機能解析を目的としたmicroRNAの検出／発現／阻害システムの確立 ～It’s a small miRNA World～

Abstract

MicroRNAs (miRNAs) are relatively short, 22nt～, and endogenous non-coding RNAs. It is known to act as a sequence specific regulator of post-transcriptional gene expression in many eukaryotes, most of the time with some sequence mismatches against target genes. Recently, a couple of miRNAs have been shown to be involved in oncogenic pathway, development, or cell differentiation. miRNAs are now getting attention to its diverse regulatory and catalytic functions.Although many of the miRNA sequences have been registered in the public databases and their target genes are predicted by computational analysis, the empirical verification has to be done. One of the difficulties to study the miRNA function will be that its structural and regulatory characteristics.In order to overcome this difficulty and utilize miRNA as an experimental tool, we investigated a system to detect, knockdown, and / or overexpress miRNAs. First, we adopted bicyclic nucleic acid, Locked Nucleic Acid (LNA), to improve affinity and single nucleotide discrimination. Next, we established the system to introduce synthetic oligos or RNAs expressed from lentiviral vectors to cells. A novel approach based on these principles on miRNA and its pathway analysis will be discussed.