Real-time functional imaging for monitoring miR-133 during myogenic differentiation

Yoshio Kato, Shigeru Miyaki, Shigetoshi Yokoyama, Shin Omori, Atsushi Inoue, Machiko Horiuchi, Hiroshi Asahara

Research output: Contribution to journalArticlepeer-review

29 Scopus citations

Abstract

MicroRNAs (miRNAs) are a class of non-coding small RNAs that act as negative regulators of gene expression through sequence-specific interactions with the 3′ untranslated regions (UTRs) of target mRNA and play various biological roles. miR-133 was identified as a muscle-specific miRNA that enhanced the proliferation of myoblasts during myogenic differentiation, although its activity in myogenesis has not been fully characterized. Here, we developed a novel retroviral vector system for monitoring muscle-specific miRNA in living cells by using a green fluorescent protein (GFP) that is connected to the target sequence of miR-133 via the UTR and a red fluorescent protein for normalization. We demonstrated that the functional promotion of miR-133 during myogenesis is visualized by the reduction of GFP carrying the miR-133 target sequence, suggesting that miR-133 specifically down-regulates its targets during myogenesis in accordance with its expression. Our cell-based miRNA functional assay monitoring miR-133 activity should be a useful tool in elucidating the role of miRNAs in various biological events.

Original languageEnglish (US)
Pages (from-to)2225-2231
Number of pages7
JournalInternational Journal of Biochemistry and Cell Biology
Volume41
Issue number11
DOIs
StatePublished - Nov 2009

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cell Biology

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