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FIGURE 2–7. Conventional gene disruption ("knockout").(A) Producing chimeric mice. First, a mutant allele is produced by replacing the coding exons of the desired gene with a neomycin (neo) cassette and transferring it into embryonic stem (ES) cells. Second, genetically altered ES cells are reintroduced into a developing blastocyst, where they contribute to the developing embryo. (B) Breeding chimeric mice. When the germ cells of the resulting chimeric mouse (chimera) are ES cell–derived (germ-line mutation), the heterozygotes (+/–) can be produced by breeding. One-half of the offspring will be heterozygous. The heterozygous animals may be bred to produce homozygous mice (–/–). TK = thymidine kinase; WT = wild type.


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