Validation of miR-21, miR-31, miR-34A, and miR-203 Changes by Quantitative Polymerase Chain Reaction in Isolated Keratinocytes.

MiRNAs play integral roles in diverse cellular functions, and their dysregulation is central to various pathological processes. Thyroid hormone (TH) is indispensable for numerous physiological processes, and it has been shown that multiple miRNAs regulate TH signaling in various tissues.This chapter describes a method for validating changes in miRNA observed through RNAseq. The method involves measuring the expression of mature miRNA levels in keratinocytes using the stem-loop real-time PCR. This method utilizes stem-loop RT primers to create cDNA for specific miRNAs in a single RT reaction. The keratinocytes used are derived from the total skin of four-day-old wild-type controls and double thyroid receptors knock-out mice (dTRKO).