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Electroporation-mediated gene transfer system applied to cultured CNS neurons

Kawabata, Izumi1; Umeda, Tatsuya1; Yamamoto, Kazuhiro2; Okabe, Shigeo1 2 3 CA


Electroporation is effective in transferring foreign genes into immature neurons in intact brain tissue. We utilized this approach to transfect genes into developing rodent hippocampi. Transfected hippocampi were subsequently dissociated and allowed to differentiate in culture. By optimizing developmental stage of the hippocampus, promoters to drive the marker cDNA, and culture conditions, neurons kept strong expression of multiple marker genes for more than two weeks after electroporation. We could also induce transient expression in mature neurons by combining electroporation of plasmids containing loxP-flanked stopper sequences and infection of Cre-producing recombinant adenoviruses. The system described here is useful in analyzing biological roles of multiple genes in specific stages of neuronal development.

1Department of Anatomy and Cell Biology, School of Medicine, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8519

2Molecular Neurophysiology Group, Neuroscience Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, Ibaraki, 305-8566

3Core Research for Evolution Science and Technology (CREST), Japan Science and Technology Corporation (JST), Kawaguchi, 332-0012, Japan

CACorresponding Author and Address:

Received 31 January 2004; accepted 6 February 2004

© 2004 Lippincott Williams & Wilkins, Inc.