HIGH THROUGHPUT DNA REPAIR CAPACITY ASSAY

高通量 DNA 修复能力测定

• 批准号: 6144486
• 负责人: KATHERINE A WOOD
• 金额: $ 9.95万
• 依托单位: TREVIGEN, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2001-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6144486
• 关键词: DNA damage; DNA repair; cell line; deoxyguanosine; genetic screening; high throughput technology; neoplasm /cancer genetics; nucleic acid sequence; oligonucleotides; technology /technique development

One commonality amongst the human cancers is that they all contain a genetic defect which directly or indirectly results in the progression of the neoplasia. These genetic defects can arise from a number of sources, including environmental damage to DNA, metabolic damage to DNA, viral infection, or faulty genes may be inherited leading to a predisposition. The accumulation of DNA damage is thought, therefore, to be at least partially responsible for the initiation of neoplastic development. When cells are subjected to extreme environments, or when there is a deficiency in part or all of the repair system, DNA damage accumulates. This Phase I SBIR proposal outlines the development of a high throughput assay for the investigation of DNA repair capacity. The assay is based upon the immobilization of short double-stranded labeled oligonucleotide DNA sequences containing specific base modifications that serve as substrates for DNA repair. The oligonucleotides are immobilized into the wells of a multiwell microplate or alternatively painted onto the surface of a microscope slide in an array format. The assay measures changes in amount of label following exposure to cell extracts. PROPOSED COMMERCIAL APPLICATIONS: The assay provides a rapid method for screening cell or patient samples for their ability to repair specific types of DNA damage. The assay has applications in the basic research, clinical and drug discovery markets. It provides a rapid screening method for DNA repair capacity and may be a useful tool for the development of new drugs that either enhance repair (maybe useful for cancer prevention) or specifically inhibit repair (useful for preventing repair during chemotherapeutic treatments). Such an assay could also allow clinical oncologists to evaluate the effectiveness of certain treatments. Trevigen estimates the commercial potential of a DNA repair capacity assay to be over $100 million annually.

人类癌症的一个共同点是，它们都含有直接或间接导致肿瘤进展的基因缺陷。这些遗传缺陷可以由多种原因引起，包括环境对DNA的破坏、对DNA的代谢损害、病毒感染或导致遗传缺陷的基因可能导致易感。因此，DNA损伤的积累被认为至少是肿瘤发生的部分原因。当细胞受到极端环境的影响时，或者当修复系统的部分或全部缺陷时，DNA损伤会累积。这份第一阶段的SBIR提案概述了高通量DNA修复能力检测方法的发展。该分析是基于固定短的双链标记的寡核苷酸DNA序列，该序列包含作为DNA修复底物的特定碱基修饰。将寡核苷酸固定到多孔微孔板的孔中，或者以阵列形式涂抹到显微镜载玻片的表面上。该检测方法测量暴露于细胞提取物后的标记量的变化。拟议的商业应用：该分析提供了一种快速筛选细胞或患者样本修复特定类型DNA损伤的能力的方法。该检测方法在基础研究、临床和药物发现市场上都有应用。它提供了一种快速筛选DNA修复能力的方法，并可能成为开发增强修复(可能用于癌症预防)或特异性抑制修复(用于预防化疗期间的修复)的新药的有用工具。这样的检测还可以让临床肿瘤学家评估某些治疗的有效性。Trevigen估计DNA修复能力检测的商业潜力每年超过1亿美元。