B6.Cg-Adora1^em1(cre)Rei/J

Stock number: 036249
Product name: Adora1-Cre
Research areas: Metabolism Research, Neurobiology Research, Research Tools

Description

Adora1-Cre knock-in mice are designed to have the endogenous Adora1 promoter/enhancer regions directing expression of Cre recombinase to multiple cell types, and in particular, providing high expression levels in neural cells. The cre knock-in results in minimal disruption to endogenous Adora1 (adenosine A1 receptor) expression. These mice are useful for studying body temperature regulation as well as cardiovascular and nervous systems.

Detailed description

Adenosine is a purine nucleoside involved in many biochemical processes including energy storage, regulation of gene expression and function, and metabolism. Adora1 (adenosine A1 receptor or A₁AR) encodes the adenosine A1 receptor (A₁AR), a G-protein coupled adenosine receptor. Adora1 is widely expressed, with high levels of expression detected in the brain, spinal cord, eye and atria. Activation of A₁AR by an agonist has inhibitory effects in multiple systems including the cardiovascular and nervous systems.

Adora1-Cre knock-in mice have a T2A peptide sequence and Cre recombinase inserted at the end of the stop codon of the Adora1 gene. The donating investigator indicates that ADORA1 and Cre are expected to translate in a 1:1 ratio, and that the added T2A sequence may reduce A₁AR function. Mice homozygous for the mutation are viable and fertile. When these Adora1-Cre mice are bred with mice containing loxP-flanked sequence, cre-mediated recombination will result in deletion of the loxP-flanked sequences. These mice are useful for studying body temperature regulation as well as cardiovascular and nervous systems.

Development

CRISPR/cas9 genome editing was used to insert a T2A peptide sequence and Cre recombinase at the Adora1 stop codon on chromosome 1. T2A sequence adds EGRGSLLTCGDVEENPG to the C terminus of A₁AR and a proline to the N terminus of cre. Single guide RNA targeting Adora1 was co-microinjected with cas9 mRNA and donor DNA construct into fertilized eggs collected from B6D2F1/J females. Embryos in the 2-cell stage were implanted into pseudopregnant foster mothers. Correctly targeted pups were identified by PCR and further bred to C57BL/6J for at least 10 generations. Note: the ampicillin resistance gene is retained in this strain. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

Allele

Symbol: Adora1^em1(cre)Rei
Name: endonuclease-mediated mutation 1, Marc L Reitman
MGI accession ID: MGI:6509134
Generation method: Endonuclease-mediated
Attributes: Recombinase-expressing
Synonyms: Adora1-Cre
Marker symbol: Adora1
Marker name: adenosine A1 receptor
Marker synonyms: A1AR; A1-AR; A1R; AA1R; ARA1; RDC7; Ri
Chromosome: 1
Strain of origin: (C57BL/6 x DBA)F1
Molecular note: CRISPR/cas9 genome editing is used to insert a T2A peptide sequence and Cre recombinase at the stop codon. T2A sequence adds EGRGSLLTCGDVEENPG to the C terminus of Adora1 and a proline to the N terminus of cre.