Overview

Also Known As: Gad2-CreER

These knock-in mice express a tamoxifen-inducible cre recombinase from the endogenous promoter/enhancer elements of the Gad2 gene. When induced, cre activity is observed in the cortex and cerebellum.

Donating Investigator

Z. Josh Huang, Cold Spring Harbor Laboratory

Genetic overview

Genetic Background	Generation

Gad2^{tm1(cre/ERT2)Zjh}

Allele Type	Gene Symbol	Gene Name
Targeted (Recombinase-expressing, Inducible)	Gad2	glutamic acid decarboxylase 2

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Research Applications

Research Tools
Neurobiology Research

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Base Price

Starting at:

$2,854.50 Domestic price Cryo Recovery

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Details

Detailed Description

The Gad2-CreERT2 knock-in allele both abolishes Gad2 gene function and expresses a CreER^T2 fusion protein (creERT2 fusion protein) from the Gad2 promoter/enhancer elements. Heterozygous mice are viable and fertile with no reported abnormalities. Homozygous mice are expected to have a knockout phenotype similar to other null mutations of this gene and may exhibit increased seizure activity (although no early death is reported). Cre-ERT2 fusion gene activity is inducible; observed following tamoxifen administration. As such, when Gad2-CreERT2 mice are bred with mice containing loxP-flanked sequence, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in the Gad2-expressing cells of the offspring.

The donating investigator reports tamoxifen-inducible Cre recombinase activity is specific and efficient (largely recapitulates the endogenous Gad2 expression pattern with highly efficient inducibility). They report tamoxifen-inducible Cre recombinase activity is observed in the cortex and cerebellum, as well as very rare incidence of Cre activity in the cortex prior to tamoxifen administration.

For characterization information, see images at the Allen Institute for Brain Science website (Gad2-CreERT2 images).

The Cre-ERT2 fusion protein consists of Cre recombinase fused to a triple mutant form of the human estrogen receptor which does not bind its natural ligand (17β-estradiol) at physiological concentrations but will bind the synthetic estrogen receptor ligands 4-hydroxytamoxifen (OHT or tamoxifen) and, with lesser sensitivity, ICI 182780. Restricted to the cytoplasm, Cre-ERT2 can only gain access to the nuclear compartment after exposure to tamoxifen. To counteract the mixed estrogen agonist effects of tamoxifen injections, which can result in late fetal abortions in pregnant mice, progesterone may be coadministered.

Development

A targeting vector was designed to insert a CreER^T2 fusion gene (Cre-ER^T2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain), an SV40 polyA signal, and an frt-flanked neo cassette into the initiation codon of the Gad2 (glutamic acid decarboxylase 2) gene. This construct was electroporated into (C57BL/6 x 129S4Sv/Jae)-derived V6.5 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric mice were bred with white C57BL/6 mice to originate the colony. Mutant mice were bred with Actin-FLPe mice (on a C57BL/6 congenic background (N10); see Stock No. 005703) to remove the neo selection cassette. These Gad2-CreERT2 mice were subsequently bred with wildtype C57BL/6 mice for at least one generation (and the FLPe transgene was removed) prior to arrival at The Jackson Laboratory. Upon arrival, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the Gad2-CreERT2 colony.

Expression Data

Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of Expression	tamoxifen-inducible Cre recombinase activity is observed in the cortex and cerebellum; very rare incidence of Cre activity is observed in the cortex prior to tamoxifen administration.

Control Suggestions

Wild-type from the colony

Approximate Controls

Additional Information

Considerations for Choosing Controls

Selected References

NIH Neuroscience Blueprint Cre Driver Network. 2009. Cre recombinase-expressing mice generated for the NIH Neuroscience Blueprint Cre Driver Network MGI Direct Data SubmissionMGI: J:151755
Taniguchi H; He M; Wu P; Kim S; Paik R; Sugino K; Kvitsani D; Fu Y; Lu J; Lin Y; Miyoshi G; Shima Y; Fishell G; Nelson SB; Huang ZJ. 2011. A resource of cre driver lines for genetic targeting of GABAergic neurons in cerebral cortex. Neuron 71(6):995-1013PubMed: 21943598 MGI: J:177261

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Genetics

Gad2^{tm1(cre/ERT2)Zjh}

Allele Symbol: Gad2^{tm1(cre/ERT2)Zjh}

Allele Name	targeted mutation 1, Z Josh Huang
Allele Type	Targeted (Recombinase-expressing, Inducible)
Allele Synonym(s)	targeted mutation 1, Z Josh Huang; Gad2^{tm1(cre/ERT2)Zjh}
Gene Symbol and Name	Gad2, glutamic acid decarboxylase 2
Gene Synonym(s)	Gad-2; GAD(65); GAD65; 6330404F12Rik; 6330404F12Rik; RIKEN cDNA 6330404F12 gene; Gad-2; gad65
Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of Expression	tamoxifen-inducible Cre recombinase activity is observed in the cortex and cerebellum; very rare incidence of Cre activity is observed in the cortex prior to tamoxifen administration.
Strain of Origin	(C57BL/6 x 129S4/SvJae)F1
Chromosome	2
General Note	s
Molecular Note	A targeting vector was designed to insert a CreERT2 fusion gene (Cre-ERT2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain), an SV40 polyA signal, and an frt-flanked neo cassette into the initiation codon of the Gad2 (glutamic acid decarboxylase 2) gene. This construct was electroporated into 129 x C57BL/6 hybrid embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric mice were bred with white C57BL/6 mice to originate the colony. Mutant mice were bred with Actin-FLPe mice (on a C57BL/6 congenic background (N10)) to remove the neo selection cassette.
Mutations Made By	Z. Josh Huang, Cold Spring Harbor Laboratory

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Research Tools
- Genetics Research
  - Tissue/Cell Markers: Cre-lox System
  - Mutagenesis and Transgenesis
  - Tissue/Cell Markers
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers: neurons
- Cre-lox System
  - Cre Recombinase Expression
  - Cre Recombinase Expression: Inducible
- Neurobiology Research
  - cell marker
Neurobiology Research
- Cre-lox System
  - Cre Recombinase expression in neural tissue

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Gad2^{tm1(cre/ERT2)Zjh}/Gad2⁺
involves: 129S4/SvJae * C57BL/6 * SJL

normal phenotype

no abnormal phenotype detected
- Normal - mice are viable and fertile

References

NIH Neuroscience Blueprint Cre Driver Network. 2009. Cre recombinase-expressing mice generated for the NIH Neuroscience Blueprint Cre Driver Network MGI Direct Data SubmissionMGI: J:151755
Taniguchi H; He M; Wu P; Kim S; Paik R; Sugino K; Kvitsani D; Fu Y; Lu J; Lin Y; Miyoshi G; Shima Y; Fishell G; Nelson SB; Huang ZJ. 2011. A resource of cre driver lines for genetic targeting of GABAergic neurons in cerebral cortex. Neuron 71(6):995-1013PubMed: 21943598 MGI: J:177261

Additional - Gad2^{tm1(cre/ERT2)Zjh} related

Technical Support

Contact Technical Support

Genotyping Protocols
- High Resolution Melting: Tg(ACTFLPe)
- Probe: Gt(rosa)26sor^Tm1Sor Probe
- Probe: Generic FLP
- Standard PCR: Gad2^{tm1(cre/ERT2)Zjh} STD PCR
- Standard PCR: Gt(ROSA)26Sor^tm(CAG)
- Standard PCR: Tg(ACTFLPe)
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred together, to wildtype siblings, or to C57BL/6J mice (Stock No. 000664).
- Additional Breeding and Husbandry Support
Citation
When using the Gad2-CreER mouse strain in a publication, please cite the originating article(s) and include JAX stock #010702 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
	Heterozygous or wildtype for Gad2<tm1(cre/ERT2)Zjh>

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.

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Also Known As: Gad2-CreER

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Gad2tm1(cre/ERT2)Zjh

Allele Symbol: Gad2tm1(cre/ERT2)Zjh

Disease Terms

Research Areas By Genotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Gad2tm1(cre/ERT2)Zjh/Gad2+involves: 129S4/SvJae * C57BL/6 * SJL

normal phenotype

References

Additional - Gad2tm1(cre/ERT2)Zjh related

Genotyping Protocols

Breeding Considerations

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

Gad2^{tm1(cre/ERT2)Zjh}

Allele Symbol: Gad2^{tm1(cre/ERT2)Zjh}

Genotype: Gad2^{tm1(cre/ERT2)Zjh}/Gad2⁺
involves: 129S4/SvJae * C57BL/6 * SJL

Additional - Gad2^{tm1(cre/ERT2)Zjh} related