Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar
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 reac...
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ftpubmed:oai:pubmedcentral.nih.gov:5431957 2023-05-15T15:32:31+02:00 Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar Zavala, Eduardo Reyes, Daniela Deerenberg, Robert Vidal, Rodrigo 2017-05-11 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5431957/ http://www.ncbi.nlm.nih.gov/pubmed/28496155 https://doi.org/10.1038/s41598-017-01970-3 en eng Nature Publishing Group UK http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5431957/ http://www.ncbi.nlm.nih.gov/pubmed/28496155 http://dx.doi.org/10.1038/s41598-017-01970-3 © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. CC-BY Article Text 2017 ftpubmed https://doi.org/10.1038/s41598-017-01970-3 2017-05-21T00:25:34Z 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. Text Atlantic salmon Salmo salar PubMed Central (PMC) Scientific Reports 7 1 |
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Article Zavala, Eduardo Reyes, Daniela Deerenberg, Robert Vidal, Rodrigo Selection of reference genes for microRNA analysis associated to early stress response to handling and confinement in Salmo salar |
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description |
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 |
Text |
author |
Zavala, Eduardo Reyes, Daniela Deerenberg, Robert Vidal, Rodrigo |
author_facet |
Zavala, Eduardo Reyes, Daniela Deerenberg, Robert Vidal, Rodrigo |
author_sort |
Zavala, Eduardo |
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 Publishing Group UK |
publishDate |
2017 |
url |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5431957/ http://www.ncbi.nlm.nih.gov/pubmed/28496155 https://doi.org/10.1038/s41598-017-01970-3 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_relation |
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5431957/ http://www.ncbi.nlm.nih.gov/pubmed/28496155 http://dx.doi.org/10.1038/s41598-017-01970-3 |
op_rights |
© The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
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CC-BY |
op_doi |
https://doi.org/10.1038/s41598-017-01970-3 |
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