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

• 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
• 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

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.

摘要 组蛋白修饰携带丰富的细胞记忆和基因调控机制的信息。单细胞 结合转录组分析组蛋白修饰可以恢复细胞身份的这一关键层 并有助于解剖复杂组织（如大脑）的细胞和分子组成。我们最近 开发了一种超高通量的单细胞多组学分析方法Paired-Tag，可以联合绘制转录组 和组蛋白的修饰。为了将这项新技术 为了推进BRAIN计划的目标，我们建议优化和进一步开发 Paired-Tag使其能够在研究社区中广泛使用。具体来说，我们将开发配对标签套件， 允许该方法在分子生物学实验室中的普遍可及性。我们将进行广泛的 在当前BRAIN倡议调查人员的实验室中对配对标签试剂盒进行现场测试，并进一步改进 基于反馈。最后，我们将使配对标签程序适应长读段测序技术 从而能够以单细胞分辨率高通量检测脑细胞中的剪接异构体。如果成功， 拟议的研究将为研究人员提供一个强大的单细胞转录组学的新工具， 表观基因组学分析