biostudies-literatureVoss GVetenskapsrådetKungliga Fysiografiska Sällskapet i LundProstatacancerfondenCancerfonden1412-1424https://www.ebi.ac.uk/biostudies/studies/S-EPMC8522694biostudies-literatureUnknown27(11)Even though microRNAs have been viewed as promising biomarkers for years, their clinical implementation is still lagging far behind. This is in part due to the lack of RT-qPCR technologies that can differentiate between microRNA isoforms. For example, A-to-I editing of microRNAs through adenosine deaminase acting on RNA (ADAR) enzymes can affect their expression levels and functional roles, but editing isoform-specific assays are not commercially available. Here, we describe RT-qPCR assays that are specific for editing isoforms, using microRNA-379 (miR-379) as a model. The assays are based on two-tailed RT-qPCR, and we show them to be compatible both with SYBR Green and hydrolysis-based chemistries, as well as with both qPCR and digital PCR. The assays could readily detect different miR-37RNA (New York, N.Y.)Quantification of microRNA editing using two-tailed RT-qPCR for improved biomarker discovery.PMC8522694CAN 2017/559VR-MH 2018-0312540526Voss GEdsjo ACeder YBjartell AfalseQuantification of microRNA editing using two-tailed RT-qPCR for improved biomarker discovery.Even though microRNAs have been viewed as promising biomarkers for years, their clinical implementation is still lagging far behind. This is in part due to the lack of RT-qPCR technologies that can differentiate between microRNA isoforms. For example, A-to-I editing of microRNAs through adenosine deaminase acting on RNA (ADAR) enzymes can affect their expression levels and functional roles, but editing isoform-specific assays are not commercially available. Here, we describe RT-qPCR assays that are specific for editing isoforms, using microRNA-379 (miR-379) as a model. The assays are based on two-tailed RT-qPCR, and we show them to be compatible both with SYBR Green and hydrolysis-based chemistries, as well as with both qPCR and digital PCR. The assays could readily detect different miR-372021-01-01T00:00:00Z2021 Nov2025-04-05T13:44:19.141Z2025-04-05T13:44:19.141ZS-EPMC85226943443363610.1261/rna.078867.121