Ultrasound-mediated gene transfer into neuronal cells

• Published in: Journal of biotechnology 2006-04, Vol.122 (4), p.393-411
• Main Authors: Fischer, Andy J., Stanke, Jennifer J., Omar, Ghezal, Askwith, Candice C., Burry, Richard W.
• Format: Article
• Language: English
• Subjects: Adenovirus; Animals; Biological and medical sciences; Biotechnology; Cell Death; Cells, Cultured; Cerebellum; Cerebellum - cytology; Cerebellum - metabolism; Chick Embryo; Chickens - metabolism; Dose-Response Relationship, Drug; Electroporation - methods; Fundamental and applied biological sciences. Psychology; Gene transfer; Gene Transfer Techniques; Hippocampus; Hippocampus - cytology; Hippocampus - metabolism; Immunohistochemistry; Mice; Neurons - cytology; Neurons - metabolism; Optic tectum; Plasmids - administration & dosage; Plasmids - metabolism; Prosencephalon - cytology; Prosencephalon - metabolism; Rats; Retina; Retina - cytology; Retina - metabolism; Sonoporation; Transfection - methods; Ultrasonics; Ultrasound
• ISSN: 0168-1656; 1873-4863
• DOI: 10.1016/j.jbiotec.2005.10.006

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.