Cre-mediated generation of a VCAM-1 null allele in transgenic mice.

Terry RW, Kwee L, Baldwin HS, Labow MA
Transgenic Res. 1997 6 (5): 349-56

PMID: 9322371

A conditional null allele for VCAM-1 was generated in mice through a one step ES cell selection procedure by flanking the proximal promoter and exons 1 and 2 with loxP sites. The ES cells were used to create chimeric mice, which were then used to produce mice homozygous for the VCAM-1 conditional null, or floxed allele. Although the PGKneo cassette was retained in the promoter, the homozygous mice produced levels of VCAM-1 transcripts similar to that seen in wild-type mice. Homozygous VCAMflox/flox mice were mated to transgenic lines of mice expressing the cre gene under control of the murine platelet endothelial cell adhesion molecule-1 (PECAM-1) promoter. Surprisingly, the VCAMflox allele in all tissues examined from mice that inherited the cre-transgene had underwent complete excision of the floxed VCAM-1 sequences. The 'deleted' VCAM-1 allele (VCAMdel) was stably inherited, even in those mice that did not inherit the cre transgene, indicating the recombination occurs at an early stage of development prior to germ cell development. Thus the cre mice can be used for ubiquitous gene rearrangement in vivo. The data also suggest a novel simplified strategy for using the Cre/loxP system in vivo, in which a single ES cell and line of mice can be used to create mice carrying either a null or conditional null allele.

MeSH Terms (19)

Alleles Animals Base Sequence Blotting, Northern Genetic Engineering Homozygote Integrases Interleukin-1 Mice Mice, Knockout Mice, Transgenic Molecular Sequence Data Platelet Endothelial Cell Adhesion Molecule-1 Sequence Deletion Stem Cells Tissue Distribution Transgenes Vascular Cell Adhesion Molecule-1 Viral Proteins

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