Broadly Accessible Technologies for Single-cell Joint Analysis of Transcriptome and Epigenome

转录组和表观基因组单细胞联合分析的广泛可用技术

基本信息

批准号：
10383385
负责人：
Bing Ren
金额：
$ 45万
依托单位：
EPIGENOME TECHNOLOGIES, INC.
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10383385
关键词：
Address Adopted Adoption Adult Alternative Splicing Antibodies Area BRAIN initiative Bar Codes Biological Assay Brain Catalogs Cell Lineage Cells Chromatin Communities Complementary DNA Complex DNA DNA Library DNA sequencing Detection Development Ensure Equipment Feedback Funding Genes Goals Hippocampus (Brain)Human Individual Institutes Joints Laboratories Lead Length Licensing Ligation Link Manuals Maps Memory Methods Modality Molecular Molecular Biology Mus Neuroglia Neurons Neurophysiology - biologic function Performance Phase Procedures Process Protein Isoforms Protocols documentation Quality Control RNA Splicing Reagent Regulator Genes Research Research Institute Research Personnel Resolution Reverse Transcription Small Business Technology Transfer Research Structure Technology Testing Tissues Transcript Translating Translations Tube Work anticancer research base brain cell brain research brain tissue cell type cognitive function design epigenome epigenomics experimental study falls field study histone modification improved innovative neurotechnologies interest multiple omics neural circuit neural network new technology operation programs single cell analysis single cell technology tool transcriptome transcriptomics

项目摘要

Abstract Histone modifications carry rich information of cellular memory and gene regulatory mechanisms. Single cell analysis of histone modification in conjunction with transcriptome could recover this critical layer of cell identity and help to dissect the cellular and molecular composition of complex tissues such as the brain. We recently developed an ultra-high throughput single cell multi-omics assay, Paired-Tag, that can jointly map transcriptome and histone modifications from up to a million single cells in parallel. In order to translate this novel technology to the marketplace and to advance the goals of the BRAIN initiative, we propose to optimize and further develop Paired-Tag to enable its broad use in the research community. Specifically, we will develop Paired-Tag kits to allow for general accessibility of the method in molecular biology laboratories. We will then carry out extensive field tests of the Paired-Tag kit in the laboratories of current BRAIN initiative investigators, and further improve it based on the feedbacks. Finally, we will adapt the Paired-Tag procedure to long-read sequencing technologies to enable high throughput detection of splicing isoforms in brain cells at single cell resolution. If successful, the proposed research would provide researchers with a powerful new tool for single cell transcriptomics and epigenomics analysis.

抽象的组蛋白修饰具有丰富的细胞记忆和基因调节机制信息。单细胞与转录组结合结合组蛋白修饰的分析可以恢复细胞身份的关键层并有助于剖析复杂组织（例如大脑）的细胞和分子组成。我们最近开发了一个超高的吞吐量单细胞多矩分析，配对标签，可以共同绘制转录组并平行地从一百万个单元格进行组蛋白修饰。为了翻译这种新技术进入市场并促进大脑计划的目标，我们建议优化并进一步发展配对标签以使其在研究界的广泛使用。具体来说，我们将开发配对的标签套件允许该方法在分子生物学实验室中的普遍可访问性。然后，我们将进行大量当前大脑倡议研究人员实验室中配对标签套件的现场测试，并进一步改进基于反馈。最后，我们将对长阅读的测序技术调整配对的程序为了使单细胞分辨率的脑细胞中剪接同工型的高吞吐量检测。如果成功，拟议的研究将为研究人员提供一个有力的新工具，用于单细胞转录组学和表观基因组学分析。