Fidelity of targeted recombination in human fibroblasts and murine embryonic stem cells.

TitleFidelity of targeted recombination in human fibroblasts and murine embryonic stem cells.
Publication TypeJournal Article
Year of Publication1991
AuthorsZheng, H, Hasty, P, Brenneman, MA, Grompe, M, Gibbs, RA, Wilson, JH, Bradley, A
JournalProc Natl Acad Sci U S A
Date Published1991 Sep 15
KeywordsAnimals, Base Sequence, Fibroblasts, Genetic Vectors, Humans, Hypoxanthine Phosphoribosyltransferase, In Vitro Techniques, Mice, Molecular Sequence Data, Oligonucleotides, Polymerase Chain Reaction, Recombination, Genetic, Stem Cells

Targeted recombination in murine embryonic stem cells promises to be a powerful tool for introducing specific mutations into target genes to study development in mice and to create animal models of human disease. Gene targeting also holds potential for correcting genetic defects as an approach to human gene therapy. To precisely modify target genes, homologous recombination must proceed with high fidelity. However, several results have suggested that targeted recombination may be highly mutagenic. To test the accuracy of gene targeting we analyzed 44 independent targeted recombinants at the hypoxanthine phosphoribosyltransferase (HPRT) locus in a human fibroblast cell line and in mouse embryonic stem cells. We surveyed 80 kilobases around the sites of recombination by using chemical cleavage of mismatches. Only two mutations were found: a T----G transversion and a thymidine deletion. Thus, gene targeting in mammalian cells can be extremely accurate. These results demonstrate the feasibility of generating precise modifications of mammalian genomes by gene targeting.

Alternate JournalProc Natl Acad Sci U S A
PubMed ID1896453
PubMed Central IDPMC52447

Similar Publications