mixedStageDrosEmbryoPEAT-TSS
A paired-end sequencing strategy to map the complex landscape of transcription
Recent high-throughput sequencing protocols have uncovered the complexity of mammalian transcription by RNA polymerase II, helping to define several initiation patterns in which transcription start sites (TSSs) cluster within both narrow and broad genomic windows. Here, we describe an improved paired-end sequencing strategy, which enables more robust mapping and characterization of capped transcripts. This strategy was applied to explore the transcription initiation landscape in the Drosophila melanogaster embryo. Extending the previous findings in mammals, we found that fly promoters exhibit distinct initiation patterns, which are linked to specific promoter sequence motifs. Furthermore, we identified a large number of 5' capped transcripts originating from coding exons; analyses support that they are unlikely the result of alternative TSSs, but rather the product of post-transcriptional modifications. Taken together, paired-end TSS analysis is demonstrated to be a powerful method to uncover the transcriptional complexity of eukaryotic genomes.