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Tg(CAG-CHRM3*,-mCitrine)1Ute
Transgene Detail
Nomenclature
Symbol: Tg(CAG-CHRM3*,-mCitrine)1Ute
Name: transgene insertion 1, Ute Hochgeschwender
MGI ID: MGI:5614052
Synonyms: CAG-LSL-Gq-DREADD, Gt(ROSA)26Sortm2(CAG-CHRM3*,-mCitrine)Ute, hM3Dq, LSL-hM3Dq, R26-hM3Dq/mCitrine, R26-LSL-Gq-DREADD, R26-LSL-hM3Dq/mCitrine
Transgene: Tg(CAG-CHRM3*,-mCitrine)1Ute  Location: unknown  
Mutation
origin
Germline Transmission:  Earliest citation of germline transmission: J:236327
Parent Cell Line:  R1 (ES Cell)
Strain of Origin:  (129X1/SvJ x 129S1/Sv)F1-Kitl+
Mutation
description
Allele Type:    Targeted (Conditional ready, No functional change)
Mutation:    Insertion
 
Mutation detailsThe 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). (J:236327)

Phenotypes
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View phenotypes and curated references for all genotypes (concatenated display).
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Mouse strains and cell lines available from the International Mouse Strain Resource (IMSR)
Carrying this Mutation:  Mouse Strains: 1 strain available      Cell Lines: 0 lines available
References
Original:  J:236327 Zhu H, et al., Cre-dependent DREADD (Designer Receptors Exclusively Activated by Designer Drugs) mice. Genesis. 2016 Aug;54(8):439-46
All:  10 reference(s)

Contributing Projects:
Mouse Genome Database (MGD), Gene Expression Database (GXD), Mouse Models of Human Cancer database (MMHCdb) (formerly Mouse Tumor Biology (MTB), Gene Ontology (GO)
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last database update
01/21/2020
MGI 6.14
The Jackson Laboratory