Mice that are homozygous null for Cd14 are viable and fertile. No Cd14 protein product is detected in thioglycollate elicited peritoneal (T-EP) macrophages by Western blot analysis. Unlike wildtype T-EP macrophages, T-EP macrophages derived from these animals fail to secrete TNF-alpha and IL-6 in response to lipopolysaccharide (LPS). Such a response is observed when Cd14 -null T-EP macrophages are exposed to whole bacteria (in the form of E. coli bioparticles), apparently by a Cd14-independent mechanism.
A targeting vector containing neomycin resistance and thymidine kinase genes was used to disrupt the entire Cd14 coding region, 4.0 kb of upstream sequence and 2.5 kb of downstream sequence. The construct was transfected into 129S4/SvJae-derived J1 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts. The resulting chimeric male animals were backcrossed to C57BL/6J females.
|Allele Name||targeted mutation 1, Mason W Freeman|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Cd14, CD14 antigen|
|Strain of Origin||129S4/SvJae|
|Molecular Note||A neomycin resistance cassette replaced a genomic fragment containing the entire coding sequences of the gene. Western blot analysis confirmed an absence of CD14 protein in macrophage lysates derived from homozygous mice.|
|Mutations Made By|| |
Mason Freeman, Massachusetts General Hospital
This strain originated on a B6;129S background and is currently being backcrossed to C57BL/6J. Coat color expected from breeding:Black
When using the B6.129S4-Cd14tm1Frm/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #003726 in your Materials and Methods section.