High-Throughput Techniques for Identifying microRNA Target Genes

MicroRNAs (miRNAs) are important regulators of cell biological processes with approximately 21–25 nucleotides in length. They are thought to suppress protein coding gene expression through translation inhibition or (and) mRNA decay through leading the RNAi-induced silencing complex (RISC) to complementary sites in the 3' untranslated region of the target mRNA. Studies have shown widespread regulation of protein levels by miRNAs in cellular and animal models. Some computeraided putative target genes have been validated using a single target gene identification method, such as luciferase reporter assays or EGFP/RFP reporter assay. However, many of other target genes are unknown and need to be experimentally validated. Recently, high-throughput multiple target screening assays that enable a large-scale, efficient and multi-level study are expected to greatly improve the target identification methods. In this review, current high-throughput technologies for identifying miRNAs target genes are introduced, together with perspectives on further developments in this field.