Mammalian NET-Seq Reveals Genome-wide Nascent Transcription Coupled to RNA Processing

Transcription is a highly dynamic process. Consequently, we have developed native elongating transcript sequencing technology for mammalian chromatin (mNET-seq), which generates single-nucleotide resolution, nascent transcription profiles. Nascent RNA was detected in the active site of RNA polymerase II (Pol II) along with associated RNA processing intermediates. In particular, we detected 5'splice site cleavage by the spliceosome, showing that cleaved upstream exon transcripts are associated with Pol II CTD phosphorylated on the serine 5 position (S5P), which is accumulated over downstream exons. Also, depletion of termination factors substantially reduces Pol II pausing at gene ends, leading to termination…

• W
  Wellcome

  Award: 091805/Z/10/Z
• KF
  Kanae Foundation for the Promotion of Medical Science
• JS
  Japan Society for the Promotion of Science
• CR
  Core Research for Evolutional Science and Technology