MicroRNA Detection by Whole-Mount In Situ Hybridization in C. elegans

Abstract

MicroRNAs (miRNAs) loaded on argonaute proteins guide RNA-induced silencing complexes to target mRNAs. An excellent method to decipher the spatiotemporal expression patterns of miRNAs is whole-mount in situ hybridization (WISH), which has been successfully used in vertebrate embryos but still remains unavailable for many animal species. Here, we describe a WISH method for miRNA detection in Caenorhabditis elegans at both embryonic and post-embryonic stages. Strategies devised for detection include fixation of animals with carbodiimide at a high temperature and subsequent partial digestion of the fixed animals with an extremely high concentration of proteinase. WISH signals are visualized by staining with a chromogenic substrate or a fluorescent dye.

MicroRNAs in Chronic Kidney Disease: Four Candidates for Clinical Application

“…Of all miR detection techniques, ISH is the only method providing spatial information. To increase specificity and stability, locked nucleic acids probes are used for this method [ 30 ], which are RNA derivative nucleotides in which the ribose ring is locked in a specific conformation.…”