title: Development of a DNA barcode tagging method for monitoring dynamic changes in gene expression by using an ultra high-throughput sequencer.
abstract: CAGE (cap analysis of gene expression) is a method for identifying transcription start sites by sequencing the first 20 or 21 nucleotides from the 5' end of capped transcripts, allowing genome-wide promoter analyses to be performed. The potential of the CAGE as a form of expression profiling was limited previously by sequencing technology and the labor-intensive protocol. Here we describe an improved CAGE method for use with a next generation sequencer. This modified method allows the identification of the RNA source of each CAGE tag within a pooled library by introducing DNA tags (barcodes). The method not only drastically improves the sequencing capacity, but also contributes to savings in both time and budget. Additionally, this pooled CAGE tag method enables the dynamic changes in promoter usage and gene expression to be monitored.
authors.affiliation: Advanced Science Institute, RIKEN Wako Main Campus, Saitama, Japan.
authors.forename: Norihiro
authors.initials: N
authors.lastname: Maeda
authors.forename: Hiromi
authors.initials: H
authors.lastname: Nishiyori
authors.forename: Mari
authors.initials: M
authors.lastname: Nakamura
authors.forename: Chika
authors.initials: C
authors.lastname: Kawazu
authors.forename: Mitsuyoshi
authors.initials: M
authors.lastname: Murata
authors.forename: Hiromi
authors.initials: H
authors.lastname: Sano
authors.forename: Kengo
authors.initials: K
authors.lastname: Hayashida
authors.forename: Shiro
authors.initials: S
authors.lastname: Fukuda
authors.forename: Michihira
authors.initials: M
authors.lastname: Tagami
authors.forename: Akira
authors.initials: A
authors.lastname: Hasegawa
authors.forename: Kayoko
authors.initials: K
authors.lastname: Murakami
authors.forename: Kate
authors.initials: K
authors.lastname: Schroder
authors.forename: Katharine
authors.initials: K
authors.lastname: Irvine
authors.forename: David
authors.initials: D
authors.lastname: Hume
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hayashizaki
authors.forename: Piero
authors.initials: P
authors.lastname: Carninci
authors.forename: Harukazu
authors.initials: H
authors.lastname: Suzuki
mesh_terms: D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000295:instrumentation*; D024363:Transcription Initiation Site