Cellular Systems Genetic Approaches to Understanding Regulatory Variation

项目总结/摘要 我实验室的首要目标是确定自然遗传变异如何影响 染色质生物学，以及最终的表型多样性。大多数与疾病相关的变异在 人类研究发生在调控元件，而不是在基因的编码区，突出了 理解调节变异在正常健康和疾病中的作用的重要性。基因转录 由顺式作用调节元件和结合它们的反式作用因子之间的相互作用控制。 调控元件内的变异可以通过破坏DNA结合位点来局部影响功能 被该位点的反式作用因子靶向，而反式作用因子的序列或表达的变化 因子可导致远端许多调节元件的功能变异。确定了监管要素 由多种染色质书写者催化的表观遗传特征。虽然我们有大量的信息， 酶，写和删除表观遗传修饰，很少有人知道的反式作用机制， 规范这些修改的位置和级别，仍然是该领域的一个决定性挑战。到 为了推动这一领域的发展，我们开发了一种新的细胞系统遗传学方法， 统计遗传学中的定量实验技术和计算方法， 全面了解监管变化，并发现广泛适用的机制， 表观遗传景观我们将利用一组独特的小鼠中固有的自然遗传变异， 胚胎干细胞，这将使我们能够利用遗传多样性，以确定关键的染色质调节，定义 它们在多种表型中的分子功能，并描绘了在不同的表型中调控变异的变化。 发展时间。这种哺乳动物遗传参考面板将大大提高统计能力， 绘制与人类直接相关的功能基因座，而细胞平台将能够识别其功能基因座。 相关基因和控制机制。使用这个系统，我们发现了多个基因组 这些位点远端控制数百个调控元件并改变基因表达。我们未来的目标 五年的时间是为了回答以下问题：什么是分子和机制背后的反式 染色质景观的调控遗传变异如何影响染色质的结构和功能？ 染色质景观的早期建立如何影响发育？该项目的成功将 描述调控元件之间的相互作用，控制它们的系统，以及表观遗传 定义他们的风景。预计这些努力将对我们的基本 了解早期发育过程中的染色质和表观遗传调控，以及我们的能力， 根据遗传背景直接定制体外细胞分化，最终影响发育生物学， 再生生物学和个性化医疗。

PROJECT SUMMARY/ABSTRACT The overarching goal of my laboratory is to determine how natural genetic variation influences chromatin biology, and, ultimately, phenotypic diversity. Most disease-associated variants identified in human studies occur in regulatory elements rather than in the coding region of genes, highlighting the importance of understanding the role of regulatory variation in normal health and disease. Gene transcription is controlled by the interplay between cis-acting regulatory elements and the trans-acting factors that bind them. Variation within a regulatory element can influence function locally through disruption of a DNA-binding site targeted by a trans-acting factor at that site, whereas variation in sequence or expression of a trans-acting factor can result in functional variation distally at many regulatory elements. Regulatory elements are identified by epigenetic features catalyzed by a variety of chromatin writers. While we have substantial information on the enzymes that write and erase epigenetic modifications, less is known about the trans-acting mechanisms that regulate the locations and levels of these modifications, remaining a defining challenge within the field. To move the field forward, we have developed a novel cellular systems genetics approach that integrates quantitative experimental techniques and computational methods in statistical genetics to create a comprehensive understanding of regulatory variation and uncover broadly applicable mechanisms that control the epigenetic landscape. We will utilize the natural genetic variation intrinsic in a unique panel of mouse embryonic stem cells that will enable us to harness genetic diversity to identify key chromatin regulators, define their molecular functionality across multiple phenotypes, and delineate changes in regulatory variation over developmental time. This mammalian genetic reference panel will greatly improve statistical power to efficiently map functional loci directly relatable to humans, whereas a cellular platform will enable the identification of their associated genes and mechanisms of control. Using this system we have discovered multiple genomic locations that distally control hundreds of regulatory elements and alter gene expression. Our goals in the next five years are to answer the following questions: What are the molecules and mechanisms underlying trans regulation of the chromatin landscape? How does genetic variation influence chromatin structure and function? How does the early establishment of the chromatin landscape impact development? Success of this project will delineate the interplay between regulatory elements, the systems that control them, and the epigenetic landscape that defines them. These efforts are expected to have broad implications in both our basic understanding of chromatin and epigenetic regulation during early development as well as on our ability to directly tailor in vitro cellular differentiation to genetic background, ultimately impacting developmental biology, regenerative biology and personalized medicine.