A robust yeast biocontainment system with two-layered regulation switch dependent on unnatural amino acid

Tiantian Chang, Weichao Ding, Shirui Yan, Yun Wang, Haoling Zhang, Yu Zhang, Zhi Ping, Huiming Zhang, Yijian Huang, Jiahui Zhang, Dan Wang, Wenwei Zhang, Xun Xu, Yue Shen () and Xian Fu ()
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Tiantian Chang: University of Chinese Academy of Sciences
Weichao Ding: University of Chinese Academy of Sciences
Shirui Yan: BGI Research
Yun Wang: BGI Research
Haoling Zhang: BGI Research
Yu Zhang: BGI Research
Zhi Ping: BGI Research
Huiming Zhang: University of Chinese Academy of Sciences
Yijian Huang: University of Chinese Academy of Sciences
Jiahui Zhang: University of Chinese Academy of Sciences
Dan Wang: Guangdong Provincial Key Laboratory of Interdisciplinary Research and Application for Data Science, BNU-HKBU United International College
Wenwei Zhang: BGI Research
Xun Xu: BGI Research
Yue Shen: University of Chinese Academy of Sciences
Xian Fu: BGI Research

Nature Communications, 2023, vol. 14, issue 1, 1-13

Abstract: Abstract Synthetic auxotrophy in which cell viability depends on the presence of an unnatural amino acid (unAA) provides a powerful strategy to restrict unwanted propagation of genetically modified organisms (GMOs) in open environments and potentially prevent industrial espionage. Here, we describe a generic approach for robust biocontainment of budding yeast dependent on unAA. By understanding escape mechanisms, we specifically optimize our strategies by introducing designed “immunity” to the generation of amber-suppressor tRNAs and developing the transcriptional- and translational-based biocontainment switch. We further develop a fitness-oriented screening method to easily obtain multiplex safeguard strains that exhibit robust growth and undetectable escape frequency (

Date: 2023
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DOI: 10.1038/s41467-023-42358-4

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