Assays for ROS-induced DNA damage

ROS 诱导的 DNA 损伤测定

• 批准号: 6550539
• 负责人: HAROLD C BOX
• 金额: $ 25万
• 依托单位: ZEPTOMETRIX CORPORATION
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-08 至 2004-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6550539
• 关键词: DNA damage; DNA repair; bioassay; chemical synthesis; formamide; free radical oxygen; guanine; high performance liquid chromatography; liquid chromatography mass spectrometry; nuclease; stable isotope; technology /technique development; thin layer chromatography; thymine

DESCRIPTION (provided by applicant):This project is a collaboration between ZeptoMetrix Corporation and Roswell Park Cancer Institute. The objective is to develop sensitive, quantitative and facile assays for measuring DNA damage caused by reactive oxygen species (ROS). ROS have been implicated in aging, cancer and several neurological diseases. Seven ROS-induced DNA lesions will be assayed, namely: the formamido remnant of pyrimidine bases, the 8-oxo-7, 9-dihydroguanine (8-oxo-guanine) lesion, the thymine glycol lesion, the 6-hydroxy-5,6-dihydrothymine lesion, the abasic lesion and two double base lesions. 32P-postlabeling or mass spectroscopic methods of detection are employed with the assays. ZeptoMetrix will make products commercially available that facilitate DNA measurements by either method. Products will include reference compounds, carriers and internal standards for assaying the aforementioned lesions. Carriers for 32P-postlabeling provide unequivocal identification of the DNA lesion of interest. Internal standards for MS measurements will be isotopically enriched to a higher mass than the DNA lesion to be quantitated. ZeptoMetrix anticipates that eventually the largest application of these products will be for evaluating the repair capabilities of cells and tissues. Products useful for repair studies will include reference compounds and duplex DNA substrates containing identified lesions. Liquid Chromatography -Mass Spectrometry (LC-MS) utilizing an electrospray interface will be developed with this application especially in mind. PROPOSED COMMERCIAL APPLICATION: Reference materials, internal standards, and carriers for measuring oxidative DNA damage will be synthesized. These products will be offered commercially for facilitating DNA damage and repair by 32P-postlabeling or by LC-MS.

描述（由申请人提供）：该项目是Zeptometrix Corporation与Roswell Park Cancer Institute之间的合作。目的是开发敏感，定量和轻松的测定，以测量由活性氧（ROS）引起的DNA损伤。 ROS与衰老，癌症和几种神经系统疾病有关。 Seven ROS-induced DNA lesions will be assayed, namely: the formamido remnant of pyrimidine bases, the 8-oxo-7, 9-dihydroguanine (8-oxo-guanine) lesion, the thymine glycol lesion, the 6-hydroxy-5,6-dihydrothymine lesion, the abasic lesion and two double base lesions.该测定法使用32p Postlabeling或质谱检测方法。 Zeptometrix将使产品在商业上可用，以通过两种方法促进DNA测量。产品将包括参考化合物，载体和内部标准，以分析上述病变。 32p固定标签的携带者提供了对DNA感兴趣病变的明确鉴定。 MS测量的内部标准标准将在同位素上富集至质量高于要定量的DNA病变。 Zeptometrix预计，最终这些产品的最大应用将用于评估细胞和组织的修复能力。有用的用于维修研究的产品将包括参考化合物和双链DNA底物，这些DNA底物含有鉴定病变。使用电喷雾界面的液相色谱 - 质谱法（LC -MS）将在此应用中尤其是考虑到该应用程序。 拟议的商业应用： 参考材料，内部标准和用于测量氧化DNA损伤的载体 将合成。 这些产品将在商业上提供以促进DNA损伤 并通过32p固定标签或LC-MS维修。