3'LIFE: A functional assay to detect miRNA targets in high-throughput

Justin M. Wolter, Kasuen Kotagama, Alexandra C. Pierre-Bez, Mari Firago, Marco Mangone

Research output: Contribution to journalArticlepeer-review

35 Scopus citations


MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene output at the post-transcriptional level by targeting degenerate elements primarily in 3 untranslated regions (3′UTRs) of mRNAs. Individual miRNAs can regulate networks of hundreds of genes, yet for the majority of miRNAs few, if any, targets are known. Misexpression of miRNAs is also a major contributor to cancer progression, thus there is a critical need to validate miRNA targets in high-throughput to understand miRNAs' contribution to tumorigenesis. Here we introduce a novel high-throughput assay to detect miRNA targets in 3 UTRs, called Luminescent Identification of Functional Elements in 3 UTRs (3'LIFE). We demonstrate the feasibility of 3'LIFE using a data set of 275 human 3′UTRs and two cancer-relevant miRNAs, let-7c and miR-10b, and compare our results to alternative methods to detect miRNA targets throughout the genome. We identify a large number of novel gene targets for these miRNAs, with only 32% of hits being bioinformatically predicted and 27% directed by non-canonical interactions. Functional analysis of target genes reveals consistent roles for each miRNA as either a tumor suppressor (let-7c) or oncogenic miRNA (miR-10b), and preferentially target multiple genes within regulatory networks, suggesting 3'LIFE is a rapid and sensitive method to detect miRNA targets in high-throughput.

Original languageEnglish (US)
Article numbere132
JournalNucleic acids research
Issue number17
StatePublished - Jun 27 2014

ASJC Scopus subject areas

  • Genetics


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