Ribosome profiling, or Ribo-Seq (also named ribosome footprinting), is an adaptation of a technique developed by Joan Steitz and Marilyn Kozak almost 50 years ago that Nicholas Ingolia and Jonathan Weissman adapted to work with next generation sequencing that uses specialized messenger RNA (mRNA) sequencing to determine which mRNAs are being actively translated.
It produces a “global snapshot” of all the ribosomes actively translating in a cell at a particular moment, known as a translatome.
[4] There are three main uses of ribosome profiling: identifying translated mRNA regions, observing how nascent peptides are folded, and measuring the amount of specific proteins that are synthesized.
[5] This allows the starting codon of the mRNAs throughout the cell lysate to be analyzed, which has been used to determine non-AUG sequences that do initiate translation.
Translation efficiency can then be computed as the ribosome occupancy of each gene while controlling for its RNA expression.
[9][10] This approach can be coupled with directed disruption of proteins that bind to RNA and using ribosome profiling to measure the difference in translation.