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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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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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. |
format | Online Article Text |
id | pubmed-7343835 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
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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