RNAscope: a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues.
By Fay Wang, J. Flanagan, N. Su, Li-chong Wang, S. Bui, Allissa Nielson, Xingyong Wu, Hong-Thuy Vo, Xiao-Jun Ma, Yuling Luo. Journal of Molecular Diagnostics, 2012
2,388 citations recorded by the source corpus.
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.
View Paper: https://www.semanticscholar.org/paper/22035a51a6749120bd24969b0a75524ee4006c0f
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Citations in the corpus
- Third-generation in situ hybridization chain reaction: multiplexed, quantitative, sensitive, versatile, robust (2018, unknown): “The principles, strengths andweaknesses underlying stages 1 and 2 of scheme C are similar to those of branched DNAmethods (bDNA), which use a four-stage protocol (Wang et al., 2012): stage 1, target detectionwith a pair of probes each carrying half of a bridge substrate; stage 2, pre-amplification w”
- Single cell transcriptomics of human and mouse lung cancers reveals conserved myeloid populations across individuals and species (2019, methodology): “RNA in situ hybridization was performed with Manual RNAscope® 2.5 HD Reagent Kit (RED) (ACD Bio, # 322350) following manufacturer’s instructions as previously described (Wang et al., 2012).”
- An introduction to spatial transcriptomics for biomedical research (2022, methodology): “Variations of this have been commercially available for some time, such as RNAscope [36].”
- Methods and applications for single-cell and spatial multi-omics (2023, background): “Whole-transcriptome DSP assays or DSP assays for >100 protein targets are possible, although coupled to image-based selection of specific regions of interest following fluorescent antibody and/or RNAscope 120 marker stainings.”