UT Austin’s 2022 iGEM team is committed to upholding safety. To conduct this year’s project safely, we have taken numerous steps in regards to institutional, lab, and project-specific safety.
UT Austin’s institutional safety guidelines, set by the Institutional Biosafety Committee, allow us to safely conduct experiments in the Barrick lab space. The Institutional Biosafety Committee at our university helps with inspecting the laboratory and reviewing experiments and protocols to ensure that they met biosafety requirements. Additionally, our lab complies with federal and state guidelines. As an added safety measure, masks are recommended in the Barrick lab so as to prevent the spread of COVID-19.
In April 2022, the UT Austin iGEM team attended a lab safety meeting and tour during which members learned the locations of important safety features such as the fire extinguisher, shower, eye wash, and glass and biohazard disposal bins. Online modules regarding hazardous waste management, compressed gases, emergency procedures, cryogen safety, fire extinguisher basics, and general lab safety were also required before members could begin lab work.
Additionally, UT Austin iGEM’s lab is Biosafety Level 2. As a result, members were required to wear close-toed shoes, long pants, and gloves at all times. In addition, sterilization techniques were used whenever working directly with microorganisms. The lab bench and work area were cleaned with ethanol before and after experimentation so as to prevent contamination. Laboratory waste was disposed of appropriately, in either biohazard bags, glass disposal bins, or sharps containers. Flammable chemicals were kept in a cabinet away from Bunsen burners.
Communication was key to upholding lab safety. Team members used Slack was to communicate on a daily basis and to delegate logistical tasks to specific people. In addition, Benchling was used to log daily experiments and to keep track of where certain organisms or DNA products are stored.
Fig. 1. A section of our strain database
Fig. 2. A section of our primer database
ARROWE’s defining feature is Acinetobacter baylyi (ADP1) as a chassis. We chose ADP1 since it is a non-pathogenic, non-white-listed, BSL-1 soil bacterium that can easily be stored in a –80 C freezer [1]. Because ADP1 is non-pathogenic, ADP1 is safe to culture in our lab space, which is BSL-2. If any ADP1 were to be released into the environment accidentally, there would be no safety risk to humans or other organisms since ADP1 lives naturally in the soil (add citation). Additionally, dual-use hazards are unlikely.
Our design for ARROWE involves the tdk/kan cassette, YFP repressor genes, and P. destructans homologies. These parts are deemed safe since they do not confer antibiotic resistance, or give the intrinsic ADP1 any dangerous or pathogenic properties.
The goal of ARROWE is to detect P. destructans, a BSL-2 fungi that causes White Nose Syndrome in bats. Because P. destructans is pathogenic and could cause contamination or pose a danger to animals if accidentally released to the environment, we chose not to work directly with environmental samples of it. Rather, we have asked Dr. Jeffrey Barrick, our PI, to culture P. destructans in a different lab space and isolate its genomic DNA for us to use. Genomic DNA from P. destructans, as well as the P. destructans itself, were ordered from ATCC. We have also discussed with Dr. Nate Fuller, a bat expert from Texas Parks and Wildlife, about managing and preventing risks associated with White Nose Syndrome within the lab space.
Working directly with bats also poses extreme danger. As discussed with Dr. Nate Fuller, bats can carry a variety of diseases and transmit them to humans. Working with bats and collecting samples from them requires specific, full-body personal protective equipment and training we do not have access to. As a result, we were not able to acquire environmental samples. Though working with environmental samples was beyond the scope of our project, we were still able to safely demonstrate the efficacy and potential of ARROWE, on our Proof of Concept and Implementation pages respectively.
[1] Metzgar, D., Bacher, J. M., Pezo, V., Reader, J., Doring, V., Schimmel, P., Marliere, P., & de Crecy-Lagard, V. (2004). Acinetobacter sp.. ADP1: An ideal model organism for genetic analysis and Genome Engineering. Nucleic Acids Research, 32(19), 5780–5790. https://doi.org/10.1093/nar/gkh881
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