Mechanisms and phenotypic consequences of structural geonomic variation

直到最近，结构基因组变异才成为人们关注的主要遗传多样性来源 人类，以及整个生物学。尽管它至关重要，但我们对它的了解仍然非常有限 导致基因组结构随时间变化的过程，以及这些过程的后果 生物有机体的大规模变化。我建议使用两种并行和综合的方法来研究这一问题 问题，利用发芽酵母模型系统中可用的独特研究工具。(1)我会 研究导致染色体断裂的作用力，以及导致染色体断裂的细胞机制 预防、测量和修复这种损害。为此，我将使用一种定制的高度敏感的分析方法来 测量基因拷贝数变异率(CNV)，并使用基因组分析工具来表征 相关的结构性变化。CNV检测有两个独特的创新特征，将允许研究 传统的实验系统从未正确解决的问题。首先，化验 使用二倍体细胞，使结果与人类发生的过程更加密切；第二， 这种分析方法可以检测到基因组中任何地方的DNA断裂引起的染色体重排， 允许对基因组变异进行更全面的采样。(2)我还会调查表型 与染色体重排相关的二倍体酵母菌株具有许多共同的 具有复杂人类基因组特征的特性。这包括高度的杂合性， 同源基因、基因冗余和CNV之间的结构染色体多态；一直 保留小的(和可管理的)酿酒酵母基因组。我将利用这种菌株进行系统性的 结构变异的功能基因组学研究。该菌株(JAY270)将被操纵以产生靶向 所有染色体中的染色体重排。然后，这批染色体突变体将被 通过一系列表型测试来建立整个基因组的结构-功能关系。我 我坚信，通过开辟这些新的和综合的调查途径，我的研究将有助于 迫切需要深入了解结构基因组变异是如何产生的，以及它如何影响生活的方方面面，从 物种进化对人类健康的影响。

Structural genomic variation has only recently come into focus as a major source of genetic diversity in humans, and in biology in general. Despite its critical importance, we still have a very limited understanding of the processes that cause the structure of genomes to change over time, and of the consequences of these large-scale changes to living organisms. I propose to use two parallel and integrated approaches to study this problem, taking advantage of the unique research tools available in the budding yeast model system. (1) I will investigate the forces that cause chromosomes to break, and the cellular mechanisms that are responsible for preventing, surveying, and repairing this damage. To do so, I will use a custom and highly sensitive assay to measure the rate of gene copy number variation (CNV), and genomic analysis tools to characterize the associated structural changes. The CNV assay has two uniquely innovative features that will allow the study of problems that have never been properly addressed by conventional experimental systems. First, the assay uses diploid cells, making the results much more germane to the processes that occur in humans; and Second, this assay can detect chromosomal rearrangements stemming from DNA breaks anywhere in the genome, allowing a more comprehensive sampling of genomic variants. (2) I will also investigate the phenotypic consequences associated with chromosomal rearrangements in a diploid yeast strain that shares many of the properties that characterize the complex human genome. These include a high degree of heterozygosity, structural chromosomal polymorphisms between homologs, gene redundancy, and CNVs; all the while retaining the small (and manageable) genome of S. cerevisiae. I will use this strain to conduct a systematic functional genomics study of structural variation. This strain (JAY270) will be manipulated to generate targeted chromosomal rearrangements in all chromosomes. This collection of chromosomal mutants will then be taken through a series of phenotypic tests to establish structure-function relationships for the entire genome. I strongly believe that by opening these new and integrated avenues of investigation, my studies will contribute much needed insight into how structural genomic variation arises and how it affects all aspects of life, from the evolution of species to human health.