Chemical Development of Fluorescent Labeling Tags for DNA Damaged Sites

DNA 损伤位点荧光标记标签的化学开发

• 批准号: 8009709
• 负责人: Peter R Andreana
• 金额: $ 7.03万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8009709
• 关键词: Achievement; Aging; Animals; Area; B-Cell Lymphomas; Base Excision Repairs; Base Pairing; Basic Science; Binding; Binding Sites; Biochemical; Biochemistry; Biological; Biological Assay; Biological Markers; Buffers; Bypass; Cancer Etiology; Cell Membrane Permeability; Cell division; Cells; Chemical Exposure; Chemicals; Chemistry; Chromosomes; Clinical; Comet Assay; DNA; DNA Adducts; DNA Damage; DNA Repair; DNA biosynthesis; DNA glycosylase; DNA lesion; DNA-(apurinic or apyrimidinic site) lyase; Deoxyribose; Detection; Development; Disease; Dose; Dyes; Equilibrium; Excision; Excision Repair; Exposure to; Fluorescent Dyes; Genetic; Goals; Human; Hydrogen Bonding; In Situ; In Vitro; Label; Laboratories; Lead; Lesion; Malignant Neoplasms; Mammalian Cell; Membrane; Methodology; Methods; Mismatch Repair; Monitor; Mutagenesis; Mutagens; Mutation; Nucleotides; Permeability; Predisposition; Probability; Process; Property; Pyrimidine Dimers; Reagent; Reporting; Research; Sampling; Site; Smoking; Stimulation of Cell Proliferation; System; Techniques; Technology; Tissue Sample; Universities; Uracil; Vertebral column; Work; base; cancer cell; carcinogenesis; cell killing; cyanine dye 5; exposed human population; functional group; human disease; human tissue; improved; innovation; inorganic phosphate; instrumentation; interest; new technology; novel; oxidative damage; repaired; sugar; tissue culture; tissue/cell culture; tool; tumor molecular fingerprint

DESCRIPTION (provided by applicant): Numerous studies have shown a strong correlation between accumulations of DNA adducts during aging and carcinogenesis. However, these correlations have not generally resulted in monitoring of DNA adducts as biomarkers for monitoring susceptibility to human diseases, in part, because of the complexities of biochemical assays for the detection of lesions in DNA. Herein, we propose to develop new chemical reagents based on well established methodologies that should greatly simplify detection of many DNA adducts in purified DNA, tissue culture cells and clinical samples. The strategy is to synthesize fluorescent dye derivatives that will react with DNA backbone following excision of DNA adducts using lesion-specific DNA glycosylases. We will demonstrate the feasibility of this approach by using synthetic, followed by biochemical approaches for detecting uracils and 8- oxoguanines in purified DNA and mammalian cells. The dyes to be developed will be considered for membrane permeability allowing the application of this technology to determining uracil content of B cell lymphoma DNA in situ. The long term goal of this project is to develop new chemical and biochemical tools to improve upon existing methodologies, to quantify and localize, at chromosome level, consequences of human exposure to DNA-damaging agents.

描述（由申请人提供）： 大量研究表明，衰老过程中 DNA 加合物的积累与致癌作用之间存在很强的相关性。然而，这些相关性通常不会导致将 DNA 加合物作为监测人类疾病易感性的生物标志物进行监测，部分原因是用于检测 DNA 损伤的生化测定的复杂性。在此，我们建议基于成熟的方法开发新的化学试剂，这将大大简化纯化 DNA、组织培养细胞和临床样本中许多 DNA 加合物的检测。该策略是合成荧光染料衍生物，在使用损伤特异性 DNA 糖基化酶切除 DNA 加合物后，该衍生物将与 DNA 主链发生反应。我们将通过使用合成方法和生化方法来检测纯化 DNA 和哺乳动物细胞中的尿嘧啶和 8-氧代鸟嘌呤，从而证明该方法的可行性。待开发的染料将考虑膜渗透性，允许应用该技术原位测定 B 细胞淋巴瘤 DNA 的尿嘧啶含量。该项目的长期目标是开发新的化学和生化工具来改进现有方法，在染色体水平上量化和定位人类暴露于 DNA 损伤剂的后果。