Pulsed Field Surface Hybridization

脉冲场表面杂交

基本信息

  • 批准号:
    1600584
  • 负责人:
  • 金额:
    $ 29.41万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
    Standard Grant
  • 财政年份:
    2016
  • 资助国家:
    美国
  • 起止时间:
    2016-07-01 至 2020-06-30
  • 项目状态:
    已结题

项目摘要

PI: Levicky, Rastislav Proposal Number: 1600584The central aim is to advance biosensing of DNA, such as the human genome, in support of personalized medicine, clinical diagnostics, and basic life science research. A technology termed pulsed-field hybridization will be developed by combining electric field control of molecular behavior with optimized sensor designs, based on synthetic DNA analogues, to eliminate errors and to significantly shorten analysis time. The project also encompasses educational goals centered on design of learning software that illustrates molecular properties and on organization of publicly accessible science lectures and demonstrations.Biosensing applications must handle complex sample matrices in which the analyte of interest needs to be distinguished against a background of other molecules. In nucleic acid diagnostics, background interference most often involves cross-hybridization from DNA or RNA molecules with a degree of similarity to the desired sequence. Cross-hybridization leads to multiple detrimental consequences including blocking binding of the perfectly matched analyte, markedly slowing down approach to equilibrium so that assay endpoints are defined by kinetic history rather than path-independent thermodynamics, and loss of precision due to contributions from the cross-hybridized species. These concerns become amplified for sequence-rich samples comprising larger genomes, mixtures of genetic material from multiple sources, or when the sequence of interest is one of several closely related sequences. This project proposes a bioanalytical method termed pulsed field hybridization (PFH) to solve these challenges. Through use of charge-neutral DNA analogues called morpholinos and modulated electric fields, PFH seeks highly efficient, field-based manipulation of hybridization to dramatically reduce equilibration timescales and to enable "distilling", through a stage-wise operation, of the purity of the hybridized product. The project will develop a thorough fundamental understanding of how surface fields influence thermodynamics and kinetics of hybridization on morpholino-derivatized sensors, and of the impact of analyte characteristics including excess nucleotides, unhybridized tails, and variously placed mismatches. These fundamental studies will be accompanied by application to complex sample matrices typical of clinical situations to demonstrate a fully functional, practical method. Project participants will in addition assist with research exhibitions open to the public, engage in organizing lectures on nanoscience, and collaborate on development of learning software to provide a fun, informative look into the behavior of molecules.
主要研究者:Levicky,Rastislav提案编号:1600584中心目标是推进DNA的生物传感,如人类基因组,以支持个性化医疗,临床诊断和基础生命科学研究。将开发一种称为脉冲场杂交的技术,将分子行为的电场控制与基于合成DNA类似物的优化传感器设计相结合,以消除错误并大大缩短分析时间。该项目还包括教育目标集中在学习软件的设计,说明分子特性和组织公众可访问的科学讲座和demonstrations.Biosensing应用程序必须处理复杂的样品矩阵,其中感兴趣的分析物需要区分对其他分子的背景。在核酸诊断中,背景干扰最常涉及来自与所需序列具有一定程度相似性的DNA或RNA分子的交叉杂交。交叉杂交导致多种有害后果,包括阻断完全匹配的分析物的结合,显著减慢接近平衡的速度,使得测定终点由动力学历史而不是路径独立的热力学定义,以及由于交叉杂交物质的贡献而导致的精度损失。对于包含较大基因组的序列丰富的样品,来自多个来源的遗传物质的混合物,或者当感兴趣的序列是几个密切相关的序列之一时,这些问题变得更加严重。该项目提出了一种称为脉冲场杂交(PFH)的生物分析方法来解决这些挑战。通过使用电荷中性的DNA类似物,称为吗啉代和调制电场,PFH寻求高效的,基于场的杂交操作,以显着减少平衡时间尺度,并使“蒸馏”,通过逐步操作,杂交产物的纯度。该项目将开发一个彻底的基本了解表面场如何影响热力学和动力学的杂交吗啉衍生的传感器,以及分析物的特性,包括过量的核苷酸,未杂交的尾巴,和各种放置的错配的影响。这些基础研究将伴随着应用到复杂的样品基质典型的临床情况,以证明一个功能齐全,实用的方法。项目参与者还将协助向公众开放的研究展览,参与组织纳米科学讲座,并合作开发学习软件,以提供有趣,信息丰富的分子行为。

项目成果

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Rastislav Levicky其他文献

Rastislav Levicky的其他文献

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{{ truncateString('Rastislav Levicky', 18)}}的其他基金

Thermodynamic Cycles and Relaxation Timescales in Surface Hybridization
表面杂交中的热力学循环和弛豫时间尺度
  • 批准号:
    1206754
  • 财政年份:
    2012
  • 资助金额:
    $ 29.41万
  • 项目类别:
    Standard Grant
MRI: Acquisition of a 500 MHz NMR Spectrometer for Teaching and Research
MRI:购买 500 MHz NMR 波谱仪用于教学和研究
  • 批准号:
    1126005
  • 财政年份:
    2011
  • 资助金额:
    $ 29.41万
  • 项目类别:
    Standard Grant
Collaborative Research: Mechanisms of Hybridization Kinetics in DNA Surface Layers
合作研究:DNA表面层杂交动力学机制
  • 批准号:
    0706170
  • 财政年份:
    2007
  • 资助金额:
    $ 29.41万
  • 项目类别:
    Continuing grant
CAREER: Genetic Diagnostics at Surfaces - Designing a Biological Polyelectrolyte System
职业:表面基因诊断 - 设计生物聚电解质系统
  • 批准号:
    0093758
  • 财政年份:
    2001
  • 资助金额:
    $ 29.41万
  • 项目类别:
    Continuing Grant

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