Team:Penn/Safety

Safety

Do the biological materials used in your lab work pose any of the following risks? Please describe.

a. Risks to the safety and health of team members or others working in the lab?
We work with SYBR^TM Safe DNA Gel Stain which is not classified as hazardous waste under US federal regulations. As far as bacteria, none of our strains are above level 1. Our team uses gloves and exercises caution when working in the lab in order to reduce risks to our health.

b. Risks to the safety and health of the general public, if released by design or by accident?
The strain of E. coli we use should not pose any risks to the safety and health of the general public' our plasmids do not produce toxic protein.

c. Risks to the environment, if released by design or by accident?
Our plasmid is made so that expression is regulated by aT7 polymerase; it should not create risks to the environment if accidentally released.

d. Risks to security through malicious misuse by individuals, groups, or countries?
We do not foresee the usefulness of our systems to individuals, groups or countries with malicious intents.

If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? (Consider the different categories of risks that are listed in parts a-d of the previous question.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? (Note: This is meant to be a somewhat open-ended discussion question.)
Since our system is an assay it should not pose risks if it becomes a widely used tool, it does however lend itself to the development of tools which could, if developed and used inappropriately, create harmful methylation patterns in organisms.

Does your project include any design features to address safety risks? (For example: kill switches, auxotrophic chassis, etc,) Note that including such features is not mandatory to participate in iGEM, but many groups choose to include them)
Yes, our main project is designing a new assay to test for off target methylation. We are designing a one plasmid system that will help ascertain whether or not off target methylation occurs and the extent to which it occurs.

What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution's safety training requirements, if available.
All lab members have gone through a rigorous Penn environmental health and safety training course and are certified to work with hazardous substance, infectious agents and human source materials.

Please provide a link to your institution biosafety guidelines.
http://www.ehrs.upenn.edu/programs/bio/

Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed project with them? Describe any concerns they raised with your project, and any changes you made to your project based on their review.
We have not discussed our project with our institution's biosafety committee.

Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible.
http ://www. absa. org/resbslinks. Html

According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? (Check the summary table on page 3, and the fuller description that starts on page 9.) If your lab does not fit neatly into category 1,2,3, or 4, please describe its safety features [see 20l3.igem.org/Safety for help].
1

What is the Risk Group of your chassis organism(s), as you stated in question l? If it does not match the BSL rating of your laboratory, please explain what additional safety measures you are taking.
1


Our team also filled out a second safety form since we are using a part from Spiroplasma sp. Strain MQ1 which is a risk group 2 organism, bellow is our response to the questions:

If you are using this organism as a chassis, write "chassis". If you are using a genetic part from the organism, give the name of the part and a brief description of what it does and why you are using it.
Name of part: M.SssI Methyltransferase Gene. This enzyme methylates CpG sites in DNA. We are using it as part of a set of targeted methylatransferase fusion proteins.

How did you physically acquire the organism or part?
We synthesized the gene from Integrated DNA Technologies.

What potential safety/health risks to team members, other people at your institution, or the general public could arise from your use of this organism/part?
The part is non toxic, it should not raise any health or safety risks.

What measures do you intend to take to ensure that your project is safe for team members, other people at your institution, and the general public?
Standard safety protocols were effectively used to ensure the containment of biologically hazardous agents, we do not work with any pathogenic bacteria in the lab.

If you are using only a part from the organism, and you believe the part by itself is not dangerous, explain why you believe it is not dangerous.
It is not dangerous because it is not toxic. Most cells have their own mechanisms for methylating DNA and it is unlikely that our gene would be transferred into other cells, but even if it were, it is difficult to imagine that it would be dangerous.

Why do you need to use this organism/part? Is there an organism/part from a less dangerous Risk Group that would accomplish the same purpose?
We chose this part because it is the most commonly and most readily available CpG Methyltransferase that has been expressed in E. coli.

Is the organism/part listed under the Australia Group guidelines, or otherwise restricted for transport? If so, how will your team ship this part to iGEM and the Jamborees?
No.

Please describe the BioSafety Level of the lab in which the team works, or description of safety features of lab (Refer to Basic Safety form, question 8. d.). If you are using organisms with a BSL level greater than you lab, please explain any additional safety precautions you are taking.
Level 1. No hazardous organisms.