Imaging of nucleolar RNA in living cells using a highly photostable deep-red fluorescent probe

Abstract

A new crescent-shape fluorescent probe (named here as CP) that selectively stains RNA in nucleoli of living cells is prepared. CP shows a deep-red emission (658 nm) and a large Stokes shift because of the introduction of rigid-conjugated coumarin moiety into the molecular structure. Cell imaging experiments indicate that CP can rapidly stain nucleoli in living cells by binding with nucleolar RNA, showing performance superior to commercially available nucleoli dye SYTO RNASelect in terms of high photostability and selectivity. More significantly, these excellent properties together with low cytotoxicity enable CP to monitor nucleolar RNA changes during mitosis, and after treating with anti-cancer drugs cisplatin, actinomycin D and α-amanitin. Thus, CP could be a potential tool for real-time, long-term visualization of the dynamic changes for nucleolar RNA and evaluation of the therapeutic effect for anti-cancer drugs that targeted RNA polymerase I (Pol I).

Keywords: Fluorescent imaging; Nucleoli; RNA; Real-time.