Dr. Chengqi Yi published a paper on Angew Chem Int Ed Engl.
Pseudouridine (Ψ) is the most abundant post-transcriptional RNA modification. Methods have been developed to achieve locus-specific Ψ detection; however, they often involve radiolabeling of RNA, require advanced experimental skills and can be time-consuming. Here we report a radiolabeling-free，qPCR-based method to rapidly detect locus-specific Ψ. This method is based on Ψ labeling adduct induced mutation/deletion during reverse transcription (RT), generating qPCR products of different melting temperatures and hence altered melting curves. We first extensively validated our method on known Ψ sites in rRNA. Next, our methodsensitively detected Ψs in lncRNA and mRNA of low abundance. Finally, we applied our method to Ψ synthase identification and show for the first time that Ψ616 in PSME2 mRNA is dependent on PUS7. Our facile and cost-effective method takes only 1.5 days to complete, and with slight adjustment it can be applied to detect other epitranscriptomic marks.
Original link: http://onlinelibrary.wiley.com/doi/10.1002/anie.201708276/full
