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Embryonic stem cells (derived from 129/Ola mice) containing a mutant hypoxanthine phosphoribosyltransferase gene that had been corrected in vitro in a planned manner by homologous recombination were injected into blastocysts obtained from C57BL/6J mice. The injected blastocysts were introduced into pseudopregnant female mice to complete their development. Eleven surviving pups were obtained. Nine were chimeras: six males and three females. Two of the males transmitted the embryonic stem cell genome containing the alteration in the hypoxanthine phosphoribosyltransferase gene to their offspring at high frequencies. These experiments demonstrate that a preplanned alteration in a chosen gene can be made in the germ line of an experimental animal by homologous recombination in an embryonic stem cell.

proceedings of the national academy of sciences of the united states of america

Germ-line transmission of a planned alteration made in a hypoxanthine phosphoribosyltransferase gene by homologous recombination in embryonic stem cells.