Gene targeting with homologous recombination in embryonic stem cells created a revolution in the analysis of the function of genes in research. 

The development of gene targeting technology, the exchange of an endogenous allele of a target gene for a mutated copy via homologous recombination, and the application of this technique to murine embryonic stem cells has made it possible to alter the germ-line of mice in a predetermined way. It’s becoming common to engineer specific gene mutations in the mouse germline by gene targeting in embryonic stem (ES) cells. This is accomplished by using a targeting vector designed to replace the corresponding endogenous gene by homologous recombination. Since it is much more common for the targeting vector to insert into a random chromosomal site than a homologous one, it’s necessary to screen colonies by Southern hybridization or polymerase chain reaction (PCR) to identify rare targeted clones.

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