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A siRNA Mediated Screen During C2C12 Myogenesis

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Myogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1889))

Abstract

Myogenesis is a multistep process taking place during pre- and postnatal stages for muscle formation, growth, and regeneration. It is a highly regulated process involving many molecular factors which act during myoblast proliferation and differentiation. To provide new insights into the molecular mechanisms and interactions behind the regulation of these different steps, RNA interference is an efficient methodology to implement. We developed a high-throughput siRNA screen in C2C12 murine myoblast cells for identification of genes relevant to signaling pathways controlling muscle growth. The proposed protocol is based on (1) the analyses of a maximum number of cells/myotubes to detect and quantify both clear and subtle phenotypes during proliferation/fusion cells and (2) the use of two cellular fluorescent markers, DAPI and myosin, decorating nuclei and myotubes respectively. Four phenotypic criteria were quantitatively assessed: cellular density, myotubes quantity, fusion index, and size and morphology of myotubes.

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Authors and Affiliations

  1. INRA, PEIRENE EA7500, USC1061 GAMAA, Université de Limoges, Limoges, France

    Alexis Parenté, Luce Pèrié, Laetitia Magnol, Khaled Bouhouche & Véronique Blanquet

Authors
  1. Alexis Parenté
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  2. Luce Pèrié
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  3. Laetitia Magnol
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  4. Khaled Bouhouche
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  5. Véronique Blanquet
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Corresponding author

Correspondence to Véronique Blanquet .

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Editors and Affiliations

  1. Nofima AS, Ås, Norway

    Sissel Beate Rønning

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Parenté, A., Pèrié, L., Magnol, L., Bouhouche, K., Blanquet, V. (2019). A siRNA Mediated Screen During C2C12 Myogenesis. In: Rønning, S. (eds) Myogenesis. Methods in Molecular Biology, vol 1889. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8897-6_13

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  • DOI: https://doi.org/10.1007/978-1-4939-8897-6_13

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8896-9

  • Online ISBN: 978-1-4939-8897-6

  • eBook Packages: Springer Protocols

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