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qPCR-based methods for expression analysis of miRNAs

Diego A Forero

Yeimy González-Giraldo

Luis J Castro-Vega

George E Barreto

Diego A Forero

*Author for correspondence:

E-mail Address: diego.forero@uan.edu.co

Laboratory of NeuroPsychiatric Genetics, Biomedical Sciences Research Group, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia

PhD Program in Health Sciences, School of Medicine, Universidad Antonio Nariño, Bogotá, Colombia

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Yeimy González-Giraldo

Departamento de Nutrición y Bioquímica, Pontificia Universidad Javeriana, Bogotá, Colombia

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Luis J Castro-Vega

INSERM, UMR970, Paris-Cardiovascular Research Center, Equipe Labellisée par la Ligue contre le Cancer, Paris, France

Université Paris Descartes, Sorbonne Paris Cité, Faculté de Médecine, Paris, France

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George E Barreto

Departamento de Nutrición y Bioquímica, Pontificia Universidad Javeriana, Bogotá, Colombia

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Published Online:6 Sep 2019https://doi.org/10.2144/btn-2019-0065

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Abstract

Several approaches for miRNA expression analysis have been developed in recent years. In this article, we provide an updated and comprehensive review of available qPCR-based methods for miRNA expression analysis and discuss their advantages and disadvantages. Existing techniques involve the use of stem–loop reverse transcriptase–PCR, polyadenylation of RNAs, ligation of adapters or RT with complex primers, using universal or miRNA-specific qPCR primers and/or probes. Many of these methods are oriented towards the expression analysis of mature miRNAs and few are designed for the study of pre-miRNAs and pri-miRNAs. We also discuss findings from articles that compare results from existing methods. Finally, we suggest key points for the improvement of available techniques and for the future development of additional methods.

Keywords:

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History

Received 28 May 2019

Accepted 4 July 2019

Published online 6 September 2019

Published in print October 2019

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Author contributions

DAF conceived the idea, reviewed the literature, contributed to the generation of tables and figures and wrote and reviewed the manuscript: YG-G conceived the idea, reviewed the literature, contributed to the generation of tables and figures and wrote and revised the manuscript; LJC-V reviewed the literature and contributed to the writing and revision of the manuscript; GEB reviewed the literature and contributed to the writing and revision of the manuscript.

Financial & competing interests disclosure

YG-G is supported by a PhD fellowship from Centro de Estudios Interdisciplinarios Básicos y Aplicados CEIBA (Rodolfo Llinás Program). DAF is supported by research grants from Colciencias and VCTI. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

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