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De novo Identification of Actively Translated Open Reading Frames with Ribosome Profiling Data
JoVE Journal
Biology
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JoVE Journal Biology
De novo Identification of Actively Translated Open Reading Frames with Ribosome Profiling Data
DOI:

08:23 min

February 18, 2022

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Chapters

  • 00:04Introduction
  • 01:01Environment Setup and RiboCode Installation
  • 01:23Data Preparation
  • 03:35Removing Ribosomal RNA Contamination
  • 03:59Aligning the Clean Reads to the Genome
  • 04:25Size Selection of Ribosome Protected Fragments, Identification of P-Sites, and De novo Annotate Translating Open Reading Frames
  • 04:59Results: Identification of the Novel Open Reding Frames (ORFs) with the Application of RiboCode
  • 07:20Conclusion

Summary

Automatic Translation

Translating ribosomes decode three nucleotides per codon into peptides. Their movement along mRNA, captured by ribosome profiling, produces the footprints exhibiting characteristic triplet periodicity. This protocol describes how to use RiboCode to decipher this prominent feature from ribosome profiling data to identify actively translated open reading frames at the whole-transcriptome level.

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