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Issue 14, 2018
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A robust split-luciferase-based cell fusion screening for discovering myogenesis-promoting molecules

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Abstract

Myogenesis-promoting chemicals are an important source of new pharmaceuticals for the treatment of skeletal muscle atrophy that impairs quality of life. This report presents a robust and quantitative bioluminescence-based assay for screening myogenesis-promoting compounds in chemical libraries. The assay system consists of two stable C2C12 myoblast cell lines, each of which expresses either an N-terminal or a C-terminal split luciferase fragment fused to a naturally split DnaE intein as an indicator for cell fusion. Cell fusion during myogenesis induces bioluminescence in the cytosol because of the reconstitution of luciferases. The luminescence intensity quantitatively represents the progress in the cell fusion and therefore indicates the extent of myogenesis. We applied this assay system to a high-throughput screening of myogenesis-promoting compouns in 1191 pharmacologically proven bioactive small molecules, which revealed two chemical compounds as myogenesis-promoting compounds: Imatinib and Doxazosin mesylate. The assay system enabled a robust and quantitative evaluation of the extent of myogenesis through simple luminescence measurements, and is expected to be widely applicable for high-throughput screening of cell fusion-promoting and inhibiting molecules.

Graphical abstract: A robust split-luciferase-based cell fusion screening for discovering myogenesis-promoting molecules

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Publication details

The article was received on 13 Feb 2018, accepted on 18 Jun 2018 and first published on 19 Jun 2018


Article type: Paper
DOI: 10.1039/C8AN00285A
Citation: Analyst, 2018,143, 3472-3480
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    A robust split-luciferase-based cell fusion screening for discovering myogenesis-promoting molecules

    Q. Li, H. Yoshimura, M. Komiya, K. Tajiri, M. Uesugi, Y. Hata and T. Ozawa, Analyst, 2018, 143, 3472
    DOI: 10.1039/C8AN00285A

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