title: Novel DNA microarray system for analysis of nascent mRNAs.
abstract: 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.
authors.affiliation: Faculty of Industrial Science and Technology, Department of Biological Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda-shi, Chiba, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Ohtsu
authors.forename: Mika
authors.initials: M
authors.lastname: Kawate
authors.forename: Masashi
authors.initials: M
authors.lastname: Fukuoka
authors.forename: Wataru
authors.initials: W
authors.lastname: Gunji
authors.forename: Fumio
authors.initials: F
authors.lastname: Hanaoka
authors.forename: Takahiko
authors.initials: T
authors.lastname: Utsugi
authors.forename: Fumitoshi
authors.initials: F
authors.lastname: Onoda
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Murakami
mesh_terms: 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*
chemical_list: D012322:RNA Precursors; D012333:RNA, Messenger