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Regulatory Non-Coding RNAs pp 39–51Cite as

Native Gel Analysis for Mammalian MicroRNPs Assembled from Pre-microRNAs

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1206))

Abstract

MicroRNAs (miRNAs) are an important class of small RNAs that regulate gene expression posttranscriptionally through the microRNP (miRNP)/RNA-induced silencing complex (RISC). The core component of miRNPs is an Argonuate protein that directly binds to a miRNA. In mammals, most miRNPs are assembled through the miRNA loading complex (miRLC), which is composed of Dicer, Ago, and TRBP. miRLC processes miRNA precursors (pre-miRNAs) into miRNA duplexes and loads miRNA duplexes to Ago. Here, we describe a native gel analysis system for detecting miRNPs assembled with pre-miRNAs from mammalian lysates that ectopically express Ago2. The methods presented here provide a powerful tool for further dissecting miRNP assembly pathways in mammals.

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Acknowledgements

Supported by NIH grant GM072777 to Z.M.

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Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104-6100, USA

    Xuhang Liu

  2. Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, 613B Stellar-Chance Laboratories, 422 Curie Blvd, Philadelphia, PA, 19104-6100, USA

    Zissimos Mourelatos

Authors
  1. Xuhang Liu
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  2. Zissimos Mourelatos
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Corresponding author

Correspondence to Zissimos Mourelatos .

Editor information

Editors and Affiliations

  1. University of Connecticut Health Center, Farmington, Connecticut, USA

    Gordon G. Carmichael

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Liu, X., Mourelatos, Z. (2015). Native Gel Analysis for Mammalian MicroRNPs Assembled from Pre-microRNAs. In: Carmichael, G. (eds) Regulatory Non-Coding RNAs. Methods in Molecular Biology, vol 1206. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1369-5_4

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  • DOI: https://doi.org/10.1007/978-1-4939-1369-5_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1368-8

  • Online ISBN: 978-1-4939-1369-5

  • eBook Packages: Springer Protocols

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