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High-throughput assessment of microRNA activity and function using microRNA sensor and decoy libraries.
Mullokandov, Gavriel; Baccarini, Alessia; Ruzo, Albert; Jayaprakash, Anitha D; Tung, Navpreet; Israelow, Benjamin; Evans, Matthew J; Sachidanandam, Ravi; Brown, Brian D.
Afiliação
  • Mullokandov G; Department of Genetics and Genomic Sciences, Mount Sinai School of Medicine, New York, New York, USA.
Nat Methods ; 9(8): 840-6, 2012 Jul 01.
Article em En | MEDLINE | ID: mdl-22751203
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.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Técnicas Biossensoriais / Biblioteca Gênica / MicroRNAs / Ensaios de Triagem em Larga Escala / Vetores Genéticos Limite: Animals / Humans Idioma: En Ano de publicação: 2012 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Técnicas Biossensoriais / Biblioteca Gênica / MicroRNAs / Ensaios de Triagem em Larga Escala / Vetores Genéticos Limite: Animals / Humans Idioma: En Ano de publicação: 2012 Tipo de documento: Article