Neuronal Transfection Using Particle-Mediated Gene Transfer

The ability to transfect differentiated neurons with DNA constructs of various sizes has been limited by the relative refractoriness of these cells, to classical calcium phosphate-DNA coprecipitation, and liposome–carrier-mediated transfection methods. Particle-mediated gene transfer (also called biolistics)represents a relatively simple solution to this problem, requiring little by the way of special technical expertise. Plasmid DNAs of unlimited sizes are used to coat approximately one micron tungsten or gold particles, and these particles are then accelerated into living cells and tissues by a blast of helium gas. A commercial device, referred to as the “Gene Gun” obtained from Bio-Rad, is available for this purpose. Biolistics has many uses, e.g., the production of antibodies after in vivo transfection of foreign DNAs into skin, ex vivo transfection of genes into tissues to be used for transplantation, and for the study of specific gene expression in distinct differentiated neurons in vitro. Cotransfection of multiple and distinct DNA constructs into single cells is easily performed, and this makes this technology of special