These Notch3d1 mutant KO mice exhibit abnormal arterial wall morphology. They may be useful in studies of the Notch pathway during development, vascular development and angiogenesis.
Madhulika Sharma, University of Kansas Medical Center
Homozygotes display a disorganized artery wall morphology, and thin vascular smooth muscle cell coat and processes. Homozygous mice are viable and fertile with no gene product (mRNA or protein) detected by Northern blot analysis of brain, lung and heart tissue or Western blot analysis of lung and brain tissues.
In an attempt to offer alleles on well-characterized or multiple genetic backgrounds, alleles are frequently moved to a genetic background different from that on which an allele was first characterized. It should be noted that the phenotype of these FVB-congenic mice could vary from that originally described on a B6;129S1 genetic background. We may modify the strain description if necessary as published results become available.
A targeting vector containing a PGK-neo cassette was used to delete 2.5kb of sequence encoding the EGF repeats 8 through 12. The construct was electroporated into 129S1/Sv-Oca2+ Tyr+ Kitl+ derived CJ7 embryonic stem (ES) cells. Correctly targeted ES cells were injected into C57BL/6J blastocysts. The resulting male chimeric animals were crossed to C57BL/6J females, and then C57BL/6J for 3 generations. These mice arrived at The Jackson Laboratory as Stock No. 010547.
After obtaining them, Dr. Madhulika Sharma from The University of Kansas Medical Center crossed these mice to B6;129S1-Notch4tm1Grid/J mice (Stock No. 010544) and double mutants were bred to FVB/NJ mice for at least six generations (see SNP note below). Backcrossed mice were sent to The Jackson Laboratory, where they were bred to FVB/NJ inbred mice (Stock No. 001800) for at least one generation to establish Stock No. 023807. The Notch4tm1Grid allele was bred out and maintained as Stock No. 023808.
In 2016, A SNP (single nucleotide polymorphism) panel analysis, markers covering most somatic chromosomes and the X chromosome, was performed on the rederived living colony at The Jackson Laboratory Repository. Two markers (one each on chromosomes 9 and 19) were not fixed for FVB allele-type (i.e., still segregating for C57BL/6 allele-type). In addition, the markers on chromosome 17 (~2 kbp away from Notch3) were heterozygous for FVB and 129 allele-type - likely to be segregating with the targeted mutation. Taken together, these data suggest the mice sent to The Jackson Laboratory Repository were incompletely backcrossed onto FVB, and they are now ~89% FVB.
|Allele Name||targeted mutation 1, Tom Gridley|
|Allele Type||Targeted (Null/Knockout)|
|Gene Symbol and Name||Notch3, notch 3|
|Strain of Origin||129S1/Sv-Oca2+ Tyr+ Kitl+|
|Molecular Note||A 2.5 kb genomic fragment containing sequence encoding EGF repeats 8 through 12 was replaced with a PGK-neo cassette inserted by homologous recombination. The absence of transcript and protein was indicated by Northern and Western blot analysis of homozygous mutant mice.|
|Mutations Made By|| |
Thomas Gridley, Maine Medical Center Research Institute
When maintaining a live colony, homozygotes may be bred together.
When using the Notch3d1 mouse strain in a publication, please cite the originating article(s) and include JAX stock #023807 in your Materials and Methods section.