B6NCrl.129P2(B6)-Apoe^tm1Unc Ccr5^tm1Blck/BlckJ

Stock number: 030894
Product name: 5AE-B
Research areas: Cardiovascular Research, Mouse/Human Gene Homologs, Research Tools

Description

5AE-B mice are C57BL/6NCrl-congenic animals harboring the ApoE knock-out allele (Apoe^tm1Unc) and the CCR5 knock-out allele (Ccr5^tm1Blck). These mice are useful for studying the pathogenesis and treatment of atherosclerosis, as well as lipoprotein/cholesterol metabolism, neurovascular function, inflammation and neurodegenerative cardiovascular disease.

Detailed description

C57BL/6N-congenic 5AE-B mice homozygous for knock-out alleles of ApoE and Ccr5 (ApoE-/-Ccr5-/-) are viable and fertile with no apparent reproductive abnormalities. Compared to ApoE-/- mice, the ApoE-/-Ccr5-/- (5AE-B) mice are protected from both high-fat and high-cholesterol diet-induced atherosclerosis (atherosclerotic lesion formation and cell accumulation). This protection is associated with a more stable plaque phenotype, reduced mononuclear cell infiltration, Th1-type immune responses and increased interleukin-10 expression. Additionally, the protective effect of CCR5-deficiency was also sustained at later time points and was observed both with a high-fat diet and with a normal chow-diet. [Braunersreuther et al. 2007 Arterioscler Thromb Vasc Biol 27:373]

Dr. Bruno Luckow has made the following C57BL/6N-congenic strains available: ApoE knock-out (AE-B; Stock No. 030891), ApoE Ccr1 double knock-out (1AE-B; Stock No. 030892), ApoE Ccr2 double knock-out (2AE-Bd; Stock No. 030893), ApoE Ccr5 double knock-out (5AE-B; Stock No. 030894), ApoE Ccr1 Ccr5 triple knock-out (15AE-B; Stock No. 030895) and ApoE Ccr2 Ccr5 triple knock-out (25AE-Bd; Stock No. 030896).

Development

These 5AE-B mice (Stock No. 030894) are C57BL/6NCrl-congenic animals harboring the ApoE knock-out allele (Apoe^tm1Unc) and the CCR5 knock-out allele (Ccr5^tm1Blck). Each mutant allele is described below.

The ApoE knock-out allele (Apoe^tm1Unc) was created in the laboratory of Dr. Nobuyo Maeda (University of North Carolina at Chapel Hill) to have a neo cassette replacing part of the coding region of the apolipoprotein E Locus on chromosome 7. In 2005, Dr. Bruno Luckow (Klinikum der Universitaet Muenchen) obtained C57BL/6J-congenic ApoE knock-out mice from The Jackson Laboratory (Stock No. 002052), bred them several generations to C57BL/6NCrl mice (Charles River Laboratories), and then used them as described below.

The CCR5 knock-out allele (Ccr5^tm1Blck) was created in the laboratory of Dr. Bruno Luckow (Klinikum der Universitaet Muenchen) to have a lacZ::neo cassette replacing most of the ORF on exon 2 (thereby deleting 342 of 354 amino acids) of the chemokine (C-C motif) receptor 5 locus on chromosome 9. After electroporation into 129P2/OlaHsd-derived E14.1 embryonic stem (ES) cells, chimeras were bred to C57BL/6NCrl mice to establish their CCR5 knock-out colony. The CCR5 knock-out colony was backcrossed 13 generations to C57BL/6NCrl, and then used as described below.

To generate the ApoE/CCR5 double mutant colony [5AE-B], Dr. Luckow bred the two mutant mouse lines together. The donating investigator reports that the 5AE-B colony was then backcrossed to C57BL/6NCrl for at least four generations. In 2017, Dr. Luckow sent 5AE-B mice to The Jackson Laboratory as Stock No. 030894. Upon arrival, sperm was cryopreserved.
Of note, the donating investigator reports that, at least once during backcrossing, a heterozygous female was bred to a C57BL/6NCrl inbred male (thus the Y chromosome of the congenic strain is of C57BL/6NCrl origin).

In 2017, a SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6N and C57BL/6J substrains, was performed on the male mice sent to The Jackson Laboratory Repository. All markers were C57BL/6 allele-type. In addition, all markers that determine C57BL/6 substrains were C57BL/6N allele-type with one exception - the marker on chromosome 6 [~107.8 Mbp] was found to be homozygous for C57BL/6J allele-type. This suggests that C57BL/6NCrl allele-type at this marker is the same as C57BL/6J allele-type (rather than C57BL/6NJ allele-type).

 

Allele

Symbol: Ccr5^tm1Blck
Name: targeted mutation 1, Bruno Luckow
MGI accession ID: MGI:3613371
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Ccr5
Marker name: chemokine (C-C motif) receptor 5
Chromosome: 9
Strain of origin: 129P2/OlaHsd
Molecular note: A lacZ-neo cassette replaced much of the single exon. Absence of transcript was confirmed by RNase-protection assay.

Allele

Symbol: Apoe^tm1Unc
Name: targeted mutation 1, University of North Carolina
MGI accession ID: MGI:1857129
Generation method: Targeted
Attributes: Null/Knockout
Marker symbol: Apoe
Marker name: apolipoprotein E
Chromosome: 7
Strain of origin: 129P2/OlaHsd
Molecular note: Insertion of a neomycin resistance cassette deleted part of exon 3 and part of intron 3 of the Apoe gene. Plasma from homozygous mutant mice gave no detectable immunoprecipitate by the Ouchterlony double immunodiffusion test using a rabbit antibody to rat APOE.