DNA Origami-Based Single-Molecule CRISPR Machines for Spatially Resolved Searching
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作者
Hao, Yaya; Li, Mingqiang; Zhang, Qian; Shi, Jiye; Li, Jiang; Li, Qian; Fan, Chunhai; Wang, Fei
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刊物名称
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
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年、卷、文献号
2022, 61, 1433-7851
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关键词
Hao, Yaya; Li, Mingqiang; Zhang, Qian; Shi, Jiye; Li, Jiang; Li, Qian; Fan, Chunhai; Wang, Fei
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摘要
Repurposing the RNA-guided endonuclease Cas9 to develop artificial CRISPR molecular machines represents a new direction toward synthetic molecular information processing. The operation of CRISPR-Cas9-based machines, nevertheless, relies on the molecular recognition of freely diffused sgRNA/Cas9, making it practically challenging to perform spatially regulated localized searching or navigation. Here, we develop a DNA origami-based single-molecule CRISPR machine that can perform spatially resolved DNA cleavage via either free or localized searching modes. When triggered at a specific site on the DNA origami with nanoscale accuracy, the free searching mode leads to searching activity that gradually decays with the distance, whereas the localized mode generates spatially-confined searching activity. Our work expands the function of CRISPR molecular machines and lays foundations to develop integrated molecular circuits and high-throughput nucleic acid detection.
