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Journal Article

High−efficiency stable gene transfection using chloroquine−treated Chinese hamster ovary cells

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Hasan, Mazahir T.
Mazahir Hasan Group, Max Planck Institute for Medical Research, Max Planck Society;
Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society;
Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society;

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Citation

Hasan, M. T., Subbaroyan, R., & Chang, T. (1991). High−efficiency stable gene transfection using chloroquine−treated Chinese hamster ovary cells. Somat. Cell Mol. Genet., 17(5), 513-517. doi: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