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Efficient Cre-IoxP-induced mitotic recombination in mouse embryonic stem cells

  1. Author:
    Liu, P. T.
    Jenkins, N. A.
    Copeland, N. G.

  2. Author Address

    NCI, Ctr Canc Res, Mouse Canc Genet Program, Frederick, MD 21702 USA. NCI, Ctr Canc Res, Mouse Canc Genet Program, Frederick, MD 21702 USA. Copeland NG NCI, Ctr Canc Res, Mouse Canc Genet Program, Frederick, MD 21702 USA.
    1. Year: 2002
  2. Journal: Nature Genetics
    1. 30
    2. 1
    3. Pages: 66-72
  4. Type of Article: Article
  1. Abstract:

    FLP/FRT-induced mitotic recombination provides a powerful method for creating genetic mosaics in Drosophila and for discerning the function of recessive genes in a heterozygous individual. Here we show that mitotic recombination can be reproducibly induced in mouse embryonic stem (ES) cells, by Cre/loxP technology, at frequencies ranging from 4.2 x 10(-5) (Snrpn) to 7.0 x 10(-3) (D7Mit178) for single allelic loxP sites, and to 5.0 x 10(-2) (D7Mit178) for multiple allelic lox sites, after transient Cre expression. Notably, much of the recombination occurs in G2 and is followed by X segregation, where the recombinant chromatids segregate away from each other during mitosis. It is X segregation that is useful for genetic mosaic analysis because it produces clones of homozygous mutant daughter cells from heterozygous mothers. Our studies confirm the predictions made from studies in Drosophila(1) that suggest that X segregation will not be limited to organisms with strong mitotic pairing, because the forces (sister-chromatid cohesion) responsible for X segregation are an elemental feature of mitosis in all eukaryotes. Our studies also show that genetic mosaic analysis in mice is feasible, at least for certain chromosomal regions.

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