in situ Epigenetic Profiling of Single Cells in Kidney

肾脏单细胞的原位表观遗传分析

基本信息

批准号：
10645605
负责人：
Joshua Vaughan
金额：
$ 22.9万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10645605
关键词：
Address Adult Aging American Blood Cells Cessation of life Chromatin Chronic Kidney Failure Complex Correlative Study Cultured Cells DNA DNA Methylation Dialysis procedure Disease Dissociation Electrolyte Balance End stage renal failure Endothelial Cells Epigenetic Process Evaluation Face Fluorescent in Situ Hybridization Generations Genome Goals Health Expenditures Heterochromatin Histones Image In Situ Individual Injury Kidney Kidney Diseases Maps Measures Methods Microscopy Molecular Molecular Conformation Mus Organ Phenotype Physiological Physiology Population Post-Translational Protein Processing Publishing Renal function Research Personnel Risk Role Specimen Stains Techniques Technology Time Tissues Transplantation Use of New Techniques Work base blood pressure regulation design epigenetic profiling field study genome-wide genomic locus glomerular basement membrane healthy aging histone modification human old age (65+)innovation insight kidney cell kidney imaging microscopic imaging molecular phenotype mouse model nanoscale novel podocyte senescence single cell analysis superresolution microscopy tool ultra high resolution wasting whole genome

项目摘要

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.

项目概要/摘要该提案的目标是开发和验证两种用于表观遗传状态原位分析的工具和肾组织中单个细胞的染色质组织。第一个工具基于我们最近开发的技术 SCEPTER（翻译后表观遗传编码的单细胞评估）建立用于培养的细胞将适合与肾组织一起使用，以便直接解析和定量组蛋白单细胞中特定 DNA 位点的原位修饰。第二个工具是一种名为 APT-FISH 的新方法（标记荧光原位杂交的扩增探针）旨在分析以下变化：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成像，以便将纳米级肾脏生理学与染色质或表观遗传状态直接联系起来，它们将被应用于肾组织衰老和衰老的研究。