High-throughput approaches for the identification of ribosome heterogeneity

Recent advances in the fields of RNA translation and ribosome biology have demonstrated the heterogeneous nature of ribosomes. This manifests not only across different cellular contexts but also within the same cell. Such variations in ribosomal composition, be it in ribosomal RNAs or proteins, can significantly influence cellular processes and responses by altering the mRNAs being translated or the dynamics of ribosomes during the translation process. Therefore, identifying this heterogeneity is crucial for unravelling the complexity of gene expression across different fields of biology. Here we provide an overview of recent advances in high-throughput techniques for identifying ribosomal heterogeneity. We cover methodologies for probing both rRNA and protein components of the ribosome and encompass the most recent next-generation sequencing and computational analyses, as well as a diverse array of mass spectrometry techniques.This article is part of the discussion meeting issue 'Ribosome diversity and its impact on protein synthesis, development and disease'.