Abstract
Single-molecule RNA fluorescent in situ hybridization (smRNA FISH) allows for the visualization, localization, and quantification of RNA transcripts within individual cells and tissues using custom-designed fluorescently labeled oligonucleotide probes. Here we describe a protocol for the preparation, imaging, and analysis of a smRNA FISH experiment that can be applied to any RNA of choice. We also provide insights as to how this powerful tool can be used to study epigenetic regulation, for example, following the epigenetic editing of genes.
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Acknowledgement
This project received funding from the European’s Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 642691 and from the Dutch Organization for Scientific Research ZonMW TOP Grant (912.12.132).
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Beckman, W., Vuist, I.M., Kempe, H., Verschure, P.J. (2018). Cell-to-Cell Transcription Variability as Measured by Single-Molecule RNA FISH to Detect Epigenetic State Switching. In: Jeltsch, A., Rots, M. (eds) Epigenome Editing. Methods in Molecular Biology, vol 1767. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7774-1_21
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DOI: https://doi.org/10.1007/978-1-4939-7774-1_21
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7774-1
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