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

Adam R. Hall; Andrew Scott; Dvir Rotem; Kunal K. Mehta; Hagan Bayley; Cees Dekker

doi:10.1038/nnano.2010.237

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

Adam R. Hall
, Andrew Scott
, Dvir Rotem
, Kunal K. Mehta
, Hagan Bayley
, Cees Dekker^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

279 Scopus citations

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 engineering ⁴, 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.

Original language	English
Pages (from-to)	874-877
Number of pages	4
Journal	Nature Nanotechnology
Volume	5
Issue number	12
DOIs	https://doi.org/10.1038/nnano.2010.237
State	Published - Dec 2010
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported by the European Union’s Seventh Framework Programme (FP7/2007-2013) under grant agreement no. 201418 (READNA), the NanoSci Eþ 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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title = "Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores",

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 engineering 4, 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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AU - Dekker, Cees

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N2 - 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 engineering 4, 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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Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores

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