Live-cell imaging of microRNA expression with post-transcriptional feedback control

Masayuki Sano*, Kana Morishita, Satoshi Oikawa, Takayuki Akimoto, Kimio Sumaru, Yoshio Kato

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

MicroRNAs (miRNAs) are small noncoding RNAs that regulate complex gene expression networks in eukaryotic cells. Because of their unique expression patterns, miRNAs are potential molecular markers for specific cell states. Although a system capable of imaging miRNA in living cells is needed to visually detect miRNA expression, very few fluorescence signal-on sensors that respond to expression of target miRNA (miR-ON sensors) are available. Here we report an miR-ON sensor containing a bidirectional promoter-driven Csy4 endoribonuclease and green fluorescent protein, ZsGreen1, for live-cell imaging of miRNAs with post-transcriptional feedback control. Csy4-assisted miR-ON (Csy4-miR-ON) sensors generate negligible background but respond sensitively to target miRNAs, allowing high-contrast fluorescence detection of miRNAs in various human cells. We show that Csy4-miR-ON sensors enabled imaging of various miRNAs, including miR-21, miR-302a, and miR-133, in vitro as well as in vivo. This robust tool can be used to evaluate miRNA expression in diverse biological and medical applications.

Original languageEnglish
Pages (from-to)547-556
Number of pages10
JournalMolecular Therapy - Nucleic Acids
Volume26
DOIs
Publication statusPublished - 2021 Dec 3

Keywords

  • Csy4
  • fluorescent protein
  • live-cell imaging
  • microRNA
  • post-transcriptional feedback control

ASJC Scopus subject areas

  • Molecular Medicine
  • Drug Discovery

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