Reduction-triggered fluorescent amplification probe for the detection of endogenous RNAs in living human cells

Kazuhiro Furukawa, Hiroshi Abe*, Kayo Hibino, Yasushi Sako, Satoshi Tsuneda, Yoshihiro Ito

*この研究の対応する著者

研究成果: Article査読

66 被引用数 (Scopus)

抄録

Oligonucleotide-templated reactions are attracting attention as a method for RNA detection in living cells. Previously, a reduction-triggered fluorescence probe has been reported that is based on azide reduction to switch fluorescence on. In this article, we report a more advanced probe, a reduction-triggered fluorescent amplification probe that is capable of amplifying a target signal. Azidomethyl fluorescein was newly synthesized and introduced into a probe. Azido-masked fluorescein on the probe showed a strong turn-on fluorescence signal upon oligonucleotide-templated Staudinger reduction. The catalytic reaction of the probe offered a turnover number of 50 as fluorescence readout within 4 h. Finally, probes were introduced into human leukemia HL-60 cells by use of streptolysin O pore-forming peptide. We successfully detected and quantitated the 28S rRNA and β-Actin mRNA signal above the background by flow cytometry. In addition, the same RNA targets were imaged by fluorescence microscopy. The data suggest that a reduction-triggered amplification probe may be a powerful tool in analyzing the localization, transcription, or processing of RNA species in living eukaryotic cells.

本文言語English
ページ(範囲)1026-1036
ページ数11
ジャーナルBioconjugate Chemistry
20
5
DOI
出版ステータスPublished - 2009 5 20

ASJC Scopus subject areas

  • バイオテクノロジー
  • バイオエンジニアリング
  • 生体医工学
  • 薬理学
  • 薬科学
  • 有機化学

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