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High-throughput assessment of microRNA activity and function using microRNA sensor and decoy libraries

机译:使用microRNA传感器和诱饵库对microRNA活性和功能进行高通量评估

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摘要

We introduce two large-scale resources for functional analysis of microRNA (miRNA): a decoy library for inhibiting miRNA function and a sensor library for monitoring microRNA activity. To take advantage of the sensor library, we developed a high-throughput assay called Sensor-seq to simultaneously quantify the activity of hundreds of miRNAs. Using this approach, we show that only the most abundant miRNAs in a cell mediate target suppression. Over 60% of detected miRNAs had no discernible activity, which indicated that the functional 'miRNome' of a cell is considerably smaller than currently inferred from profiling studies. Moreover, some highly expressed miRNAs exhibited relatively weak activity, which in some cases correlated with a high target-to-miRNA ratio or increased nuclear localization of the miRNA. Finally, we show that the miRNA decoy library can be used for pooled loss-of-function studies. These tools are valuable resources for studying miRNA biology and for miRNA-based therapeutics.
机译:我们介绍了两个用于microRNA(miRNA)功能分析的大型资源:一个用于抑制miRNA功能的诱饵库和一个用于监视microRNA活性的传感器库。为了利用传感器库的优势,我们开发了一种高通量测定方法,称为Sensor-seq,可同时定量数百种miRNA的活性。使用这种方法,我们表明只有细胞中最丰富的miRNA才能介导靶标抑制。超过60%的检测到的miRNA没有明显的活性,这表明细胞的功能性“ miRNome”比目前从谱研究中推断的小得多。此外,一些高表达的miRNA表现出相对较弱的活性,这在某些情况下与高的靶标与miRNA比率或miRNA的核定位增加有关。最后,我们证明了miRNA诱饵文库可用于合并的功能丧失研究。这些工具是研究miRNA生物学和基于miRNA的疗法的宝贵资源。

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