Overview

The Calb2-CreERT2 (CR-CreER or CR-CreERT2) knock-in allele was designed to both abolish Calb2 (calbindin 2; also called calretinin or CR) gene function and direct CreER^T2 fusion protein expression to calretinin positive interneurons in the brain by the endogenous Calb2 promoter/enhancer elements. Cre-ERT2 fusion gene activity is inducible by tamoxifen administration.

Donating Investigator

Z. Josh Huang, Cold Spring Harbor Laboratory

Genetic overview

Genetic Background	Generation

Calb2^{tm2.1(cre/ERT2)Zjh}

Allele Type	Gene Symbol	Gene Name
Targeted (Recombinase-expressing, Inducible)	Calb2	calbindin 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 Calb2-CreERT2 (CR-CreER or CR-CreERT2) knock-in allele was designed to both abolish Calb2 (calbindin 2; also called calretinin or CR) gene function and expresses CreER^T2 fusion protein from the Calb2 promoter/enhancer elements. Cre-ERT2 fusion gene activity is inducible; observed following tamoxifen administration. As such, when CR-CreERT2 mice are bred with mice containing loxP-flanked sequences, tamoxifen-inducible Cre-mediated recombination will result in deletion of the floxed sequences in the CR-expressing cells of the offspring.

Heterozygous mice are viable, fertile, normal in size and do not display any gross physical or behavioral abnormalities. The donating investigator reports that the phenotype of CR-CreER homozygous mice has not been assessed. However, CR-CreER homozygotes may be expected to exhibit the same phenotype as mice homozygous for other null mutations of this gene on a similar genetic background (impaired motor coordination, abnormal nervous system electrophysiology, and impaired long-term potentiation induction in dentate gyrus). Calretinin mRNA or protein expression from the CR-CreER mutant allele was not determined. The donating investigator also reports tamoxifen-inducible Cre recombinase activity recapitulates the endogenous calretinin expression pattern, but with moderate efficiency of induction. In olfactory bulb, the induction efficiency is higher. Tamoxifen-inducible Cre recombinase activity is not observed prior to tamoxifen treatment. Following tamoxifen administration, Cre recombinase activity is observed in calretinin positive interneurons in the brain. The donating investigators did not examine cre expression in tissues other than brain.

For characterization information, see images at the Allen Institute for Brain Science website (Calb2-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 Calb2 (calbindin 2; also called calretinin or CR) locus. This construct was electroporated into C57BL/6-derived Bruce4 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric mice were bred with white C57BL/6 mice (harboring the Tyr^c-2J mutation) 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 Calb2-CreERT2 (CR-CreER or CR-CreERT2) mice were subsequently bred to white C57BL/6 mice for a few additional generations (and the FLPe transgene was removed) prior to sending to The Jackson Laboratory Repository. Upon arrival, mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the Calb2-CreERT2 colony. These Calb2-CreERT2 mice may still be segregating at the Tyr locus.

Expression Data

Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of Expression	Following tamoxifen administration, Cre recombinase activity is observed in calretinin positive interneurons in the brain.

Control Suggestions

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

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

Calb2^{tm2.1(cre/ERT2)Zjh}

Allele Symbol: Calb2^{tm2.1(cre/ERT2)Zjh}

Allele Name	targeted mutation 2.1, Z Josh Huang
Allele Type	Targeted (Recombinase-expressing, Inducible)
Allele Synonym(s)	targeted mutation 2.1, Z Josh Huang; Calb2^{tm2.1(cre/ERT2)Zjh}
Gene Symbol and Name	Calb2, calbindin 2
Gene Synonym(s)	CAB29; CAL2; CR; calretinin
Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of Expression	Following tamoxifen administration, Cre recombinase activity is observed in calretinin positive interneurons in the brain.
Strain of Origin	C57BL/6
Chromosome	8
Molecular Note	A targeting vector was designed to insert a CreERT2 fusion gene, an SV40 polyA signal, and anFRT-flanked neo cassette into the initiation codon of the Calb2 (calbindin 2; also called calretinin or CR) locus. This construct was electroporated into C57BL/6-derived embryonic stem (ES) cells. Mutant mice were bred with Actin-FLPe mice to remove the neo 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: Calb2^{tm2.1(cre/ERT2)Zjh}/Calb2^{tm2.1(cre/ERT2)Zjh}
involves: C57BL/6 * SJL

behavior/neurological phenotype

impaired coordination
- homozygotes are expected to show defects similar to those in mice with null mutations in Calb2, such as impaired motor coordination

nervous system phenotype

abnormal nervous system electrophysiology
- homozygotes are expected to show defects similar to those in mice with null mutations in Calb2

reduced long term potentiation
- homozygotes are expected to show defects similar to those in mice with null mutations in Calb2, including impaired induction of LTP in the dentate gyrus

References

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 - Calb2^{tm2.1(cre/ERT2)Zjh} related

Technical Support

Contact Technical Support

Genotyping Protocols
- Probe:Gt(rosa)26sor^Tm1SorProbe
- Separated MCA:Calb2^{tm2.1(cre/ERT2)Zjh}
- Separated PCR:Calb2^{tm2.1(cre/ERT2)Zjh}
- 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). The donating investigator has not characterized the homozygous phenotype to date (December 2010).
- Additional Breeding and Husbandry Support
Citation
When using the B6(Cg)-Calb2^{tm2.1(cre/ERT2)Zjh}/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #013730 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 Calb2<tm2.1(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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The Jackson Laboratory has rigorous genetic quality control and mutant gene genotyping programs to ensure the genetic background of JAX® Mice strains as well as the genotypes of strains with identified molecular mutations. JAX® Mice strains are only made available to researchers after meeting our standards. However, the phenotype of each strain may not be fully characterized and/or captured in the strain data sheets. Therefore, we cannot guarantee a strain's phenotype will meet all expectations. To ensure that JAX® Mice will meet the needs of individual research projects or when requesting a strain that is new to your research, we suggest ordering and performing tests on a small number of mice to determine suitability for your particular project.

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Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Selected References

Calb2tm2.1(cre/ERT2)Zjh

Allele Symbol: Calb2tm2.1(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: Calb2tm2.1(cre/ERT2)Zjh/Calb2tm2.1(cre/ERT2)Zjhinvolves: C57BL/6 * SJL

behavior/neurological phenotype

nervous system phenotype

References

Additional - Calb2tm2.1(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

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

Calb2^{tm2.1(cre/ERT2)Zjh}

Allele Symbol: Calb2^{tm2.1(cre/ERT2)Zjh}

Genotype: Calb2^{tm2.1(cre/ERT2)Zjh}/Calb2^{tm2.1(cre/ERT2)Zjh}
involves: C57BL/6 * SJL

Additional - Calb2^{tm2.1(cre/ERT2)Zjh} related