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


ABSTRACT: 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.

SUBMITTER: Mullokandov G 

PROVIDER: S-EPMC3518396 | biostudies-literature | 2012 Jul

REPOSITORIES: biostudies-literature

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

Mullokandov Gavriel G   Baccarini Alessia A   Ruzo Albert A   Jayaprakash Anitha D AD   Tung Navpreet N   Israelow Benjamin B   Evans Matthew J MJ   Sachidanandam Ravi R   Brown Brian D BD  

Nature methods 20120701 8


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 activit  ...[more]

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