High-efficiency stable gene transfection using chloroquine-treated Chinese hamster ovary cells
- 1 September 1991
- journal article
- research article
- Published by Springer Nature in Somatic Cell and Molecular Genetics
- Vol. 17 (5) , 513-517
- https://doi.org/10.1007/bf01233175
Abstract
We describe a highly efficient stable gene transfection procedure for Chinese hamster ovary (CHO) cells using a modification of the calcium phosphate-DNA coprecipitation method. We have found that treatment of CHO cells with chloroquine increases the efficiency of gene transfer by up to 20-fold (from approx. 0.01% to approx. 0.2%) when transfection is done using the pSV2-neo plasmid. The optimized transfection procedure requires that CHO cells to be incubated with calcium phosphate-DNA coprecipitate and chloroquine (100 µM) for a total of 16 h. By using high-molecular-weight human genomic DNA as a DNA source for transfection, we show that this procedure is equally efficient for stably transferring a much larger gene, such as the 49-kb human hypoxanthine phosphoribosyltransferase gene.Keywords
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