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Oogenesis pp 217-230 | Cite as

Quantitative Microinjection of Morpholino Antisense Oligonucleotides into Mouse Oocytes to Examine Gene Function in Meiosis-I

  • Shoma Nakagawa
  • Greg FitzHarrisEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1457)

Abstract

Specific protein depletion is a powerful approach for assessing individual gene function in cellular processes, and has been extensively employed in recent years in mammalian oocyte meiosis-I. Conditional knockout mice and RNA interference (RNAi) methods such as siRNA or dsRNA microinjection are among several approaches to have been applied in this system over the past decade. RNAi by microinjection of Morpholino antisense Oligonucleotides (MO), in particular, has proven highly popular and tractable in many studies, since MOs have high specificity of interaction, low cell toxicity, and are more stable than other microinjected RNAi molecules. Here, we describe a method of MO microinjection into the mouse germinal vesicle-stage (GV) oocyte followed by a simple immunofluorescence approach for examination of gene function in meiosis-I.

Key words

Morpholino antisense oligonucleotides Microinjection Mouse oocyte Meiosis-I Immunofluorescence 

Notes

Acknowledgements

Works in GFs lab is supported by CIHR, NSERC, CFI, and Fondation Jean-Louis Lévesque. Elements of the described procedure were learned from labs of John Carroll, Karl Swann, Jay Baltz, and Tomohiro Kono. We thank Jenna Haverfield, Cayetana Vázquez-Diez, and Angus MacCaulay for critical reading of the manuscript.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Centre de Recherche du Centre Hospitalier de l’Université de MontréalMontrealCanada
  2. 2.Department of Obstetrics and GynecologyUniversité de MontréalMontréalCanada

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