Experimental Validation of MicroRNA Targets: Luciferase Reporter Assay

  • Luisa TomaselloEmail author
  • Landon Cluts
  • Carlo M. Croce
Part of the Methods in Molecular Biology book series (MIMB, volume 1970)


MicroRNAs (miRNAs) are small noncoding RNAs of 22–25 nucleotides that control gene expression at the posttranscriptional level through the degradation of mRNAs or translational repression. In the last 15 years, the study of these small molecules helped elucidate their role in the regulation of many cellular processes and the onset and development of several diseases. Therefore, many computational tools based on algorithms for target prediction have been developed to identify potential miRNA–target interactions. The improvement of experimental approaches to more easily and quickly confirm in silico predictions has become essential for the study of these small RNAs and their molecular functions. In this chapter, we summarized the principal steps of one of the most used techniques for the validation of microRNA targets, the Luciferase assay, thus explaining the underlying principles and the procedures to apply it best.

Key words

MicroRNA target validation Luciferase assay Cloning In vitro techniques 


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Luisa Tomasello
    • 1
    Email author
  • Landon Cluts
    • 1
  • Carlo M. Croce
    • 1
  1. 1.Department of Cancer Biology and Medical Genetics, Comprehensive Cancer CenterThe Ohio State UniversityColumbusUSA

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