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MicroRNA Protocols pp 117–127Cite as

Evaluating the MicroRNA Targeting Sites by Luciferase Reporter Gene Assay

Part of the Methods in Molecular Biology book series (MIMB,volume 936)

Abstract

MicroRNAs are post-transcriptional regulators that control mRNA stability and the translation efficiency of their target genes. Mature microRNAs are approximately 22-nucleotide in length. They mediate post-transcriptional gene regulation by binding to the imperfect complementary sequences (a.k.a. microRNA regulatory elements, MRE) in the target mRNAs. It is estimated that more than one-third of the protein-coding genes in the human genome are regulated by microRNAs. The experimental methods to examine the interaction between the microRNA and its targeting site(s) in the mRNA are important for understanding microRNA functions. The luciferase reporter gene assay has recently been adapted to test the effect of microRNAs. In this chapter, we use a previously identified miR-138 targeting site in the 3′-untranslated region (3′-UTR) of the RhoC mRNA as an example to describe a quick method for testing the interaction of microRNA and mRNA.

Key words

  • MicroRNA
  • MicroRNA targeting sequence
  • MicroRNA regulatory element
  • Luciferase reporter gene assay
  • miR-138
  • RhoC

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Acknowledgment

This work was supported in part by NIH PHS grants (CA135992, CA139596, DE014847) and supplementary funding from UIC CCTS (UL1RR029879). Y.J. is supported by PHS T32DE018381 from NIDCR. We thank Ms. Katherine Long for her editorial assistance.

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Authors and Affiliations

  1. Center for Molecular Biology of Oral Diseases, University of Illinois at Chicago, Chicago, IL, USA

    Yi Jin & Xiqiang Liu

  2. Department of Anatomy and Cell Biology, Rush University Medical Center, Chicago, IL, USA

    Zujian Chen

  3. Center for Molecular Biology of Oral Diseases, College of Dentistry, University of Illinois at Chicago, Chicago, IL, USA

    Xiaofeng Zhou

Authors
  1. Yi Jin
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  2. Zujian Chen
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  3. Xiqiang Liu
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  4. Xiaofeng Zhou
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Corresponding author

Correspondence to Xiaofeng Zhou .

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Editors and Affiliations

  1. Keck School of Medicine, Dept. of Cell and Neurobiology, University of Southern California, San Pablo St. 1333, Los Angeles, 90033, California, USA

    Shao-Yao Ying

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Jin, Y., Chen, Z., Liu, X., Zhou, X. (2013). Evaluating the MicroRNA Targeting Sites by Luciferase Reporter Gene Assay. In: Ying, SY. (eds) MicroRNA Protocols. Methods in Molecular Biology, vol 936. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-083-0_10

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  • DOI: https://doi.org/10.1007/978-1-62703-083-0_10

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  • Publisher Name: Humana Press, Totowa, NJ

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  • Online ISBN: 978-1-62703-083-0

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