Article

DNA Origami Gatekeepers for Solid-State Nanopores

Walter Schottky Institute, Technische Universität München, Munich, Germany.
Angewandte Chemie International Edition (Impact Factor: 11.26). 05/2012; 51(20):4864-7. DOI: 10.1002/anie.201200688
Source: PubMed

ABSTRACT

DNA has it covered: DNA origami gatekeeper nanoplates convert nanopores in solid-state membranes into versatile devices for label-free macromolecular sensing applications. The custom apertures in the nanoplates can be chemically addressed for sequence-specific detection of DNA.

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    • "This approach paves the way for the fabrication and sensing applications of the single-molecule nanopore sensors based on DNA nanostructures. At the same time, another research group also reported the synthesis of DNA origami gatekeepers and the application as solid-state nanopores[181]. The fabricated electrical nanopore sensor can be utilized for the label-free sensing of DNA molecules. Linear virus can be served as the excellent 1D template to connect the electrodes for the fabrication of electrical sensors. "
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    • "DNA nanotechnology is leading to various applications in structural biology (17), biophysics (18–20), biosensing (21–24) and therapeutic delivery (25–28), many of which entail the preparation of micrograms to milligrams of enriched high-quality DNA nanostructures. Traditional electrophoresis-based methods excel at separation resolution but often fall short in scalability. "
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    ABSTRACT: Most previously reported methods for purifying DNA-origami nanostructures rely on agarose-gel electrophoresis (AGE) for separation. Although AGE is routinely used to yield 0.1–1 µg purified DNA nanostructures, obtaining >100 µg of purified DNA-origami structure through AGE is typically laborious because of the post-electrophoresis extraction, desalting and concentration steps. Here, we present a readily scalable purification approach utilizing rate-zonal centrifugation, which provides comparable separation resolution as AGE. The DNA nanostructures remain in aqueous solution throughout the purification process. Therefore, the desired products are easily recovered with consistently high yield (40–80%) and without contaminants such as residual agarose gel or DNA intercalating dyes. Seven distinct three-dimensional DNA-origami constructs were purified at the scale of 0.1–100 µg (final yield) per centrifuge tube, showing the versatility of this method. Given the commercially available equipment for gradient mixing and fraction collection, this method should be amenable to automation and further scale up for preparation of larger amounts (e.g. milligram quantities) of DNA nanostructures.
    Full-text · Article · Nov 2012 · Nucleic Acids Research
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    • "What of the future? In the near term, we are likely to see new developments such as the further implementation of gaps, apertures and pores formed from DNA origami [20] [21], which should be at least as amenable to refined manipulations of structure as protein nanopores. In the longer term, by using solid-state pores, a main theme of Wanunu's review [1], it may be possible to read DNA sequences at microseconds rather than milliseconds per base by using tunneling currents [22] [23] or other characteristics of the DNA bases for which graphene with its unusual electronic properties might after additional development provide a superior substrate [24] . . . "

    Preview · Article · May 2012 · Physics of Life Reviews
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