Human centromere variation and function

项目总结/摘要 细胞分裂过程中染色体的均等分离，保证了遗传物质的准确遗传 信息.异常的染色体分离可以导致染色体数目的不平衡，或非整倍性， 这可能导致自然流产和出生缺陷，是癌症的主要原因。所需的轨迹 使染色体平均分离的是着丝粒。在人类中，着丝粒由 在每条染色体上跨越几个兆碱基的重复α-卫星序列。的重复性质 这些区域已经挑战了使用短读来确定它们的序列、结构和变异的努力。 测序数据。因此，我们对着丝粒的自然变异的了解非常有限， 这种变异对生命至关重要的细胞生物学过程的影响。在这份提案中，我旨在解决 通过对不同人类的着丝粒进行测序和组装， 长读段测序技术和新型计算组装工具（Aim 1; K99阶段）。此外，本发明还 我建议从遗传学、表观遗传学和转录学的角度来评估人类着丝粒的自然变异 水平使用创新的计算方法和多组测序方法，最终建立一个模型 人着丝粒变异（Aim 2; K99期）。最后，我建议确定 着丝粒影响细胞分裂过程中染色体的准确分离，使用基于细胞的分析，长期 读段测序和多组测序方法（Aim 3; R 00阶段）。这项工作将提供 第一次全面评估人类着丝粒变异，并揭示这种变异如何影响 细胞中的着丝粒功能。K99阶段开发的工具、资源和技能将应用于 R 00期，以确定着丝粒变异的功能后果及其在人类健康中的作用， 疾病我的目标是建立一个独立的研究计划，跨越基因组学和基因组学之间的差距。 着丝粒生物学我将从我的导师（埃文Eichler博士），共同接受必要的跨学科培训- 我的导师（苏·比金斯博士）和博士后顾问委员会的其他成员（黛博拉·尼克森博士、安德鲁· Stergachis和Kelley Harris）。此外，我还将参加通过 华盛顿大学和弗雷德·哈奇森癌症研究中心。一起，我的研究训练， 导师，咨询委员会和学术环境将为我过渡到一个独立的 作为一名学术科学家。

Project Summary/Abstract The equal segregation of chromosomes during cell division ensures the accurate inheritance of genetic information. Aberrant chromosome segregation can cause an imbalance in chromosome number, or aneuploidy, which can result in spontaneous abortion and birth defects and is a major cause of cancer. The locus required for the equal segregation of chromosomes is the centromere. In humans, centromeres are comprised of repetitive α-satellite sequences that span several megabases on each chromosome. The repetitive nature of these regions has challenged efforts to determine their sequence, structure, and variation using short-read sequencing data. As a result, we have a very limited understanding of the natural variation of centromeres and the impact of this variation on essential cell biological process critical for life. In this proposal, I aim to address this gap in knowledge by sequencing and assembling centromeres from diverse humans using a combination of long-read sequencing technologies and novel computational assembly tools (Aim 1; K99 phase). Additionally, I propose to assess the natural variation of human centromeres at the genetic, epigenetic, and transcriptional level using innovative computational methods and multiomic sequencing approaches, ultimately building a model of human centromere variation (Aim 2; K99 phase). Finally, I propose to determine how variation among centromeres affects the accurate segregation of chromosomes during cell division using cell-based assays, long- read sequencing, and multiomic sequencing approaches (Aim 3; R00 phase). Together, this work will provide the first comprehensive assessment of human centromere variation and reveal how this variation affects centromere function in cells. The tools, resources, and skills developed in the K99 phase will be applied in the R00 phase to determine the functional consequences of centromere variation and its role in human health and disease. My goal is to build an independent research program that spans the gap between genomics and centromere biology. I will receive the necessary interdisciplinary training from my mentor (Dr. Evan Eichler), co- mentor (Dr. Sue Biggins), and the rest of my postdoctoral advisory committee (Drs. Deborah Nickerson, Andrew Stergachis, and Kelley Harris). In addition, I will participate in career development activities offered through the University of Washington and the Fred Hutchison Cancer Research Center. Together, my research training, mentors, advisory committee, and academic environment will prepare me well as I transition to an independent position as an academic scientist.