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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Bibliographic Details
Published in:Scientific reports 2020-07, Vol.10 (1), p.11211, Article 11211
Main Authors: Tan, Minjie, van Tol, Helena T. A., Mokry, Michal, Stout, Tom A. E., Roelen, Bernard A. J.
Format: Article
Language:English
Subjects:
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Animals
Cattle
Female
Gene expression
Gene Expression Regulation, Developmental
Gene Knockdown Techniques - adverse effects
Gene Knockdown Techniques - methods
Humanities and Social Sciences
Injection
Microinjection
Microinjections - adverse effects
multidisciplinary
Oocytes
Oocytes - metabolism
Protein transport
RNA, Messenger - genetics
RNA, Small Interfering - administration & dosage
RNA, Small Interfering - metabolism
RNA-Seq
Science
Science (multidisciplinary)
Single-Cell Analysis
siRNA
Transcriptome - genetics
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Summary: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.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-67603-4