Genomic targeting of a bicistronic DNA fragment by Cre-mediated site-specific recombination

Andreas Kolb, S G Siddell

    Research output: Contribution to journalArticle

    13 Citations (Scopus)

    Abstract

    The Cre-recombinase of bacteriophage P1 catalyses site-specific recombination between DNA fragments containing loxP sites. Targeting of predefined genomic loci can be achieved by Cre-mediated linkage of a promoterless resistance marker gene to a flexed promoter pre-existing in the genome. In order to avoid the introduction of plasmid sequences into the host genome, we have constructed a series of plasmids in which the DNA segment to be integrated is flanked by two loxP sites. We show here that this flexed targeting fragment is reliably and effectively separated from the vector backbone and integrated into genomic loxP sites by Cre-mediated site-specific recombination in mammalian cells. We also demonstrate that by using this approach two convergent, promoterless coding regions can simultaneously be linked to two independent promoter elements at a pre-existing genomic loxP site. This methodology will be particularly useful for genomic targeting experiments in transgenic animals. (C) 1997 Elsevier Science B.V.

    Original languageEnglish
    Pages (from-to)209-216
    Number of pages8
    JournalGene
    Volume203
    Issue number2
    DOIs
    Publication statusPublished - 12 Dec 1997

    Keywords

    • genetic engineering
    • cloning vectors
    • loxP site
    • transgenic animals
    • transcriptional interference
    • mammalian-cells
    • gene-expression
    • integration

    Cite this

    Genomic targeting of a bicistronic DNA fragment by Cre-mediated site-specific recombination. / Kolb, Andreas; Siddell, S G .

    In: Gene, Vol. 203, No. 2, 12.12.1997, p. 209-216.

    Research output: Contribution to journalArticle

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