Studies on gene transfer and reversion to UV resistance in xeroderma pigmentosum cells

Roger A. Schultz, Dina P. Barbis, Errol C. Friedberg

Research output: Contribution to journalArticle

27 Scopus citations

Abstract

We have examined several parameters which address the feasibility of complementing the UV-sensitive phenotype of xeroderma pigmentosum (XP) fibroblasts by gene transfer. We present a comparative study which demonstrates that, relative to immortalized cells, human diploid cells are poor recipients for gene transfer. As measured by both transient and stable expression assays, diploid fibroblasts were completely refractory to DNA transfer by calcium phosphate coprecipitation and exhibited substantially reduced levels of expression following gene transfer by fusion with E. coli protoplasts. We also examined the significance of reversion of the phenotype of UV sensitivity in SV40-immortalized XP-A cell lines. In addition to confirming a previous report of reversion to wild-type levels of UV resistance at a frequency of ∼10-7, we have attempted to facilitate the identification of XP-A cells complemented with genomic DNA by employing less stringent selection schemes and cotransfection of a selectable marker. Under these conditions, we observed an increased frequency of reversion and were unable to identify true transfectants.

Original languageEnglish (US)
Pages (from-to)617-624
Number of pages8
JournalSomatic Cell and Molecular Genetics
Volume11
Issue number6
DOIs
StatePublished - Nov 1 1985

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ASJC Scopus subject areas

  • Genetics
  • Cell Biology

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