Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Subscribe

Sci. STKE, 22 July 2003
Vol. 2003, Issue 192, p. pl11
[DOI: 10.1126/stke.2003.192.pl11]

PROTOCOLS

Biolistic Transfection of Cultured Myotubes

Christian Antolik1, Patrick G. De Deyne2, and Robert J. Bloch1*

1Departments of Physiology, University of Maryland School of Medicine, 655 W. Baltimore St., Baltimore, MD 21201 USA.
2Departments of Physical Therapy and Rehabilitation Sciences, University of Maryland School of Medicine, 655 W. Baltimore St., Baltimore, MD 21201 USA.

Abstract: Transfection of cells in culture with cDNA constructs is a powerful tool in cell biology, but postmitotic cells, including myotubes, can be hard to transfect with classic methods. Biolistics provides an alternative. We have used this biolistic technique to introduce cDNAs into cultured rat, chick, and C2C12 myotubes. This protocol results in efficient (20 to 70%, depending on cell type) transfection of myotubes, high levels of cDNA expression in individual myotubes, and little cellular damage. Using this procedure, we have expressed different muscle-specific cDNAs as green fluorescent protein (GFP) fusions. This technique is rapid, reliable, uses minimal amounts of reagent per transfection, and yields high transfection rates in a previously hard-to-transfect cell type. Its efficiency and reliability are high, regardless of plasmid size or epitope tag. Muscle cell biologists may now perform experiments in mature myotubes rather than relying on transfection of myoblast cultures or heterologous expression systems.

*Corresponding author. Telephone, 410-706-3020; e-mail, rbloch{at}umaryland.edu

Citation: C. Antolik, P. G. De Deyne, R. J. Bloch, Biolistic Transfection of Cultured Myotubes. Sci. STKE 2003, pl11 (2003).

Read the Full Text

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Muscle-specific RING finger-2 (MURF-2) is important for microtubule, intermediate filament and sarcomeric M-line maintenance in striated muscle development.
A. S. McElhinny, C. N. Perry, C. C. Witt, S. Labeit, and C. C. Gregorio (2004)
J. Cell Sci. 117, 3175-3188
   Abstract »    Full Text »    PDF »

To Advertise     Find Products


Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882