Calcium phosphate transfection of DNA into neurons in primary culture.
Calcium phosphate/DNA coprecipitation is a widely used method for the introduction of foreign DNA into cells. DNA and calcium phosphate are allowed to form a precipitate that is then added to cells in culture. The cells internalize the DNA, leading to the expression of the transfected genes in the cell. Despite the simplicity of this method, it has not been used very often for primary neurons because of its potential to cause neuronal toxicity. However, low toxicity and reasonably high transfection efficiency (0.5% to 5%) can be achieved by optimization of the transfection parameter, combined in some cases with the use of inhibitors of neuronal activity. This unit describes a very easy and inexpensive method for neuronal gene delivery that can be used with standard eukaryotic expression vectors for the gene of interest.
Dudek
H
H
Ontogeny, Inc., Cambridge, Massachusetts, USA.
Ghosh
A
A
Greenberg
M E
ME
D000818:Animals; D002130:Calcium Phosphates / Q000378:metabolism / Q000494:pharmacology*; D018929:Cell Culture Techniques / Q000379:methods*; D004247:DNA / Q000235:genetics*; D018014:Gene Transfer Techniques; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D014162:Transfection / Q000379:methods*
D002130:Calcium Phosphates; D004247:DNA; C020243:calcium phosphate