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Contenu archivé le 2024-05-14

Microdiagene: nucleic acid sequencing in a diamond-silicon chip

Objectif



The "DiaGene" project will demonstrate an innovatory system for the sequencing of nucleic acids on a single semiconductor chip. This technology combines four key technologies already devised, developed and led by members of our collaboration. We fit squarely within biotech area 2..
We will demonstrate a novel sequencing system based on the intrinsic absorption in nucleic acids of ultraviolet (UV) light. The demonstration step will comprise our producing nucleic acid sequences in four research laboratories, all well known as specialists for different sequences. Our specific goal is to demonstrate these existing macroscopic technologies - developed in the last two years and in the process of being commercialized - at the chip level. 'DiaGene' can in principle also accept conventionally labelled nucleic acids to allow in situ cross checking. Our target sequences include: Arabidopsis, Lactococus Lactis, HIV, and an analysis of the spectrum of mutations in Cystic Fibrosis, X-Linked Adrenal Hypoplasia, and Crigler-Lajjar Syndrome.
The "DiaGene" system is based on an intrinsic absorption approach to molecular imaging for nucleic acid and protein sequencing and mapping, a technique developed at IC over the last three years and in the process of being commercialized. Its advantages are:
- the complete removal of mutagenic, toxic and carcinogenic chemi-,bioluminescent and radio labels with associated benefits in running costs,health and safety, operational and training and disposal imperatives; - an order of magnitude speeding of a typical sequence: a 6 hour (say) 500 Bp sequence electrophoresis can be achieved in 10 minutes in "DiaGene". This saving in time comes because the image of bands identified by conventional labels is smeared by the isotropic emission of those labels (whether a radioactive label, or a laser-induced fluorescent detection (CE-LIF) system)while in the absorption technique we gain a factor of ten since the image of the nucleic acid band is the same size as the band (smeared slightly by diffraction). This can be made very small with muTAS technology. This is anew paradigm in sequencing.
- improved sensitivities and signal-to-noise in sub-micro liter sequencing and imaging.
"DiaGene" demonstrates the synergistic power of our combining our individual technologies:
- intrinsic absorption imaging and sequencing of nucleic acids; - the methods of analysis on a chip as pioneered and demonstrated at IC Centre for Analytical Sciences under in TAS (Total Analysis Systems) programme; - the development of UV-sensitized silicon pixel detectors, first applied in biomedical imaging and taken to its highest level of development at University of Cagliari (CagF) and
- the use of micro ridge sensor topologies (devised at IC).
Our team includes the foremost company involved in the manipulation of nano liter quantities of biological materials, Evotec GmbH., who also provide a user-driven edge.
Results are intrinsically digital, and given the exceptional sensitivities, rapid turn-around possible and high through-puts attainable, the "DiaGene" technologies are ideally suited to the Human Genome research networks. Since the "DiaGene" system allows pre-programmed sequence recognition masks to accompany any installation, in principle a "yes-no" answer could be obtained by relatively untrained end-users. A sequence triage system results, which coupled to the extremely low cost of this technology, can result in the wide dissemination of "DiaGene" systems,

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Coordinateur

Imperial College of Science, Technology and Medicine
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Adresse
Blackett Laboratory Prince Consort Road
SW7 2BZ London
Royaume-Uni

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