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siRNA Design pp 291–314Cite as

Production and Application of Long dsRNA in Mammalian Cells

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 942))

Abstract

Double-stranded RNA (dsRNA) is involved in different biological processes. At least three different pathways can respond to dsRNA in mammals. One of these pathways is RNA interference (RNAi) where long dsRNA induces sequence-specific degradation of transcripts carrying sequences complementary to dsRNA. Long dsRNA is also a potent trigger of the interferon pathway, a sequence-independent response that leads to global suppression of translation and global RNA degradation. In addition, dsRNA can be edited by adenosine deamination, which may result in nuclear retention and degradation of dsRNA or in alteration of RNA coding potential. Here, we provide a technical review summarizing different strategies of long dsRNA usage. While the review is largely focused on long dsRNA-induced RNAi in mammalian cells, it also provides helpful information on both the in vitro production and in vivo expression of dsRNAs. We present an overview of currently available vectors for dsRNA expression and provide the latest update on oocyte-specific transgenic RNAi approaches.

Katerina Chalupnikova and Jana Nejepinska contributed equally to this article.

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Acknowledgements

Authors thank Radek Malik for help with preparation and Camille Du Roure for text revision of this manuscript. This work was supported by the GACR 204/09/0085, EMBO SDIG program #1488, and the Purkynje Fellowship to PS. JN is supported in part by Faculty of Science, Charles University in Prague.

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Authors and Affiliations

  1. Institute of Molecular Genetics AS CR, Prague, Czech Republic

    Katerina Chalupnikova, Jana Nejepinska & Petr Svoboda

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  1. Katerina Chalupnikova
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  2. Jana Nejepinska
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  3. Petr Svoboda
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Correspondence to Katerina Chalupnikova .

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Editors and Affiliations

  1. , Lineberger Comprehensive Cancer Center, University of North Carolina, West Drive 450, Chapel Hill, 27599, North Carolina, USA

    Debra J. Taxman

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Chalupnikova, K., Nejepinska, J., Svoboda, P. (2013). Production and Application of Long dsRNA in Mammalian Cells. In: Taxman, D. (eds) siRNA Design. Methods in Molecular Biology, vol 942. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-119-6_16

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  • DOI: https://doi.org/10.1007/978-1-62703-119-6_16

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-118-9

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