AIEgen-Conjugated Phase-Separating Peptides Illuminate Intracellular RNA through Coacervation-Induced Emission

Intrinsically disordered peptides drive dynamic liquid-liquid phase separation (LLPS) in membraneless organelles and encode cellular functions in response to environmental stimuli. Engineering design on phase-separating peptides (PSPs) holds great promise for bioimaging, vaccine delivery, and disease theranostics. However, recombinant PSPs are devoid of robust luminogen or suitable cell permeability required for intracellular applications. Here, we synthesize a peptide-based RNA sensor by covalently connecting tetraphenylethylene (TPE), an aggregation-induced emission luminogen (AIEgens), to tandem peptide repeats of (RRASL)_n (n = 1, 2, 3). Interestingly, the conjugation of TPE luminogen promotes liquid−liquid phase separation of the peptide repeats, and the minimum coacervation concentration (MCC) of TPE-(RRASL)_n is decreased by an order of magnitude, compared to that of the untagged, TPE-free counterparts. Moreover, the luminescence of TPE-(RRASL)_n is enhanced by up to 700-fold with increasing RNA concentration, which is attributed to the constricted rotation of the TPE moiety as a result of peptide/RNA coacervates within the droplet phase. Besides, at concentrations above MCC, TPE-(RRASL)_n can efficiently penetrate through human gallbladder carcinoma cells (SGC-996), translocate into the cell nucleus, and colocalize with intracellular RNA. These observations suggest that AIEgen-conjugated PSPs can be used as droplet-based biosensors for intracellular RNA imaging through a regime of coacervation-induced emission. © 2023 American Chemical Society.