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

基本信息

批准号：
10657586
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
项目类别：
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/10657586
关键词：
Affect Behavior Biochemical Biology Bromodomains and extra-terminal domain inhibitor Categories Cell Line Cells Chemicals Chromatin Chromatin Modeling Communities Data Data Set Dependence Development Disparate EZH2 gene Enzymes Fluorescence Microscopy Gene Expression Gene Expression Profiling Genes Genetic Genetic Transcription Genome Genomics Goals Heterochromatin Histone Deacetylase Inhibitor Human Image In Vitro Individual Introns Link Liquid substance Maps 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 cell fixing cellular imaging chromatin remodeling computerized tools dosage induced pluripotent stem cell inhibitor 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 technology platform tool transcriptome transcriptome sequencing

项目摘要

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.

项目概要最近对几种染色质调节因子的相分离能力的发现催生了新的模型用于染色质组织。直接测试相分离机制的因果效应需要一种能力定量调节内源染色质调节剂的相分离能力并同时进行测试用于基因表达和细胞命运的变化。为了这个目标，我们将开发两个工具包：（1）相位转盘以增强或以定量和受控的方式抑制内源蛋白质的相分离潜力，以及 (2) 一个强大且可转移的技术平台，将相凝结的实时成像与单细胞水平的基因表达谱。利用这些工具，我们将研究监管和影响干细胞分化过程中异染色质形成和转录激活的研究。