Quantitative approaches for small RNA expression analysis via multi-step RT-qPCR

Key roles small RNA play in the field of biomarker discovery and current approaches to best quantify these targets and their expression using modern multi-step RT-qPCR methodology. Recent biomedical research has been punctuated by the surprising discovery of numerous non-coding small RNAs species encoded by genomes spanning every species including microRNAs (miRNAs), piwi-interacting RNAs (piRNAs), repeat-associated siRNAs (RasiRNAs), 21U endo-siRNAs, and small nucleolar RNAs (snoRNAs). These small RNAs are involved in myriad aspects of cellular functions via direct or indirect interplay with genomic DNA, RNAs and proteins. Functional studies on these small RNAs have continued to expand and some preliminary findings have suggested that they are involved in regulating genome stability, epigenetic marking, transcription, translation, and protein functions. Detection and quantification of miRNA and other small RNAs, approximately 20-30 nucleotides in length, has provided key insights into understanding their biological functions, particularly functions in regulating gene expression and RNA silencing. Additionally, recent research has identified and elucidated the importance of miRNAs as key targets, potential novel therapeutic biomarkers and how dysregulation of these small RNAs can lead to the onset of various cancers and other pathologies. That said, quantitative analyses of small RNAs have been challenging because of their small sizes (~20 nucleotides (nt) for miRNAs, ~30 nt for piRNAs and 60-200 nt for snoRNas). In this webinar, we will examine small RNAs, all that they impact, and the current state of quantifying these novel targets in multi-step RT-qPCR analysis.