High-resolution, genome-scale mapping of natural variation between reproductively isolated individuals

生殖隔离个体之间自然变异的高分辨率、基因组规模图谱

• 批准号: 9319010
• 负责人: Rachel Beth Brem
• 金额: $ 39.52万
• 依托单位: BUCK INSTITUTE FOR RESEARCH ON AGING
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-15 至 2021-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9319010
• 关键词: Aging; Alleles; Animals; Architecture; Biological Assay; Caenorhabditis; Caenorhabditis elegans; Catalogs; Cells; Complement; Dissection; Distant; Employee Strikes; Environment; Eukaryota; Frequencies; Gene Expression; Gene Transfer Techniques; Genes; Genetic; Genomics; Goals; High temperature of physical object; Human; Hybrids; Individual; Invertebrates; Lipids; Longevity; Mammals; Maps; Measures; Methods; Modeling; Modernization; Molecular; Molecular Genetics; Mutagenesis; Nematoda; Orthologous Gene; Oxidative Stress; Parents; Partner in relationship; Pattern; Phenotype; Plants; Resistance; Resolution; Role; Saccharomyces cerevisiae; Sister; Sterility; Stress; Surveys; System; Testing; Tissues; Variant; Virulence; Work; Yeasts; age effect; anti aging; base; experimental study; fitness; genome-wide; interest; lipid metabolism; male; member; mutant; novel; parental influence; pathogen; reproductive; species difference; trait

SUMMARY One of the primary goals of genetics is to understand how and why naturally varying individuals differ in phenotype. Mapping the molecular basis of this trait variation can be straightforward, but only in individuals that can be interbred. We have found that a little-studied worm species, Caenorhabditis latens, has a median lifespan 50% longer than related nematodes, and we have observed robust, conserved thermotolerance in Saccharomyces cerevisiae, the only species of its clade that can act as an opportunistic pathogen. In the current proposal, we dissect these species divergences using a massively parallel version of the reciprocal hemizygote test. We create a genomic complement of hemizygote mutants, by generating viable, sterile F1 hybrids between species and subjecting them to transposon mutagenesis. We pool the hemizygotes, measure their longevity and fitness in sequencing-based assays, and test for differences in frequency between clones of the pool bearing the two parents' alleles of a given gene. The result is a catalog of loci at which variants between species influence the trait of interest. In our proof of principle using yeast (Aim 1) and worm (Aim 2), we will uncover alleles that have arisen in wild species to boost healthspan and stress resistance. These results will stand in contrast to the alleles weakening fitness that are often mapped in intra-specific studies. Orthologs of our yeast loci will be of immediate interest as candidates for virulence genes in prevalent fungal pathogens, and orthologs of our worm lifespan factors will be well-suited to analysis for anti-aging effects in mammals. This effort will be the first-ever comprehensive survey of the genetic architecture and molecular genetics of trait variation between reproductively isolated individuals. And the methods we pioneer will enable genetic dissection in any eukaryotic system in which banks of F1 hybrid individuals or their tissues are available.

概括 遗传学的主要目标之一是了解如何以及为什么自然变化的个体不同 表型。映射这种性状变化的分子基础可以很简单，但只有 可以交织。我们发现，有点研究的蠕虫物种，caenorhabditis latens，有一个中位数 寿命比相关线虫长50％，我们观察到了坚固的，保守的温耐 酿酒酵母，其进化枝的唯一物种可以充当机会性病原体。在 当前的提案，我们使用互惠的大量平行版本剖析了这些物种差异 半合子测试。我们通过产生可行的无菌F1来创建半合子突变体的基因组补体 物种之间的杂种并使它们经过转座子诱变。我们汇集半合子，测量 它们在基于测序的测定中的寿命和适应性，并测试 带有给定基因的两个父母等位基因的池。结果是一个基因座目录，其中变体 物种之间影响感兴趣的特征。在我们使用酵母（AIM 1）和蠕虫（AIM 2）的原理证明中， 我们将发现在野生物种中出现的等位基因，以增强健康范围和抗压力。这些 结果将与通常在特异性研究中映射的等位基因相反。 我们的酵母基因座的直系同源物将立即成为普遍真菌中毒力基因的候选者 病原体和蠕虫寿命因素的直系同源物将非常适合分析抗衰老作用 哺乳动物。这项工作将是对遗传结构和分子的首次全面调查 生殖分离个体之间性状变化的遗传学。我们开拓的方法将启用 在任何真核系统中的遗传解剖，其中F1杂种个体或其组织的库是 可用的。