B6N.129S6-Kera^tm1Cyl/J

Stock number: 025568
Research areas: Developmental Biology Research, Immunology, Inflammation and Autoimmunity Research, Sensorineural Research

Description

Kera KO mice may be useful for studying the chemokines involved in corneal inflammation.

Detailed description

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.

Development

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

Symbol: Kera^tm1Cyl
Name: targeted mutation 1, Chia-Yang Liu
MGI accession ID: MGI:2664936
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Kera
Marker name: keratocan
Chromosome: 10
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.