4.4 Article

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

Journal

POLYMER JOURNAL
Volume 53, Issue 5, Pages 667-675

Publisher

SPRINGERNATURE
DOI: 10.1038/s41428-020-00454-y

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Funding

  1. RIKEN-TIT International Program Associate

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Using PCR-mediated recombination, researchers prepared single RNA chains containing bifunctional RNA sequences for substrate binding and phosphorescent signaling. Structural and functional analyses confirmed that the conjugated RNAs maintained the binding abilities and functions of the original aptamers, making them suitable as phosphorescent RNA probes for protein cognates. This one-step drawing strategy shows promise as a biological method for generating phosphorescent RNA probes without loss of function from the original aptamers.
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.

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