Site-specific control of human gene regulation for therapeutically applicable mechanistic insights

人类基因调控的位点特异性控制以获得治疗上适用的机制见解

• 批准号: 10488643
• 负责人: Isaac Hilton
• 金额: $ 37.71万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10488643
• 关键词: Affect; Cells; Chromatin; Chromatin Remodeling Factor; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; DNA Methylation; Disease; Equilibrium; Gene Expression; Gene Expression Regulation; Genes; Goals; Health; Histones; Human; Human Genome; Knowledge; Modification; Post-Translational Protein Processing; Process; Proteins; Proteomics; Regulator Genes; Regulatory Element; Research Personnel; Resolution; Site; System; Therapeutic; Widespread Disease; base; cell behavior; cell type; chromatin modification; epigenome editing; epigenomics; genomic locus; human disease; improved; insight; interdisciplinary approach; novel therapeutics; nuclease; programs; protein complex; spatiotemporal; tool; transcription factor

PROJECT SUMMARY Dysregulated gene expression is a widespread and disease-agnostic driver of human illness. Therefore, the ability to understand and precisely control gene expression has the potential to revolutionize the therapeutic landscape. The expression of human genes is naturally controlled by an elegant convergence of regulatory forces, including the physical compaction of chromatin, post-translational modifications (PTMs) to histone proteins, DNA methylation, and the dynamic engagement between transcription factors and chromatin modifying proteins and the human genome. Although this coordinated control safeguards the balance between health and disease, our mechanistic understanding of how these regulatory forces unite to drive human gene expression, and how they can be predictably redesigned for new therapies, remains limited. Programmable epigenome editing tools based upon nuclease null CRISPR/Cas-based systems have recently emerged and enable new ways to control endogenous human gene expression and covalent modifications to native chromatin. Despite this exciting progress, major technological and conceptual gaps remain. For instance, it is mechanistically unclear how the expression levels of specific genes can be precisely tuned over wide ranges in human cells. In addition, it is incompletely understood why the same transcription factors and chromatin modifiers have different effects when localized to specific regulatory elements in different human cell types. Further, the spatiotemporal stability and functional durations of transcription factors and chromatin modifiers at different genomic loci are not well defined. The goal of this MIRA project is to overcome these conceptual and technological gaps by developing and combining diverse CRISPR/Cas-based transcription factors and chromatin modifiers with bulk and single cell epigenomics and sensitive proteomics. We will use this multidisciplinary approach to answer fundamental questions about human gene regulatory mechanisms including: (1) How can human gene expression be site- specifically controlled at the resolution observed in health and disease? (2) What are the causal functions and operational stabilities of diverse chromatin modifications? (3) To what extent does chromatin compaction, modification state, protein complex composition, and spatial proximity affect the function of transcription factors and chromatin modifiers? Altogether, our proposal has great potential to uncover and enable control over pivotal mechanisms with broad and significant importance to human health.

项目总结 基因表达失调是一种广泛存在的、与疾病无关的人类疾病驱动因素。因此， 理解和精确控制基因表达的能力可能会给治疗带来革命性的变化 风景。人类基因的表达自然而然地受到调控机制的巧妙融合 作用力，包括染色质的物理压缩，组蛋白的翻译后修饰(PTM) 蛋白质、DNA甲基化、转录因子与染色质修饰的动态结合 蛋白质和人类基因组。尽管这种协调控制保护了健康和健康之间的平衡 疾病，我们对这些调控力量如何联合起来驱动人类基因表达的机械性理解， 以及如何可以预测地为新的疗法重新设计它们，仍然有限。 基于核酸酶零CRISPR/CAS系统的可编程表观基因组编辑工具最近已经 出现了控制内源性人类基因表达和共价修饰的新方法 天然染色质。尽管取得了这一令人振奋的进展，但在技术和概念上仍存在重大差距。例如, 目前还不清楚如何在大范围内精确调节特定基因的表达水平。 在人类细胞中。此外，人们还不完全理解为什么相同的转录因子和染色质 当修饰物定位于不同类型的人类细胞中的特定调控元件时，具有不同的效果。 此外，转录因子和染色质修饰物的时空稳定性和作用持续时间 不同的基因组座位没有很好的定义。 该Mira项目的目标是通过开发和开发 多种基于CRISPR/Cas的转录因子和染色质修饰物与块状和单细胞相结合 表观基因组学和敏感蛋白质组学。我们将使用这种多学科方法来回答基本问题 关于人类基因调控机制的问题包括：(1)人类基因表达是如何定位的？ 具体控制在观察到的健康和疾病的决议？(2)什么是因果函数和 不同染色质修饰的操作稳定性？(3)染色质紧凑到什么程度， 修饰状态、蛋白质复合体组成和空间邻近影响转录因子的功能 染色质修饰剂呢？总而言之，我们的提案具有发现和控制Pivotal的巨大潜力 对人类健康具有广泛和重大意义的机制。