B6N;129-Tg(CAG-CHRM3*, -mCitrine)1Ute/J

Cat. No.: CEMM-07250591

Common Name: R26-hM3Dq/mCitrine

The CAG-LSL-Gq-DREADD allele (formerly called R26-LSL-Gq-DREADD) allows Cre recombinase-inducible expression of a CAG promoter-driven HA-hM3Dq-pta-mCitrine. Following cre-mediated removal of an upstream floxed-STOP cassette, expression of HA-tagged hM3Dq and mCitrine yellow fluorescent protein is observed. hM3Dq is a mutant G protein-coupled receptor (GPCR) that induces the canonical Gq pathway specifically following administration of the pharmacologically inert molecule clozapine-N-oxide (CNO). These CAG-LSL-Gq-DREADD mice may be useful for chemogenetic/pharmacogenetic studies to express an activating DREADD that effectively induces firing of neurons. Note on allele, transgene insertion and copy number: This was originally designed as a targeted insertion into the Gt(ROSA)26Sor locus.

Inquiry

Status	Live Mouse Frozen Embryo
Age	4 weeks 12 weeks Customized Age
Sex	Male Female

GENETICS
Allele Symbol	Tg(CAG-CHRM3*,-mCitrine)1Ute show more close
Allele Name	transgene insertion 1 show more close
Allele Attributes	Conditional ready (e.g. floxed); No functional change show more close
Gene Symbol	Tg(CAG-CHRM3*,-mCitrine)1Ute show more close
Gene Name	transgene insertion 1 show more close
Chromosome	14 show more close
Expressed Genes	YFP, Yellow Fluorescent Protein, jellyfish; CHRM3, cholinergic receptor muscarinic 3, human show more close
MGI Accession ID	5614052 show more close
Site of Expression	Yellow fluorescent protein variant mCitrine and the hemagglutinin epitope-tagged, mutant G protein-coupled receptor hM3Dq (modified CHRM3 sequence) will be expressed in tissues expressing Cre recombinase. show more close
Strain of Origin	(129X1/SvJ x 129S1/Sv)F1-Kitl+ show more close
Molecular Note	The construct was designed with (from 5' to 3') a CMV-IE enhancer/chicken beta-actin/rabbit beta-globin hybrid promoter (CAG), an FRT site, a loxP-flanked STOP cassette (with stop codons in all 3 reading frames and a triple polyA signal), a hemagglutinin epitope tag (HA), the hM3Dq sequence, a porcine teschovirus (PTA) cleavage site, the mCitrine fluorescent protein, a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE; to enhance the mRNA transcript stability), a BGH polyA signal, and an attB/attP-flanked PGK-FRT Neo-polyA cassette. The hM3Dq sequence is a Gq-coupled human M3 muscarinic DREADD (designer receptor exclusively activated by designer drug). To Create the hM3Dq sequence, the wildtype human muscarinic 3 receptor (CHRM3) sequence was modified via site-directed mutagenesis to harbor two amino acid substitutions (Y149C<3.33>/A239G<5.46>) that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small drug-like molecule clozapine-N-oxide (CNO).Citrine is a yellow-green variant of Citrine YFP additionally harboring the A206K mutation. mCitrine is a ~5.7 pKa monomer with 516nm excitation and 529nm emmission spectra. Compared to wildtype GFP, Citrine harbors the S65G, V68L, Q69M, S72A and T203Y mutations, and mCitrine additionally has the A206K mutation (enhancing its ability to be a stable monomer).Aanalysis of the transgene integration site revealed a single (or partial) transgene copy integrated on chromosome 14 (chr14:41,019,438-41,024,312 [mm10]) in a manner suggestive of complex structural variation surrounding the integration site. Specifically, there is duplication of nearby genome sequences involving the genes Tspan14, Prxl2a, Dydc2, Dydc1, Mat1a, Sftpa1 and Mbl1 show more close

HUSBANDRY
Suggested Controls	C57BL/6NJ show more close
Breeding Considerations	When maintaining a live colony, hemizygous mice may be bred together, to wildtype (non-carrier) mice from the colony or to C57BL/6NJ inbred animals. While homozygous mice are expected to be viable and fertile, the generation of homozygous mice has not been attempted to date. show more close
Breeding Strategy	Hemizygote x Noncarrier; Noncarrier x Hemizygote show more close

For Research Use Only.