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Direct selection of hepatoma cell variants deficient in α1-antitrypsin gene expression

Somatic Cell and Molecular Genetics volume 18, pages 361–370 (1992)Cite this article

Abstract

Expression plasmids containing the human α1-antitrypsin (α1 AT) promoter fused to either adenine phosphoribosyltransferase (aprt) or xanthine-guanine phosphoribosyltransferase (gpt) coding sequences were sequentially introduced into APRT− HPRT− rat hepatoma cells. Stable transfectants expressing both transgenes were isolated and characterized. Nonexpressing variants were subsequently obtained by selecting against expression of one or both transgenes. Variants isolated by selecting against expression of either transgene alone generally displayed deficiency phenotypes incis, as only three of 20 clones tested were affected for expression of α1AT mRNA. In contrast, double selection yielded predominantlytrans effects: 12 of 14 lines tested showed impaired ability to express their chromosomal α1AT genes. Furthermore, expression of several other liver genes, including the gene encoding the HNF-1trans-activator, was repressed in many of the variant lines. Thus, double selection using chimeric transgenes is a useful approach for generating variant cell lines deficient in expression of specific mammalian genes.

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Authors and Affiliations

  1. Department of Molecular Medicine, Fred Hutchinson Cancer Research Center, 1124 Columbia Street, 98104, Seattle, Washington

    Gary A. Bulla & R. E. K. Fournier

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  1. Gary A. Bulla
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  2. R. E. K. Fournier
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Bulla, G.A., Fournier, R.E.K. Direct selection of hepatoma cell variants deficient in α1-antitrypsin gene expression. Somat Cell Mol Genet 18, 361–370 (1992). https://doi.org/10.1007/BF01235759

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  • DOI: https://doi.org/10.1007/BF01235759

Keywords

  • Hepatoma Cell
  • Direct Selection
  • Stable Transfectants
  • Impaired Ability
  • Mammalian Gene