Transcriptome analysis using fluorescence-labeled oligonucleotide.
In our previous studies, the fluorescence intensity at 480 nm of the bispyrene-labeled 2'-O-methyl-oligoribonucleotide (OMUpy2) was drastically enhanced only when it was hybridized with its complementary RNA. In this study, we demonstrated the potential of the OMUpy2 for transcriptome analysis in the cell free system and in the living cell. For the analysis in the cell free system, the micro-chamber was designed and used to evaluate the detection limit of the target RNA. The detection limit of oligo-RNA was approximately 15 fmol. For the analysis of the living cells, C-OMUpy2 was introduced to the cervical carcinoma cells, C4II, and the fluorescence from C-OMUpy2 was monitored. The fluorescence from the cells began to be observed 5 min after the serum stimulation, and the fluorescence was decreased after 20 min. These results seemed to be consistent with the results from the RT-PCR analysis and suggested that OMUpy2 could be a useful tool for the transcriptome analysis.
Shimada
Naohiko
N
Department of Polymer Science and Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyoku, Kyoto 606-8585, Japan.
Mahara
Atsushi
A
Sakamoto
Takashi
T
Yamaoka
Tetsuji
T
Murakami
Akira
A
D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D008856:Microscopy, Fluorescence; D009843:Oligoribonucleotides / Q000378:metabolism*; D018075:RNA, Complementary; D014158:Transcription, Genetic
D005456:Fluorescent Dyes; D009843:Oligoribonucleotides; D018075:RNA, Complementary