Ribosome Profiling in Trypanosomatids
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 wordsDeep sequencing Gene expression mRNA levels Translation efficiency Protein abundance Translational control Ribosome positions CDSs
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).