Real-time monitoring of mRNAs with fluorescence-modified RNA probes in living cells.
In our previous study, we reported that bispyrene-modified RNA probes (OMUpy2) were useful for RNA detection in homogeneous physiological media(1-2). The aim of this study is to establish in situ monitoring system to detect mRNAs in living cells by OMUpy2. We chose c-fos mRNA as a target RNA which is known as one of the immediate-early genes. The real-time moni- toring of mRNAs was carried out in living C4II cells. After transfection of OMUpy2 into the cells which had been incubated with serum-free medium, cells were stimulated by media with serum and the resulting expression of mRNA was monitored by a fluorescence microscope. In the case of OMUpy2-CF3S, which was complementary to c-fos mRNA, fluorescence emission around 480 nm was observed obviously. These results suggest that mRNA expressed by the serum stimulation was successfully visualized in a real-time manner.
Waki
Reiko
R
Department of Biomolecular Engineering, Kyoto Institute of Technology, Kyoto 606-8585, Japan.
Ueda
Takako
T
Yamayoshi
Asako
A
Kobori
Akio
A
Murakami
Akira
A
D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D016760:Proto-Oncogene Proteins c-fos / Q000032:analysis / Q000235:genetics; D015347:RNA Probes / Q000737:chemistry*; D012333:RNA, Messenger / Q000032:analysis*; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D016760:Proto-Oncogene Proteins c-fos; D015347:RNA Probes; D012333:RNA, Messenger