RIKEN Center for Life Science Technologies, Division of Genomics Technologies, Yokohama 230-0045, Japan.
RIKEN Omics Science Center, Yokohama 230-0045, Japan.
Sci Data. 2017 Oct 3;4:170147. doi: 10.1038/sdata.2017.147.
The FANTOM5 expression atlas is a quantitative measurement of the activity of nearly 200,000 promoter regions across nearly 2,000 different human primary cells, tissue types and cell lines. Generation of this atlas was made possible by the use of CAGE, an experimental approach to localise transcription start sites at single-nucleotide resolution by sequencing the 5' ends of capped RNAs after their conversion to cDNAs. While 50% of CAGE-defined promoter regions could be confidently associated to adjacent transcriptional units, nearly 100,000 promoter regions remained gene-orphan. To address this, we used the CAGEscan method, in which random-primed 5'-cDNAs are paired-end sequenced. Pairs starting in the same region are assembled in transcript models called CAGEscan clusters. Here, we present the production and quality control of CAGEscan libraries from 56 FANTOM5 RNA sources, which enhances the FANTOM5 expression atlas by providing experimental evidence associating core promoter regions with their cognate transcripts.
FANTOM5 表达图谱是对近 20 万个启动子区域在近 2000 种不同的人类原代细胞、组织类型和细胞系中的活性进行的定量测量。该图谱的生成得益于 CAGE 技术的应用,这是一种通过测序经 cDNA 转化后的加帽 RNA 的 5' 端,以单核苷酸分辨率定位转录起始位点的实验方法。虽然 50% 的 CAGE 定义的启动子区域可以被明确地关联到相邻的转录单元,但近 100000 个启动子区域仍然是基因孤儿。为了解决这个问题,我们使用了 CAGEscan 方法,其中随机引物 5'-cDNA 进行配对末端测序。从同一区域开始的配对被组装成称为 CAGEscan 簇的转录本模型。在这里,我们展示了 56 个 FANTOM5 RNA 来源的 CAGEscan 文库的生产和质量控制,这通过提供将核心启动子区域与其同源转录本相关联的实验证据,增强了 FANTOM5 表达图谱。