High-throughput Single Cell Co-assay of Histone Modifications andTranscriptome

组蛋白修饰和转录组的高通量单细胞联合分析

基本信息

批准号：
10698374
负责人：
Christopher Hartl
金额：
$ 113.31万
依托单位：
EPIGENOME TECHNOLOGIES, INC.
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：
2023-07-03 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10698374
关键词：
Address Adoption Antibodies Architecture Automation Bar Codes Binding Bioinformatics Biological Biological Assay Biopsy Cancerous Cell Nucleus Cells Chemicals Chromium Chromosomes Collection Computer software Contract Services Custom DNA DNA Crosslinking DNA Methylation DNA-Binding Proteins Data Data Analyses Data Set Disease Ensure Epigenetic Process Equipment Gene Expression Gene Expression Profile Gene Expression Regulation Genes Genetic Fingerprintings Genetic Materials Genetic Transcription Goals Heritability Heterogeneity Hour Image Individual Informatics Infrastructure Intestines Investments Joints Laboratories Laboratory Research Libraries Management Information Systems Manuals Memory Methodology Methods Microfluidics Molecular Monitor Neurons Nuclear Organism Performance Permeability Persons Phase Population Preparation Protocols documentation Publishing Reagent Regulator Genes Regulatory Element Renal Tissue Reproducibility Research Research Institute Running Sampling Scanning Services Signal Transduction Small Business Innovation Research Grant System Technology Testing Tissue Sample Tissues Tn5 transposase Tube Work analysis pipeline cell type chromatin immunoprecipitation combinatorial computerized data processing cost crosslink data complexity design developmental disease epigenetic profiling epigenome experimental study histone modification human error improved insight multiple omics new technology next generation preservation programs protein expression relational database segregation single cell technology tool transcription factor transcriptome sequencing tumor tumorigenesis virtual web site

项目摘要

Summary Disruption of gene regulation is a major causal factor in heritable disease, developmental disorders, and oncogenesis. While chromatin immunoprecipitation followed by sequencing (ChIP-seq) and Tn5- transposase based tagging (Cut&Tag) enable analysis of transcription factor binding and epigenetic state profiling in bulk tissue samples and tumor biopsies, they produce only population average signals. Yet regulatory networks and perturbations are heterogeneous between cell classes and types. Single-cell technologies can overcome the challenge of cellular heterogeneity and provide deeper insight into cell type-specific gene regulatory programs in healthy, diseased, and cancerous tissues. In prior work, we developed a single-cell joint assay of histone modification and RNA expression (Paired-Tag), enabling cell-type-stratified epigenetic profiling from bulk samples. This technology has attracted customers in both academic and nonprofit research. In the proposed study, we will develop automated protocols and effect laboratory informatics systems to establish a Paired-Tag services laboratory, we will refine our protocol to improve experimental throughput and reduce cost, and we will develop a Paired-Tag “TF” protocol for profiling transcription factor binding profiles. If successful, the research would enable next- generation multi-omic analysis of tumor or disease samples at comparable cost to single-omic technologies.

概括基因调节的破坏是可遗传疾病，发育障碍，和肿瘤发生。而染色质免疫沉淀，然后进行测序（chip-seq）和TN5- 基于转座酶的标记（剪切和标签）启用转录因子结合和表观遗传状态的分析在散装组织样品和肿瘤活检中进行分析，仅产生人口平均信号。然而调节网络和扰动是细胞类别和类型之间的异质性。单细胞技术可以克服细胞异质性的挑战，并为细胞提供更深入的洞察力在健康，解散和取消组织中的类型特异性基因调节程序。在先前的工作中，我们开发了组蛋白修饰和RNA表达（配对标签）的单细胞关节测定法来自散装样品的细胞类型分层的表观遗传分析。这项技术吸引了客户学术和非营利研究。在拟议的研究中，我们将制定自动化协议，并影响实验室信息系统建立配对的服务实验室，我们将完善我们的改善实验吞吐量并降低成本的协议，我们将开发一个配对的标签“ TF” 用于分析转录因子结合曲线的协议。如果成功，这项研究将使下一步肿瘤或疾病样品的多摩尼克生成分析，其成本与单摩克技术。