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Polycistronic Expression of Interfering RNAs from RNA Polymerase III Promoters

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Functional Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 815))

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Abstract

In many RNA silencing applications, there is a benefit to expressing multiple interfering RNAs simultaneously. This can be achieved by using a single RNA polymerase II promoter to express multiple micro(mi)RNA-formatted interfering RNAs that are arranged in a polycistronic cluster, mimicking the organization of naturally clustered, endogenous miRNAs. While RNA pol III promoters are often used to express individual short hairpin (sh) RNAs, we have recently shown that pol III promoters can also be used to drive polycistronic expression of miRNA-formatted interfering RNAs. Here, we present methods for the assembly of polycistronic miRNA expression vectors that use pol III promoters. In addition, we present methods for testing the potency and the level of expression of each of the individual miRNAs encoded in the construct.

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Author information

Authors and Affiliations

  1. Department of Microbiology and Immunology, Institute for Molecular Medicine and Infectious Disease, Drexel University College of Medicine, 245 North 15th Street, Philadelphia, PA, 19102, USA

    Laura F. Steel & Viraj R. Sanghvi

Authors
  1. Laura F. Steel
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  2. Viraj R. Sanghvi
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Corresponding author

Correspondence to Laura F. Steel .

Editor information

Editors and Affiliations

  1. School of Medicine, Witten/Herdecke University, Group Stockumer Str. 10 50, Witten, 58448, Germany

    Michael Kaufmann

  2. School of Medicine, Private Universität Witten/Herdecke, Witten/Herdecke University 50, Witten, 58448, Germany

    Claudia Klinger

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Steel, L.F., Sanghvi, V.R. (2012). Polycistronic Expression of Interfering RNAs from RNA Polymerase III Promoters. In: Kaufmann, M., Klinger, C. (eds) Functional Genomics. Methods in Molecular Biology, vol 815. Springer, New York, NY. https://doi.org/10.1007/978-1-61779-424-7_26

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  • DOI: https://doi.org/10.1007/978-1-61779-424-7_26

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-61779-423-0

  • Online ISBN: 978-1-61779-424-7

  • eBook Packages: Springer Protocols

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