Mutational mechanisms underlying mouse mosaicism

小鼠嵌合体的突变机制

• 批准号: RGPIN-2016-04437
• 负责人: Hill, Kathleen
• 金额: $ 2.77万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=653578
• 关键词: Mutational; mechanisms; underlying; mouse; mosaicism

A genome is often called a blueprint, but this image of a single plan fails to capture the continuous change occurring in both the structure and the sequence of a genome within an organism over a lifespan. Recent evidence in mutation research reveals that de novo mutations occur at higher rates and with greater impact on the genome landscape than thought previously. Development is a critical period of mutagenesis during which individual tissues and cells are affected at different times and to different degrees. My research team seeks to understand the origins and mechanisms of mutation characteristic of normal development by investigating the load of de novo mutations in tissues and cells of the adult mouse. Mice provide an excellent model of the mutational mechanisms in mammalian genomes and enable study of any tissue in the context of well studied genetic backgrounds. My students and I have expertise using a genomic platform to examine point mutations at 493,290 polymorphic sites in the mouse genome and to detect deletions and duplications through assay of copy number at 915,519 sites distributed across the genome. We used this genomics platform to detect single nucleotide variants and copy number variants genome wide in an evolutionary study of extensive genetic diversity in Mus musculus. We now propose to examine the load of new mutations in a panel of tissues in individuals (parents and three sons) of three families of C57BL/6J (B6) inbred mice, a reference genetic background. We selected somatic tissues with different developmental histories and we included analysis of male germ cells at two points in sperm development. We propose to repeat our study for three families of hybrid B6D2 mice predicted to have higher rates of mutation associated with heterozygosity. Since our genetics platform is biased in analyzing point mutations in nongenic regions of the genome, we propose whole exome sequencing to detect de novo mutations in protein coding regions of the genome. Our research will document the number and nature of new mutations in individual tissues of adult mice with specific examination of the influences of genomic heterozygosity, developmental history and coding sequence. Our data provide a baseline of the degree and nature of somatic mosaicism arising with normal mouse development that will be used in biomonitoring studies for understanding mutational mechanisms associated with environmental mutagens and in comparisons with mouse genetic backgrounds with compromised genome integrity to understand mechanisms of genome maintenance. Our research is fundamental to understanding mechanisms contributing to the normal genetic diversity that arises within an individual. Graduate and undergraduate students will gain technical and computational expertise in analysis of mouse genome variation valuable to research in academic institutions, research institutes and government agencies.

基因组通常被称为蓝图，但这种单一计划的图像无法捕捉生物体内基因组结构和序列在生命周期中发生的连续变化。突变研究的最新证据表明，从头突变的发生率更高，对基因组景观的影响比以前认为的更大。发育是诱变的关键时期，在此期间，个体组织和细胞在不同时间和不同程度上受到影响。我的研究团队试图通过研究成年小鼠组织和细胞中的从头突变负荷来了解正常发育特征突变的起源和机制。小鼠提供了哺乳动物基因组中突变机制的极好模型，并且使得能够在充分研究的遗传背景的背景下研究任何组织。我和我的学生拥有使用基因组平台检查小鼠基因组中493，290个多态性位点的点突变的专业知识，并通过分析分布在基因组中的915，519个位点的拷贝数来检测缺失和重复。我们使用该基因组学平台在小家鼠广泛遗传多样性的进化研究中检测全基因组范围内的单核苷酸变异和拷贝数变异。我们现在建议检查一组组织中的新突变的负荷在个人（父母和三个儿子）的三个家庭的C57 BL/6 J（B6）近交系小鼠，一个参考遗传背景。我们选择了具有不同发育历史的体细胞组织，并在精子发育的两个时间点对雄性生殖细胞进行了分析。我们建议重复我们的研究三个家庭的杂交B6 D2小鼠预测有较高的突变率与杂合性。由于我们的遗传学平台在分析基因组的非基因区域中的点突变时存在偏差，因此我们提出全外显子组测序来检测基因组的蛋白质编码区中的从头突变。 我们的研究将记录成年小鼠个体组织中新突变的数量和性质，并具体检查基因组杂合性，发育历史和编码序列的影响。 我们的数据提供了一个基线的程度和性质的体细胞嵌合体所产生的正常小鼠的发展，将用于生物监测研究，了解与环境诱变剂相关的突变机制，并在与小鼠遗传背景的比较与受损的基因组完整性，以了解基因组的维护机制。 我们的研究对于理解个体内产生的正常遗传多样性的机制至关重要。 研究生和本科生将获得对学术机构，研究机构和政府机构的研究有价值的小鼠基因组变异分析的技术和计算专业知识。