C57BL/6-Vipr2^em1Litt/J

Stock number: 035395
Product name: Vipr2^fl
Research areas: Neurobiology Research, Research Tools

Description

Exons 3 and 4 of the mouse Vipr2 (vasoactive intestinal peptide receptor 2) gene are flanked by loxP sites in these CRISPR-generated Vipr2^fl mice. Cre recombinase-mediated excision of the floxed region results in a knock-out allele.

Detailed description

During food consumption, the activation of vasoactive intestinal peptide-producing (VIPergic) neurons in the intestinal mucosa enhances the growth of segmented filamentous bacteria associated with the epithelium, and increases lipid absorption. Projections of VIPergic neurons in the lamina propria are in close proximity to clusters of type-3 innate lymphoid cells (ILC3s) that selectively express VIP receptor type 2 (VIPR2; also known as VPAC2).

Exons 3 and 4 of the mouse Vipr2 (vasoactive intestinal peptide receptor 2) gene are flanked by loxP sites in these CRISPR-generated Vipr2^fl mice. Cre recombinase-mediated excision of the floxed region results in a knock-out allele.

Circulating plasma triglycerides (12 hours after feeding) are substantially reduced in the progeny derived from crosses with Rorc(t)^cre mice (Stock No. 022791) to knock out VIPR2 expression in RORγt⁺ cells. Unlike wildtype controls, segmented filamentous bacteria (SFB) do not form segmented filaments even after 12 hours of feeding in Rorc(t)^creVipr2^fl/fl knock-out mice.

Development

The Vipr2^fl mutation was created using CRISPR-Cas9 technology. Guide RNAs, Cas9 mRNA, and ssDNA donor templates were injected into C57BL/6J embryos to introduce loxP sites on either side of the exon 3/exon 4 segment. Resultant mice were sequenced to identify mice homozygous for both loxP insertions. Founders were backcrossed to C57BL/6J for at least one generation, then backcrossed to C57BL/6NTac for 4 generations by the donating laboratory.

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. While the 43 markers throughout the genome suggested a C57BL/6 genetic background, 4 of the 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating, including the rd8 recessive mutation associated with retinal degeneration on Chromosome 1 within the Crb1 locus. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Allele

Symbol: Vipr2^em1Litt
Name: endonuclease-mediated mutation 1, Dan R Littman
MGI accession ID: MGI:6436298
Generation method: Endonuclease-mediated
Attributes: Conditional ready (e.g. floxed)
Synonyms: Vipr2^fl
Marker symbol: Vipr2
Marker name: vasoactive intestinal peptide receptor 2
Marker synonyms: C16DUPq36.3; DUP7q36.3; PACAP-R3; PACAP-R-3; VIP receptor subtype 2; Vip2; VIP-R-2; VPAC2; VPAC2R; VPCAP2R
Chromosome: 12
Strain of origin: Not Specified
Molecular note: CRISPR/Cas9 genome editing is used to introduce loxP sites on either side of the exons 3-4. Resultant mice were sequenced to identify mice homozygous for both loxP insertions.