DNA sequencing using nanopore-nanoelectrode devices for sensing and manipulation

使用纳米孔-纳米电极装置进行 DNA 测序以进行传感和操作

• 批准号: 7676235
• 负责人: Marija Drndic
• 金额: $ 27.48万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-19 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676235
• 关键词: Ablation; Achievement; Address; Arts; Caliber; Communities; Complement; Complex; Computers; DNA; DNA Sequence; Development; Devices; Diagnosis; Electric Capacitance; Electrodes; Electron Beam; Electrons; Electrostatics; Fingers; Genetic; Genome; Goals; Hemolysin; Individual; Length; Measurement; Measures; Medical; Membrane; Metals; Methodology; Microfluidic Microchips; Microfluidics; Mole the mammal; Motion; Nanostructures; Noise; Operative Surgical Procedures; Patients; Pattern; Performance; Process; Research; Resolution; Running; Seeds; Signal Transduction; Solutions; Speed; Staging; Structure; Study Section; Symptoms; Techniques; Technology; Time; Validation; Vestibule; Work; base; cost; disorder prevention; ds-DNA; electric impedance; fluorescence microscope; improved; lithography; nano; nanochannel; nanofluidic; nanometer; nanopore; nanoscale; prototype; research study; sensor; silicon nitride; simulation; single molecule; solid state; transmission process

DESCRIPTION (provided by applicant): The aim of this research effort is to leverage the unique capabilities of our recently developed sub-nanometer precision transmission electron beam ablation lithography (TEBAL) to demonstrate that the precise integration of solid-state nanopores with nanoelectrodes, nanochannels and other nanostructures will address key obstacles that must be overcome to achieve nanopore-based low-cost high-speed sequencing of chromosomal length DNA molecules, and the resultant medical and scientific benefits enabled by this technology. Fast and low cost full genome DNA sequencing will allow, for example, major improvements in the understanding, diagnosis, treatment and prevention of disease, and significant advances in evolutionary research and the understanding of cellular operation. This project will build on the remarkable progress towards nanopore-based DNA sequencing over the past decade, and it is planned to continue the tradition of sharing results, techniques and nanopore devices with the research community so that the work carried out will make the maximal contribution to advancing the state of the art. It is expected that the nanopore-nanoelectrode devices produced will seed further work by other groups on a variety of transverse electrode sensing methodologies and nanoelectrode-based single DNA molecule manipulation, which will contribute to the achievement of a nanopore-based "$1000 genome sequencer". This development (R21) project will begin work on the long term goals described above by demonstrating the improvements that can be achieved using nanopore devices with integrated nanoelectrodes and nanochannels. Beyond developing reliable nanopore-nanoelectrode devices, the unique aspects of the proposed work include the integration of geometrically more complex electrode patterns to manipulate DNA motion, and the integration of these devices with microfluidics and a fluorescent microscope setup to allow tracking of DNA molecules, so that they can be actively transported to the nanopore. The specific tasks are to show that: * DNA molecule length can be measured more accurately by transverse sensing with nanoelectrodes * the translocation speed of double stranded DNA can be reduced by applying forces via nanoelectrodes * by constraining DNA molecules, length measurement resolution improves for longer molecules * individual DNA molecules can be selected, transported to a nanopore and translocated through it These objectives will be accomplished in several steps. The required nanopore-nanoelectrode, nanopore-nanochannel and microfluidics devices will first be fabricated and characterized (some of this has already been achieved). Next, experiments with these devices will be conducted to evaluate their performance and identify problems. Finally, several cycles of device refinement and further experiments will resolve these problems and improve device performance to optimal levels, so that achievement of the objectives can be demonstrated. PROJECT HEALTH RELEVANCE This research aims to achieve much faster and lower-cost DNA sequencing by developing a nanotechnological sensor. This sensor works just like picking out the knots on a string by running it through one's fingers, except the string is a million times thinner! It will enable major improvements in the understanding, diagnosis, treatment and prevention of disease, by allowing us to determine the underlying genetic causes and symptoms, detect these rapidly and accurately in patients, and treat them appropriately.

描述（由申请人提供）：这项研究的目的是利用我们最近开发的亚纳米精密透射电子束烧蚀光刻（TEBAL）的独特能力，证明固体纳米孔与纳米电极、纳米通道和其他纳米结构的精确集成，将解决必须克服的关键障碍，以实现基于纳米孔的染色体长度DNA分子的低成本高速测序。以及由此带来的医学和科学效益。例如，快速和低成本的全基因组DNA测序将使对疾病的理解、诊断、治疗和预防方面取得重大进展，并在进化研究和对细胞运作的理解方面取得重大进展。