2026/04/13 by Jakob Trendel, Polina Prokofeva, Zhuo Chen +6 · 1 voice
Agricultural and Biological Sciences · Chemistry · Materials Science · #Biomolecule #Click Chemistry and Applications #Cysteine #Irradiation #Light effects on plants #Photochromic and Fluorescence Chemistry #Phototoxicity #Proteome #Ultraviolet #Ultraviolet radiation
paper · doi:10.1093/nar/gkag339
published in Nucleic Acids Research 54(7) (Oxford University Press)
openalex publication_date 2026/04/13 · openalex created_date 2026/04/22 · openalex updated_date 2026/08/06
The activation of chemical reactions in living cells using ultraviolet (UV) light enables the interrogation of biomolecules in their native environment with photoreactive probes or crosslinking reagents. Although numerous photo-crosslinking approaches have been successfully employed, they often suffer from common limitations, including low reaction yields, the need for long exposure times, and irradiation-induced cellular damage from heat, desiccation, or side reactions. We recently showed that 365 nm light-emitting diodes enable rapid, biorthogonal protein-DNA crosslinking in living cells, incurring minimal photodamage. Here, we generalize this approach and demonstrate that high-intensity, longwave UV light reduces the irradiation time for in-cell photo-crosslinking reactions by up to 1000-fold, allowing protein-drug, protein-protein, protein-DNA, and protein-RNA interactions to be fixed within seconds. Benchmarking this rapid photo-activation for the analysis of RNA-interacting proteomes responding to RNA-binding drugs or UV-induced RNA damage, we demonstrate both qualitative and quantitative advantages of controlled, high-intensity UV irradiation, uncovering emergent experimental opportunities that were previously inaccessible to light-activated chemistry in intact cells and tissues.