Circular single-stranded DNA as a programmable vector for gene regulation in cell-free protein expression systems

Zhijin Tian, Dandan Shao, Linlin Tang, Zhen Li, Qian Chen, Yongxiu Song, Tao Li, Friedrich C. Simmel and Jie Song ()
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Zhijin Tian: University of Science & Technology of China
Dandan Shao: Shanghai Jiao Tong University
Linlin Tang: Chinese Academy of Sciences
Zhen Li: Shanghai Jiao Tong University
Qian Chen: Northeast Forestry University
Yongxiu Song: Chinese Academy of Sciences
Tao Li: University of Science & Technology of China
Friedrich C. Simmel: Technische Universität München
Jie Song: Chinese Academy of Sciences

Nature Communications, 2024, vol. 15, issue 1, 1-11

Abstract: Abstract Cell-free protein expression (CFE) systems have emerged as a critical platform for synthetic biology research. The vectors for protein expression in CFE systems mainly rely on double-stranded DNA and single-stranded RNA for transcription and translation processing. Here, we introduce a programmable vector - circular single-stranded DNA (CssDNA), which is shown to be processed by DNA and RNA polymerases for gene expression in a yeast-based CFE system. CssDNA is already widely employed in DNA nanotechnology due to its addressability and programmability. To apply above methods in the context of synthetic biology, CssDNA can not only be engineered for gene regulation via the different pathways of sense CssDNA and antisense CssDNA, but also be constructed into several gene regulatory logic gates in CFE systems. Our findings advance the understanding of how CssDNA can be utilized in gene expression and gene regulation, and thus enrich the synthetic biology toolbox.

Date: 2024
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DOI: 10.1038/s41467-024-49021-6

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