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Single-Molecule Fluorescence Energy Transfer Assays for the Characterization of Reaction Pathways of miRNA-Argonaute Complex

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Drug Target miRNA

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1517))

Abstract

Argonaute proteins are key components of the microRNA-induced silencing complexes (miRISCs) that mediate the posttranscriptional gene silencing of microRNAs and small interfering RNA (siRNAs). The complex reaction mechanism of miRISC is expected to be characterized by tracing the reaction pathways of miRISC at the single-molecule level in real time. In this chapter, we describe single-molecule fluorescence resonance energy transfer (FRET) assays to observe the target binding and reaction pathways of miRISC composed of a recombinant Argonaute and a small RNA.

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Acknowledgment

This work was supported by Creative Research Initiatives (Physical Genetics Laboratory, 2009-0081562) to S.H.

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Authors and Affiliations

  1. Department of Physics and Astronomy, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul, 08826, Republic of Korea

    Myung Hyun Jo & Sungchul Hohng

  2. Institute of Applied Physics, Seoul National University, Seoul, 08826, Republic of Korea

    Myung Hyun Jo & Sungchul Hohng

  3. National Center of Creative Research Initiatives, Seoul National University, Seoul, 08826, Republic of Korea

    Myung Hyun Jo & Sungchul Hohng

  4. Department of Biophysics and Chemical Biology, Seoul National University, Seoul, 08826, Republic of Korea

    Sungchul Hohng

Authors
  1. Myung Hyun Jo
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  2. Sungchul Hohng
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Corresponding author

Correspondence to Sungchul Hohng .

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Editors and Affiliations

  1. BEROCEUTICA GmbH, Potsdam, Germany

    Marco F. Schmidt

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Jo, M.H., Hohng, S. (2017). Single-Molecule Fluorescence Energy Transfer Assays for the Characterization of Reaction Pathways of miRNA-Argonaute Complex. In: Schmidt, M. (eds) Drug Target miRNA. Methods in Molecular Biology, vol 1517. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6563-2_21

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  • DOI: https://doi.org/10.1007/978-1-4939-6563-2_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6561-8

  • Online ISBN: 978-1-4939-6563-2

  • eBook Packages: Springer Protocols

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