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

Our project uses strains and reagents that are used in biosafety level 1 (BSL1) laboratories. No special precautions are required, other than those specified for work in laboratories at that level, i.e. no food and drinks permitted in the laboratory, gloves and other appropriate personal protective equipment worn during experiments and any toxic solvents used to be disposed of in accordance with Environmental Protection Agency regulations.

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

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. We are also using the cellulose producing strain Acetobacter xylinum and the naturally occurring fungus, Ganoderma lucidum. These organisms 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 bacterial strains of E. coli K-12, Acetobacter xylinum and the eukaryotic fungus, Ganoderma lucidum. Clearly, 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 bio-polymers, such as cellulose or chitin based derivatives, within a fermenter-type vessel, with subsequent harvesting of the bio-polymers. 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 or A. xylinum, nor is there any data to suggest that they pose any detrimental effects on the environment. Furthermore, since the known uses of cellulose based bio-polymers is primarily as structural material used in biodegradable wound dressings and construction purposes, 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, Acetobacter or humans. The only known function of these expressed enzymes is to catalyze the formation of chitin based bio-polymers.

Threats to environmental quality?

None of the parts or devices that we have used during our project pose any known threats to environmental quality. The end products are also biodegradable.

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?

Genspace, where sub-cloning and culturing is performed, adheres 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
• 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 BioBrick construction, the construction of plasmids containing sequenced required for chitin biosynthesis and other non-pathogenic sequences and the production of biodegradable biopolymers. This is within the project parameters recommended by our Advisory Board.

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 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 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 could 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, if the organisms are intended to be released into the environment.