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. 1987 Oct;84(19):6820–6824. doi: 10.1073/pnas.84.19.6820

Accurate modification of a chromosomal plasmid by homologous recombination in human cells.

K Y Song 1, F Schwartz 1, N Maeda 1, O Smithies 1, R Kucherlapati 1
PMCID: PMC299176  PMID: 2821545

Abstract

We have examined the consequences of modifying mammalian cellular DNA sequences by homologous recombination. A plasmid carrying a 248-base-pair deletion in the neomycin phosphotransferase (neo) gene was introduced into hamster and human cells. The integrated, defective neo gene was used as a target for modification by a second round of transfection with a plasmid carrying a different (283-base-pair) deletion in the neo gene. Recombinants resulting in an intact neo gene were selected by their G418 resistance phenotype. The best ratio of homologous to nonhomologous recombination events was about 1:80. Analyses of the functional neo genes in various independent cell lines establish that simple crossovers (single and double) generated the wild-type neo genes.

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Selected References

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