5-Hydroxyuridine is primarily used in biochemical and molecular biology research as a modified nucleoside. It plays a role in studying RNA structure, function, and dynamics due to its ability to mimic natural nucleosides while introducing specific modifications. This compound is valuable in investigating RNA-protein interactions, enzymatic processes, and the effects of oxidative stress on RNA molecules. Additionally, it serves as a tool in the development of therapeutic strategies targeting RNA-related diseases, such as viral infections and certain cancers. Its incorporation into RNA sequences helps researchers understand the impact of oxidative damage on RNA stability and function, providing insights into cellular mechanisms and potential therapeutic interventions.