mRNA Display for Identifying Peptide Substrates of Enzymes

Among display technologies, mRNA display has emerged as a powerful approach to identify de novo ligands for proteins of interest. Applying the same methods to studying the substrates of protein/peptide-modifying enzymes has received much less attention, but progress in this area has accelerated rapidly over the last 5 years. In this article, we review the published literature to date (up to December 2025) of dedicated efforts to identify and understand enzyme substrate preferences using mRNA- or cDNA-display. We include observations of trends from these reports over time and reflections on where the area may go in the future. We hope this will serve as a useful primer for researchers in this and related areas.