from other chemical waste.

Bunsen burners are used in conjunction with flammable 70% ethanol solutions for the purpose of sterilizing certain laboratory implements such as plate spreaders and inoculation pics; such methods are capable of causing burns if misused.

b. Recombinant strains of E.Coli containing resistance to various antibiotics are likely the greatest threat to the general public, as antibiotic resistance can potentially spread to pathogenic wild-type bacteria through horizontal gene transfer. Antibiotics commonly used in the lab include ampicillin and kanamycin, both of which are used in public health to treat infections. The strains of E.Coli used in this process are non-pathogenic and pose little risk to those handling them, let alone public health

Hydrochloric acid in the concentrations used is potentially dangerous to the public if mishandled. HCl possesses both caustic and reactive properties and is capable of evolving gaseous chlorine on contact with certain compounds, as well as producing certain harmful inorganic salts such as copper (I) chloride.

c. Rich media such as LB, if released in the environment in large quantities, has the ability to disrupt ecosystems through the facilitation of abnormal bacterial and fungal growth.

Hydrochloric acid, if released into the environment, poses little long-term hazard due to its reactivity and subsequent short half-life. However, it still possess the potential to alter ecosystems by changing the chemical makeup of soils and groundwater if released in sufficient quantities.

Ethidium bromide is utilized in minuscule quantities and poses little environmental danger at its functional concentration.

d. Large amounts of concentrated hydrochloric acid can potentially be used to produce more toxic compounds such as gaseous chlorine and mustard gas and be used as weapons, if the individuals or states in question possess sufficient knowledge and equiptment. Bacteria in the lab are non-pathogenic and cannot be used to make bioweapons without extensive modification.

Please explain your responses (whether yes or no) to these questions.

Specifically, are any parts or devices in your project associated with(or known to cause):

- pathogenicity, infectivity, or toxicity?

- threats to environmental quality?

- security concerns?

Due to our level of lab safety, we are unable to work with strains that are associated with pathogenicity, infectivity, and toxicity. The lab strains we used, DH5a and BL21DE3, are nonpathogenic.

Although our project involves cells binding to DNA, there is very little risk of double-stranded DNA located in the outside binding to our cells. Thus, threats to environmental quality are low.

Because our strains are nonpathogenic and do not pose a serious threat to the environment, we believe that our project does not pose any security concerns.

2. If your response to any of the questions above is yes:

a. Explain how you addressed these issues in project design and while conducting laboratory work.

b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry.

3. Under what biosafety provisions will / do you operate?

a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible.

b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review.

c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.

d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.

4. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

1. Would any of your project ideas raise safety issues in terms of:

• researcher safety,
• public safety, or
• environmental safety?

Overall, our project is very safe. If we wanted to allow the public to look at our cell patterns, we would have to take precautions to ensure that any viewers would be safe. We have discussed this with the company we get our microarrays from, and we are sure that the microarray slides themselves are safe to work with.

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?

There are no major safety issues involved with our part. Even though the zinc finger binding domain binds to double-stranded DNA, it will not mutate the DNA in any shape or form (unlike ethidium bromide that goes between the bases of DNA). Additionally our protein that contains the zinc finger is inert, no nuclease activity.

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?

We take safety seriously on our team. All of our members are required to attend training sessions provided by the Occupational Safety and Environmental Health [http://www.oseh.umich.edu/] administration. These sessions deal with General Lab Safety and Autoclave Training. Our team has had meetings throughout the year with medical faculty to discuss biosafety. We follow very closely to the guidelines in the Biosafety in Microbiological and Biomedical Laboratories 5th Edition [http://books.google.com/books?hl=en&lr=&id=QFHUzRoDFt4C&oi=fnd&pg=PA1&dq=Biosafety+in+Microbiological+and+Biomedical+Laboratories+&ots=iW_LtVAF73&sig=m6ke00oT800OJYpcJ5V4c5Nv1qY#v=onepage&q&f=false] Moreover, we adhere to the OSEH guidelines [http://www.oseh.umich.edu/] tailored specially to the University of Michigan.

4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

All iGEM teams should go through mandatory lab safety training at a minimum. In order to make parts safer, we could require that each part pass the approval of a “safety committee.” This is not ideal however, because it would slow down the cloning process and would be expensive in terms of time and cost to implement. We could also prepare tests that each part could go through to ensure that they meet safety standards.