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Leishmania pp 109-122 | Cite as

Ribosome Profiling in Trypanosomatids

  • Amelie J. Kraus
  • Raúl O. CosentinoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1971)

Abstract

Ribosomes are the machinery responsible for reading mRNAs and translating them into proteins. The ribosome profiling approach is based on high-throughput sequencing of ribosome-protected mRNAs. RNAs not harboring ribosomes are removed by nuclease digestion leaving the so-called ribosome “footprints.” The purified “footprint” RNA molecules are processed into DNA libraries and their individual abundance is determined by deep sequencing. Ribosome profiling reveals the portion of transcripts which are actually protein-coding and can be used for differential gene expression analysis addressing rates of protein synthesis, and translational control and efficiency.

Key words

Deep sequencing Gene expression mRNA levels Translation efficiency Protein abundance Translational control Ribosome positions CDSs 

Notes

Acknowledgments

We thank T. Nicolai Siegel and Juan José Vasquez for establishing the ribosome profiling protocol in Trypanosoma brucei. This work was funded by a European Research Council Starting Grant (3D_Tryps 715466). R.O.C was supported by a Georg Forster Fellowship (Humboldt Foundation).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Veterinary Sciences, Experimental ParasitologyLudwig-Maximilians-Universität MünchenMunichGermany
  2. 2.Department of Physiological Chemistry, Biomedical Center MunichLudwig-Maximilians-Universität MünchenPlanegg-MartinsriedGermany
  3. 3.Research Center for Infectious DiseasesUniversity of WürzburgWürzburgGermany

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