Advisor: Corey J. Wilson, Ph.D. (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology)

Committee:

John Blazeck, Ph.D. (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology)

Hang Lu, Ph.D. (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology)

Ravi Kane, Ph.D. (School of Chemical and Biomolecular Engineering, Georgia Institute of Technology)

Richard M. Murray, Ph.D. (Division of Biology and Biological Engineering, California Institute of Technology)

Next-Generation Biosecurity: Advancing Tools for Multi-Layer Security

The rapid expansion of synthetic biology from laboratory spaces to industrial and healthcare sectors has brought about concerns of unwanted release of genetically modified organisms and protection of biological intellectual property. Current physical and operational security measures for these assets can be comprised by theft or accidental release. This thesis proposes the designing, building, and testing of a next-generation biosecurity system that is intrinsic to the engineered organism. While prior studies have demonstrated basic intrinsic security measures, there is considerable value in advancing the state of the art with novel tools, multi-layer security platforms, and the introduction of penetration testing. First, an 8-digit biological keypad will be developed in Escherichia coli for 2-input and 3-input passcodes to protect a biological asset. This technology will be combined with toxin-based penalties to design a first-of-its-kind Biohackathon challenge. Furthermore, a novel ‘Command Center’ for editing biological programs will be shown in situ using an engineered E. coli strain. The Command Center would be a promising tool for biosecurity applications. Finally, an authentication system will be engineered through synthetic auxotrophy, and will be deployed with other biosecurity measures to a design a second Biohackathon challenge. Together, these technologies will establish a foundation for integrating multi-layer biosecurity into biotechnology.