Nanopore detection of haplotype

单倍型的纳米孔检测

• 批准号: 6541529
• 负责人: DANIEL none BRANTON
• 金额: $ 52.33万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2005-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6541529
• 关键词: DNA; biotechnology; electrophoresis; high throughput technology; membrane channels; nanotechnology; nucleic acid quantitation /detection; nucleic acid sequence; nucleic acid structure; polynucleotides; protein structure; rapid diagnosis; single cell analysis; single nucleotide polymorphism; technology /technique development

DESCRIPTION (provided by applicant): I will develop the technology and basic science needed to use nanopores for high speed SNP detection and haplotyping. The technology depends on driving single polynucleotide molecules through a nanopore coupled to sensitive single channel recording electronics that can provide a direct read-out of the polymer's characteristics. The development of these new molecular diagnostic methods takes advantage of three recent discoveries: (1) A membrane channel, or nanopore, can be used as a high-throughput device that detects and probes single molecules as they translocate through the nanopore; (2) The ionic current through the nanopore is sensitive to local changes in the cross-sectional area of the translocating molecule; (3) A new, planar fabrication method - ion beam sculpting - that allows us to create a single digit nanoscale pore of a desired dimension in robust solid state insulating membranes. Our work will optimize: (a) zinc-finger protein labeling of DNA; (b) the electrical readout of DNA length; (c) the minimum fragment length and number of different zinc-finger proteins needed to achieve reliable SNP identification and high-speed haplotyping. The proposed technologies will impact basic research areas such as development and cancer that must deal with complex sets of genes and mutations, and where existing methods to rapidly examine the linkages between a large number of polymorphic sites on multiple chromosomes in a large number of individuals are limiting. The tools and basic research proposed will open new possibilities for the future development of high sensitivity, information rich diagnostic methods that are critical for early disease detection.

描述（由申请人提供）：我将开发使用纳米孔进行高速SNP检测和单倍分型所需的技术和基础科学。该技术取决于驱动单个多核苷酸分子通过纳米孔连接到敏感的单通道记录电子产品，该电子产品可以直接读取聚合物的特性。这些新分子诊断方法的开发利用了三个最近的发现：（1）膜通道或纳米孔可用作高通量设备，在通过纳米孔转移时检测和探测单分子； （2）通过纳米孔的离子电流对易位分子的横截面区域的局部变化敏感； （3）一种新的平面制造方法 - 离子束雕刻 - 使我们能够在坚固的固态绝缘膜中创建一个所需尺寸的单位纳米级孔。我们的工作将优化：（a）DNA的锌指蛋白标记； （b）DNA长度的电读数； （c）实现可靠的SNP识别和高速单倍型所需的不同锌指蛋白的最小片段长度和数量。所提出的技术将影响必须处理复杂的基因和突变集的基础研究领域，例如开发和癌症，以及在其中迅速检查大量个体多染色体上大量多态性位点之间的联系的现有方法。提出的工具和基础研究将为未来的高灵敏度，信息丰富的诊断方法开发新的可能性，这对于早期疾病检测至关重要。