Increased ratio of targeted to random integration after transfection of chicken B cell lines

Jean Marie Buerstedde, Shunichi Takeda

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    Abstract

    Constructs of four different genetic loci were transfected into the avian leukosis virus-induced chicken B cell line DT40, which continues diversification of its rearranged light chain immunoglobulin gene by gene conversion. Analysis of stable transfectants revealed an unexpectedly high frequency of targeted integration into the homologous gene loci of DT40. Transcriptional activity of the target gene locus is not required, since a construct of the untranscribed ovalbumin gene also integrated predominantly by homologous recombination. A construct derived from the β-actin locus was transfected into other chicken cell lines to determine the cell type specificity of the phenomenon. Targeted integration still occurred at high frequency in two other B cell lines that do not have the gene conversion activity. However, the ratios of targeted to random integration were reduced by at least one order of magnitude in three non-B cell lines.

    Original languageEnglish (US)
    Pages (from-to)179-188
    Number of pages10
    JournalCell
    Volume67
    Issue number1
    DOIs
    StatePublished - Oct 4 1991

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    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)

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