Single-cell sequencing and in situ mapping of RNA transcripts in human brains

人脑 RNA 转录本的单细胞测序和原位作图

• 批准号: 8688362
• 负责人: Kun Zhang
• 金额: $ 200.36万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-19 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8688362
• 关键词: Address; Area; Biological Markers; Brain; Cell Nucleus; Cells; Chromosome Mapping; Complex; Custom; Data; Data Set; Development; Disease; Female; Fingerprint; Functional RNA; Funding; Gene Expression Profile; Generations; Genes; Genetic Transcription; Genome; Genomics; Goals; Grant; Heterogeneity; Human; Image; In Situ; Individual; Label; Maps; Messenger RNA; Methods; MicroRNAs; Mosaicism; Neuroglia; Neurons; Noise; Ploidies; Production; Protocols documentation; RNA; RNA Sequences; Research Personnel; Resolution; Sorting - Cell Movement; Structure; TNFRSF5 gene; Technology; Testing; Three-Dimensional Imaging; Tissues; Transcript; United States National Institutes of Health; Variant; Visual; Visual Cortex; base; brain disorder diagnosis; cognitive function; cost; design; extrastriate visual cortex; instrument; instrumentation; new technology; novel; outcome forecast; piRNA; public health relevance; submicron; transcriptome sequencing; white matter

DESCRIPTION (provided by applicant): A comprehensive characterization of transcriptional diversity and heterogeneity of the human cortex is crucial to understand its functions in healthy and disease conditions. The diversity and cellular states of the densely packed cellular network in the cortex can be accurately captured by the transcriptional activities of individual cells. An overarching goal is to establish a high-resolution three dimensional map of all transcriptional activities in the human cortex. In this project we will generate 10,000 sets of full transcriptome data on single cells and nuclei from three areas (visual, temporal, prefrontal) of the human cortex, using a new RNA sequencing method developed by Illumina that can capture all mRNA, miRNA, piRNA and other non-coding RNA species in single cells. In addition, we will develop a novel RNA in situ sequencing method, and apply it to cortex sections to map and quantify at least 500 transcripts directly within the tissue at a spatial resolution of single cells. Using the spatial information of these ~500 transcripts as fingerprints, we will computationally map the additional transcripts in the 10,000 full transcriptome data sets to the cortex sections at the single-cell resolution, which will yield a highly comprehensive map of transcriptional activities i the human cortex in an unprecedented level of resolution.

描述（由申请人提供）：人类皮质的转录多样性和异质性的全面表征对于理解其在健康和疾病条件下的功能至关重要。皮质中密集的细胞网络的多样性和细胞状态可以通过单个细胞的转录活动准确地捕获。一个首要目标是建立一个高分辨率的三维地图的所有转录活动在人类大脑皮层。在这个项目中，我们将使用Illumina开发的一种新的RNA测序方法，从人类皮层的三个区域（视觉，颞叶，前额叶）的单细胞和细胞核中生成10，000组全转录组数据，该方法可以捕获单细胞中的所有mRNA，miRNA，皮尔纳和其他非编码RNA物种。此外，我们将开发一种新的RNA原位测序方法，并将其应用于皮质切片，以单细胞的空间分辨率直接在组织内映射和量化至少500个转录本。使用 将这约500个转录物的空间信息作为指纹，我们将以单细胞分辨率将10，000个全转录组数据集中的额外转录物计算映射到皮层切片，这将以前所未有的分辨率水平产生人类皮层中转录活性的高度全面的映射。