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: 10640172
• 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/10640172
• 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; Shapes; Site; System; Technology; Therapeutic; cell behavior; cell type; chromatin modification; design; epigenome editing; epigenomics; genomic locus; human disease; improved; insight; interdisciplinary approach; novel therapeutics; nuclease; 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.

项目摘要 基因表达失调是人类疾病的广泛且敏锐的驱动力。因此， 理解和精确控制基因表达的能力有可能彻底改变治疗性 景观。人类基因的表达由调节的优雅收敛自然控制 力，包括染色质的物理压实，翻译后修饰（PTMS）对组蛋白 蛋白质，DNA甲基化以及转录因子和染色质修饰之间的动态参与 蛋白质和人类基因组。尽管这种协调的控制措施保护了健康与 疾病，我们对这些调节力如何团结起来驱动人类基因表达的机械理解， 并且如何可以预测地重新设计新疗法，仍然有限。 基于Null CRISPR/CAS的系统，可编程的表观基因组编辑工具最近已有 出现并启用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。尽管取得了令人兴奋的进步，但仍然存在重大的技术和概念差距。例如， 在机械上不清楚特定基因的表达水平如何精确地在宽范围内调节 在人类细胞中。另外，它不完全理解为什么相同的转录因子和染色质 在不同人类细胞类型的特定调节元件上，修饰符具有不同的影响。 此外，转录因子的时空稳定性和功能持续时间 不同的基因组基因座没有很好地定义。 这个MIRA项目的目的是通过发展和 将各种CRISPR/CAS基于CRISPR/CAS的转录因子和染色质修饰符与散装和单细胞结合 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 有关人类基因调节机制的问题，包括：（1）人类基因表达如何位置 - 专门控制在健康和疾病中观察到的决议？ （2）什么是因果功能和 各种染色质修饰的操作稳定性？ （3）染色质压实在多大程度上， 修饰状态，蛋白质复合物组成和空间近端影响转录因子的功能 和染色质修饰符？总的来说，我们的建议具有探索和控制关键的巨大潜力 对人类健康具有广泛和重要性的机制。