ArticleJournal of visualized experiments : JoVE2015
Detection of miRNA Targets in High-throughput Using the 3'LIFE Assay.
Article in Journal of visualized experiments : JoVE, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
6 citing papers in PubMed.
- MicroRNA Expression Profiling, Target Identification, and Validation in Chondrocytes.Methods in molecular biology (Clifton, N.J.) · 2021Article
- The tRNA pseudouridine synthase TruB1 regulates the maturation of let-7 miRNA.The EMBO journal · 2020Article
- Identification of targets of tumor suppressor microRNA-34a using a reporter library system.Proceedings of the National Academy of Sciences of the United States of America · 2017Article
- RESA identifies mRNA-regulatory sequences at high resolution.Nature methods · 2017Article
- Evolutionary patterns of metazoan microRNAs reveal targeting principles in the let-7 and miR-10 families.Genome research · 2017Article
- Article
Corrections and comments
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Authors and funding
4 authors.
Funding
Abstract
Luminescent Identification of Functional Elements in 3'UTRs (3'LIFE) allows the rapid identification of targets of specific miRNAs within an array of hundreds of queried 3'UTRs. Target identification is based on the dual-luciferase assay, which detects binding at the mRNA level by measuring translational output, giving a functional readout of miRNA targeting. 3'LIFE uses non-proprietary buffers and reagents, and publically available reporter libraries, making genome-wide screens feasible and cost-effective. 3'LIFE can be performed either in a standard lab setting or scaled up using liquid handling robots and other high-throughput instrumentation. We illustrate the approach using a dataset of human 3'UTRs cloned in 96-well plates, and two test miRNAs, let-7c and miR-10b. We demonstrate how to perform DNA preparation, transfection, cell culture and luciferase assays in 96-well format, and provide tools for data analysis. In conclusion 3'LIFE is highly reproducible, rapid, systematic, and identifies high confidence targets.
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Registered trials
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