Analysis of Pro-Inflammatory Gene Activation and RNA Processing by RNAseq of Nascent Transcripts I. Mus musculus

We examined pro-inflammatory gene activation in activated murine macrophages by performing RNA-Seq with fractionated chromatin-associated, nucleoplasmic, and cytoplasmic transcripts. This experimental strategy allowed a global, high-resolution analysis of the transcriptional regulation of diverse classes of co-expressed genes, with a direct comparison to transcript processing and the transit of RNA from the chromatin to the nucleoplasm and the cytoplasm. The results provide quantitative insights into the relationship between transcription and distinct promoter and chromatin properties, as well as genome-scale insights into RNA processing and dynamics. Overall design: RNA-seq of subcelluar fractions from five stimulation time-points