Overview

Also Known As: K5-CreER^T2 knock-in

Krt5-CreERT2 knock-in mice have both endogenous gene and tamoxifen-inducible Cre recombinase expression directed to basal epithelial cells by the endogenous promoter/enhancer elements of the keratin 5 locus. These mice may be useful in generating conditional alleles for studying basal cells, cancer and epidermolysis bullosa.

Donating Investigator

Brigid LM Hogan, Duke University Medical Center

Genetic overview

Genetic Background	Generation
	?+pN2F5 (2019-03-01 00:00:00)

Krt5^{tm1.1(cre/ERT2)Blh}

Allele Type	Gene Symbol	Gene Name
Targeted (Recombinase-expressing, Inducible)	Krt5	keratin 5

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Research Applications

Cancer Research
Research Tools

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Base Price

Starting at:

$255.00 Domestic price for female 4-week

333.51 Domestic price for breeder pair

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Details

Detailed Description

The keratin 5 gene (Krt5) encodes cytokeratin-5 or K5, a type II cytokeratin specifically expressed in basal epithelial cells (including epidermis, salivary gland and digestive tract). K5 is a biomarker for several different types of cancer (including breast and lung).

The Krt5-CreERT2 knock-in allele has an IRES sequence and a tamoxifen-inducible Cre recombinase (CreER^T2) inserted in the 3' UTR of Krt5. As such, Krt5-CreERT2 mice have both endogenous gene and CreER^T2 expression directed to basal epithelial cells by the endogenous promoter/enhancer elements of the keratin 5 locus. CreER^T2 fusion gene activity is inducible; observed following tamoxifen administration. When Krt5-CreERT2 mice are bred with mice containing loxP-flanked sequences, tamoxifen-inducible Cre-mediated recombination will result in deletion of floxed sequences in the Krt5-expressing cells of the double mutant offspring.

Specifically, tamoxifen-inducible Cre recombinase activity recapitulates the endogenous Krt5 expression pattern. Recombination efficiency is reported to be high in the basal layer of the epidermis, prostate and esophagus, but only 25% in the basal cells of the trachea (which may reflect the lower Krt5 expression in this tissue). The donating investigator did not characterize if Cre recombinase is expressed prior to tamoxifen exposure. Heterozygotes are viable and fertile with no reported abnormalities. To date (May 2016), it has not been attempted to make this strain homozygous.

For example, when Krt5-CreERT2 mice are bred to have a cre-conditional knockout of suppressor of cytokine signals 3 (Socs3^flox; Stock No. 010944) and fluorescent cre-reporter (R26-LSL-EYFP; Stock No. 006148), the resulting triple mutant mice allow tamoxifen-inducible deletion of Socs3 in basal cells and activation of YFP expression as a lineage tracer. Such animals are useful for studying negative feedback regulation / inhibition of the JAK/STAT3 pathway.

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

The Krt5-CreERT2 knock-in allele (K5-CreER, K5-CreER^T2 or K5CREER) was created by the laboratory of Dr. Brigid LM Hogan (Duke University Medical Center) to have an IRES-CreER^T2 inserted into the 3' UTR of the keratin 5 gene (Krt5) on chromosome 15. The specific details are below.

The targeting vector contained, from 5' to 3', an internal ribosome entry site (IRES), 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) and an frt-flanked neomycin selection cassette. This construct was electroporated into 129S6/SvEvTac-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with B6;SJL-Tg(ACTFLPe)9205Dym/J females (Stock No. 003800) to remove the neo selection cassette. The resulting Krt5-CreERT2 colony was maintained by breeding to C57BL/6NCrl wildtype mice for at least eight generations (and the FLPe transgene was removed) prior to sending male mice to The Jackson Laboratory Repository in 2016. Upon arrival, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6N oocytes (Stock No. 005304).

Expression Data

Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of Expression	Tamoxifen-inducible Cre recombinase is expressed in basal epithelial cells.

Control Suggestions

Wild-type from the colony

Approximate Controls

005304 C57BL/6NJ

Additional Information

Considerations for Choosing Controls

Selected References

Van Keymeulen A; Rocha AS; Ousset M; Beck B; Bouvencourt G; Rock J; Sharma N; Dekoninck S; Blanpain C. 2011. Distinct stem cells contribute to mammary gland development and maintenance. Nature 479(7372):189-93PubMed: 21983963 MGI: J:180048

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Genetics

Krt5^{tm1.1(cre/ERT2)Blh}

Allele Symbol: Krt5^{tm1.1(cre/ERT2)Blh}

Allele Name	targeted mutation 1.1, Brigid L Hogan
Allele Type	Targeted (Recombinase-expressing, Inducible)
Allele Synonym(s)	targeted mutation 1.1, Brigid L Hogan; Krt5^{tm1.1(cre/ERT2)Blh}
Gene Symbol and Name	Krt5, keratin 5
Gene Synonym(s)	Tfip8; K5; keratin complex 2, basic, gene 5; expressed sequence AW146334; RIKEN cDNA 3300001P10 gene; 3300001P10Rik; Krt2-5; Krt2-5; KRT5A; EBS2; Sak57; CK5; tuftelin interacting protein 8; Tfip8; 3300001P10Rik; AW146334; DDD; DDD1
Expressed Gene	cre/ERT2, Cre recombinase and estrogen receptor 1 (human) fusion gene,
Site of Expression	Tamoxifen-inducible Cre recombinase is expressed in basal epithelial cells.
Strain of Origin	129S6/SvEvTac
Chromosome	15
Molecular Note	An IRES-cre/ERT2 fragment was inserted into the 3'UTR of the Krt5 gene along with an FRT flanked neomycin selection cassette. The selection cassette was subsequenty excised from properly targeted mice using flp recombinase.

Disease/Phenotype

Disease Terms

Research Areas By Genotype

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

Cancer Research
- Cre-lox System
  - Cre recombinase expression
  - Cre Recombinase expression: inducible
Research Tools
- Cancer Research
  - Cre-lox System
- Cre-lox System
  - Cre Recombinase Expression
  - Cre Recombinase Expression: Inducible
- Developmental Biology Research
  - Cre-lox System
- Genetics Research
  - Mutagenesis and Transgenesis: Cre-lox System
  - Tissue/Cell Markers
  - Tissue/Cell Markers: Cre-lox System

Mammalian Phenotype Terms by Genotype

References

Van Keymeulen A; Rocha AS; Ousset M; Beck B; Bouvencourt G; Rock J; Sharma N; Dekoninck S; Blanpain C. 2011. Distinct stem cells contribute to mammary gland development and maintenance. Nature 479(7372):189-93PubMed: 21983963 MGI: J:180048

Additional - Krt5^{tm1.1(cre/ERT2)Blh} related

Technical Support

Contact Technical Support

Genotyping Protocols
- Separated PCR:Krt5^{tm1.1(cre/ERT2)Blh}
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred together, to wildtype 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
- Wild-type x Heterozygote
- Heterozygote x Wild-type
Citation
When using the K5-CreER^T2 knock-in mouse strain in a publication, please cite the originating article(s) and include JAX stock #029155 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX18 (Maximum)

Pricing & Availability

Available

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Breeder Pair

Sex

Genotype

Price

Female
Male

Cryorecovery - Pricing

Service	Genotype	Price
	Heterozygous or wildtype for Krt5<tm1.1(cre/ERT2)Blh>

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.

Related Products and Services

Frozen Mouse Embryo	B6N.129S6(Cg)-Krt5<tm1.1(cre/ERT2)Blh>/J	$2595.00
Frozen Mouse Embryo	B6N.129S6(Cg)-Krt5<tm1.1(cre/ERT2)Blh>/J	$2595.00
Frozen Mouse Embryo	B6N.129S6(Cg)-Krt5<tm1.1(cre/ERT2)Blh>/J	$3373.50
Frozen Mouse Embryo	B6N.129S6(Cg)-Krt5<tm1.1(cre/ERT2)Blh>/J	$3373.50

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Also Known As: K5-CreERT2 knock-in

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Krt5tm1.1(cre/ERT2)Blh

Allele Symbol: Krt5tm1.1(cre/ERT2)Blh

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 - Krt5tm1.1(cre/ERT2)Blh related

Genotyping Protocols

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Breeder Pair

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

Also Known As: K5-CreER^T2 knock-in

Krt5^{tm1.1(cre/ERT2)Blh}

Allele Symbol: Krt5^{tm1.1(cre/ERT2)Blh}

Additional - Krt5^{tm1.1(cre/ERT2)Blh} related