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

Risks to the safety and health of team members or others in the lab?

Cadmium is a toxic heavy metal and so all solutions, media, bacteria, etc. containing cadmium must be disposed of as hazardous waste. In the first part of our project the E. coli must be incubated in cadmium-containing media in order to form quantum dots. This part of the project will be performed at Cooper Union in the laboratory for biomedical engineering. The laboratory at the Engineering School at Cooper Union contracts a licensed chemical waste disposal service that is handled by a designated Hazardous Waste Disposal Coordinator/Chemistry Stockroom Technician, who is responsible for scheduling and overseeing the collection of wastes (see link in section 3a). The proper disposal of media containing cadmium salts and the use of personal protective gear, such as nitrile gloves, safety goggles and laboratory coats, will greatly minimize the risks posed to team members.
One major advantage of synthesizing quantum dots biologically, as opposed to chemically, is that it allows the quantum dots to be encapsulated so that the toxic cadmium salts are not interacting directly with the environment. However they are still contain cadmium and the quantum dots must be disposed of as hazardous chemical waste. Therefore, if the resulting cadmium containing quantum dots are used in consumer products, such as electronics items, proper disposal or recycling of such products at the end of their service life must be observed. We are attempting to solve this problem by using other less toxic metals, such as zinc, to synthesize quantum dots rather than the toxic cadmium. If the research is successful, and we succeed in making quantum dots using zinc in bacteria, we will have made the process safer and more ecologically friendly.

Risks to the safety and health of the general public if released by design or accident?

Since all heavy metals will be disposed of using a hazardous waste disposal service, there will be no possibility of cadmium release. Our project utilizes non-pathogenic derivatives of E. coli strain K-12, so in the event of accidental or otherwise release, the health risks are minimal. E. coli K-12 and derivative strains have an outstanding track record regarding human health and safety. They are classified as non-pathogenic and can be safely used in a BSL1 facility.

Risks to environmental quality if released by design or accident?

The risks upon accidental environmental risk are minimal, as described previously, concerning the uses of strains of E. coli K-12. Regarding the expressed peptides, the future deployment of a further disabled strain of bacteria expressing similar proteins, that nucleate the formation of crystals containing toxic metals within the cytoplasm of cells, may be used to bio-remediate contaminated soils. So in that sense, release of the strain may actually improve environmental quality! However, for any project contemplating intentional release of genetically modified organisms, proper assessments regarding long-term safety and subsequent approval by the United States Department of Agriculture, Food and Drug Administration and/or the Environmental Protection Agency, must be adhered to. In our project, the new organism we are engineering will be used for manufacturing quantum dots within a fermenter-type vessel, with subsequentl lysis of the organism in order to purify the quantum dots. Thus, we do not intend to release the viable engineered organism as part of our project.

Risks to security through malicious misuse by individuals, groups or states?

Our BioBricks are not designed to confer any pathogenic attributes to E. coli, nor is there any data to suggest that they pose any detrimental effects on the environment. Furthermore, since the sole known uses of the quantum dot nucleating peptides are to assist in catalyzing the crystallization of salts of cadmium and zinc, we do not anticipate any risk of malicious misuse of this technology by individual, groups or states.

Are any parts or devices in your project associated with(or known to cause):

Pathogenicity, infectivity, or toxicity?

None of the parts or devices that we have used during our project confer any known pathogenicity, infectivity or toxicity to E. coli or humans. The only known function of these expressed peptides is to nucleate the formation of quantum dot forming salts.

Threats to environmental quality?

None of the parts or devices that we have used during our project pose any known threats to environmental quality. Instead, the nucleating peptides are designed to sequester toxic metals.

Security concerns?

None of the parts or devices that we have used during our project raise any known security concerns as they are not designed to confer any known pathogenicity, infectivity, toxicity or environmental damage.

Under what biosafety provisions will / do you operate?

We are operating in laboratories that are designed to be fully compliant with bio-safety level one (BSL1) recommendations, as described in �Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition�.

Does your institution have its own biosafety rules and if so what are they?

Both Cooper Union, where characterization assays are to be performed, and Genspace, where sub-cloning is performed, adhere to biosafety rules that are in accordance with established guidelines found in the publication �Biosafety in Microbiological and Biomedical Laboratories (BMBL) 5th Edition�.

• Genspace Biosafety Rules
• Biosafety and Chemical Disposal Rules for the Kanbar Center for Biomedical Engineering at Cooper Union
• Interim Lab Safety Rules

Does your institution have an Institutional Biosafety Committee or equivalent group?

Genspace has an Advisory Board consisting of experts in biosafety, biosecurity, and genetic engineering who hold positions in academia, industry and government. They serve as our Institutional Biosafety Committee. The portions of the project to be carried out at Genspace are strictly BioBrick construction and the construction of plasmids containing metal-binding peptides and other non-pathogenic sequences. This is within the project parameters recommended by our Advisory Board. Cooper Union does not have an Institutional Biosafety Committee, but the project has been reviewed by David Wootton, Ph.D., Director of The Maurice Kanbar Center for Biomedical Engineering and is being supervised by Jody Grapes, Campus-Wide Safety Officer for Cooper Union.

Describe any concerns or changes that were made based on this review.

No additional concerns or changes to our project were raised.

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

Students participating in this project received safety training (general/chemical/biological) either at Genspace or Cooper Union prior to beginning the project. The safety training consisted of a presentation covering the various aspects of safety found in molecular biology laboratories; i.e. proper microbiological techniques, safe disposal of recombinant organisms, etc. Upon completion of the presentation, students were shown the location of all safety equipment i.e. eye wash stations, first aid kits, fire extinguishers and safety exits. Students were also supervised by iGEM instructors at Genspace or Cooper Union throughout the duration of the project.

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

Guidelines for national bio-safety regulations in the United States via the National Institutes of Health and the Center for Disease Control

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?

Genetically engineered bacteria have the ability to encapsulate harmful contaminants such as heavy metals. They have the ability to separate them from the researcher, the public, and the environment. If we were using this system for bio-remediation rather than for the sole purpose of quantum dot formation, we would potentially require the incorporation of redundant genetic pathways that can function as a �kill-switch� to prevent the unchecked replication of the genetically modified organism in the wild.
Manufacturing processes will be made more �green� by the use of bacteria to manufacture nanotechnology such as quantum dots because the current manufacturing processes rely on various toxic solvents. In addition, the particular issues raised by cadmium containing quantum dots can be mitigated by the substitution of other less toxic metals.