Mechanism of Chromatin Accessibility, 3D Chromosome Organization, and Their Functions in Gene Regulation

项目摘要 父母补助金R35 GM139654的主要重点之一是研究机制， 先锋因子（PFs）。先锋因子（Pioneer factors，PFs）可侵入核小体， 染色质可及性附近的结合位点，因此发挥关键作用，在基因 调控PFs的错误调节与癌症和发育疾病密切相关。尽管 本文综述了PFs的基本功能、PFs侵入核小体的机制及其在核小体形成中的作用。 基因调控需要进一步阐明。特别与本补充申请相关的是， 我们希望对先驱因子如何影响基因表达有定量的了解， 水平，噪音，和动态信号的存在。为此，我们计划 干扰先锋因子并测量单细胞中基因表达的后果 通过延时荧光成像。在这里，我们要求资金购买显微镜 这个系统可以让我们进行长期的活细胞成像。该设备将满足一个关键的 需要我的实验室，以及宾夕法尼亚州立大学的基因调控中心，我的实验室是其中的一部分 的.

Project Summary One of the major focus of the parental grant, R35 GM139654, is to study the mechanism and function of pioneer factors (PFs). Pioneer factors (PFs) can invade nucleosome and increase chromatin accessibility near their binding sites and therefore play critical roles in gene regulation. Mis-regulation of PFs is highly linked to cancer and developmental disease. Despite their essential functions, the mechanism of nucleosome invasion by PFs and their function in gene regulation require further elucidation. Particularly relevant to this supplement application, we want to have a quantitative understanding of how pioneer factors affect gene expression level, noise, and dynamics in the presence of signaling. Towards this purpose, we plan to perturb pioneer factors and measure the consequences in gene expression in single cells through time-lapse fluorescence imaging. Here, we request funding to purchase a microscope system that allows us to perform long-term live cell imaging. This equipment will fulfill a critical need for my lab, as well as the Center of Gene Regulation at Penn State, which my lab is part of.