PAP: a technology platform

PAP：一个技术平台

• 批准号: 7110242
• 负责人: STEVE Seev SOMMER
• 金额: $ 62.55万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7110242
• 关键词: DNA footprinting; alleles; biotechnology; chromatin; clinical research; gene expression; gene mutation; genomic imprinting; human data; human tissue; method development; methylation; microarray technology; natural gene amplification; nucleic acid quantitation /detection; nucleic acid sequence; oligonucleotides; polymerase chain reaction; polymerization

DESCRIPTION (provided by applicant): Pyrophosphorolysis activated polymerization (PAP) offers a novel approach for retrieving multiple types of information from nucleic acids. The exceptional specificity of PAP derives from an inactive dideoxy terminated oligonucleotide (P*). P* is activated by pyrophosphorolysis of the 3? terminal nucleotide, followed by extension of the activated oligonucleotide by DNA polymerization. The initial studies established proof of principle and recently the efficiency of PAP was improved greatly with genetically engineered polymerases that have a high affinity for dideoxy nucleotides. Herein, we propose to demonstrate that PAP is a platform technology. Three novel methods will be developed on the platform: i) allele-specific detection of a single nucleotide change in the presence of 10expl6-10exp9 normal alleles (PAP-A): ii) microarray-based resequencing (PAP-R) and iii) analysis of in vivo chromatin structure (LM-PAP). These methods have many applications including: i) ultra sensitive detection of minimal residual disease or measurement of mutation load (PAP-A); ii) molecular epidemiological analysis of sequence variants that predispose to complex disease or rapid molecular diagnosis (PAP-R); and iii) analysis of chromatin structure as a function of imprinting, X-inactivation and gene expression or quantitation of the level of methylation (LM-PAP).

描述（由申请人提供）： 焦磷酸解析激活聚合（PAP）为一种新颖的方法提供了新的方法 从核酸检索多种类型的信息。异常 帕普的特异性源自无活跃的二氧基终止寡核苷酸 （p*）。 p*通过3的焦磷酸解析而激活？末端核苷酸， 然后通过DNA聚合扩展活性寡核苷酸。 最初的研究建立了原理证明，最近效率 PAP的基因组合酶可大大改善 对甲氧基核苷酸的高亲和力。在此，我们建议证明 该帕普是一种平台技术。将开发三种新颖的方法 平台：i）等位基因特异性检测单核苷酸变化 10EXPL6-10EXP9普通等位基因的存在（PAP-A）：ii）基于微阵列的 重新陈述（PAP-R）和III）分析体内染色质结构 （LM-PAP）。这些方法具有许多应用程序，包括：i）超敏感 检测最小残留疾病或突变负荷的测量（PAP-A）； ii）易感性变体的分子流行病学分析 复杂疾病或快速分子诊断（PAP-R）； iii）分析 染色质结构是印迹，X灭活和基因的函数 甲基化水平（LM-PAP）的表达或定量。

项目成果

Microarray-based DNA resequencing using 3' blocked primers.

使用 3 封闭引物进行基于微阵列的 DNA 重测序。

• DOI: 10.1016/j.ab.2007.10.044
• 发表时间: 2008
• 期刊: Analytical biochemistry
• 影响因子: 2.9
• 作者: Sram,Jakub;Sommer,SteveS;Liu,Qiang
• 通讯作者: Liu,Qiang

Multiplex dosage pyrophosphorolysis-activated polymerization: application to the detection of heterozygous deletions.

多剂量焦磷酸解激活聚合：应用于检测杂合缺失。

• DOI: 10.2144/000112164
• 发表时间: 2006
• 期刊: BioTechniques
• 影响因子: 2.7
• 作者: Liu,Qiang;Nguyen,VuQ;Li,Xuemin;Sommer,SteveS
• 通讯作者: Sommer,SteveS