Gene expression is the process by which genetic information is used to make the proteins which carry out the biological functions essential to all life. Interrogation of this process at a global level facilitates early-stage drug discovery and allows signatures of disease susceptibility to be elucidated. This type of analysis is typically carried out at the level of transcription using RNA-seq, where the focus is on the difference in abundance of mRNA transcripts between conditions. However, a significant weakness of this approach is its inability to provide any information at the level of translation. Thus, an entire layer of regulation, pivotal to determining the ultimate level of protein expression, is therefore neglected.
On the other hand, Ribosome Profiling (an integrated analysis of RNA-seq and Ribo-seq data) allows for a multi-layered categorisation of expression changes. The dual layers of information allow for a high degree of confidence in identified transcriptionally driven responses which otherwise would require extensive follow-up experimentation to validate changes at the protein level. Crucially, consolidation of the two technologies also allows for translationally driven responses and buffered responses (where RNA levels increase without a corresponding increase in protein levels) to be identified – phenomena which are beyond the scope of RNA-seq alone. Extensive expertise is required to interpret the different expression categories arising from the integrated analysis and obtain relevant insights into the processes underlying the changes observed between experimental conditions.
