Utilization of microhomologous recombination in yeast to generate targeting constructs for mammalian genes.
We have developed a new procedure utilizing microhomologous recombination in yeast to generate targeting constructs for producing targeted mutations in mice. This procedure is rapid and efficient, and should be directly applicable to all mammalian genes. Moreover, only minimal information about the locus being targeted is required. The feasibility of this approach was demonstrated by producing another allele of the mouse Tg737 polycystic kidney gene.
Khrebtukova
I
I
Life Sciences Division, Oak Ridge National Laboratory, Oak Ridge, TN 37831, USA.
Michaud
E J
EJ
Foster
C M
CM
Stark
K L
KL
Garfinkel
D J
DJ
Woychik
R P
RP
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D002678:Chimera; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers; D005091:Exons; D018390:Gene Targeting; D005822:Genetic Vectors; D015698:Genomic Library; D008322:Mammals; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D016296:Mutagenesis; D007690:Polycystic Kidney Diseases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic
D017931:DNA Primers