A method for detecting single mRNA molecules in Arabidopsis thaliana.

A method for detecting single mRNA molecules in Arabidopsis thaliana.
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DOI:
10.1186/s13007-016-0114-x
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发表时间:
2016
期刊:
影响因子:
5.1
通讯作者:
Rosa S
Rosa S
中科院分区:
生物学2区
文献类型:
--
作者:
Duncan S;Olsson TSG;Hartley M;Dean C;Rosa S

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尽管在其他模式生物中取得了进展,但目前还没有技术来探索植物单细胞水平上RNA分子的细胞间变异和亚细胞定位。在这里,我们描述了一种利用多个单标记寡核苷酸探针成像拟南芥根细胞中单个mRNA分子的方法。我们展示了管家基因蛋白磷酸酶2A的mRNA和新生转录物的检测。我们的图像分析管道也可以量化mrna,揭示每个细胞转录本的频率分布在群体平均水平之下。这种方法使原位单分子RNA成为研究植物基因调控的有力工具。本文的在线版本(doi:10.1186/s13007-016-0114-x)包含补充材料,仅供授权用户使用。
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. Here we describe a method for imaging individual mRNA molecules in Arabidopsis thaliana root cells using multiple singly labeled oligonucleotide probes. We demonstrate detection of both mRNA and nascent transcripts of the housekeeping gene Protein Phosphatase 2A. Our image analysis pipeline also enables quantification of mRNAs that reveals the frequency distribution of transcripts per cell underlying the population mean. This method allows single molecule RNA in situ to be exploited as a powerful tool for studying gene regulation in plants. The online version of this article (doi:10.1186/s13007-016-0114-x) contains supplementary material, which is available to authorized users.
DOI: 10.1038/nmeth.2069
发表时间: 2012-06-03
期刊: NATURE METHODS
影响因子: 48
作者:
Lubeck, Eric;Cai, Long
通讯作者: Cai, Long
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发表时间: 2008-10
期刊: NATURE METHODS
影响因子: 48
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