A multi-cycle signal amplification-mediated single quantum dot nanosensor for PIWI-interacting RNA detection

Abstract

We report a catalytic single QD nanosensor for simple, selective, and highly sensitive quantification of piRNA.

Accurate identification of 8-oxoguanine in RNA with single-nucleotide resolution using ligase-dependent qPCR

“…We utilized RNA-1-G and RNA-1-o 8 G as templates to assess the substantial selectivity of the ligases towards o 8 G. Previous research has indicated that ribosylation of the two bases at the 3′ end of a probe can enhance both the efficiency and specificity of the ligation reaction. 39 Consequently, we synthesized the control DNA probe R, which has the same sequence as the RNA probe R but lacks the ribonucleotide modifications at the 3′ end. Probe L and DNA probe R were hybridized with RNA-1-G and RNA-1-o 8 G and then ligated using various ligases.…”

Construction of a Spatial-Confined Self-Stacking Catalytic Circuit for Rapid and Sensitive Imaging of Piwi-Interacting RNA in Living Cells

“…In summary, we have demonstrated the construction of a spatial-confined self-stacking catalytic circuit for rapid and sensitive imaging of piRNA in living cells based on intramolecular and intermolecular hybridization-accelerated CHA. Compared with the reported piRNA assays (Table S3), − this strategy has distinct advantages: (1) this strategy enables one-pot, single-tube isothermal analysis of piRNAs; (2) the enzyme-free reaction patterns of this strategy eliminate the need for careful optimization of various enzymic reaction conditions (e.g., pH, temperature, ion concentration, and strength), simplifying the experimental procedures; (3) the assembled 3WJ probe leads to the spatial proximity of two CHA reactant hairpins, which increases the local concentrations of CHA reactants and accelerates reaction kinetics of intramolecular CHA; (4) the protruding terminus resulting from the intramolecular CHA reaction can serve as a claw to agglutinate the unreacted 3WJ probes, reducing the turnaround time of recycled targets and facilitating the initiation of next rounds of CHA reactions; and (5) only when the target is present do the 3WJ probes come close to each other and assemble into a nanowire, which not only provides a track for target movement, but also efficiently avoids the risk of background signal leakage caused by the cross-entanglement of preassembled probes. This strategy achieves good specificity and high sensitivity with shortened assay time.…”