These floxed mutant mice possess loxP sites flanking exon 2 of the Picalm gene. This strain may be useful for generating conditional mutations in applications related to clathrin-dependent endocytosis.
Takahiro Maeda, Brigham and Women's Hospital/ Harvard Medical School
The Picalm gene encodes for a clathrin adaptor protein important in regulation of clathrin-coated vesicle formation at the plasma membrane as well as regulation of amyloid-β blood-brain barrier transcytosis and transferrin receptor endocytosis in erythroblasts. Polymorphisms of the human PICALM gene are associated with the pathogenesis of Alzheimer's Disease. These mice possess loxP sites on either side of exon 2 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 2 deleted in the cre-expressing tissues.
When bred to a strain with widespread Cre recombinase expression in (see Stock No. 003724 for example), this mutant mouse strain may be useful in studies of clathrin-dependent endocytosis and autophagy.
A targeting vector containing a FRT site flanked PGK-neo cassette was utilized in the construction of this mutant. This selection cassette, in reverse orientation, and a 3’ loxP site were inserted downstream of exon 2 of the targeted gene, and another loxP site was inserted upstream of exon 2. This construct was electroporated into B6.Cg-Thy1a derived Bruce4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into MB6 albino blastocysts. The resulting chimeric animals were crossed to albino B6(Cg)-Tyrc-2J/J (see Stock no. 000058).
PicalmNeo/+ mice were crossed to transgenic mice (on the B6;SJL genetic background) expressing FLP1 recombinase under the control of the human ACTB promoter (see Stock no. 003800) to excise the selection cassette. Mice that retained the loxP site flanked exon 2 were then bred to C57BL/6 mice to remove the FLPe transgene.
Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
|Allele Name||targeted mutation 1.1, Takahiro Maeda|
|Allele Type||Targeted (Conditional ready (e.g. floxed), No functional change)|
|Gene Symbol and Name||Picalm, phosphatidylinositol binding clathrin assembly protein|
|Strain of Origin||B6.Cg-Thy1a|
|Molecular Note||The targeting vector contains an FRT-flanked PGK-neo cassette, in reverse orientation, and a 3' loxP site downstream of exon 2, and another loxP site inserted upstream of exon 2. Flp-mediated recombination removed the FRT-flanked neo cassette leaving exon 2 floxed.|
When maintaining a live colony, these mice can be bred as homozygotes.
When using the Picalmflox mouse strain in a publication, please cite the originating article(s) and include JAX stock #027393 in your Materials and Methods section.