Development of a DNA barcode tagging method for monitoring dynamic changes in gene expression by using an ultra high-throughput sequencer
Development of a DNA barcode tagging method for monitoring dynamic changes in gene expression by using an ultra high-throughput sequencer
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DOI：
10.2144/000112814
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发表时间：
2008-07-01
期刊：
影响因子：
2.7
通讯作者：
Suzuki, Harukazu
中科院分区：
文献类型：
--
作者：
Maeda, Norihiro;Nishiyori, Hiromi;Suzuki, Harukazu
CAGE（基因表达的帽分析）是一种通过对加帽转录物5'端的前20或21个核苷酸进行测序来鉴定转录起始位点的方法，允许进行全基因组启动子分析。CAGE作为表达谱分析的一种形式的潜力以前受到测序技术和劳动密集型方案的限制。在这里，我们描述了一种改进的CAGE方法，用于下一代测序仪。这种改进的方法允许通过引入DNA标签（条形码）来鉴定汇集的文库内每个CAGE标签的RNA来源。该方法不仅大大提高了测序能力，而且有助于节省时间和预算。此外，这种合并的CAGE标签方法能够监测启动子使用和基因表达的动态变化。
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.