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In Vitro Silencing of lncRNA Expression Using siRNAs

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Long Non-Coding RNAs in Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2348))

Abstract

Recent advances in sequencing technologies have uncovered the existence of thousands of long noncoding RNAs (lncRNAs) with dysregulated expression in cancer. As a result, there is burgeoning interest in understanding their function and biological significance in both homeostasis and disease. RNA interference (RNAi) enables sequence-specific gene silencing and can, in principle, be employed to silence virtually any gene. However, when applied to lncRNAs, it is important to consider current limitations in their annotation and current principles regarding lncRNA regulation and function when assessing their phenotype in cancer cell lines. In this chapter we describe the analysis of lncRNA splicing variant expression, including subcellular localization, transfection of siRNAs in cancer cell lines, and validation of gene silencing by quantitative PCR and single molecule in situ hybridization. All protocols can be performed in a laboratory with essential equipment for cell culture, molecular biology, and imaging.

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Acknowledgments

We thank Philip Allan Seymour and Mohamed Osman, members of the Arnes laboratory, for technical assistance and critical reading of the manuscript. The Novo Nordisk Foundation Center for Stem Cell Biology is supported by grant number NNF17CC0027852.

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Authors and Affiliations

  1. Biotech Research & Innovation Centre (BRIC), Novo Nordisk Foundation Center for Stem Cell Biology (DanStem), University of Copenhagen, Copenhagen, Denmark

    Meike S. Thijssen, Jennifer Bintz & Luis Arnes

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  1. Meike S. Thijssen
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  2. Jennifer Bintz
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Correspondence to Luis Arnes .

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  1. Faculty of Medicine and Health Sciences, University of Barcelona, Barcelona, Spain

    Alfons Navarro

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Thijssen, M.S., Bintz, J., Arnes, L. (2021). In Vitro Silencing of lncRNA Expression Using siRNAs. In: Navarro, A. (eds) Long Non-Coding RNAs in Cancer. Methods in Molecular Biology, vol 2348. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1581-2_9

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  • DOI: https://doi.org/10.1007/978-1-0716-1581-2_9

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  • Print ISBN: 978-1-0716-1580-5

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