| RRC ID |
19275
|
| Author |
Furukawa K, Abe H, Hibino K, Sako Y, Tsuneda S, Ito Y.
|
| Title |
Reduction-triggered fluorescent amplification probe for the detection of endogenous RNAs in living human cells.
|
| Journal |
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.
|
| Volume |
20(5)
|
| Pages |
1026-36
|
| Published |
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
|
| Times Cited |
66
|
|
WOS Category
|
CHEMISTRY, ORGANIC
BIOCHEMICAL RESEARCH METHODS
BIOCHEMISTRY & MOLECULAR BIOLOGY
CHEMISTRY, MULTIDISCIPLINARY
|
| Resource |
| Human and Animal Cells |
HL60(RCB0041) |
