ArticleThe Journal of reproduction and development2018
Lipofection of siRNA into bovine 8-16-cell stage embryos using zona removal and the well-of-the-well culture system.
Article in The Journal of reproduction and development, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed.
- miRNAs in Follicular and Oviductal Fluids Support Global DNA Demethylation in Early-Stage Embryos.International journal of molecular sciences · 2024Article
- Back to the future: optimised microwell culture of individual human preimplantation stage embryos.Journal of assisted reproduction and genetics · 2021Article
- Embryonic MTHFR contributes to blastocyst development.Journal of assisted reproduction and genetics · 2020Article
- The effects of electroporation on viability and quality of in vivo-derived bovine blastocysts.The Journal of reproduction and development · 2019Article
- microRNAs associated with the quality of follicular fluids affect oocyte and early embryonic development.Reproductive medicine and biologyArticle
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bovine preimplantation embryos exhibit dramatic biological changes between before and after the 8-16-cell stage. Here we report a simple lipofection method to transfect siRNA into bovine 8-16-cell stage embryos using zona removal and the well-of-the-well (WOW) culture system. Bovine one-cell embryos produced in vitro were freed from the zona pellucida and cultured up to the 8-16-cell stage in WOW dishes. The 8-16-cell embryos were lipofected with siRNA and the transfection efficiency was assessed at 48 h of transfection. Lipofection with a red fluorescent non-targeting siRNA revealed the importance of zona removal for transfection of siRNA into embryos. Using this method, we knocked down the methionine adenosyltransferase 2A (MAT2A) gene, achieving a significant reduction in MAT2A expression (P < 0.05) concomitant with the marked inhibition of blastocyst development. Our proposed method, tentatively named 'Octo-lipofection', may be useful to analyze gene functions in bovine preimplantation embryos without expensive equipment and skill-intensive techniques.
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