Fig. (4).

Summary of methodological workflow for studying miRNA function. The workflow summarizes the emerging high-throughput experimental approaches for the study of miRNA gene regulatory networks in aging. For miRNA discovery, high-throughput methods such as massively parallel sequencing or microarray can be utilized to identify differentially expressed miRNAs in a variety of aging models. Target sites of aging-related miRNAs are identified either by in silico analysis or experimental approaches such as HITS-CLIP and PAR-CLIP. Target sites are then validated by 3’ URT reporter assays or by assessing the anti-correlation between a miRNA and its target gene/protein levels in transfection experiments with miRNA minics, antagomir, or target protectors. Biological pathway analysis of target genes will provide insights into aging gene regulatory networks.