“One stroke drawing” of poly(ribonucleic acids) with different aptamer functions for sensing probes

Using polymerase chain reaction (PCR)-mediated recombination, single ribonucleic acid (RNA) chains containing bifunctional RNA sequences involving substrate binding and phosphorescent signaling were prepared. For substrate binding and phosphorescent labeling, thrombin- or lysozyme-binding and ruthenium complex-binding RNA aptamer sequences were used. It was demonstrated that the structural properties of the conjugated RNAs showed similar characteristics to the original aptamers using circular dichroism (CD) spectrometry. Furthermore, electrophoretic mobility shift assays of the proteins and phosphorescence measurements of the ruthenium complexes suggested that the binding abilities of the conjugated RNAs maintained the original aptamer functions. Finally, it was established that the conjugated RNA sequences were suitable as phosphorescent RNA probes for protein cognates. Therefore, this “one stroke drawing” strategy is proposed as a promising biological method for the generation of phosphorescent RNA probes that do not culminate in a loss of function of the original aptamers. Using polymerase chain reaction (PCR)-mediated recombination, we prepared single ribonucleic acid (RNA) chains containing bifunctional RNA sequences involving substrate binding and phosphorescent signaling. The prepared RNAs largely maintained the functionalities of the original aptamers even after bifunctionalization. We demonstrated that the bifunctionalized RNAs can be used as phosphorescent detection probes for the target protein. Therefore, we suggested that the PCR-mediated “one stroke drawing” is a promising strategy for the preparation of RNA detection probes.