Ultrasound-mediated gene transfer into neuronal cells.
A new field of gene transfer is emerging as a simple, effective means to drive the expression foreign genes in cells: ultrasound-mediated gene transfer or sonoporation. We report here that sonoporation is an effective means of gene transfer for cultured neurons, a cell type that has been difficult to transfect. Neuronal cell types that are effectively sonoporated include chick retinal neurons, chick dorsal forebrain, chick optic tectum, PC12 cells, rat cerebellar neurons and mouse hippocampal neurons. Depending on the type of cell and conditions of sonoporation the transfection efficacy was as high as 20%. Sonoporation of plasmid DNA was effective for cells adherent to a substrate and for free-floating cells that were freshly dissociated. In the free-floating preparations, between 60 and 95% of the cells that were transfected were neuronal, as much as 90% higher than that observed for other methods of gene transfer including adenovirus and lipid-based transfection methods. We conclude that sonoporation is a simple, effective and inexpensive means by which to preferentially transfect DNA into neuronal cells.
Fischer
Andy J
AJ
Department of Neuroscience, The Ohio State University, College of Medicine and Public Health, 3020 Graves Hall, 333 W. 10th Avenue, Columbus, OH 43210-1239, USA. fischer.412@osu.edu
Stanke
Jennifer J
JJ
Omar
Ghezal
G
Askwith
Candice C
CC
Burry
Richard W
RW
D000818:Animals; D016923:Cell Death; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism; D002642:Chick Embryo; D002645:Chickens / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage / Q000378:metabolism; D016548:Prosencephalon / Q000166:cytology / Q000378:metabolism; D051381:Rats; D012160:Retina / Q000166:cytology / Q000378:metabolism; D014162:Transfection / Q000379:methods*; D014465:Ultrasonics