Detection of microRNAs in cultured cells and paraffin-embedded tissue specimens by in situ hybridization.

Determination of gene expression is essential for understanding the role of a given gene in normal cell growth or disease processes. Recently, newly described microRNAs have been shown to play a key role in the regulation of gene expression; in particular, deregulation of microRNAs is often associated with a variety of human disorders including cancer. Although microRNAs are small RNA molecules with about 20-23 nucleotides in length and detection of their expression is believed to be challenging, with the introduction of modified nucleotides such as locked nucleic acid, the specificity and sensitivity of detection have been greatly improved. There are many methods developed for microRNA detection, but our focus in this chapter is on in situ hybridization (ISH) detection of microRNAs. We have successfully used ISH to detect several microRNAs in paraffin-embedded tumor specimens or cells-cultured in vitro.