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Detection of Reporter Gene Activity in Cell Cultures and Murine Epidermis After Helios Gene Gun-Mediated Particle Bombardment

Andreas Henke and Eunike Grohmann, Institute of Virology, Medical Center, Friedrich Schiller University Jena, Winzerlaer Str. 10, 07745 Jena, Germany


Summary
The Helios gene gun-mediated DNA transfer can be used as a fast and efficient method to transfect living cells. In this study we describe the preparation of DNA-coated microcarriers and conditions used for plasmid inoculations. Expression from these plasmids was demonstrated by localization of -galactosidase activity in HeLa cell monolayers and murine epidermal cells and by the detection of HeLa cells positive for the green fluorescent protein (GFP).


Introduction
The application of gene transfer by particle bombardment was first described by Sanford et al., (1987) and was shown to be an efficient method for transformation of many different organisms (Klein et al., 1992). One of the newest technical systems used for the delivery of DNA-coated gold microcarriers by helium gas shock wave is the Helios gene gun developed by Bio-Rad (Hercules, CA, USA). With the gene gun, a certain amount of DNA-coated microcarriers is placed onto the inner surface of small Teflon tubes. After cutting the Teflon tube, up to 12 cartridges can be placed in the cartridge holder of the Helios gene gun at the same time. A high velocity helium gas stream causes a detachment of the microcarriers from the surfaces. After that, the DNA-coated gold microcarriers are fast enough to penetrate and transform living cells under in vitro and in vivo conditions. Advantages of this technology are

Any cell, tissue, or organ that can be made accessible for the gene gun can be transformed.

No vacuum is necessary during the application.

Convenient and ef
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