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Nat Methods. 2012 Jul 1;9(8):840-6. doi: 10.1038/nmeth.2078.

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

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1
Department of Genetics and Genomic Sciences, Mount Sinai School of Medicine, New York, New York, USA.

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.

PMID:
22751203
PMCID:
PMC3518396
DOI:
10.1038/nmeth.2078
[Indexed for MEDLINE]
Free PMC Article
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