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

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Oogenesis

Part of the book series: Methods in Molecular Biology ((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.

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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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Authors and Affiliations

  1. Centre de Recherche du Centre Hospitalier de l’Université de Montréal, 900 Rue St. Denis, Montreal, QC, Canada, H2X 0A9

    Shoma Nakagawa & Greg FitzHarris

  2. Department of Obstetrics and Gynecology, Université de Montréal, 3175, Ch. Côte-Sainte-Catherine, Montréal, QC, Canada, H3T 1C5

    Greg FitzHarris

Authors
  1. Shoma Nakagawa
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  2. Greg FitzHarris
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Correspondence to Greg FitzHarris .

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Editors and Affiliations

  1. School of Life Sciences, University of Warwick, Coventry, United Kingdom

    Ioannis P. Nezis

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Nakagawa, S., FitzHarris, G. (2016). Quantitative Microinjection of Morpholino Antisense Oligonucleotides into Mouse Oocytes to Examine Gene Function in Meiosis-I. In: Nezis, I. (eds) Oogenesis. Methods in Molecular Biology, vol 1457. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3795-0_16

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  • DOI: https://doi.org/10.1007/978-1-4939-3795-0_16

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3793-6

  • Online ISBN: 978-1-4939-3795-0

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