Circular single-stranded DNA as switchable vector for gene expression in mammalian cells

Linlin Tang, Zhijin Tian, Jin Cheng, Yijing Zhang, Yongxiu Song, Yan Liu, Jinghao Wang, Pengfei Zhang, Yonggang Ke (), Friedrich C. Simmel () and Jie Song ()
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Linlin Tang: Shanghai Jiao Tong University
Zhijin Tian: Chinese Academy of Sciences
Jin Cheng: Shanghai Jiao Tong University
Yijing Zhang: Chinese Academy of Sciences
Yongxiu Song: Chinese Academy of Sciences
Yan Liu: Shanghai Jiao Tong University
Jinghao Wang: Chinese Academy of Sciences
Pengfei Zhang: Chinese Academy of Sciences
Yonggang Ke: Georgia Institute of Technology and Emory University
Friedrich C. Simmel: Technische Universität München
Jie Song: Shanghai Jiao Tong University

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

Abstract: Abstract Synthetic gene networks in mammalian cells are currently limited to either protein-based transcription factors or RNA-based regulators. Here, we demonstrate a regulatory approach based on circular single-stranded DNA (Css DNA), which can be used as an efficient expression vector with switchable activity, enabling gene regulation in mammalian cells. The Css DNA is transformed into its double-stranded form via DNA replication and used as vectors encoding a variety of different proteins in a wide range of cell lines as well as in mice. The rich repository of DNA nanotechnology allows to use sort single-stranded DNA effectors to fold Css DNA into DNA nanostructures of different complexity, leading the gene expression to programmable inhibition and subsequently re-activation via toehold-mediated strand displacement. The regulatory strategy from Css DNA can thus expand the molecular toolbox for the realization of synthetic regulatory networks with potential applications in genetic diagnosis and gene therapy.

Date: 2023
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DOI: 10.1038/s41467-023-42437-6

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