Mouse radiation and chemical induced deletion mutagensis

小鼠辐射和化学诱导缺失诱变

• 批准号: 6916186
• 负责人: WEI-WEN CAI
• 金额: $ 18.81万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2007-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6916186
• 关键词: DNA damage; artificial chromosomes; biotechnology; chlorambucil; functional /structural genomics; gamma radiation; gene deletion mutation; genetic mapping; genetically modified animals; germ cells; laboratory mouse; method development; microarray technology; nucleic acid sequence; polymerase chain reaction; site directed mutagenesis

DESCRIPTION (provided by applicant): Mouse deletion stocks are valuable resources for functional dissection of the mouse genome. Several strategies have been developed for generating mice with chromosome deletion. But all these methods are currently applied to recover deletions only around specific loci. Generation of random deletions in the genome using chemical or radiation-induced deletion directly in the mouse germline can be a powerful strategy for generating mice with inheritable deletion. However, application of this mutation strategy to genome-wide was not possible because of a lack of powerful mapping technologies. The high resolution and high throughput mouse genome scanning technology we recently developed promises to solve this mapping problem. Our mouse whole genome BAC arrays contain more than 19,000 overlapping BACs, providing a tiling path to cover the entire mouse genome. Using these arrays, the mouse genome can be scanned to reveal chromosome deletion or gain larger than 200 kb in any region of the genome by simple hybridizations without the need for breeding and phenotypic tracking as required in conventional meiotic mapping strategy. In this pilot study, we propose to apply our whole genome BAC array technology to reevaluate the feasibility of generating genome-wide mouse deletion stocks. The goal of this study is to define the optimal conditions for chemical and/or radiation induced deletion mutagenesis methods for generating mice carrying inheritable deletions.

描述（由申请人提供）：小鼠缺失库存是用于小鼠基因组功能解剖的宝贵资源。已经开发出几种策略来产生染色体缺失的小鼠。但所有这些方法目前仅用于恢复特定基因座周围的删除。直接在小鼠种系中使用化学或辐射诱导的缺失来产生基因组中的随机缺失可能是产生具有可遗传缺失的小鼠的强大策略。然而，由于缺乏强大的作​​图技术，将这种突变策略应用于全基因组是不可能的。我们最近开发的高分辨率和高通量小鼠基因组扫描技术有望解决这一绘图问题。我们的小鼠全基因组 BAC 阵列包含超过 19,000 个重叠的 BAC，提供覆盖整个小鼠基因组的平铺路径。使用这些阵列，可以扫描小鼠基因组，通过简单的杂交来揭示基因组任何区域的染色体缺失或大于 200 kb 的增益，而不需要传统减数分裂作图策略所需的育种和表型跟踪。在这项试点研究中，我们建议应用我们的全基因组 BAC 阵列技术来重新评估生成全基因组缺失小鼠种群的可行性。本研究的目的是确定化学和/或辐射诱导的缺失诱变方法的最佳条件，以产生携带可遗传缺失的小鼠。