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）： 焦磷酸解激活聚合（PAP）提供了一种新方法 从核酸中检索多种类型的信息。非凡的 PAP 的特异性源自无活性的双脱氧终止寡核苷酸 （P*）。 P* 通过 3? 的焦磷酸解被激活。末端核苷酸， 随后通过 DNA 聚合延伸激活的寡核苷酸。 初步研究建立了原理证明，最近的效率 基因工程聚合酶大大提高了 PAP 的质量 对双脱氧核苷酸有高亲和力。在此，我们建议证明 PAP是一种平台技术。将开发三种新方法 该平台：i) 对单个核苷酸变化进行等位基因特异性检测 10expl6-10exp9 正常等位基因 (PAP-A) 的存在：ii) 基于微阵列 重测序 (PAP-R) 和 iii) 体内染色质结构分析 （LM-PAP）。这些方法有许多应用，包括： i) 超灵敏 检测微小残留病或测量突变负荷（PAP-A）； ii) 对易诱发的序列变异进行分子流行病学分析 复杂疾病或快速分子诊断（PAP-R）； iii) 分析 染色质结构作为印记、X 失活和基因的函数 甲基化水平的表达或定量（LM-PAP）。

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