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Updated: Jun 8, 2026

MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
Identification of targets of specific miRNAs by selective amplification of Ago2-associated mRNA
Botao Zhao1, Yi Shi, Wenmin Qin
1State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences Shanghai, China.
Abstract:
To study the function of a miRNA, it is necessary to identify its target genes. The most common methods to reveal miRNA target genes rely on ectopically expressed tagged Ago2 and nonphysiological overexpression or inhibition of the miRNA of interest. To uncover the natural association between miRNAs and their target genes, we isolated endogenous Ago2 protein followed by a selective strategy, which only amplified target genes of the selected miRNA from the purified RNA-induced silencing complex by miRNA specific primers. This enabled us to identify the mRNAs regulated by miRNAs of interest. Our data demonstrated that this strategy is effective and highly credible. Moreover, our results showed the evidence of efficient miRNA target sites in 5' untranslated regions and open reading frames of target mRNAs.
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