Ribosome Profiling: Principles and Variations

Abstract Ribosome profiling provides a snapshot of ribosome positions and density across the transcriptome at a sub‐codon resolution. By sequencing the entire set of ribosome‐protected mRNA fragments, this powerful approach has been successfully used to measure ribosome dynamics and reveal the hidden coding potential of transcriptome. Since its conceptual inception, ribosome profiling has evolved into a versatile method with many innovative variations. It has been applied to study translation in diverse cell types, adapted to capture specific subsets of ribosomes and further improved to address translational regulation in multicellular organisms. The continuous development of ribosome profiling technologies over the coming decade promises a broad view of translational regulation of gene expression. Key Concepts Ribosome profiling reveals a global translation snapshot. Ribosome profiling is applicable to diverse cell types. Profiling of specific ribosomes permits analysis of nascent chains. Profiling of initiating ribosomes allows the identification of alternative start codon. Ribosome profiling can be adapted to probe translation in vivo .