Abstract

The MS2 system has been widely adopted for live-cell single molecule imaging of mRNA, but is of limited applicability for RNAs present in the cell nucleus due to excess nuclear-localised fluorescent capsid protein. Here we describe Nuclear-BRITE, an improved methodology that reduces nuclear background signal, enabling fast live-cell imaging of a wide range of endogenously tagged nuclear RNAs at single-molecule resolution without perturbing their abundance, localisation or function.