論文 - 詳細
| RRC ID | 19275 |
|---|---|
| 著者 | Furukawa K, Abe H, Hibino K, Sako Y, Tsuneda S, Ito Y. |
| タイトル | Reduction-triggered fluorescent amplification probe for the detection of endogenous RNAs in living human cells. |
| ジャーナル | Bioconjug Chem |
| Abstract |
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 beta-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. |
| 巻・号 | 20(5) |
| ページ | 1026-36 |
| 公開日 | 2009-5-20 |
| DOI | 10.1021/bc900040t |
| PMID | 19374406 |
| MeSH | Azides / chemistry Cell Survival Coumarins / chemistry DNA / chemistry Drug Design Flow Cytometry Fluorescein / chemical synthesis Fluorescein / chemistry Fluorescent Dyes / chemical synthesis Fluorescent Dyes / chemistry* HL-60 Cells Humans Kinetics Microscopy, Fluorescence Molecular Imaging / methods* Oxidation-Reduction RNA / metabolism* RNA, Messenger / metabolism RNA, Ribosomal, 28S / metabolism Spectrometry, Fluorescence |
| IF | 4.031 |
| 引用数 | 66 |
| WOS 分野 | CHEMISTRY, ORGANIC BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY CHEMISTRY, MULTIDISCIPLINARY |
| オルトメトリクス指標 |
オルトメトリクス指標項目
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| 最多言及媒体 | Patent(IFI CLAIMS) |
| 各媒体での言及数の合計 | 2 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HL60(RCB0041) |