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

Hasan, M. T., Subbaroyan, R., & Chang, T. (1991). High-efficiency stable gene transfection using chloroquine-treated Chinese hamster ovary cells. Somatic Cell and Molecular Genetics, 17(5), 513-517. doi:10.1007/BF01233175.

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Item Permalink: https://hdl.handle.net/21.11116/0000-0000-6303-D Version Permalink: https://hdl.handle.net/21.11116/0000-0000-6304-C
Genre: Journal Article

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SomatCellMolGenet_17_1991_513.pdf (Any fulltext), 495KB
 
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 Creators:
Hasan, Mazahir T.1, 2, 3, Author           
Subbaroyan, Ram, Author
Chang, Ta−Yuan, Author
Affiliations:
1Mazahir Hasan Group, Max Planck Institute for Medical Research, Max Planck Society, ou_1497726              
2Department of Molecular Neurobiology, Max Planck Institute for Medical Research, Max Planck Society, ou_1497704              
3Department of Biomedical Optics, Max Planck Institute for Medical Research, Max Planck Society, ou_1497699              

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Free keywords: Gene Transfer; Chloroquine; Chinese Hamster Ovary Cell; Ovary Cell; Chinese Hamster Ovary
 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 microM) 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.

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Language(s): eng - English
 Dates: 1991-04-041991-07-171991-09
 Publication Status: Issued
 Pages: 5
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1007/BF01233175
URI: https://www.ncbi.nlm.nih.gov/pubmed/1763389
 Degree: -

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Title: Somatic Cell and Molecular Genetics
  Other : Somatic Cell and Molecular Genetics
Source Genre: Journal
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Publ. Info: New York, NY : Kluwer Academic Publishers
Pages: - Volume / Issue: 17 (5) Sequence Number: - Start / End Page: 513 - 517 Identifier: ISSN: 0740-7750
CoNE: https://pure.mpg.de/cone/journals/resource/954927608797