An siRNA-based screen in C2C12 myoblasts identifies novel genes involved in myogenic differentiation

Journal Article · Sun Oct 01 04:00:00 EDT 2017 · Experimental Cell Research

DOI:https://doi.org/10.1016/J.YEXCR.2017.07.037· OSTI ID:22738167

Alwan, Rayan ^[1]; Bruel, Ange-Line ^[2]; Da Silva, Anne ^[1]; Blanquet, Véronique ^[1]; Bouhouche, Khaled ^[1]

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1. University of Limoges, INRA, UMR 1061, UGMA, F- 87000 Limoges (France)
2. University of Burgundy, INSERM, UMR 1231, GAD, F-1079, Dijon (France)

Myogenesis is a highly regulated multi-step process involving myoblast proliferation and differentiation. Although studies over the last decades have identified several factors governing these distinct major phases, many of them are not yet known. In order to identify novel genes, we took advantage of the C2C12 myoblastic line to establish a functional siRNA screen combined with quantitative-imaging analysis of a large amount of differentiated myoblasts. We knocked down 100 preselected mouse genes without a previously characterized role in muscle. Using image analysis, we tracked gene-silencing phenotypes by quantitative assessment of cellular density, myotube quantity, myotube morphology and fusion index. Our results have revealed six genes involved in both stages of C2C12 myogenesis and 13 genes specific to the differentiation stage. These findings prove that our RNAi-based screen could be an efficient tool to detect clear and subtle phenotypes allowing the identification of new myogenic regulators in mammals. - Highlights: • A new optimized protocol to perform a functional siRNA based screen. • 100 preselected genes knocked-down in C2C12 myoblastic cell line. • 19 candidates genes identified as novel myogenic actors.

• OSTI ID: 22738167
• Journal Information: Experimental Cell Research, Journal Name: Experimental Cell Research Journal Issue: 1 Vol. 359; ISSN 0014-4827; ISSN ECREAL
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
CELL DIFFERENTIATION
GENES
IMAGE PROCESSING
MYOBLASTS
PARTICLE TRACKS
SCREENING