An improved method of preparing microcarriers for biolistic transfection.
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.
O'Brien
John
J
Neurobiology Division, MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.
Lummis
Sarah C R
SC
D001426:Bacterial Proteins; D019470:Biolistics / Q000295:instrumentation* / Q000379:methods*; D002451:Cell Compartmentation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004247:DNA; D005822:Genetic Vectors; D017979:Gold Colloid; D006801:Humans; D008164:Luminescent Proteins; D014162:Transfection / Q000295:instrumentation / Q000379:methods*
D001426:Bacterial Proteins; D017979:Gold Colloid; D008164:Luminescent Proteins; C054562:yellow fluorescent protein, Bacteria; D004247:DNA