Issue 14, 2018

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

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

Supplementary files

Article information

Article type
Paper
Submitted
13 Feb 2018
Accepted
18 Jun 2018
First published
19 Jun 2018

Analyst, 2018,143, 3472-3480

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

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements