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Chromogenic In Situ Hybridization Methods for microRNA Biomarker Monitoring of Drug Safety and Efficacy

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Drug Safety Evaluation

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1641))

Abstract

Disease research and treatment development have turned to the impact and utility of microRNA. The dynamic and highly specific expression of these molecular regulators can be used to predict and monitor disease progression as well as therapeutic treatment efficacy and safety, thus aiding decisions in patient care. In situ hybridization (ISH) of biopsy material has become a routine clinical pathology procedure for monitoring gene structure, expression, and sample characterization. For ribonucleic acid (RNA), determining cell source and level of expression of these biomarkers gives insight into the cellular function and physiopathology. Identification and monitoring of microRNA biomarkers are made possible through locked nucleic acid (LNA)™-based detection probes. LNA™ enhances the sensitivity and specificity of target binding, most profoundly so for the short, highly similar, microRNA sequences. We present a robust 1-day ISH protocol for formalin-fixed, paraffin-embedded (FFPE) tissue sections based on microRNA-specific LNA™ detection probes which can be labeled with digoxigenin (DIG) or 6-carboxyfluorescein (FAM) and detected through enzyme-linked specific antibodies that catalyze substrates into deposited chromogen products at the target RNA site. The variety of haptens and detection reagents in combination with LNA™ chemistry offer flexibility and ease to multiple target assessment of therapeutic response.

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Acknowledgments

The authors would like to thank Trine Møller at Bioneer for technical expertise as well as Marie-Louise Lunn at Exiqon for her support in the preparation of this manuscript. Exiqon is a QIAGEN company.

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Authors and Affiliations

  1. Exiqon Inc., 12 Gill St. Suite 1650, Woburn, MA, 01801, USA

    Barbara R. Gould

  2. Exiqon A/S, Vedbæk, Denmark

    Tina Damgaard

  3. Bioneer A/S, Hørsholm, Denmark

    Boye Schnack Nielsen

Authors
  1. Barbara R. Gould
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  2. Tina Damgaard
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  3. Boye Schnack Nielsen
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Corresponding author

Correspondence to Barbara R. Gould .

Editor information

Editors and Affiliations

  1. Preclinical Safety, Sanofi, R&D, Vitry-sur-Seine, France

    Jean-Charles Gautier

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Gould, B.R., Damgaard, T., Nielsen, B.S. (2017). Chromogenic In Situ Hybridization Methods for microRNA Biomarker Monitoring of Drug Safety and Efficacy. In: Gautier, JC. (eds) Drug Safety Evaluation. Methods in Molecular Biology, vol 1641. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7172-5_22

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  • DOI: https://doi.org/10.1007/978-1-4939-7172-5_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7170-1

  • Online ISBN: 978-1-4939-7172-5

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