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An improved method of preparing microcarriers for biolistic transfection.

John O'Brien·Sarah C R Lummis
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Abstract

The hand-held gene gun uses a pulse of helium to fire small gold particles coated with desiccated DNA (microcarriers) at target cells. This method of biolistic transfection is becoming increasingly popular as an effective means of rapid gene delivery into mammalian tissue. Current methods of microcarrier preparation, however, are slow (up to 2 days) and can result in variations in transfection efficiency due to a number of problems including shearing of DNA, agglomeration and adhesion of gold particles. Here we describe an improved, more rapid method of microcarrier preparation. To evaluate the new procedure we have used DNA encoding yellow fluorescent protein (EYFP), a modified version of the green fluorescent protein, which we have transfected into HEK293 cells. The data show that transfection by the new method results in high levels of transfection efficiency and low variability compared to an alternative method.

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Bacterial ProteinsBiolisticsCell CompartmentationCells, CulturedDNAGenetic VectorsGold ColloidHumansLuminescent ProteinsTransfectionJournal Article

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John O'Brien, Sarah C R Lummis. 2002. An improved method of preparing microcarriers for biolistic transfection.. https://doi.org/10.1016/s1385-299x(02)00175-7

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