These floxed mutant mice possess loxP sites flanking exon 5 of the Afap1 gene. This strain may be useful for generating conditional mutations in applications related to Src signaling and actin filament integrity.
Jess Cunnick, The Commonwealth Medical College
Youngjin Cho, The Commonwealth Medical College
The protein encoded by the Afap1 gene is a Src binding partner. It may represent a potential modulator of actin filament integrity in response to cellular signals, and may function as an adaptor protein by linking Src family members and/or other signaling proteins to actin filaments. These mice possess loxP sites on either side of exon 5 of the targeted gene. Mice that are homozygous for this allele are viable and fertile. When these mutant mice are bred to mice that express Cre recombinase, resulting offspring will have exon 5 deleted in the cre-expressing tissues. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
When bred to a strain with Cre recombinase expression in all tissues (see Stock No. 006054 for example), this mutant mouse strain may be useful in studies of the Src signaling pathway.
A targeting vector containing a FRT site flanked PGK-Neo selection cassette was utilized in the construction of this mutant. This selection cassette and a loxP site was inserted downstream of exon 5 of the targeted gene, and another loxP site was inserted upstream of exon 5. This construct was electroporated into B6.Cg-Thy1a derived Bruce4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. Resulting chimeric animals were backcrossed to wildtype C57BL/6J mice for 4 generations. The mice were then crossed to OzFlpE mice, on the C57BL/6J genetic background, expressing FLPe recombinase. Mice that retained the loxP site flanked exon 5 were then bred to C57BL/6J mice for 5 generations, and the FlpE allele/transgene removed. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
|Allele Name||targeted mutation 1.1, Youngjin Cho|
|Allele Type||Targeted (Conditional ready (e.g. floxed))|
|Gene Symbol and Name||Afap1, actin filament associated protein 1|
|Strain of Origin||B6.Cg-Thy1a|
|Molecular Note||Exon 5 was floxed. Flp-mediated recombination removed the FRT-flanked selection cassette inserted downstream of exon 5.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Afapfl mouse strain in a publication, please cite the originating article(s) and include JAX stock #026214 in your Materials and Methods section.