N-cad-CreERT knock-in/knock-out mice express cre/ERT in N-cadherin (Cdh2) expressing cells. They are suitable for use in applications related to the study of development and cartilage and bone formation.
Linheng Li, Stowers Institute for Medical Research
The targeted Cdh2 gene encodes a calcium-dependent cell adhesion protein. The N-cad-CreERT knock-in allele consists of cre/ERT, a non-functional mCherry, BGH polyadenylation sequence and an FRT-site flanked neo inserted into exon 1 (replacing the start codon) of the Cdh2 gene. cre/ERT is expressed in N-cadherin expressing cells in bone marrow and endosteal cells. When bred with mice containing loxP-flanked sequence, tamoxifen-inducible, cre-mediated recombination will result in deletion of the floxed sequences. Heterozygotes are viable and fertile. The donating investigator has not observed homozygotes, which are not expected to be viable. During backcrossing, the Y chromosome was not fixed to the C57BL/6 genetic background.
A targeting vector containing cre/ERT-IRES-mCherry-BGH polyA-FRT-Neo-FRT was inserted into exon 1 of the Cdh2 gene, replacing endogenous start codon. The construct was electroporated into (C57BL/6 x 129X1/SvJ)F1 derived ASE-9005 (Applied Stem Cell) embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were backcrossed to C57BL/6 for 6 generations by the donating lab (see SNP note below). Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).
In 2020, a 48 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/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. Two of the 43 markers, on Chromosomes 4 and 11, were segregating with 129 suggesting an incomplete backcross.
|Allele Name||targeted mutation 1, Linheng Li|
|Allele Type||Targeted (Recombinase-expressing, Inducible)|
|Gene Symbol and Name||Cdh2, cadherin 2|
|Strain of Origin||(C57BL/6 x 129X1/SvJ)F1|
|Molecular Note||A targeting vector containing cre/ERT-IRES-mCherry-BGH polyA-FRT-Neo-FRT is inserted into exon 1 of the gene, replacing endogenous start codon. MCherry expression cannot detected by direct fluorescence or by antibodies.|
When maintaining a live colony, heterozygous mice may be bred to wildtype siblings, or to C57BL/6J inbred mice (Stock No. 000664). The donating investigator has not observed homozygotes, which are not expected to be viable.
When using the N-cad-CreERT , Ncad-CreER mouse strain in a publication, please cite the originating article(s) and include JAX stock #033109 in your Materials and Methods section.