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单细胞中多个基因的蛋白和 RNA 定量。

Protein and RNA quantification of multiple genes in single cells.

机构信息

Centre for Research in Neuroscience, Research Institute of the McGill University Health Centre, Montréal, Québec, H3G 1A4, Canada.

Departments of Medicine and Neurology & Neurosurgery, McGill University, Montréal, Québec, H3G 1A4, Canada.

出版信息

Biotechniques. 2019 Jan;66(1):15-21. doi: 10.2144/btn-2018-0130. Epub 2018 Oct 18.

Abstract

Single-cell analysis overcomes the problems of cellular heterogeneity by revealing the individual differences between cells in tissue. The current tools used to profile gene expression at the single-cell level are arduous and often require specialized equipment. We have previously developed a technique to quantify protein expression levels in single living cells. Here, we combine quantification of protein expression with absolute measurement of mRNA amounts of the same gene in the same cell, to profile the expression of genes at the transcriptional and translational levels. We show that high heterogeneity exists at both the mRNA and protein levels for multiple genes, even among monoclonal cells. We demonstrate a rapid, straightforward approach to single-cell profiling of RNA and protein production.

摘要

单细胞分析通过揭示组织中细胞之间的个体差异来克服细胞异质性的问题。目前用于单细胞水平基因表达分析的工具繁琐,通常需要专门的设备。我们之前开发了一种定量单个活细胞中蛋白质表达水平的技术。在这里,我们将蛋白质表达的定量与同一细胞中同一基因的 mRNA 量的绝对测量相结合,以对转录和翻译水平的基因表达进行分析。我们表明,即使是在单克隆细胞中,多个基因的 mRNA 和蛋白质水平都存在高度异质性。我们展示了一种快速、直接的单细胞 RNA 和蛋白质产生分析方法。

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