The safety page is where users can see the safety precautions we took in lab. This includes sterilization techniques, lab rules, and biosafety ratings.

Safety Questions

1. Description of the Chassis organism(s) we used for this project

We used three different strains of ''E. coli'' including BL21, XL-Blue, and BM2.5. These were all Risk Group 1. ''E. coli'' are a human pathogen which causes irritation if it makes contact with skin or eyes. If the bacteria is injected, it can also cause irritation to the respiratory tract and kidneys. Below is the link for the risk assessment of our bacteria:

http://www.absa.org/riskgroups/bacteriasearch.php?genus+&species+coli

2. Our highest Risk Group used was Group 1

3. List of all new coding regions used in this project:

	Part Number	Where did you get the physical DNA for this part?	What species does this part originally come from?	What is the Risk Group of the species?	What is the function of this part, in its parent species?
1	BBa_K1225000	IDT	E. coli	1	Promoter
2	BBa_K1225001	IDT	E. coli	1	Promoter
3	BBa_K1225002	IDT	E. coli	1	Promoter
4	BBa_K1225003	IDT	E. coli	1	Promoter
5	BBa_K1225004	IDT	E. coli	1	Promoter

4. Do the biological materials used in your lab work pose any of the following risks?
1. Risks to the safety and health of team members or others working in the lab?
1. None of the biological materials pose risk to members. We worked only with GFP as a gene of interest with regulatory parts that do not function in isolation. We are not adding any viral DNA, toxins, or carcinogens that would pose a threat to the researchers. GFP has been used repeatedly and is well documented without evidence of risk to the safety of those who work with it directly.
2. Risks to the safety and health of the general public, if released by design or by accident?
1. None of the regulatory parts would be functional in isolation. The bacteria expressing these genes do not show viability changes and are not viable outside of the lab environment. GFP has not been shown to cause a safety or health risk to the general public, and our regulatory DNA has not shown any indication that this would change.
3. Risks to the environment, if released by design or by accident?
1. The primary risk to the environment would be the uptake of the DNA by other bacteria; however, because our gene of interest is GFP, this is not a cause for concern as the result would be the bacteria fluorescing green. The use of GFP has not been shown in the past to pose a risk to the environment by design or accident. If the regulatory genes were released, the result would be eventual degradation of the DNA as they do not function in isolation.
4. Risks to security through malicious misuse by individuals, groups, or countries?
1. The misuse of our parts would have to include the individuals, groups or countries inserting their own gene of interest as no toxins, viral DNA, or carcinogens are currently present. The parts submitted can be used as any other regulatory part found in the Registry by anyone with access.
5. 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.)
1. GFP would not be the commercial/industrial product. The parts would be used to regulate the expression of the product by the bacteria. There would be little to no risk to the environment, workers, or general public based solely on the regulatory parts; however, the new gene of interest would be consistently expressed at a given level based on the part selected. This could cause problems if released as there is no current “off” or kill switch in the regulatory machinery. Based on the new gene of interest, the risk level would have to be assessed. Other factors to be considered in a scale-up is the viability of the bacteria outside of the lab/production center, the safety of individuals using bacteria in large quantities that can cause irritation if contacted or ingested, and how the introduced GOI affects the viability and pathogenicity of the bacteria.
6. 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.
1. No, we do not have any design features to address safety risks.
7. 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.
1. To work in our lab, Bindley Bioscience Center, we all had to go through Biological Safety Training, Laboratory Chemical Safety Training, as well as Personal Protective Equipment Training. The link for all three is:http://www.purdue.edu/rem/home/files/training.htm We also had to go through individual training for each of the lab equipment that was used such as centrifuges, autoclaving, and PCR machine.
8. Under what biosafety provisions do you work?
1. Please provide a link to your institution biosafety guidelines.
1. Purdue University has a biosafety manual that includes all of the guidelines that researchers must follow. It can be found at:http://www.purdue.edu/rem/home/booklets/bioman.pdf
2. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you made to your project plan based on their review.
1. Yes, Purdue University has an Institutional Biosafety Committee or IBC. All research proposals and experiments must be approved by the IBC if it includes “working with an organism or product of an organism that presents a risk to humans.” (http://www.purdue.edu/research/vpr/rschadmin/rschoversight/rdna/)Because our project did not include either, we did not discuss our project with them.
3. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible.
1. Yes, there are two main national biosafety regulations and guidelines to follow. One is NIH, or National Institute of Health, as well as the CDC, or Center for Disease Control. The link for NIH is: http://oba.od.nih.gov/oba/rac/Guidelines/NIH_Guidelines.htm. The link for CDC is: http://www.cdc.gov/biosafety/.
4. 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 2013.igem.org/Safety for help].
1. Our lab is a BioSafety Level of 1.
5. What is the Risk Group of your chassis organism(s), as you stated in question 1? If it does not match the BSL rating of your laboratory, please explain what additional safety measures you are taking.
1. Our chassis organisms are all Risk Group 1.

Researcher safety

For the majority of the project that we worked on there was limited safety issues that came up while working in lab. The parts that we worked with were parts that had previously been submitted in the parts registry particularly when working on the Robustness and standardization projects. The parts that we selected were chosen based on their availability and usefulness. These parts were primarily promoters, RBS, and terminators which are naturally found in E. coli. The reporter of interest was green fluorescent protein which although is not native in E. coli; however, it has been used for years in E. coli. When utilizing the BCDs, there was no immediate threat to the students and researchers. These were parts that were only used to regulate the amount of red fluorescent protein that was being expressed. As far as safety for our team while carrying out lab procedures, everyone was properly trained by an expert before using any and all laboratory equipment. Extensive training was performed before any team members were allowed to work in lab (see training certification). This ensured that everyone was aware of safety procedures such as handling biosafety level 1 material, disposal of waste materials, and proper lab equipment. Because we work in a shared lab space, special caution was taken to label every reagent, chemical, and solution made in lab. Placement of such materials were specified and labeled as well. This was in the case that something was dropped, misplaced, or handled by a researcher not in our lab group; they would be able to react appropriately. A buddy system was also put into place so that no one worked in lab by themselves during or after lab hours. This ensured that if an accident were to occur, help would be at hand.

Public safety

Because the projects that this year’s team took on do not have a direct link to the public, it was difficult to analyze the safety impact on the public. We focused on creating means to make engineering biology more of a reality; however, because synthetic biology is still at a young stage in this field’s history, the direct impact of what we have worked on cannot yet be estimated. The human practices aspect of our project delves into the deeper impact that we hope this project will have. However, as far as a direct impact in the near future, there is not a foreseeable public safety issue.

Environmental Safety

The biggest issues that came up over the course of this project that directly impacted the environment was the disposal of lab materials and chemicals. This was done through autoclaving waste and disposal through REM specifications. As far as the project outcomes affecting the environment, just as with the public safety issue there are no short term foreseeable safety issues that need to be addressed. Any issue that would arise would be an indirect use of our project to advance the field of synthetic biology. This was also covered in our human practices portion of our project.