Journal
ACS SYNTHETIC BIOLOGY
Volume 6, Issue 11, Pages 2108-2117Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.7b00148
Keywords
biofirewall; orthogonal ribosome; genetically modified organisms; biosafety; biocontainment; synthetic biology
Categories
Funding
- Ministry of Science and Technology of China [2014CB745100]
- International S&T Cooperation Program of China [2015DFA00960]
- National Natural Science Foundation of China [21390203, 21621004]
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Biocontainment systems are crucial for preventing genetically modified organisms from escaping into natural ecosystems. Here, we describe the orthogonal ribosome biofirewall, which consists of an activation circuit and a degradation circuit. The activation circuit is a genetic AND gate based on activation of the encrypted pathway by the orthogonal ribosome in response to specific environmental signals. The degradation circuit is a genetic NOT gate with an output of I-SceI homing endonuclease, which conditionally degrades the orthogonal ribosome genes. We demonstrate that the activation circuit can be flexibly incorporated into genetic circuits and metabolic pathways for encryption. The plasmid-based encryption of the deoxychromoviridans pathway and the genome-based encryption of lacZ are tightly regulated and can decrease the expression to 7.3% and 7.8%, respectively. We validated the ability of the degradation circuit to decrease the expression levels of the target plasmids and the orthogonal rRNA (O-rRNA) plasmids to 0.8% in lab medium and 0.76% in nonsterile soil medium, respectively. Our orthogonal ribosome biofirewall is a versatile platform that can be useful in biosafety research and in the biotechnology industry.
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