Novel DNA microarray system for analysis of nascent mRNAs.
Transcriptional activation and repression are a key step in the regulation of all cellular activities. The development of comprehensive analysis methods such as DNA microarray has advanced our understanding of the correlation between the regulation of transcription and that of cellular mechanisms. However, DNA microarray analysis based on steady-state mRNA (total mRNA) does not always correspond to transcriptional activation or repression. To comprehend these transcriptional regulations, the detection of nascent RNAs is more informative. Although the nuclear run-on assay can detect nascent RNAs, it has not been fully applied to DNA microarray analysis. In this study, we have developed a highly efficient method for isolating bromouridine-labeled nascent RNAs that can be successfully applied to DNA microarray analysis. This method can linearly amplify small amounts of mRNAs with little bias. Furthermore, we have applied this method to DNA microarray analysis from mouse G2-arrested cells and have identified several genes that exhibit novel expression profiles. This method will provide important information in the field of transcriptome analysis of various cellular processes.
Ohtsu
Masaya
M
Faculty of Industrial Science and Technology, Department of Biological Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba, Japan.
Kawate
Mika
M
Fukuoka
Masashi
M
Gunji
Wataru
W
Hanaoka
Fumio
F
Utsugi
Takahiko
T
Onoda
Fumitoshi
F
Murakami
Yasufumi
Y
D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods*; D051379:Mice; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012322:RNA Precursors / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis*
D012322:RNA Precursors; D012333:RNA, Messenger