RDI Calculator: An Analysis Tool to Assess RNA Distributions in Cells.
RDI Calculator: An Analysis Tool to Assess RNA Distributions in Cells.
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RDI 计算器：评估细胞中 RNA 分布的分析工具。
DOI：
10.1038/s41598-019-44783-2
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发表时间：
2019
影响因子：
4.6
通讯作者：
Mili,Stavroula
中科院分区：
文献类型：
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作者：
Stueland,Michael;Wang,Tianhong;Park,HyeYoon;Mili,Stavroula
RNA定位于不同的亚细胞目的地已成为一种广泛使用的机制，它调节极化细胞中很大比例的转录本。已经开发了许多方法，允许在单分子分辨率下检测和成像RNA。然而，定量描述RNA分布的方法是有限的。这种方法通常依赖于作为参照点的细胞质和核边界的确定。在这里，我们描述了一个自动化的交互式图像分析程序，它有助于从单个细胞准确地生成细胞轮廓，并随后计算量化RNA分子群体如何分布在细胞质中的指标。我们将这一分析应用于老鼠和人类细胞中的mRNAs，以演示这些指标如何突出不同RNA物种分布模式的差异。我们将进一步讨论实际使用该工具的注意事项。该程序为机械和功能研究提供了一种促进和加速亚细胞RNA定位分析的方法。
Localization of RNAs to various subcellular destinations has emerged as a widely used mechanism that regulates a large proportion of transcripts in polarized cells. A number of methodologies have been developed that allow detection and imaging of RNAs at single-molecule resolution. However, methodologies to quantitatively describe RNA distributions are limited. Such approaches usually rely on the identification of cytoplasmic and nuclear boundaries which are used as reference points. Here, we describe an automated, interactive image analysis program that facilitates the accurate generation of cellular outlines from single cells and the subsequent calculation of metrics that quantify how a population of RNA molecules is distributed in the cell cytoplasm. We apply this analysis to mRNAs in mouse and human cells to demonstrate how these metrics can highlight differences in the distribution patterns of distinct RNA species. We further discuss considerations for the practical use of this tool. This program provides a way to facilitate and expedite the analysis of subcellular RNA localization for mechanistic and functional studies.