Modulation and functional characterization of protein condensation in chromatin organization

染色质组织中蛋白质凝聚的调节和功能表征

• 批准号: 10264161
• 负责人: Bo Huang
• 金额: $ 108.66万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10264161
• 关键词: Affect; Behavior; Biochemical; Biochemical Genetics; Biology; Categories; Cell Line; Cells; Chemicals; Chromatin; Chromatin Modeling; Communities; Data; Data Set; Dependence; Development; EZH2 gene; Enzymes; Fluorescence Microscopy; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Genome; Genomics; Goals; Heterochromatin; Histone Deacetylase Inhibitor; Human; Image; In Vitro; Individual; Introns; Link; Liquid substance; Measurement; Measures; Modeling; Mutation; Nature; Nuclear; Output; Pathway interactions; Phase; Phenotype; Physical condensation; Process; Property; Protein Dynamics; Protein Engineering; Protein Overexpression; Proteins; RNA; Regulation; Role; System; Testing; Transcription Coactivator; Transcriptional Activation; base; cellular imaging; chromatin remodeling; computerized tools; dosage; induced pluripotent stem cell; inhibitor/antagonist; instrument; live cell imaging; live cell microscopy; multidisciplinary; novel; overexpression; pluripotency; programs; protein protein interaction; reconstitution; single molecule; single-cell RNA sequencing; stem cell differentiation; synergism; tool; transcriptome; transcriptome sequencing

Project Summary Recent discoveries of the phase-separation abilities of several chromatin regulators have led to novel models for chromatin organization. Directly testing the causal effects of phase-separation mechanisms requires an ability to quantitatively tune the phase-separation ability of an endogenous chromatin regulator and simultaneously test for gene expression and cell fate changes. For this goal, we will develop two tool kits: (1) a phase-dial to enhance or suppress the phase-separation potential of endogenous proteins in a quantitative and controlled manner, and (2) a robust and transferrable technological platform to correlate live imaging of phase condensates with the profiling of gene expression at the single cell level. Utilizing these tools, we will study the regulation and impact of heterochromatin formation and transcription activation during stem cell differentiation.

项目摘要 最近发现的相分离能力的几个染色质调节导致了新的模型 对于染色质组织。直接测试相分离机制的因果效应需要一种能力 为了定量调节内源性染色质调节剂的相分离能力， 基因表达和细胞命运的改变。为了实现这一目标，我们将开发两个工具包：（1）一个相位拨号，以提高 或以定量和受控的方式抑制内源性蛋白质的相分离潜力，和 (2)一个强大的和可转移的技术平台，将相凝析物的实时成像与 在单细胞水平上的基因表达谱分析。利用这些工具，我们将研究监管和影响 异染色质的形成和转录激活过程中干细胞分化。