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Microinjection induces changes in the transcriptome of bovine oocytes

Gene knockdown techniques are widely used to examine the function of specific genes or proteins. While a variety of techniques are available, a technique commonly used on mammalian oocytes is mRNA knockdown by microinjection of small interfering RNA (siRNA), with non-specific siRNA injection used as...

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Autores principales: Tan, Minjie, van Tol, Helena T. A., Mokry, Michal, Stout, Tom A. E., Roelen, Bernard A. J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7343835/
https://www.ncbi.nlm.nih.gov/pubmed/32641751
http://dx.doi.org/10.1038/s41598-020-67603-4
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author Tan, Minjie
van Tol, Helena T. A.
Mokry, Michal
Stout, Tom A. E.
Roelen, Bernard A. J.
author_facet Tan, Minjie
van Tol, Helena T. A.
Mokry, Michal
Stout, Tom A. E.
Roelen, Bernard A. J.
author_sort Tan, Minjie
collection PubMed
description Gene knockdown techniques are widely used to examine the function of specific genes or proteins. While a variety of techniques are available, a technique commonly used on mammalian oocytes is mRNA knockdown by microinjection of small interfering RNA (siRNA), with non-specific siRNA injection used as a technical control. Here, we investigate whether and how the microinjection procedure itself affects the transcriptome of bovine oocytes. Injection of non-specific siRNA resulted in differential expression of 119 transcripts, of which 76 were down-regulated. Gene ontology analysis revealed that the differentially regulated genes were enriched in the biological processes of ATP synthesis, molecular transport and regulation of protein polyubiquitination. This study establishes a background effect of the microinjection procedure that should be borne in mind by those using microinjection to manipulate gene expression in oocytes.
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spelling pubmed-73438352020-07-10 Microinjection induces changes in the transcriptome of bovine oocytes Tan, Minjie van Tol, Helena T. A. Mokry, Michal Stout, Tom A. E. Roelen, Bernard A. J. Sci Rep Article Gene knockdown techniques are widely used to examine the function of specific genes or proteins. While a variety of techniques are available, a technique commonly used on mammalian oocytes is mRNA knockdown by microinjection of small interfering RNA (siRNA), with non-specific siRNA injection used as a technical control. Here, we investigate whether and how the microinjection procedure itself affects the transcriptome of bovine oocytes. Injection of non-specific siRNA resulted in differential expression of 119 transcripts, of which 76 were down-regulated. Gene ontology analysis revealed that the differentially regulated genes were enriched in the biological processes of ATP synthesis, molecular transport and regulation of protein polyubiquitination. This study establishes a background effect of the microinjection procedure that should be borne in mind by those using microinjection to manipulate gene expression in oocytes. Nature Publishing Group UK 2020-07-08 /pmc/articles/PMC7343835/ /pubmed/32641751 http://dx.doi.org/10.1038/s41598-020-67603-4 Text en © The Author(s) 2020 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/.
spellingShingle Article
Tan, Minjie
van Tol, Helena T. A.
Mokry, Michal
Stout, Tom A. E.
Roelen, Bernard A. J.
Microinjection induces changes in the transcriptome of bovine oocytes
title Microinjection induces changes in the transcriptome of bovine oocytes
title_full Microinjection induces changes in the transcriptome of bovine oocytes
title_fullStr Microinjection induces changes in the transcriptome of bovine oocytes
title_full_unstemmed Microinjection induces changes in the transcriptome of bovine oocytes
title_short Microinjection induces changes in the transcriptome of bovine oocytes
title_sort microinjection induces changes in the transcriptome of bovine oocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7343835/
https://www.ncbi.nlm.nih.gov/pubmed/32641751
http://dx.doi.org/10.1038/s41598-020-67603-4
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