Kera KO mice may be useful for studying the chemokines involved in corneal inflammation.
Semin Goins, University of Cincinnati
Chia-Yang Liu, University of Cincinnati
Kera KO mice have a neo cassette replacing exon 1 and part of exon 2 of the keratocan (Kera) gene, abolishing gene expression. Keratocan is a cornea-specific keratan sulfate proteoglycan expressed in ocular surface tissues including cornea and eyelids during morphogenesis. Keratocan is involved in maintaining corneal transparency and for the structure of the stromal matrix to ensure normal vision. It has been shown to bind to the neutrophil chemokine CXCL1, which facilitates neutrophil migration into the corneal stroma in corneal inflammation models. Mice homozygous for this allele are viable and fertile. These mice display normal corneal transparency at 12 months of age with subtle structural alterations of collagenous matrix. They have a thinner corneal stroma and a narrower cornea-iris angle of the anterior segment than WT mice. They also have larger stromal fibril diameters and less organized packing of collagen fibrils in stroma than those of wild type. In a model of corneal inflammation, LPS treated Kera KO mice, corneas have a decreased number of neutrophils in the cornea.
A targeting vector was designed to replace exon 1 and part of exon 2 encoding the keratocan (Kera) gene with a neomycin resistance (neo) cassette in reverse orientation to the gene. The construct was electroporated into 129S6/SvEvTac-derived Duffy embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts and the resulting chimeric males were bred to C57BL/6NJ females. These mice were bred to C57BL/6NJ mice (Stock No. 005304) for at least 15 generations. Upon arrival at The Jackson Laboratory, mice were bred to C57BL/6NJ mice for at least one generation to establish the colony.
|Allele Name||targeted mutation 1, Chia-Yang Liu|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Kera, keratocan|
|Strain of Origin||129S6/SvEvTac|
|Molecular Note||About 1 kb of intron 1 and all but the last 0.6kb of exon 2 were replaced with a neomycin-resistance gene inserted in antisense orientation. In-situ hybridization indicated no mRNA in the cornea of homozygous mutant mice and protein was undetectable by immunohistochemical staining or by Western blot analysis.|
When maintaining a live colony, homozygous mice may be bred together.
When using the B6N.129S6-Keratm1Cyl/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #025568 in your Materials and Methods section.