High-throughput assessment of microRNA activity and function using microRNA sensor and decoy libraries.
High-throughput assessment of microRNA activity and function using microRNA sensor and decoy libraries.
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DOI：
10.1038/nmeth.2078
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发表时间：
2012-07-01
期刊：
影响因子：
48
通讯作者：
Brown, Brian D.
中科院分区：
文献类型：
--
作者：
Mullokandov, Gavriel;Baccarini, Alessia;Ruzo, Albert;Jayaprakash, Anitha D.;Tung, Navpreet;Israelow, Benjamin;Evans, Matthew J.;Sachidanandam, Ravi;Brown, Brian D.
我们介绍了两个用于microRNA功能分析的大型资源--一个用于抑制microRNA功能的诱骗/海绵库和一个用于监测microRNA活性的传感器文库。为了利用传感器文库，我们开发了一种称为传感器序列的高通量分析方法，它允许同时定量数百个microRNAs的活性。使用这种方法，我们表明只有细胞内最丰富的microRNAs才能调节显著的靶标抑制。超过60%的检测到的microRNAs没有可识别的活性，这表明一个细胞的功能‘miRNome’比目前从图谱研究中推断的要小得多。此外，一些高表达的microRNA表现出相对较弱的活性，在某些情况下，这与高靶标与microRNA的比率或microRNA的核定位增加有关。最后，我们证明了microRNA诱骗文库可以用于功能丧失的联合研究。这些工具为研究microRNA生物学和基于microRNA的疗法提供了宝贵的资源。
We introduce two large-scale resources for functional analysis of microRNA—a decoy/sponge library for inhibiting microRNA 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, which permits the activity of hundreds of microRNAs to be quantified simultaneously. Using this approach, we show that only the most abundant microRNAs within a cell mediate significant target suppression. Over 60% of detected microRNAs had no discernible activity, indicating that the functional ‘miRNome’ of a cell is considerably smaller than currently inferred from profiling studies. Moreover, some highly expressed microRNAs exhibit relatively weak activity, which in some cases correlated with a high target-to-microRNA ratio or increased nuclear localization of the microRNA. Finally, we show that the microRNA decoy library can be used for pooled loss-of-function studies. These tools provide valuable resources for studying microRNA biology and for microRNA-based therapeutics.
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影响因子：
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