Current experimental strategies for intracellular target identification of microRNA
Tóm tắt
Intracellular target identification of microRNA (miRNA), which is essential for understanding miRNA-involved cellular processes, is currently the most challenging task in miRNA-related studies. Although bioinformatic methods have been developed as the most efficient strategy for miRNA target identification, high-throughput experimental strategies are still highly demanded. In this review paper, we summarize and compare current experimental strategies for miRNA target identification, including gene expression profiling, immunoprecipitation and pull-down methods. Gene expression profiling methods mainly rely on the measurement of target gene expression through overexpression or inhibition of specific miRNA, which are indirect strategies to unveil miRNA targets. Immunoprecipitation methods use specific antibody to isolate RISC and bound mRNAs, followed by analysis with high-throughput techniques and bioinformatics to reveal miRNA-mRNA interactions. Pull-down methods use tagged miRNA mimics as probes to isolate associated target genes through affinity purification, which directly indicate miRNA-mRNA interactions after analysis of isolated target genes. Each method has its own advantages and limitations, which will be summarized and discussed in details. Overall, this review paper aims to provide a brief outline of recent achievements at experimental strategies for miRNA target identification. With the further development or improvement, we envision these experimental strategies will ultimately contribute a lot to the research on miRNA and miRNA-targeted biomedicine.
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