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Comprehensive Measurement of Gene Silencing Involving Endogenous MicroRNAs in Mammalian Cells

  • Masashi Fukuoka
  • Hirohiko Hohjoh
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1733)

Abstract

MicroRNAs (miRNAs) are functional small noncoding RNAs that work as mediators in gene silencing and that play important roles in gene regulation. A number of miRNAs have been found and their expression profiles have been examined by means of various microarray systems and real-time polymerase chain reaction (PCR) systems. Conventional microarrays as well as real-time PCR are able to detect existing miRNAs, in which inactive miRNAs that hardly contribute to gene silencing may be also contained. Here, we describe a comprehensive miRNA bioassay system with reporter genes for the detection of active miRNAs that are present in the RNA-induced silencing complexes, and actually working as mediators in gene silencing.

Key words

Active miRNAs Comprehensive bioassay Luciferase reporter gene psiCHECK2 vector Heat shock 

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.National Institute of Neuroscience, NCNPTokyoJapan

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