High-fidelity gene targeting in embryonic stem cells by using sequence replacement vectors.
Mutations were targeted to the Hprt locus in murine embryonic stem cells by using sequence replacement vectors. When the vector was designed such that the mutated sequences were flanked on both sides by several kilobases of DNA homologous to the target locus, replacement of chromosomal sequences with the exogenous DNA occurred with precision. If, on the other hand, the target-homologous DNA on one arm of the vector was reduced to below 1 kb in length, the fidelity of recombination was diminished.
Thomas
K R
KR
HHMI Research Laboratories, Eccles Institute of Human Genetics, University of Utah Medical Center, Salt Lake City 84112.
Deng
C
C
Capecchi
M R
MR
D000818:Animals; D004351:Drug Resistance; D015321:Gene Rearrangement / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D005839:Gentamicins / Q000494:pharmacology; D051379:Mice; D016297:Mutagenesis, Site-Directed / Q000235:genetics*; D011995:Recombination, Genetic / Q000235:genetics*; D012641:Selection, Genetic; D013234:Stem Cells / Q000378:metabolism*; D013866:Thioguanine / Q000494:pharmacology; D014170:Transformation, Genetic
D005839:Gentamicins; C010680:antibiotic G 418; D013866:Thioguanine