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

• Type of Material: Article
• Authors: Linlin Tang, Shanghia 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, Emory University; Friedrich C. Simmel, Technische Universität München; Jie Song, Shanghia Jiao Tong University
• Language: English
• Date: 2023-10-20
• Publisher: Springer Nature
• Publication Version: Final Published Version
• Copyright Statement: © The Author(s) 2023
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): https://doi.org/10.1038/s41467-023-42437-6
• Title of Journal or Parent Work: Nature Communications
• Volume: 14
• Start Page: 6665
• Grant/Funding Information: The authors are grateful for the financial support from the National Natural Science Foundation of China (Nos. 22161132008, to J.S.), the National Key Research and Development Program of China (Grant No. 2021YFF1200200, to J.S.), and the Starry Night Science Fund of Zhejiang University Shanghai Institute for Advanced Study (SN-ZJU-SIAS-006, to J.S.). F.C.S. acknowledges support through the Deutsche Forschungsgemeinschaft (DFG SI 761/5-1) and the Max Planck School Matter to Life.
• Keywords: Gene regulation; Synthetic biology; DNA nanotechnology; Expression systems
• Research Categories: Biology, Genetics; Biology, Molecular

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.