Chemically Tuned Silicon Nitride Nanopores for Nucleic Acid Sequencing

用于核酸测序的化学调谐氮化硅纳米孔

• 批准号: 10162635
• 负责人: Jason Rodger Dwyer
• 金额: $ 25.19万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-11 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10162635
• 关键词: Affect; Alkenes; Alkynes; Automobile Driving; Biological Assay; Biopolymers; Caliber; Cations; Characteristics; Charge; Chemicals; Chemistry; Complex; DNA; DNA Sequence; DNA sequencing; Devices; Diagnosis; Dimensions; Disease; Electrophoresis; Electrostatics; Elements; Environment; Film; Foundations; Funding; Genetic; Genotype; Interfacial Phenomena; Label; Length; Measurement; Medical; Membrane; Methods; Modification; Molecular; Motion; Nature; Noise; Nucleic Acids; Nucleic acid sequencing; Oligonucleotides; Organic Chemicals; Organic Chemistry; Oxides; Performance; Plant Roots; Polymers; Procedures; Property; Proteins; RNA; Reagent; Research Personnel; Resolution; Rest; Role; Sampling; Science; Signal Transduction; Silanes; Silicon; Speed; Structure of molecular layer of cerebellar cortex; Surface; Technology; Testing; Thick; Thinness; Time; Traction; Work; base; chemical property; chemical synthesis; cost; design; driving force; experience; experimental study; functional group; improved; insight; materials science; molecular film; molecular scale; monolayer; nanofabrication; nanopore; sequencing platform; silicon nitride; solid state; surface coating; tool; transcriptome sequencing

PROJECT SUMMARY This project aims to improve DNA and RNA sequencing technology by at least an order of magnitude by dramatically improving the ability to control silicon nitride nanopore surface chemistry and to modify silicon nitride nanopore size. While silicon nitride is a conventional material for nanopore sequencing applications, its complex charged native surface chemistry can present a challenging and complicated environment for a charged nucleic acid biopolymer passing through a nanopore not much larger than itself. Highly desirable long read lengths heighten the need for chemical control over the nanopore surface. Coating the nanopore surface with even a single molecular layer will change the nanopore diameter, which thus also provides for molecular-scale tuning of nanopore dimensions. Broadly, chemically tuned nanopore surface chemistry affords control over motion of (native or labelled) nucleic acid polymers through the pore through electrostatics, specific chemical interactions, and electrokinetics (e.g. electroosmosis). It offers the potential for passivation against fouling in complex matrices, thereby supporting more minimal sample processing. It also affords control over interfacial phenomena that can affect nanopore current noise. Thin-film silicon nitride is a widely used nanofabrication material with widespread commercial utility, so that its continued use in a host of nanopore sequencing implementations is warranted in spite of its often challenging surface chemistry. But efforts to control and improve its surface chemistry using silane chemistry have not gained traction in the field, in significant part because the chemistry is inherently challenging to implement, the more so given the variability of the silicon nitride oxide coating. We thus propose to develop a radically different type of surface chemical modification strategy that is simple to implement, produces highly reliable results, and that can be used to install surface coatings with a wide variety of chemical properties and sizes. We propose to test the surface coatings through their effect on the nanopore conductance and current noise, and on the sequence-specific signal characteristics when sensing well-defined sequences of DNA. The project will be implemented by an interdisciplinary team that combines more than 20 years of Principal and Co-Investigator experience in physical organic chemistry and chemical synthesis (MK); materials science (MK&JRD); and nanofabrication (JRD), with a decade of experience in nanopore science begun in nanopore genotyping (JRD), including a specialization in nanopore surface chemistry modification and characterization.

项目摘要 该项目旨在通过至少一定的顺序提高DNA和RNA测序技术 通过显着提高控制氮化硅纳米孔表面的能力来大小 化学和修饰氮化硅纳米孔的大小。而氮化硅是常规的 纳米孔测序应用的材料，其复杂的充电天然表面化学 可以为带电的核酸生物聚合物带来具有挑战性且复杂的环境 穿过纳米孔不大的纳米孔。高度理想的长读长度 在纳米孔表面上增加了化学控制的需求。覆盖纳米孔 表面甚至具有单个分子层也会改变纳米孔直径，因此也会改变 提供了纳米孔尺寸的分子尺度调整。广泛，化学调节 纳米孔表面化学可控制（天然或标记）核酸的运动 通过孔通过静电，特定的化学相互作用和 电动学（例如电流）。它提供了反对犯规的潜力 复杂的矩阵，从而支持更多最小的样品处理。它还提供控制权 界面现象可能影响纳米孔电流噪声。 薄膜硝化硅是一种广泛使用的纳米化材料，广泛商业 实用程序，以便必须在许多纳米孔测序实现中使用它 尽管经常具有挑战性的表面化学。但是努力控制和改善其表面 使用硅烷化学的化学反应尚未在田间受到关注，在很大程度上是因为 化学本质上是具有挑战性的，鉴于 氮化硅氧化物涂料。因此，我们建议开发出根本不同类型的表面 化学修改策略，易于实施，产生高度可靠的结果，并 可用于安装具有多种化学性质和尺寸的表面涂料。 我们建议通过对纳米孔电导的影响测试表面涂层，并 当前噪声以及序列特异性信号特性时感应明确定义时 DNA的序列。 该项目将由一个跨学科团队实施，该团队结合了20年以上 在物理有机化学和化学方面的主要和共同评估者的经验 合成（MK）;材料科学（MK＆JRD）;和纳米制造（JRD），有十年 纳米孔科学的经验始于纳米孔基因分型（JRD），包括专业化 在纳米孔表面化学修饰和表征中。