Massively parallel DNA nanopores using hybrid DNA origami for faster Sequencing

A sensing system developed at Cambridge is being commercialised in the UK for use in rapid, low-cost DNA sequencing, which would make the prediction and diagnosis of disease more efficient, and individualised treatment more affordable. Researchers have developed a system which combines a solid-state nanopore with DNA origami, for use in DNA sequencing, protein sensing and other applications. The technology has been licensed for development and commercialisation to UK-based company Oxford Nanopore.

Currently, there are two main types of nanopores in use: solid state nanopores constructed by fabricating tiny holes in silicon or graphene with electron beam equipment; and biological nanopores made by inserting pore-forming proteins into a biological membrane such as a lipid bilayer.

Biological nanopores are cheap and easy to manufacture in large quantities of identical pores. It is possible through genetic engineering to define their structure at the atomic level, varying the pores for the analysis of different target molecules. However, they are only suitable for a limited range of applications, and may be replaced over time by solid-state nanopores. At present, solid-state nanopores are difficult to manufacture and are not as sensitive as biological nanopores, as it is difficult to position specific chemical groups on the surface.

In collaboration with researchers at Ludwig Maximilian University in Munich, Dr Keyser and his team have developed a hybrid nanopore which combines a solid-state material, such as silicon or graphene, and DNA origami – small, well-controlled shapes made of DNA.

Since complementary sequences of DNA can bind to one another, the origami structures can be customised so that functional groups, fluorescent compounds and other molecular adapters can be added to the DNA strands with sub-nanometre precision, improving sensitivity and reliability. Additionally, hundreds of billions of self-assembling origami structures can be produced at the same time, with yields of up to 90 per cent.

Recent research by the team, published in the journal Lab on a Chip, has shown that up to 16 measurements can be taken simultaneously, allowing for much higher data throughput and screening of different DNA origami structures.

Lab on a chip –
Multiplexed ionic current sensing with glass nanopores

ABSTRACT – We report a method for simultaneous ionic current measurements of single molecules across up to 16 solid state nanopore channels. Each device, costing less than $20, contains 16 glass nanopores made by laser assisted capillary pulling. We demonstrate simultaneous multichannel detection of double stranded DNA and trapping of DNA origami nanostructures to form hybrid nanopores.

SOURCES – Lab on a chip, University of Cambridge

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