Fluorescent biosensors based on the biological macromolecule are convenient tools for investigating the event occurring in the living cell. As for one of the candidates of such biosensors, we have reported a fluorescent sensor by utilizing a ribonucleopeptide (RNP) framework. Fluorescent RNP sensors are obtained from the fluorescent RNP library constructed by the combination of the RNA subunit, a substrate recognition unit selected by in vitro selection, and a fluorophore-modified peptide subunit. By taking the advantage of the noncovalent nature of fluorescent RNP complexes, RNP sensors with desired optical sensing properties are selected in a high-throughput manner. However, the noncovalent nature of the fluorescent RNP sensor is not suitable for practical applications. We report here a strategy to generate stable covalently linked RNP sensors and demonstrate a multiple ligands sensing system by using the covalently linked RNP sensors to detect biologically active ligands.
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