Статья

An estimate of the total number of true human miRNAs

Julia AllesInstitute of Human Genetics, Saarland University, 66421 Homburg, GermanyTobias FehlmannChair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, GermanyUlrike FischerInstitute of Human Genetics, Saarland University, 66421 Homburg, GermanyChristina BackesChair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, GermanyValentina GalataChair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, GermanyMarie MinetChair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, GermanyMartin HartInstitute of Human Genetics, Saarland University, 66421 Homburg, GermanyMasood Abu‐HalimaInstitute of Human Genetics, Saarland University, 66421 Homburg, GermanyFriedrich A. GrässerInstitute of Virology, Saarland University Medical School, 66421 Homburg, GermanyHans‐Peter LenhofAndreas KellerChair for Clinical Bioinformatics, Saarland University, 66123 Saarbrücken, GermanyEckart MeeseInstitute of Human Genetics, Saarland University, 66421 Homburg, Germany

2019en

ABI

Аннотация

While the number of human miRNA candidates continuously increases, only a few of them are completely characterized and experimentally validated. Toward determining the total number of true miRNAs, we employed a combined in silico high- and experimental low-throughput validation strategy. We collected 28 866 human small RNA sequencing data sets containing 363.7 billion sequencing reads and excluded falsely annotated and low quality data. Our high-throughput analysis identified 65% of 24 127 mature miRNA candidates as likely false-positives. Using northern blotting, we experimentally validated miRBase entries and novel miRNA candidates. By exogenous overexpression of 108 precursors that encode 205 mature miRNAs, we confirmed 68.5% of the miRBase entries with the confirmation rate going up to 94.4% for the high-confidence entries and 18.3% of the novel miRNA candidates. Analyzing endogenous miRNAs, we verified the expression of 8 miRNAs in 12 different human cell lines. In total, we extrapolated 2300 true human mature miRNAs, 1115 of which are currently annotated in miRBase V22. The experimentally validated miRNAs will contribute to revising targetomes hypothesized by utilizing falsely annotated miRNAs.

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Идентификаторы

DOI: 10.1093/nar/gkz097

Цитирования и источники

Цитирований: 2Использованных источников: 0

Показатели — AkademScholar