B6.Cg-Agtr2^em1(cre)Adk/J

Stock number: 036825
Product name: AT2R-Cre
Research areas: Neurobiology Research, Research Tools

Description

AT2R-Cre knock-in mice have the endogenous Agtr2 promoter/enhancer sequences directing expression of Cre recombinase. These mice are a Cre-lox tool allowing Cre recombination in Agtr2-expressing cells/tissues (including neurons GABAergic neurons in the nucleus of the solitary tract), and may be useful in studying hypertension.

Detailed description

The Agtr2 gene encodes the angiotensin type-2 receptor (AT2R) which plays a role in cardiovascular and renal function as a regulator of blood pressure and of water and electrolyte balance.

The AT2R-Cre knock-in allele (also called AT2R-Cre) has a T2A sequence and a Cre recombinase inserted into the 3' region of the endogenous angiotensin II receptor, type 2 gene. This is designed to have the endogenous Agtr2 promoter/enhancer elements directing cre expression primarily in GABAergic neurons of the nucleus of the solitary tract, as well as to retain endogenous Agtr2 expression.

The donating investigator reports that AT2R-Cre mice have Cre recombinase activity that recapitulates the endogenous Agtr2 expression pattern - observed mainly in GABAergic neurons of the nucleus of the solitary tract, with some widespread expression expected in the brain and other organs (Mohammed et al., 2021). This knock-in allele is designed to avoid disrupting expression of the endogenous protein; although Agtr2 expression from the AT2R-Cre allele has not been evaluated.

Homozygous females and hemizygous males are viable and fertile with no reported gross physical or behavioral abnormalities.

Development

The Agtr2^cre knock-in allele (also called AT2R-Cre) was created in the laboratory of Dr. Annette de Kloet (University of Florida) using CRISPR/Cas9 endonuclease-mediated genome editing. Guide RNAs were selected to target the 3' region of the angiotensin II receptor, type 2 gene (Agtr2) on the X chromosome. Double stranded oligonucleotide donor DNAs were created containing a viral 2A oligopeptide sequence (T2A; mediates ribosomal skipping), an SV40 nuclear localization signal, and a Cre recombinase gene. These sequences and Cas9 nuclease were introduced into single cell C57BL/6J X SJL hybrid zygotes and transferred to pseudopregnant females. Progeny were screened by DNA sequencing to identify correctly targeted pups, which were then bred to C57BL/6J mice (Stock No. 000664) for germline transmission. The resulting At2r-Cre colony was then backcrossed to C57BL/6J mice for at least 10 generations prior to sending males to The Jackson Laboratory Repository. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes.

Allele

Symbol: Agtr2^em1(cre)Adk
Name: endonuclease-mediated mutation 1, Annette D de Kloet
MGI accession ID: MGI:6834781
Generation method: Endonuclease-mediated
Attributes: Recombinase-expressing
Synonyms: Agtr2^Cre; AT2R-Cre
Marker symbol: Agtr2
Marker name: angiotensin II receptor, type 2
Marker synonyms: AT^sub2^/subR; AT2; AT2 receptor; AT2R; AT2-R; ATGR2; MRX88
Chromosome: X
Strain of origin: C57BL/6J x SJL
Molecular note: Using CRISPR/Cas9 endonuclease-mediated genome editing, a viral 2A oligopeptide sequence (T2A; mediates ribosomal skipping), an SV40 nuclear localization signal, and a Cre recombinase gene were inserted into the Agtr2 gene 3'UTR.