RNAscope A Novel in Situ RNA Analysis Platform for Formalin-Fixed, Paraffin-Embedded Tissues
RNAscope A Novel in Situ RNA Analysis Platform for Formalin-Fixed, Paraffin-Embedded Tissues
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DOI：
10.1016/j.jmoldx.2011.08.002
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发表时间：
2012-01-01
影响因子：
4.1
通讯作者：
Luo, Yuling
中科院分区：
文献类型：
--
作者：
Wang, Fay;Flanagan, John;Luo, Yuling
生物标志物的原位分析在分子病理学中是非常理想的，因为它允许在临床标本的组织病理学背景下检查生物标志物状态。免疫组织化学和DNA原位杂交（ISH）被广泛用于临床环境中，分别评估蛋白质和DNA生物标志物，但临床使用原位RNA分析是罕见的。当考虑到通过全基因组表达谱发现的RNA生物标志物的丰度时，这种差异尤其显著。这在很大程度上是由于目前RNA ISH技术的高度技术复杂性和不足的灵敏度和特异性。在这里，我们描述了RNAscope，一种新的RNA ISH技术，具有独特的探针设计策略，允许同时进行信号放大和背景抑制，以实现单分子可视化，同时保留组织形态。RNAscope与常规福尔马林固定、石蜡包埋的组织标本兼容，可以使用常规显色染料进行明场显微镜检查，也可以使用荧光染料进行多重分析。与实时RT-PCR等研磨结合RNA分析方法不同，RNAscope为RNA生物标志物带来了原位分析的好处，并可能使基于RNA ISH的分子诊断测定快速发展。（J Mol Diagn 2012，14：22-29; DOI：10.1016/j.j.moldx.2011.08.002）
In situ analysis of biomarkers is highly desirable in molecular pathology because it allows the examination of biomarker status within the histopathological context of clinical specimens. Immunohistochemistry and DNA in situ hybridization (ISH) are widely used in clinical settings to assess protein and DNA biomarkers, respectively, but clinical use of in situ RNA analysis is rare. This disparity is especially notable when considering the abundance of RNA biomarkers discovered through whole-genome expression profiling. This is largely due to the high degree of technical complexity and insufficient sensitivity and specificity of current RNA ISH techniques. Here, we describe RNAscope, a novel RNA ISH technology with a unique probe design strategy that allows simultaneous signal amplification and background suppression to achieve single-molecule visualization while preserving tissue morphology. RNAscope is compatible with routine formalin-fixed, paraffin-embedded tissue specimens and can use either conventional chromogenic dyes for bright-field microscopy or fluorescent dyes for multiplex analysis. Unlike grind-and-bind RNA analysis methods such as real-time RT-PCR, RNAscope brings the benefits of in situ analysis to RNA biomarkers and may enable rapid development of RNA ISH-based molecular diagnostic assays. (J Mol Diagn 2012, 14:22-29; DOI: 10.1016/j.j.moldx.2011.08.002)