MicroRNAs (miRNAs) comprise a class of small non-coding RNAs that regulate the stability and/or translatability of most protein-coding transcripts. Steady-state levels of mature miRNAs can be controlled through mechanisms that influence their biogenesis and/or decay rates. Pathways that mediate mature miRNA decay are less well understood than those that mediate miRNA biogenesis. We recently described Tudor-staphylococcal/micrococcal-like nuclease (TSN)-mediated miRNA decay (TumiD) as a cellular pathway that promotes the sequence-specific endonucleolytic decay of miRNAs that harbor a CA and/or UA dinucleotide. Here, we describe an in vitro assay for evaluating the susceptibility of AGO2-loaded miRNAs to degradation by different classes of nucleases. This in vitro approach can be used to complement in vivo studies that aim to identify novel miRNA decay factors.

中文翻译：

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体外评估载有 AGO2 的 microRNA 对核酸酶降解的敏感性

MicroRNAs (miRNAs) 包含一类小的非编码 RNA，它们调节大多数蛋白质编码转录物的稳定性和/或翻译性。成熟 miRNA 的稳态水平可以通过影响其生物发生和/或衰减率的机制来控制。与介导 miRNA 生物发生的途径相比，对介导成熟 miRNA 衰变的途径了解较少。我们最近将 Tudor 葡萄球菌/微球菌样核酸酶 (TSN) 介导的 miRNA 衰减 (TumiD) 描述为一种细胞途径，可促进含有 CA 和/或 UA 二核苷酸的 miRNA 的序列特异性核酸内切衰减。在这里，我们描述了一种体外测定，用于评估加载 AGO2 的 miRNA 对不同类别核酸酶降解的敏感性。