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Combined MicroRNA In Situ Hybridization and Immunohistochemical Detection of Protein Markers

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Target Identification and Validation in Drug Discovery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1953))

Abstract

MicroRNAs are short (18–23 nucleotides) noncoding RNAs involved in posttranscriptional regulation of gene expression through their specific binding to the 3′UTR of mRNAs. MicroRNAs can be detected in tissues using specific locked nucleic acid (LNA)-enhanced probes. The characterization of microRNA expression in tissues by in situ detection is often crucial following a microRNA biomarker discovery phase in order to validate the candidate microRNA biomarker and allow better interpretation of its molecular functions and derived cellular interactions. The in situ hybridization data provides information about contextual distribution and cellular origin of the microRNA. By combining microRNA in situ hybridization with immunohistochemical staining of protein markers, it is possible to precisely characterize the microRNA-expressing cells and to identify the potential microRNA targets. This combined technology can also help to monitor changes in the level of potential microRNA targets in a therapeutic setting. In this chapter, we present a fluorescence-based detection method that allows the combination of microRNA in situ hybridization with immunohistochemical staining of one and, in this updated version of the paper, two protein markers detected with primary antibodies raised in the same host species.

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Acknowledgments

We thank Trine Møller for excellent technical assistance and the Danish Ministry of Science, Innovation and Technology, for financial funding.

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Authors and Affiliations

  1. Bioneer A/S, Hørsholm, Denmark

    Boye Schnack Nielsen & Kim Holmstrøm

Authors
  1. Boye Schnack Nielsen
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  2. Kim Holmstrøm
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Corresponding author

Correspondence to Kim Holmstrøm .

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Editors and Affiliations

  1. Pharmacology and Translational Research, Boehringer Ingelheim RCV GmbH & Co KG, Vienna, Austria

    Jürgen Moll

  2. Department of Cancer Cell Signaling, Boehringer Ingelheim RCV GmBH & Co KG, Vienna, Austria

    Sebastian Carotta

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Nielsen, B.S., Holmstrøm, K. (2019). Combined MicroRNA In Situ Hybridization and Immunohistochemical Detection of Protein Markers. In: Moll, J., Carotta, S. (eds) Target Identification and Validation in Drug Discovery. Methods in Molecular Biology, vol 1953. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9145-7_17

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  • DOI: https://doi.org/10.1007/978-1-4939-9145-7_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9144-0

  • Online ISBN: 978-1-4939-9145-7

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