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Identifying RISC Components Using Ago2 Immunoprecipitation and Mass Spectrometry

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mRNA Decay

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1720))

Abstract

Complex immunoprecipitation (Co-IP) is a powerful technique for precipitating an intact protein complex out of solution and cell lysates using an antibody that specifically binds to a particular protein in a large complex of proteins. Mass spectrometry (MS) is used to identify, sequence, and quantify proteins. RNA-induced silencing complexes (RISCs), Ago2 centered protein assemblies, are essential for miRNA mediated RNA decay and gene expression regulation; however, the complete list of RISCs is unknown. Here we describe methods used to combine IP and MS to identify new components of RISCs.

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Authors and Affiliations

  1. Department of Biological Chemistry and Molecular Pharmacology of Harvard Medical School, Boston, MA, USA

    Tingfang Yi

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  1. Tingfang Yi
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Correspondence to Tingfang Yi .

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Editors and Affiliations

  1. Murdoch Children’s Research Institute, The Royal Children’s Hospital, Parkville, Victoria, Australia

    Shireen R. Lamandé

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Yi, T. (2018). Identifying RISC Components Using Ago2 Immunoprecipitation and Mass Spectrometry. In: Lamandé, S. (eds) mRNA Decay. Methods in Molecular Biology, vol 1720. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7540-2_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7540-2_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7539-6

  • Online ISBN: 978-1-4939-7540-2

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