Modulation and functional characterization of protein condensation in chromatin organization

项目摘要 最近对几种染色质调节剂的相分离能力的发现导致了新的模型 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 定量调节内源性染色质调节剂的相分离能力并同时测试 基因表达和细胞命运的改变。为此，我们将开发两个工具包：(1)相位转盘，以增强 或以定量和可控的方式抑制内源蛋白质的相分离潜力，以及 (2)一个强大的、可转移的技术平台，将相凝析油的实时成像与 在单细胞水平上的基因表达谱。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的形成和转录激活。

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.