Intron-mediated RNA interference, intronic microRNAs, and applications

Abstract

Nearly 97% of the human genome is non-coding DNA. The intron occupies most of it around the gene-coding regions. Numerous intronic sequences have been recently found to encode microRNAs (miRNAs), responsible for RNA-mediated gene silencing through RNA interference (RNAi)-like pathways. miRNAs, small single-stranded regulatory RNAs capable of interfering with intracellular messenger RNAs (mRNAs) that contain either complete or partial complementarity, are useful for the design of new therapies against cancer polymorphism and viral mutation. This flexible characteristic differs from double-stranded siRNAs (small interfering RNAs) because more rigid complementarity is required for siRNA-induced RNAi gene silencing. miRNAs were firstly discovered in Caenorhabditis elegans as native RNA fragments that modulate a wide range of genetic regulatory pathways during embryonic development. Currently, varieties of miRNAs are widely reported in plants, animals, and even microorganisms. Intronic miRNA is a new class of miRNAs derived from the processing of gene introns. The intronic miRNAs differ from previously described intergenic miRNAs due to the requirement of type II RNA polymerases (Pol-II) and spliceosomal components for their biogenesis. Several kinds of intronic miRNAs have been identified in C. elegans, mouse, and human cells. However, neither function nor application has been reported. Here, we show that, for the first time, intron-derived miRNAs are able to induce RNA interference not only in human and mouse cell lines but also in zebrafish, chicken, and mouse, which demonstrates the evolutionary preservation of the intron-mediated gene silencing through miRNA functionality in cell and in vivo. Based on this novel mechanism, numerous biomedical applications have been developed, including cosmetic skin whitening, transgenic animal generation, anti-viral vaccination and therapy, and somatic cell reprogramming into induced pluripotent stem (iPS) cells. These findings suggest an important miRNA-mediated gene regulatory system, which fine-tunes a variety of cellular and developmental events through the mechanism of RNAi-like gene silencing.