Design of 365 nm-Light-triggered Fluorogenic Crosslinking via a Genetically Encoded 3-CF3-p-phenylenediamine-Lysine Featuring Red-shifted Photo-defluorination

Abstract

We introduce 3-CF₃-p-phenylenediamine-L-lysine, a genetically encodable photo-Defluorination-Acyl Fluoride Exchange ncAA. It features red-shifted activation (311 → 365 nm) for faster photo-defluorination relative to 1^st generation photo-DAFEx ncAA, and subsequent Ac-F crosslinking toward R-NH₂ with fluorogenic emission at visible 498 nm. This facilitates capturing and imaging of protein-protein interactions in living systems.