一种用于检测……中单个信使核糖核酸分子的方法。 （原文中“in.”后面内容缺失）
A method for detecting single mRNA molecules in .
作者信息
Duncan Susan, Olsson Tjelvar S G, Hartley Matthew, Dean Caroline, Rosa Stefanie
机构信息
Department of Computational and Systems Biology, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH UK.
Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park, Norwich, NR4 7UH UK.
出版信息
Plant Methods. 2016 Aug 5;12:13. doi: 10.1186/s13007-016-0114-x. eCollection 2016.
BACKGROUND
Despite advances in other model organisms, there are currently no techniques to explore cell-to-cell variation and sub-cellular localization of RNA molecules at the single-cell level in plants.
RESULTS
Here we describe a method for imaging individual mRNA molecules in root cells using multiple singly labeled oligonucleotide probes. We demonstrate detection of both mRNA and nascent transcripts of the housekeeping gene . Our image analysis pipeline also enables quantification of mRNAs that reveals the frequency distribution of transcripts per cell underlying the population mean.
CONCLUSION
This method allows single molecule RNA in situ to be exploited as a powerful tool for studying gene regulation in plants.
背景
尽管在其他模式生物方面取得了进展，但目前尚无技术可在植物的单细胞水平上探索RNA分子的细胞间变异和亚细胞定位。
结果
在此，我们描述了一种使用多个单标记寡核苷酸探针在根细胞中对单个mRNA分子进行成像的方法。我们展示了对管家基因的mRNA和新生转录本的检测。我们的图像分析流程还能够对mRNA进行定量，从而揭示群体平均值背后每个细胞转录本的频率分布。
结论
该方法使单分子RNA原位技术成为研究植物基因调控的有力工具。
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