microRNAs (miRNAs) are small noncoding RNAs that regulate gene expression at the posttranscriptional level by binding to sites within the 3' untranslated regions of messenger RNA (mRNA) transcripts. The discovery of this completely new mechanism of gene regulation necessitated the development of a variety of techniques to further characterize miRNAs, their expression, and function. In this chapter, we will discuss techniques currently used in the miRNA field to detect, express and inhibit miRNAs, as well as methods used to identify and validate their targets, specifically with respect to the miRNAs encoded by human cytomegalovirus.
Keywords: Human cytomegalovirus (HCMV); Locked nucleic acids (LNA); Luciferase assay; Northern Blot; Photoactivatable ribonucleoside-enhanced cross-linking and Immunoprecipitation (PAR CLIP); RISC-immunoprecipitation (RISC-IP); Stem-loop real-time PCR; microRNA (miRNA).