Optimized protocol for biolistic transfection of brain slices and dissociated cultured neurons with a hand-held gene gun

DNA-transfer into postmitotic neurons or neuronal tissues has been a major problem in neurobiology. For this aim different methods have been used such as viral infection, microinjection, lipofection or calcium phosphate preci pitation. However, using these techniques, very poor transfection efficienc y was achieved except for virus-mediated gene transfer. Though viral infect ions are very efficient, this method is expensive and labor-intensive, espe cially when recombination is used to prepare viral vectors. Biolistic gene transfer of neurons represents another promising transfection technique. Th is technique was originally used to transfect plant cells and has been furt her developed for gene transfer into neurons or neuronal tissues. Up to now , only a few reports are available where successful biolistic gene transfer into neurons or neuronal tissues could be shown. Transfection efficiencies were only about 2%. Most of the previously published experiments were carr ied out under vacuum conditions using in-chamber gene gun types. Here we de scribe an improved method for efficient neuronal cell transfection using a hand-held gene gun. Expression vectors could be successfully transferred in to dissociated cultured hippocampal neurons, PC12 cells, cultured cerebella r granule cells and cerebellar brain slices. In cerebellar granule cells an d hippocampal neurons, transfection efficiencies of about 10% were reached. (C) 1999 Elsevier Science B.V. All rights reserved.