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Angew Chem Int Ed Engl. 2019 Dec 27. doi: 10.1002/anie.201915555. [Epub ahead of print]

DNA Origami Post-Processing by CRISPR-Cas12a.

Author information

1
Department of Cell Biology & Nanobiology Institute, Yale University, 850 West Campus Drive, West Haven, CT, 06516, USA.

Abstract

Customizable nanostructures built through the DNA-origami technique hold tremendous promise in nanomaterial fabrication and biotechnology. Despite the cutting-edge tools for DNA-origami design and preparation, it remains challenging to separate structural components of an architecture built from-thus held together by-a continuous scaffold strand, which in turn limits the modularity and function of the DNA-origami devices. To address this challenge, here we present an enzymatic method to clean up and reconfigure DNA-origami structures. We target single-stranded (ss) regions of DNA-origami structures and remove them with CRISPR-Cas12a, a hyper-active ssDNA endonuclease without sequence specificity. We demonstrate the utility of this facile, selective post-processing method on DNA structures with various geometrical and mechanical properties, realizing intricate structures and structural transformations that were previously difficult to engineer. Given the biocompatibility of Cas12a-like enzymes, this versatile tool may be programmed in the future to operate functional nanodevices in cells.

KEYWORDS:

CRISPR-Cas; DNA nanotechnology; DNA origami; molecular devices; self-assembly

PMID:
31883145
DOI:
10.1002/anie.201915555

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