Accurate quantification of 3′-terminal 2′-O-methylated small RNAs by utilizing oxidative deep sequencing and stem-loop RT-qPCR

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

“…Notably, the 2′-O-methyl modification at the 3′ terminal of piRNAs can inhibit enzymatic ligation and polymerization reactions, posing great challenges for simple and sensitive quantification of piR-NAs. 15,16 Available technologies for the detection of piRNAs include Northern blotting, 17,18 microarrays, 12,19 and quantitative reverse transcription-polymerase chain reaction (qRT-PCR), 20 but they inevitably involve hazardous radioactive labeling, cumbersome operating procedures, and poor sensitivity. To circumvent these limitations, several isothermal amplification approaches (e.g., three-way junction-based strand displacement amplification, 21 target-induced interstrand ligation reaction, 22 and duplex-specific nuclease-assisted cycling amplification 23 ) have been introduced for piRNA detection.…”

Identification of 3′‐terminal 2′‐O‐methylated miRNA in plasma as a novel diagnostic biomarker of NSCLC

“…Furthermore, we conducted initial screening of 22 miR-NAs using mixed plasma samples from NSCLC patients and control subjects by stem-loop reverse transcript quantitative PCR (RT-qPCR) and poly(A)-tailing RT-qPCR as previously described. 7,8 The 3′t-2′Ome levels of nine miR-NAs were significantly elevated in NSCLC samples (Table S1). Subsequently, 3′t-2′Ome levels of these nine miRNAs were measured in 10 NSCLC patients and 10 control subjects.…”