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Transcription Elongation Factors02:35

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Related Experiment Video

Updated: Jan 19, 2026

Transcription Elongation Factors & Pausing of RNA Polymerase
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Analysis of Mammalian Native Elongating Transcript sequencing (mNET-seq) high-throughput data.

Pedro Prudêncio1, Kenny Rebelo2, Ana Rita Grosso3

  • 1Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina, Universidade de Lisboa, Lisboa, Portugal; Center for Biomedical Research, Universidade do Algarve, Faro, Portugal.

Methods (San Diego, Calif.)
|September 8, 2019
PubMed
Summary
This summary is machine-generated.

Mammalian Native Elongating Transcript sequencing (mNET-seq) offers genome-wide maps of nascent RNA polymerase II (Pol II) transcripts. This method profiles Pol II CTD phosphorylation and RNA processing with single-nucleotide resolution.

Keywords:
Co-transcriptional splicingNascent RNARNA polymerase IImNET-seq

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Area of Science:

  • Molecular Biology
  • Genomics
  • Transcriptomics

Background:

  • Understanding gene expression regulation requires precise mapping of nascent RNA transcripts.
  • RNA polymerase II (Pol II) elongation complexes play a crucial role in transcription and RNA processing.
  • Existing methods may lack the resolution or specificity to capture dynamic transcriptional events.

Purpose of the Study:

  • To describe the Mammalian Native Elongating Transcript sequencing (mNET-seq) pipeline.
  • To enable genome-wide profiling of nascent transcripts and Pol II CTD phosphorylation states.
  • To provide insights into coupled RNA processing events during transcription.

Main Methods:

  • Isolation of ternary transcription complexes (Pol II, DNA, nascent RNA) via immunoprecipitation.
  • Antibody-based recognition of specific Pol II C-terminal domain (CTD) phosphorylation states.
  • High-throughput sequencing to identify the 3' end coordinates of nascent RNA.

Main Results:

  • Generation of global, bidirectional maps of Pol II CTD phosphorylation-specific nascent transcripts.
  • Single nucleotide resolution of nascent transcription and coupled RNA processing.
  • Detailed description of the data preparation and analysis pipeline for mNET-seq.

Conclusions:

  • mNET-seq is a powerful technique for studying transcription dynamics and RNA processing.
  • The described pipeline facilitates the application and analysis of mNET-seq data.
  • This method enhances our understanding of gene expression regulation at the transcript level.