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

摘要 组蛋白修饰携带丰富的细胞记忆信息和基因调控机制。单细胞 结合转录组分析组蛋白修饰可以恢复这一关键的细胞身份层 并有助于剖析大脑等复杂组织的细胞和分子组成。我们最近 建立了一种可以联合定位转录组的超高通量单细胞多组学分析方法--配对标签 和组蛋白修饰从多达一百万个单细胞并行。为了将这项新技术 为了市场和推进大脑计划的目标，我们建议优化和进一步发展 配对标签，使其能够在研究界广泛使用。具体地说，我们将开发成对标记工具包来 允许该方法在分子生物学实验室中普遍可用。然后我们将广泛开展 在当前脑活动研究人员的实验室中对配对标签试剂盒进行现场测试，并进一步改进它 根据反馈信息。最后，我们将使成对标签程序适应长阅读测序技术 以实现在单细胞分辨率下高通量检测脑细胞中的剪接异构体。如果成功，则 拟议的研究将为研究人员提供一种强大的新工具，用于单细胞转录和 表观基因组学分析。

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.