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Multiplexed targeted profiling of single-cell proteomes and transcriptomes in a single reaction

机译:在单个反应中对单细胞蛋白质组和转录组进行多重靶向分析

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摘要

We present a scalable, integrated strategy for coupled protein and RNA detection from single cells. Our approach leverages the DNA polymerase activity of reverse transcriptase to simultaneously perform proximity extension assays and complementary DNA synthesis in the same reaction. Using the Fluidigm C1™ system, we profile the transcriptomic and proteomic response of a human breast adenocarcinoma cell line to a chemical perturbation, benchmarking against in situ hybridizations and immunofluorescence staining, as well as recombinant proteins, ERCC Spike-Ins, and population lysate dilutions. Through supervised and unsupervised analyses, we demonstrate synergies enabled by simultaneous measurement of single-cell protein and RNA abundances. Collectively, our generalizable approach highlights the potential for molecular metadata to inform highly-multiplexed single-cell analyses.Electronic supplementary materialThe online version of this article (doi:10.1186/s13059-016-1045-6) contains supplementary material, which is available to authorized users.
机译:我们提出了一种可扩展的,整合的策略,用于从单个细胞中检测蛋白质和RNA。我们的方法利用逆转录酶的DNA聚合酶活性,在同一反应中同时进行邻近延伸分析和互补DNA合成。使用Fluidigm C1™系统,我们分析了人乳腺腺癌细胞系对化学扰动的转录和蛋白质组学反应,对原位杂交和免疫荧光染色以及重组蛋白,ERCC Spike-Ins和群体裂解物稀释液进行了标定。通过监督和非监督分析,我们证明了通过同时测量单细胞蛋白质和RNA丰度实现的协同作用。总的来说,我们的通用方法强调了分子元数据为高度多元化的单细胞分析提供信息的潜力。电子补充材料本文的在线版本(doi:10.1186 / s13059-016-1045-6)包含补充材料,可用于授权用户。

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