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

基本信息

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
项目类别：
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

项目摘要

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）：人类皮层的转录多样性和异质性的全面表征对于了解其在健康和疾病状况中的功能至关重要。皮质中密集填充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性准确地捕获。一个总体目标是建立人类皮质中所有转录活动的高分辨率三维图。在这个项目中，我们将使用Illumina开发的新RNA测序方法，从三个区域（视觉，时间，前额叶）中生成10,000组完整的转录组数据（视觉，时间，前额叶，前额叶），可以捕获单个细胞中的所有mRNA，miRNA，PIRNA和其他非编码RNA物种。此外，我们将开发一种新型的RNA原位测序方法，并将其应用于皮层切片，以在单个细胞的空间分辨率下直接在组织中直接绘制和量化500个转录本。使用这些约500个笔录作为指纹的空间信息，我们将按单个单细胞分辨率在计算上绘制10,000个完整的转录组数据集中的其他成绩单为皮层截面，这将产生一个非常全面的转录活动图，I人类皮质在未经预言的分辨率水平中。