STAR Protocols (Sep 2022)

End-to-end pipeline for differential analysis of pausing in ribosome profiling data

  • Keegan Flanagan,
  • Wanxin Li,
  • Ethan J. Greenblatt,
  • Khanh Dao Duc

Journal volume & issue
Vol. 3, no. 3
p. 101605

Abstract

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Summary: Ribosome profiling is a powerful technique which maps the distribution of ribosomes along mRNAs to analyze translation genome-wide. Ribosome density can be affected by multiple factors, such as changes to translation initiation or elongation rates. We describe the application of a metric for identifying genes rate-limited by these rates by analyzing the relative distribution of ribosome footprints along transcripts. This protocol also details two sample analyses comparing gene translation efficiencies and the distribution of ribosome densities on downloadable datasets.For complete details on the use and execution of this protocol, please refer to Flanagan et al. (2022). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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