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A Biochemical Approach to Identify Direct MicroRNA Targets

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Regulatory Non-Coding RNAs

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

Abstract

We have recently developed a biochemical approach to isolate miRNA-bound mRNAs and have used this method to identify the genome-wide mRNAs regulated by the tumor suppressor miRNA miR-34a. This method involves transfection of cells with biotinylated miRNA mimics, streptavidin pulldown, RNA isolation, and qRT-PCR. The protocol in this chapter describes these steps and the issues that should be considered while designing such pulldown experiments.

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Acknowledgements

This work was supported by the Intramural Research Program of the National Institutes of Health, National Cancer Institute, Center for Cancer Research.

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Authors and Affiliations

  1. Genetics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, USA

    Murugan Subramanian, Xiao Ling Li & Toshifumi Hara

  2. Regulatory RNAs and Cancer Section, Genetics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Building 37, Room 6134, 37 Convent Drive, Bethesda, MD, 20892, USA

    Ashish Lal Ph.D.

Authors
  1. Murugan Subramanian
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  2. Xiao Ling Li
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  3. Toshifumi Hara
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  4. Ashish Lal Ph.D.
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Corresponding author

Correspondence to Ashish Lal Ph.D. .

Editor information

Editors and Affiliations

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

    Gordon G. Carmichael

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© 2015 Springer Science+Business Media New York

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Subramanian, M., Li, X.L., Hara, T., Lal, A. (2015). A Biochemical Approach to Identify Direct MicroRNA Targets. 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_3

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  • DOI: https://doi.org/10.1007/978-1-4939-1369-5_3

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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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