C57BL/6-Ndfip2^tm1.1Pmo/J

Stock number: 033321
Product name: Ndfip2 knockout/GFP knock-in mice
Research areas: Immunology, Inflammation and Autoimmunity Research, Research Tools

Description

Ndfip2 knock-out/EGFP knock-in mice may be useful in studies related to inflammation and T cell activation.

Detailed description

The targeted Ndfip2 gene encodes an activator of Nedd4 family ligases. These Ndfip2 knock-out/EGFP knock-in mice carry a knock-out allele in which GFP deletes part of exon 2 and all of exons 3 and 4, resulting in a null allele, with a resulting frame shift of the downstream exons. Mice that are homozygous for the targeted mutation are viable and fertile. EGFP expression is detected in splenic cells (with the highest EGFP signal observed in T cells) from heterozygotes by flow cytometry analysis. No gene product (mRNA) is detected by qPCR analysis of samples from homozygotes.

When bred to Ndfip1 floxed mice (Stock No. 033320) crossed to a strain with Cre recombinase expression in T cells, the resulting NDFIP deficient mutant mice exhibit increased levels of inflammation in lung, esophagus and skin, as well as an increased number of activated CD4+ T cells.

Development

A targeting vector containing an EGFP reporter followed by polyadenylation signal and an F3-flanked puromycin resistance cassette was inserted into exon 2, resulting in a partial deletion of exon 2 and deletion of exons 3 and 4. The construct was electroporated into C57BL/6N Tac embryonic stem (ES) cells. Correctly targeted ES cells were injected into blastocysts. The resulting chimeric animals were tested for germline transmission. The mice were then crossed to FLP-deleter mice, C57BL/6-Tg(CAG-Flpe)2Arte, to remove the selection cassette. 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 2019 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. One of the 43 markers, on Chromosome 12 was segregating with an unknown source. Also, 2 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Ndfip2^tm1.1Pmo
Name: targeted mutation 1.1, Paula M Oliver
MGI accession ID: MGI:6392820
Generation method: Targeted
Attributes: Reporter; Null/Knockout
Marker symbol: Ndfip2
Marker name: Nedd4 family interacting protein 2
Marker synonyms: 0710001O20Rik; 9130207N19Rik; N4WBP5A
Chromosome: 14
Strain of origin: C57BL/6N
Molecular note: A targeting vector containing an EGFP reporter followed by polyadenylation signal and an F3-flanked puromycin resistance cassette was inserted into exon 2, resulting in a partial deletion of exon 2 and deletion of exons 3 and 4. Flp-mediated recombination removed the FRT-flanked puro cassette.