Variation Scanning with Mismatch Repair Detection (MRD)

使用错配修复检测 (MRD) 进行变异扫描

• 批准号: 6804479
• 负责人: Thomas D. WILLIS
• 金额: $ 21.42万
• 依托单位: PARALLELE BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-21 至 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6804479
• 关键词: DNA primers; DNA repair; autism; biotechnology; genetic polymorphism; genetic screening; genotype; high throughput technology; human genetic material tag; microarray technology; nucleic acid hybridization; nucleic acid quantitation /detection; nucleic acid sequence

DESCRIPTION (provided by applicant): One widely considered approach to identify susceptibility alleles is through linkage disequilibrium (LD) between these alleles and a universal set of relatively frequent SNPs distributed in all or most ethnic populations. This approach, however, may be unrealistic when variant alleles responsible for disease susceptibility are infrequent or are specific to a particular population. In such cases, identifying susceptibility alleles may require comprehensive sequence comparison between patients and controls. There is a paucity of techniques for high throughput scanning for unknown variations. Mismatch Repair Detection (MRD)'s potential for high throughput scanning can be used to comprehensively compare sequences between patients and controls. MRD has been described previously and it utilizes a bacterial mismatch repair system in vivo to detect sequence variants in human DNA samples. Many fragments can be introduced into a specific bacterial strain Mutation Sorter (MS) that is engineered to sort these fragments to two pools: those carrying variations and those that do not. The problem of DNA variation detection in then reduced to the problem of identification of the fragment content of the two pools, and that can be done using a microarray hybridization. In phase II of this grant we aim to apply MRD multiplex scanning ability to test about 1,000 candidate exons in hundreds of patients and controls to identify susceptibility alleles of a complex trait. In the long term, we see applying MRD on a whole genomic scale to scan all the exons of patients and controls to elucidate the genetic basis of complex disease.

描述（由申请人提供）：一种广泛考虑的鉴定易感性等位基因的方法是通过这些等位基因与分布在所有或大多数种族人群中的一组通用的相对频繁的 SNP 之间的连锁不平衡 (LD)。然而，当导致疾病易感性的变异等位基因不常见或特定于特定人群时，这种方法可能不切实际。在这种情况下，识别易感性等位基因可能需要患者和对照之间进行全面的序列比较。缺乏用于高通量扫描未知变化的技术。错配修复检测 (MRD) 的高通量扫描潜力可用于全面比较患者和对照之间的序列。 MRD 之前已经描述过，它利用体内细菌错配修复系统来检测人类 DNA 样本中的序列变异。许多片段可以被引入特定的细菌菌株突变分选机（MS）中，该菌株被设计为将这些片段分选为两个池：携带变异的和不携带变异的。 DNA变异检测的问题就简化为鉴定两个池的片段内容的问题，并且这可以使用微阵列杂交来完成。在本次资助的第二阶段，我们的目标是应用 MRD 多重扫描能力来测试数百名患者和对照的约 1,000 个候选外显子，以确定复杂性状的易感性等位基因。从长远来看，我们看到在整个基因组规模上应用 MRD 来扫描患者和对照的所有外显子，以阐明复杂疾病的遗传基础。