Electroporation of non-viral plasmid DNA is a valuable tool to alter gene expression in the adult central nervous system. It offers a number of advantages over viral gene delivery as non-viral plasmids can integrate larger inserts and reduce the risk of inducing unintended immunological responses. Generally, electroporation of the adult brain is accomplished in rodents by applying high-amplitude voltage-controlled pulses through the entire brain with plate electrodes surrounding the animal's head. Here, we describe an alternative electroporation protocol making use of current-controlled low-amplitude pulses that are delivered locally by means of needlelike electrodes in the brain of adult mice. This allows altering gene expression in very-well-defined areas of the brain while inducing minimal tissue damage.