MOUSE MODEL FOR OXIDATIVE DNA DAMAGE REPAIR

用于氧化 DNA 损伤修复的小鼠模型

• 批准号: 6489385
• 负责人: JEFFREY H MILLER
• 金额: $ 30.55万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-01-30 至 2004-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6489385
• 关键词: DNA repair; cell growth regulation; disease /disorder model; embryo /fetus tissue /cell culture; fibroblasts; gene mutation; gene targeting; genetically modified animals; laboratory mouse; neoplasm /cancer genetics; oncoproteins; oxidative stress; p53 gene /protein

The long term goal of this research program is to uncover and understand mutational pathways, the DNA repair processes that counteract these pathways, and the consequences of repair defects. This proposal is an extension of previous work that defined new "mutator strains" in bacteria that have higher mutation rates than wild-type and that led to the elucidation of the GO system for oxidative damage. The human mutt gene, an essential component of this system, has been cloned. This work has now been extended to include targeted gene knockout mice. This proposal seeks to engineer and characterize a set of knockout mice lacking oxidative repair, as well as mice lacking different combinations of repair enzymes. Lacking certain repair systems can lead to cancer susceptibilities. It is therefore important to determine whether the absence of repair systems for oxidative damage leads to increased cancer rates or other abnormalities, such as accelerated aging. Specifically, the proposed research seeks to do the following. 1. Generate a double knockout mouse of mMYH with a DNA repair gene mOGG1. The OGG1 gene encodes a protein with a function similar to the bacterial MutM protein. Double knockouts might be expected to lack the ability to repair certain types of oxidative damage to the DNA. 2. Characterize mMYH knockout mice and mMYH-/- mOGG1-/- double knockout mice, analyze their phenotype and screen for abnormalities and cancer susceptibilities that might result from the reduced ability to repair oxidative damage. 3. Characterize primary mouse embryonic fibroblasts derived from both MYH-/- and MYH-/- OGG1-/- mice. 4. Generate and characterize additional combinations of mMYH knockout mice with other repair gene or tumor suppressor gene knockouts, such as MSH2 and p53.

这项研究计划的长期目标是揭示和理解突变途径，抵消这些途径的DNA修复过程，以及修复缺陷的后果。该建议是先前工作的延伸，该工作定义了细菌中具有比野生型更高突变率的新“突变菌株”，并导致氧化石墨烯系统氧化损伤的阐明。这个系统的重要组成部分——人类突变基因已经被克隆出来。这项工作现在已经扩展到包括靶向基因敲除小鼠。该提案旨在设计和表征一组缺乏氧化修复的基因敲除小鼠，以及缺乏不同修复酶组合的小鼠。缺乏某些修复系统会导致癌症易感性。因此，确定氧化损伤修复系统的缺失是否会导致癌症发病率增加或其他异常，如加速衰老，是很重要的。具体而言，拟议的研究旨在做到以下几点。1. 产生具有DNA修复基因mOGG1的mMYH双敲除小鼠。OGG1基因编码一种功能类似于细菌MutM蛋白的蛋白质。双敲除可能缺乏修复某些类型的DNA氧化损伤的能力。2. 对mMYH基因敲除小鼠和mMYH-/- mOGG1-/-双基因敲除小鼠进行表征，分析其表型，筛选可能由于氧化损伤修复能力降低而导致的异常和癌症易感性。3. 表征来自MYH-/-和MYH-/- OGG1-/-小鼠的原代小鼠胚胎成纤维细胞。4. 生成并表征mMYH敲除小鼠与其他修复基因或肿瘤抑制基因敲除的其他组合，如MSH2和p53。