This PlauGFDhu/GFDhu targeted mutation strain carries a mutant protein unable to bind to the urokinase plasminogen activator receptor. These mutant mice may be useful in studies of chronic tissue inflammation, and fibrin deposition and elimination.
Dr. Thomas Bugge, NIH/NIDCR
These mice carry 6 nucleotide substitutions in exon 4 that result in 4 amino acid substitution mutations (Y23N, R28N, R30H, and R31W) and abolish binding interaction of the protein with its receptor: plasminogen activator, urokinase receptor (uPAR). Homozygous mutant mice express a mutant urokinase plasminogen activator protein with a 400 fold reduction in affinity to the mouse urokinase plasminogen activator receptor and a binding affinity to human urokinase plasminogen activator receptor that is similar to endogenous human PLAU affinity. The mutant protein is functional, with homozygotes exhibiting normal skin wound healing. Mice that are homozygous for the targeted mutation are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. The nucleotide substitutions were confirmed by sequence analysis. Levels of gene product (mRNA and protein) expression are normal as detected by qPCR or Western blot analysis. Homozygotes exhibit 2.5 fold more leukocytic infiltrate foci in their livers than wildtype controls, with some homozygotes also displaying mild to moderate inflammation of the trachea. Fibrin deposit elimination is impaired leading to chronic inflammation. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
A targeting vector containing nucleotide substitutions encoding the amino acid substitutions Y23N, R28N, R30H, and R31W was used to replace exon 4. The construct was electroporated into C57BL/6 derived Bruce 4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts. The resulting chimeric animals were crossed to Oz_Cre deleter mice (on the C57BL/6 background) to remove the floxed neo selection cassette, and then backcrossed to C57BL/6J for 7 generations. During backcrossing, the Y chromosome may not have been fixed to the C57BL/6J genetic background.
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, Thomas H Bugge|
|Allele Type||Targeted (Not Applicable)|
|Allele Synonym(s)||PlauGFDhu; uPAY23N-R28N-R30H-R31W|
|Gene Symbol and Name||Plau, plasminogen activator, urokinase|
|Promoter||Plau, plasminogen activator, urokinase, mouse, laboratory|
|Strain of Origin||B6.Cg-Thy1a|
|Molecular Note||Exon 4 was replaced with one in which nucleotide substitutions (resulting in the amino acid substitutions Y23N, R28N, R30H, and R31W) abolish the ligand receptor interacting domain governed by the burial of the beta-hairpin of the growth factor domain (GFD). The overal structure and catalytic function of the protein is preserved. A floxed neo cassette inserted downstream of the modified exon 4 was removed by cre mediated recombination.|
|Mutations Made By|| |
Dr. Thomas Bugge, NIH/NIDCR
When maintaining a live colony, these mice can be bred as homozygotes.
When using the C57BL/6-Plautm1.1Bug/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #014557 in your Materials and Methods section.