Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
Abstract MicroRNAs are key non-coding RNA molecules that play a relevant role in the regulation of gene expression through translational repression and/or transcript cleavage during normal development and physiological adaptation processes like stress. Quantitative reverse transcription polymerase c...
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ftdoajarticles:oai:doaj.org/article:a0d66163efbd49b69fcfcd67af706e35 2023-05-15T15:32:32+02:00 Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar Eduardo Zavala Daniela Reyes Robert Deerenberg Rodrigo Vidal 2017-05-01T00:00:00Z https://doi.org/10.1038/s41598-017-01970-3 https://doaj.org/article/a0d66163efbd49b69fcfcd67af706e35 EN eng Nature Portfolio https://doi.org/10.1038/s41598-017-01970-3 https://doaj.org/toc/2045-2322 doi:10.1038/s41598-017-01970-3 2045-2322 https://doaj.org/article/a0d66163efbd49b69fcfcd67af706e35 Scientific Reports, Vol 7, Iss 1, Pp 1-10 (2017) Medicine R Science Q article 2017 ftdoajarticles https://doi.org/10.1038/s41598-017-01970-3 2022-12-31T07:10:11Z Abstract MicroRNAs are key non-coding RNA molecules that play a relevant role in the regulation of gene expression through translational repression and/or transcript cleavage during normal development and physiological adaptation processes like stress. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) has become the approach normally used to determine the levels of microRNAs. However, this approach needs the use of endogenous reference. An improper selection of endogenous references can result in confusing interpretation of data. The aim of this study was to identify and validate appropriate endogenous reference miRNA genes for normalizing RT-qPCR survey of miRNAs expression in four different tissues of Atlantic salmon, under handling and confinement stress conditions associated to early or primary stress response. Nine candidate reference normalizers, including microRNAs and nuclear genes, normally used in vertebrate microRNA expression studies were selected from literature, validated by RT-qPCR and analyzed by the algorithms geNorm and NormFinder. The results revealed that the ssa-miR-99-5p gene was the most stable overall and that ssa-miR-99-5p and ssa-miR-23a-5p genes were the best combination. Moreover, the suitability of ssa-miR-99-5p and ssa-miR-23a-5p as endogeneuos reference genes was demostrated by the expression analysis of ssa-miR-193-5p gene. Article in Journal/Newspaper Atlantic salmon Salmo salar Directory of Open Access Journals: DOAJ Articles Scientific Reports 7 1 |
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Medicine R Science Q |
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Medicine R Science Q Eduardo Zavala Daniela Reyes Robert Deerenberg Rodrigo Vidal Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar |
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Medicine R Science Q |
description |
Abstract MicroRNAs are key non-coding RNA molecules that play a relevant role in the regulation of gene expression through translational repression and/or transcript cleavage during normal development and physiological adaptation processes like stress. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) has become the approach normally used to determine the levels of microRNAs. However, this approach needs the use of endogenous reference. An improper selection of endogenous references can result in confusing interpretation of data. The aim of this study was to identify and validate appropriate endogenous reference miRNA genes for normalizing RT-qPCR survey of miRNAs expression in four different tissues of Atlantic salmon, under handling and confinement stress conditions associated to early or primary stress response. Nine candidate reference normalizers, including microRNAs and nuclear genes, normally used in vertebrate microRNA expression studies were selected from literature, validated by RT-qPCR and analyzed by the algorithms geNorm and NormFinder. The results revealed that the ssa-miR-99-5p gene was the most stable overall and that ssa-miR-99-5p and ssa-miR-23a-5p genes were the best combination. Moreover, the suitability of ssa-miR-99-5p and ssa-miR-23a-5p as endogeneuos reference genes was demostrated by the expression analysis of ssa-miR-193-5p gene. |
format |
Article in Journal/Newspaper |
author |
Eduardo Zavala Daniela Reyes Robert Deerenberg Rodrigo Vidal |
author_facet |
Eduardo Zavala Daniela Reyes Robert Deerenberg Rodrigo Vidal |
author_sort |
Eduardo Zavala |
title |
Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar |
title_short |
Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar |
title_full |
Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar |
title_fullStr |
Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar |
title_full_unstemmed |
Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar |
title_sort |
selection of reference genes for microrna analysis associated to early stress response to handling and confinement in salmo salar |
publisher |
Nature Portfolio |
publishDate |
2017 |
url |
https://doi.org/10.1038/s41598-017-01970-3 https://doaj.org/article/a0d66163efbd49b69fcfcd67af706e35 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
Scientific Reports, Vol 7, Iss 1, Pp 1-10 (2017) |
op_relation |
https://doi.org/10.1038/s41598-017-01970-3 https://doaj.org/toc/2045-2322 doi:10.1038/s41598-017-01970-3 2045-2322 https://doaj.org/article/a0d66163efbd49b69fcfcd67af706e35 |
op_doi |
https://doi.org/10.1038/s41598-017-01970-3 |
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Scientific Reports |
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7 |
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1 |
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1766363035310489600 |