Targeted integration of the Ren-1D locus in mouse embryonic stem cells.
We have introduced a Ren-1D targeting vector into embryonic stem cells containing the two highly homologous mouse renin genes Ren-1D and Ren-2. Using a polymerase chain reaction (PCR) screen designed to detect targeted integration at Ren-1D and Ren-2, we isolated 15 targeted embryonic stem cell clones, all of which had undergone a gene conversion event at the Ren-1D locus. We did not isolate any clones in which the incoming DNA had recombined with Ren-2. Over the region encompassed by our transgene, Ren-1D and Ren-2 display greater than 95% homology. Our results suggest that the machinery driving gene targeting by means of homologous recombination in mammalian cells is capable of distinguishing between these two sequences. Construction of transgenic mice with the embryonic stem cells reported here carrying a mutated renin gene will permit a greater understanding of the functions of the Ren-1D and Ren-2 gene products and their relative contribution to cardiovascular homeostasis.
Miller
C C
CC
Biotechnology Department, ICI Pharmaceuticals, Macclesfield, Cheshire, United Kingdom.
McPheat
J C
JC
Potts
W J
WJ
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002478:Cells, Cultured; D005785:Gene Conversion; D005796:Genes; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011995:Recombination, Genetic; D012083:Renin / Q000235:genetics*; D013234:Stem Cells; D014162:Transfection
D009838:Oligodeoxyribonucleotides; D012083:Renin