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


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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Article: DNA Origami Gatekeepers for Solid-State Nanopores

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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.
    Nucleic Acids Research 11/2012; 41(2). DOI:10.1093/nar/gks1070 · 9.11 Impact Factor
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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] . . . "

    Physics of Life Reviews 05/2012; 9(2):161-3; discussion 174-6. DOI:10.1016/j.plrev.2012.05.015 · 7.48 Impact Factor
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    ABSTRACT: Many processes in biology rely fundamentally on the relative position and orientation of interacting molecules. It is notoriously difficult to observe, let alone control, the position and orientation of molecules because of their small size and the constant thermal fluctuations that they experience. Molecular self-assembly with DNA origami enables building customshaped nanometer-scale objects with molecular weights in the megadalton regime. It provides a unique route for placing molecules and constraining their fluctuations in user-defined ways and thus opens up completely new avenues for scientific exploration.
    BioSpektrum 05/2012; 18(3). DOI:10.1007/s12268-012-0176-x
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