Electroporation-mediated gene transfer system applied to cultured CNS neurons.
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.
Kawabata
Izumi
I
Department of Anatomy and Cell Biology, School of Medicine, Tokyo Medical and Dental University, 1-5-45, Yushima, Bunkyo-ku, Tokyo, 113-8519, Japan.
Umeda
Tatsuya
T
Yamamoto
Kazuhiro
K
Okabe
Shigeo
S
D000818:Animals; D002478:Cells, Cultured; D002490:Central Nervous System / Q000737:chemistry / Q000166:cytology; D018274:Electroporation / Q000295:instrumentation / Q000379:methods*; D004622:Embryo, Mammalian; D018014:Gene Transfer Techniques / Q000295:instrumentation; D006624:Hippocampus / Q000737:chemistry / Q000166:cytology*; D051379:Mice; D009474:Neurons / Q000737:chemistry / Q000166:cytology*