Cdc42loxP/loxP floxed mice may be useful for studying the role of CDC42 in cellular adhesion and migration, and cell cycle progression.
Yi Zheng, Cincinnati Children's Hospital Medical Center
Cdc42loxP/loxP floxed mice possess loxP sites flanking exon 2 of the cell division cycle 42 (Cdc42) gene. CDC42 is a Rho-family GTPase that controls cadherin-based intercellular junctions and cell polarity, as well as cellular adhesion, migration, endocytosis, and cell cycle progression. Homozygotes are viable and fertile. When bred to mice that express tissue-specific Cre recombinase, resulting offspring will have exon 2 deleted in the cre-expressing tissues.
For example, when bred to B6.Cg-Tg(Mx1-cre)1Cgn/J transgenic mice (Stock No. 003556) expressing Cre recombinase in hematopoietic stem cells (HSCs) after administration of poly(IC), CDC42 is undetectable in bone marrow (BM) five days after administration of the third poly(IC) dose. These mice show increased number and frequency of the stem/progenitor cells in the BM. Cdc42 deficiency also causes impaired adhesion, homing, lodging, and retention of HSCs, leading to massive egress of HSCs from BM to distal organs and peripheral blood and to an engraftment failure.
A targeting vector was designed to insert (from 5’ to 3’) a loxP site, a frt site, a neomycin resistance (neo) cassette, and a second loxP site followed by a second frt site upstream of exon 2 of the cell division cycle 42 (Cdc42) gene. A third loxP site was placed downstream of exon 2. The construct was electroporated into 129S6/SvEvTac-derived iTL1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts and resulting chimeric mice were bred with C57BL/6J females. Subsequently, resulting offspring were bred to Flp transgenic mice to delete the neo cassette. Progeny were crossed to remove the Flp-expressing transgene, and resulting Cdc42loxP/loxP mice were maintained on a mixed background. Upon arrival, mice were bred to C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
|Allele Name||targeted mutation 1, Yi Zheng|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Allele Synonym(s)||cdc42flox; Cdc42loxP|
|Gene Symbol and Name||Cdc42, cell division cycle 42|
|Strain of Origin||129S6/SvEvTac|
|Molecular Note||The targeting construct introduced a neomycin resistance cassette (neor), flanked on each side by a loxP followed by a frt site, upstream of exon 2 and a single loxP site downstream of exon 2. The neor was deleted by FLP recombinase, leaving a single frt site in intron 1 and loxP sites flanking exon 2, which contains the translation initiation site.|
When maintaining a live colony, homozygous mice may be bred together.
When using the Cdc42loxP mouse strain in a publication, please cite the originating article(s) and include JAX stock #027576 in your Materials and Methods section.