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Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores

Nature Nanotechnology volume 5, pages 874–877 (2010)Cite this article

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Abstract

Most experiments on nanopores have concentrated on the pore-forming protein α-haemolysin (αHL)1 and on artificial pores in solid-state membranes2. While biological pores offer an atomically precise structure3 and the potential for genetic engineering4, solid-state nanopores offer durability, size and shape control5, and are also better suited for integration into wafer-scale devices. However, each system has significant limitations: αHL is difficult to integrate because it relies on delicate lipid bilayers for mechanical support, and the fabrication of solid-state nanopores with precise dimensions remains challenging. Here we show that these limitations may be overcome by inserting a single αHL pore into a solid-state nanopore. A double-stranded DNA attached to the protein pore is threaded into a solid-state nanopore by electrophoretic translocation. Protein insertion is observed in 30–40% of our attempts, and translocation of single-stranded DNA demonstrates that the hybrid nanopore remains functional. The hybrid structure offers a platform to create wafer-scale device arrays for genomic analysis, including sequencing6.

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Figure 1: Molecular construct and experimental setup.
Figure 2: Directed αHL insertion.
Figure 3: DNA translocation through a hybrid αHL/SS nanopore.

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Acknowledgements

This work was supported by the European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 201418 (READNA), the NanoSci E+ program and the European Research Council. K.M. was supported by a Whitaker Foundation fellowship. The authors wish to thank M. Salichou for assistance with engineering of the αHL proteins.

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  1. Adam R. Hall

    Present address: Present address: Joint School of Nanoscience and Nanoengineering, University of North Carolina Greensboro, 2901 East Lee Street, Greensboro, North Carolina 27401, USA,

Authors and Affiliations

  1. Kavli Institute of Nanoscience, Delft University of Technology, Lorentzweg 1, CJ Delft, 2628, The Netherlands

    Adam R. Hall, Andrew Scott & Cees Dekker

  2. Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Mansfield Road, Oxford, OX1 3TA, UK

    Dvir Rotem, Kunal K. Mehta & Hagan Bayley

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  1. Adam R. Hall
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Contributions

A.R.H., H.B. and C.D. designed the experiment and wrote the manuscript. D.R. and K.M made the engineered αHL pores. A.R.H and A.S. performed the measurements and analysed data.

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Correspondence to Cees Dekker.

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The authors declare no competing financial interests.

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Hall, A., Scott, A., Rotem, D. et al. Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores. Nature Nanotech 5, 874–877 (2010). https://doi.org/10.1038/nnano.2010.237

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