Overview

Also Known As: β-GsD Tg (or β-R-s Tg)

These β-GsD Tg (or β-R-s Tg) mice express the G protein-coupled receptor DREADD GsD (also called rM3Ds or R-s) directed to pancreatic β-cells by the rat insulin 2 promoter. GsD induces the canonical Gα_s signaling pathway specifically following administration of the pharmacologically inert molecule clozapine-N-oxide (CNO). These mice may be useful for chemogenetic/pharmacogenetic applications to express an activating DREADD for studying G protein regulation of β-cell function.

Donating Investigator

Jürgen Wess, National Institute of Diabetes and Digestive and Kidney Diseases (NIH-NIDDK)

Genetic overview

Genetic Background	Generation

Tg(Ins2-Chrm3*)SJwe

Allele Type
Transgenic (Inserted expressed sequence)

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Research Applications

Neurobiology Research
Research Tools
Immunology, Inflammation and Autoimmunity Research
Diabetes and Obesity Research
Cancer Research
Cell Biology Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

The β-GsD (or β-R-s) transgene has the rat insulin 2 promoter directing expression of a hemagglutinin epitope-tagged, mutant G protein-coupled receptor (GsD [also called rM3Ds or R-s]) to pancreatic β-cells.
The GsD is a G_s-coupled rat muscarinic 3 DREADD modified to have its second and third intracellular loops replaced with the corresponding turkey β1-adrenergic receptor sequences. Two amino acid substitutions (Y148C and A238G) have been added that abolish receptor affinity for the native ligand, acetylcholine (ACh), but allow receptor binding and subsequent activation by the small pharmacologically inert molecule clozapine-N-oxide (CNO). CNO binding to GsD induces the selective activation of the canonical Gα_s signaling pathway, effectively increasing intracellular cAMP levels.

Mice that are hemizygous for the transgene (β-GsD Tg or β-R-s Tg) are viable and fertile. The donating investigator reports they have not tried to generate homozygous mice (May 2016). The phenotype described below compares β-GsD Tg with a similarly created transgenic mouse that expresses a rat insulin 2 promoter-driven rhM3Dq (β-rhM3Dq Tg ; Stock No. 028964).

The transgenic mouse lines β-rhM3Dq Tg (Stock No. 028964) and β-GsD Tg (Stock No. 028966) express similar numbers of rhM3Dq and GsD in their pancreatic islets, respectively. RT-PCR studies confirmed that rhM3Dq and GsD transcripts were not detectable in tissues other than pancreatic islets.

In the absence of CNO, both transgenic lines have normal growth, insulin sensitivity, plasma insulin levels and glucagon levels. While freely-fed β-rhM3Dq Tg have normal blood glucose levels, freely-fed β-GsD Tg have decreased blood glucose levels. Taken together, this indicates the rhM3Dq receptor is devoid of ligand-independent signaling in vivo, whereas the GsD receptor retains some degree of agonist-independent signaling in vivo.

CNO treatment of transgenic mice leads to the activation of β-cell G_q/11 or Gα_s signaling pathways, respectively, in a conditional and β-cell-selective fashion.
Specifically, following CNO administration in β-rhM3Dq Tg, the canonical G_q/11 pathway is activated in rhM3Dq-expressing pancreatic β-cells. This leads to significant increases in both first- and second-phase insulin release, greatly improved glucose tolerance in obese, insulin-resistant mice, and elevated β-cell mass, associated with pathway-specific alterations in islet gene expression levels. For β-GsD Tg, CNO-induced activation of the canonical Gα_s pathway in GsD-expressing pancreatic β-cells results in qualitatively similar in vivo metabolic effects.

Development

The β-R-s transgene was designed in the laboratory of Dr. Jürgen Wess (NIH-NIDDK) to have (from 5prime to 3prime) a 650 bp rat insulin 2 (Ins2; RIP II) promoter fragment, a hemagglutinin epitope tag (HA), the GsD coding sequence (R-s; described below), and untranslated human growth hormone sequences (containing transcriptional termination, polyadenylation and splicing signals).
The 4.2 kbp transgene was microinjected into the pronuclei of ova from fertilized C57BL/6NTac mice. Founder mice were bred with C57BL/6NTac mice, and a germline transmitting founder line was established (default line designation S). The β-GsD Tg (or β-R-s Tg) colony was backcrossed onto C57BL/6NTac for seven generations, and then frozen as sperm. In 2016, β-GsD Tg sperm was sent to The Jackson Laboratory Repository. Upon arrival, our β-GsD Tg transgenic colony may be maintained by breeding hemizygous mice together, to wildtype (noncarrier) mice from the colony, or to C57BL/6NJ inbred mice (Stock No. 005304).

The GsD sequence is a Gα_s-coupled rat M3 DREADD (designer receptor exclusively activated by designer drug), also referred to as R-s, rM3Ds, rM3/turkey β1AR chimer DREADD, Gs-DREADD, Gs-D, or D-s. To create the rM3Ds sequence, the wildtype rat muscarinic 3 receptor (Chrm3; M3R) sequence was first modified to replace the second and third intracellular loops with the corresponding turkey β1-adrenergic receptor (ADRB1 or β1AR) sequences. The second modification introduced, via site-directed mutagenesis, the Y148C and A238G amino acid substitutions 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).

Expression Data

Expressed Gene	Chrm3, cholinergic receptor, muscarinic 3, rat
Expressed Gene	HA tag, HA tag,
Site of Expression	Pancreatic Beta-cells.

Control Suggestions

Noncarrier

Approximate Controls

005304 C57BL/6NJ

Additional Information

Considerations for Choosing Controls

Selected References

Guettier JM; Gautam D; Scarselli M; de Azua IR; Li JH; Rosemond E; Ma X; Gonzalez FJ; Armbruster BN; Lu H; Roth BL; Wess J. 2009. A chemical-genetic approach to study G protein regulation of beta cell function in vivo. Proc Natl Acad Sci U S A 106(45):19197-202PubMed: 19858481 MGI: J:154657

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Genetics

Tg(Ins2-Chrm3*)SJwe

Allele Symbol: Tg(Ins2-Chrm3*)SJwe

Allele Name	transgene insertion S, Jurgen Wess
Allele Type	Transgenic (Inserted expressed sequence)
Allele Synonym(s)	Tg(Ins2-Chrm3*)SJwe; transgene insertion S, Jurgen Wess
Gene Symbol and Name	Tg(Ins2-Chrm3*)SJwe, transgene insertion S, Jurgen Wess
Gene Synonym(s)
Promoter	Ins2, insulin 2, rat
Expressed Gene	Chrm3, cholinergic receptor, muscarinic 3, rat
Expressed Gene	HA tag, HA tag,
Site of Expression	Pancreatic Beta-cells.
Strain of Origin	C57BL/6NTac
Chromosome	UN
Molecular Note	The transgene was designed to have (from 5' to 3') a 650 bp rat insulin 2 (Ins2; RIP II) promoter fragment, a hemagglutinin epitope tag (HA), the GsD coding sequence (R-s; described below), and untranslated human growth hormone sequences (containing transcriptional termination, polyadenylation and splicing signals). The GsD sequence is a Galpha>s<-coupled rat M3 DREADD (designer receptor exclusively activated by designer drug). To create the rM3Ds sequence, the wildtype rat muscarinic 3 receptor (Chrm3; M3R) sequence was first modified to replace the second and third intracellular loops with the corresponding turkey beta1-adrenergic receptor (ADRB1 or beta1AR) sequences. The second modification introduced, via site-directed mutagenesis, the Y148C and A238G amino acid substitutions 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).

Disease/Phenotype

Disease Terms

Research Areas By Genotype

This mouse can be used to support research in many areas including:

Neurobiology Research
- Optogenetic and Chemogenetic tools
  - Chemogenetic tools
- Receptor Defects
  - G protein-coupled receptor
Research Tools
- Diabetes and Obesity Research
Immunology, Inflammation and Autoimmunity Research
- Intracellular Signaling Molecules
- Growth Factors/Receptors/Cytokines
Diabetes and Obesity Research
- Type 1 Diabetes (IDDM)
- Islet Transplantation Studies
- Type 2 Diabetes (NIDDM)
- Insulin Receptors and Growth Factors
Cancer Research
- Growth Factors/Receptors/Cytokines
Cell Biology Research
- Signal Transduction

Mammalian Phenotype Terms by Genotype

References

Guettier JM; Gautam D; Scarselli M; de Azua IR; Li JH; Rosemond E; Ma X; Gonzalez FJ; Armbruster BN; Lu H; Roth BL; Wess J. 2009. A chemical-genetic approach to study G protein regulation of beta cell function in vivo. Proc Natl Acad Sci U S A 106(45):19197-202PubMed: 19858481 MGI: J:154657

Additional - Tg(Ins2-Chrm3*)SJwe related

Technical Support

Contact Technical Support

Genotyping Protocols
- MELT: Tg(Ins2-Chrm3*)SJwe
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, hemizygous mice may be bred together, to wildtype (noncarrier) mice from the colony, or to C57BL/6NJ inbred mice (Stock No. 005304). To date (May 2016), it has not been attempted to make this strain homozygous.
- Additional Breeding and Husbandry Support
Mating System
- Noncarrier x Hemizygote
- Hemizygote x Noncarrier
Citation
When using the β-GsD Tg (or β-R-s Tg) mouse strain in a publication, please cite the originating article(s) and include JAX stock #028966 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Pricing & Availability

Cryo Recovery

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service	Genotype	Price
	Hemizygous or non carrier for Tg(Ins2-Chrm3*)SJwe

We will fulfill your order by providing at least two carriers for each strain ordered. The total number, sex, and genotypes provided will vary, although typically 8 or more animals are provided. Please check genotypes which will be recovered. While the genotypes of all animals produced will be communicated to you prior to scheduling shipment, the genotypes of animals provided may not reflect the mating scheme and genotypes described in the strain description. Animals are typically ready to ship in 11-14 weeks. If a second recovery is required to produce the minimum number of animals, then delivery time would increase to approximately 25 weeks. If we fail to produce animals of the correct genotype, you will not be charged. We cannot guarantee the reproductive success of mice shipped to your facility. If the mice are lost after the first three days (post-arrival) or do not produce progeny at your facility, a new order and fee will be necessary.

Cryorecovery to establish a Dedicated Supply for greater quantities of mice. Mice recovered can be used to establish a dedicated colony to contractually supply you mice according to your requirements. Price by quotation.

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Also Known As: β-GsD Tg (or β-R-s Tg)

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Tg(Ins2-Chrm3*)SJwe

Allele Symbol: Tg(Ins2-Chrm3*)SJwe

Disease Terms

Research Areas By Genotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

References

Additional - Tg(Ins2-Chrm3*)SJwe related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

The Jackson Laboratory's Genotype Promise

Terms of Use

Additional Use Restrictions Apply

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability