in situ Epigenetic Profiling of Single Cells in Kidney

项目摘要/摘要 这项提案的目标是开发和验证两种工具，用于表观遗传状态和表观遗传状态的原位分析 肾组织中单个细胞的染色质组织。第一个工具是基于我们最近开发的 为翻译后表观遗传编码建立的单细胞评估 用于肾组织的培养细胞，以便直接分解和定量组蛋白 单细胞中特定DNA位点的原位修饰。第二个工具是一种称为APT-FISH的新方法 (来自标记荧光原位杂交的扩增探针)，旨在描述 打开染色质。这些工具中的每一个都将与超分辨率扩展显微镜同时使用 为了将纳米级的肾脏生理与染色质或表观遗传状态直接联系起来进行成像，它们将 应用于肾组织衰老和衰老的研究。

Project Summary/Abstract The goal of this proposal is to develop and validate two tools for in situ analysis of epigenetic state and chromatin organization in single cells in kidney tissue. The first tool is based on our recently developed technology SCEPTRE (Single-Cell Evaluation of Post TRanslational Epigenetic encoding) established for cultured cells that will be adapted for use with kidney tissues in order to directly resolve and quantify histone modifications at specific DNA loci in situ in single cells. The second tool is a new method called APT-FISH (Amplified Probes from Tagmentation Fluorescence in situ Hybridization) that is designed to profile changes to open chromatin. Each of these tools will be used concurrently with super-resolution expansion microscopy imaging in order to directly relate nanoscale renal physiology to chromatin or epigenetic state and they will be applied to the study of aging and senescence in kidney tissue.