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Microinjection induces changes in the transcriptome of bovine oocytes
Minjie Tan1, Helena T A van Tol1, Michal Mokry2
1Department of Population Health Sciences, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands.
Scientific Reports
|July 10, 2020
Summary
Microinjection of non-specific small interfering RNA (siRNA) into bovine oocytes alters gene expression. This procedure impacts crucial biological processes, including ATP synthesis and molecular transport, highlighting a necessary control for gene function studies.
Area of Science:
- Reproductive biology
- Molecular genetics
- Gene expression analysis
Background:
- Gene knockdown is essential for studying gene and protein functions.
- Messenger RNA (mRNA) knockdown via small interfering RNA (siRNA) microinjection is common in mammalian oocytes.
- Non-specific siRNA injection serves as a technical control for microinjection procedures.
Purpose of the Study:
- To investigate the impact of the microinjection procedure itself on the transcriptome of bovine oocytes.
- To identify specific transcriptomic changes induced by non-specific siRNA microinjection.
Main Methods:
- Microinjection of non-specific siRNA into bovine oocytes.
- Transcriptome analysis to detect differential gene expression.
- Gene ontology analysis to identify enriched biological pathways.
Main Results:
- Microinjection of non-specific siRNA led to the differential expression of 119 transcripts in bovine oocytes.
- A significant portion (76) of these transcripts were down-regulated.
- Enriched gene ontology terms included ATP synthesis, molecular transport, and regulation of protein polyubiquitination.
Conclusions:
- The microinjection procedure itself causes significant background transcriptomic changes in oocytes.
- These changes affect fundamental cellular processes, including energy production and protein regulation.
- Researchers using microinjection for gene expression manipulation in oocytes must consider this procedural background effect.
