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| Heterozygous Lgr5-EGFP-IRES-creERT2 mice harbor a Lgr5-EGFP-IRES-CreERT2 "knock-in" allele that both abolishes Lgr5 (leucine rich repeat containing G protein coupled receptor 5) gene function and expresses EGFP and CreERT2 fusion protein from the Lgr5 promoter/enhancer elements. EGFP fluorescence is observed in crypt base columnar cells in small intestine (aka stem cells of the small intestine) and colon. Cre-ERT2 fusion gene activity is inducible; observed in the same cells only following taxomifen administration. These Lgr5-EGFP-IRES-creERT2 mice may be useful for lineage-tracing or marking Lgr5-expressing stem cells of the small intestine (and other Lgr5-expressing cells including pre-malignant mouse adenomas, colon cancer cells, epithelial stem cells of the stomach gland, basal epithelial layer stem cells of the mammary glands, and hair follicle stem cells). | |||||||||||||||
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Description
Strain Information
Type Congenic; Targeted Mutation; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Additional information on Congenic nomenclature. Mating System +/+ sibling x Heterozygote (Female x Male) 11-AUG-09 Species laboratory mouse Donating Investigator Hans Clevers, Hubrecht Institute Important Note
The donating investigator reports variegated expression of the Lgr5-EGFP-IRES-CreERT2 transgene in the small intestine and colon (something which may happen often with genes that are expressed early during intestinal development). This variegated expression is advantageous for performing clonal lineage tracing and sorting intestinal stem cells, but may have limitations for more quantitative studies such as Lgr5-Cre driven knockout strategies.Description
While homozygous mice are not viable, heterozygous Lgr5-EGFP-IRES-CreERT2 mice are viable and fertile; harboring a Lgr5-EGFP-IRES-creERT2 "knock-in" allele that both abolishes Lgr5 (Gpr49) gene function and expresses EGFP and CreERT2 fusion protein from the Lgr5 promoter/enhancer elements. EGFP fluorescence is observed in crypt base columnar cells in small intestine (aka stem cells of the small intestine) and colon. Cre-ERT2 fusion gene activity is inducible; observed in the same cells only following taxomifen administration. EGFP or inducible CreERT2 expression may also be observed in other Lgr5-expressing cell types (including pre-malignant mouse adenomas, colon cancer cells, epithelial stem cells of the stomach gland, basal epithelial layer stem cells of the mammary glands, and hair follicle stem cells).The donating investigator reports variegated expression of the Lgr5-EGFP-IRES-CreERT2 transgene in the small intestine and colon (something which may happen often with genes that are expressed early during intestinal development). This variegated expression is advantageous for performing clonal lineage tracing and sorting intestinal stem cells, but may have limitations for more quantitative studies such as Lgr5-Cre driven knockout strategies.
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. As such, when Lgr5-EGFP-IRES-creERT2 mice are bred with mice containing loxP-flanked sequence of interest, tamoxifen-inducible, Cre-mediated recombination will result in deletion of the floxed sequences in the Lgr5-expressing cells of the offspring.
Development
The EGFP-IRES-creERT2 targeting construct was designed to insert an enhanced green fluorescent protein sequence (EGFP), an internal ribosome entry site (IRES), a CreERT2 fusion gene (Cre-ERT2; Cre recombinase fused to a G400V/M543A/L544A triple mutation of the human estrogen receptor ligand binding domain), a polyA signal, and a loxP-flanked neo cassette into the first ATG codon of the targeted gene. This construct was inserted into the targeted gene via electroporation into male 129P2/OlaHsd-derived IB10/E14IB10 embryonic stem (ES) cells. Correctly targeted ES cells were injected into recipient blastocysts and chimeric males were bred with C57BL/6 females. Mutant mice were then crossed to EIIa-cre mice (C57BL/6 genetic background; see Stock No. 003724) to remove the neo selection cassette. The resulting Lgr5-EGFP-IRES-creERT2 mice (floxed-neo removed) were then backcrossed to C57BL/6J for at least 10 generations before arriving at The Jackson Laboratory. Upon arrival the mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony.
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
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Additional Web Information
Fluorescent Proteins/lacZ Systems
Introduction to Cre-lox technology
Phenotype
Phenotype Information
assigned by genotype
The following phenotype information may relate to a genetic background differing from this JAX® Mice strain.
Lgr5tm1(cre/ESR1)Cle/Lgr5tm1(cre/ESR1)Cle
involves: 129P2/OlaHsd * C57BL/6 * FVB/N
- no phenotypic analysis
- *normal* no phenotypic analysis (MGI Ref ID J:127123)
This mouse can be used to support research in many areas including:
Cancer Research
Other
Research Tools
Cancer Research
Cre-lox System
Cre Recombinase Expression: Inducible
Developmental Biology Research
Cre-lox System
transplantation marker for embryonic and adult tissue
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers: Cre-lox System
Tissue/Cell Markers: multiple
Tissue/Cell Markers: transplantation marker for embryonic and adult tissue
Toxicology Research
xenograft/transplant host
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Lgr5tm1(cre/ESR1)Cle | ||
|---|---|---|---|
| Allele Name | targeted mutation 1, Hans Clevers | ||
| Allele Type | Targeted (knock-in) | ||
| Common Name(s) | Lgr5 EGFP-IRES-creERT2; Lgr5-cre; | ||
| Strain of Origin | 129P2/OlaHsd | ||
| ES Cell Line Name | IB10/E14IB10 | ||
| ES Cell Line Strain | 129P2/OlaHsd | ||
| Gene Symbol and Name | Lgr5, leucine rich repeat containing G protein coupled receptor 5 | ||
| Chromosome | 10 | ||
| Gene Common Name(s) | FEX; G protein-coupled receptor 49; GPR49; GPR67; GRP49; Gpr49; HG38; MGC117008; | ||
| Inducible Note | Lgr5 | ||
| Molecular Note | An EGFP-IRES-creESR1 cassette was inserted into exon 1 of the gene along with a loxP-flanked neomycin cassette. The neomycin gene was then excised in vivo by crossing with cre-deleter strain. GFP expression is driven by the endogenous promoter. Cre expression is induced in Lgr5 expressing cells by tamoxofen treatment. [MGI Ref ID J:127123] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Lgr5tm1(cre/ESR1)Cle STD PCR, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
Barker N; van Es JH; Kuipers J; Kujala P; van den Born M; Cozijnsen M; Haegebarth A; Korving J; Begthel H; Peters PJ; Clevers H. 2007. Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449(7165):1003-7. [PubMed: 17934449] [MGI Ref ID J:127123]
Additional References
Lgr5tm1(cre/ESR1)Cle relatedBarker N; Ridgway RA; van Es JH; van de Wetering M; Begthel H; van den Born M; Danenberg E; Clarke AR; Sansom OJ; Clevers H. 2009. Crypt stem cells as the cells-of-origin of intestinal cancer. Nature 457(7229):608-11. [PubMed: 19092804] [MGI Ref ID J:144216]
Jaks V; Barker N; Kasper M; van Es JH; Snippert HJ; Clevers H; Toftgard R. 2008. Lgr5 marks cycling, yet long-lived, hair follicle stem cells. Nat Genet 40(11):1291-9. [PubMed: 18849992] [MGI Ref ID J:143598]
van der Flier LG; van Gijn ME; Hatzis P; Kujala P; Haegebarth A; Stange DE; Begthel H; van den Born M; Guryev V; Oving I; van Es JH; Barker N; Peters PJ; van de Wetering M; Clevers H. 2009. Transcription factor achaete scute-like 2 controls intestinal stem cell fate. Cell 136(5):903-12. [PubMed: 19269367] [MGI Ref ID J:148715]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, heterozygous mice may be bred together, to wildtype siblings, or to C57BL/6J inbred mice (Stock No. 000664). Homozygous mice are not viable. Mating System +/+ sibling x Heterozygote (Female x Male) 11-AUG-09 Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
| Pricing for USA, Canada and Mexico shipping destinations |
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Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $243.50 Female or Male Heterozygous for Lgr5tm1(cre/ESR1)Cle
Pairs /Price (US dollars $) Pair Genotype $297.85 Heterozygous for Lgr5tm1(cre/ESR1)Cle x Wild-type for Lgr5tm1(cre/ESR1)Cle $297.85 Wild-type for Lgr5tm1(cre/ESR1)Cle x Heterozygous for Lgr5tm1(cre/ESR1)Cle
Additional Supply Details
| Pricing for International shipping destinations |
|
Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $316.60 Female or Male Heterozygous for Lgr5tm1(cre/ESR1)Cle
Pairs /Price (US dollars $) Pair Genotype $387.30 Heterozygous for Lgr5tm1(cre/ESR1)Cle x Wild-type for Lgr5tm1(cre/ESR1)Cle $387.30 Wild-type for Lgr5tm1(cre/ESR1)Cle x Heterozygous for Lgr5tm1(cre/ESR1)Cle
Additional Supply Details
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Supply Details
| Standard Supply | Repository-Live. A collection of over 1000 strains maintained as live colonies. Individual colonies are sized to meet current customer demand. Delivery for orders of 10 mice or less ranges on average from one to eight weeks; mice are generally shipped between four to six weeks of age with a maximum shipping age of approximately nine weeks. Colony sizes do not generally support stringent age specifications for large volumes of mice; however custom orders and larger quantities of mice are easily arranged. Estimated ship dates for all orders provided within two business days following order placement. |
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| Supply Notes |
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| Important Note | |
| The donating investigator reports variegated expression of the Lgr5-EGFP-IRES-CreERT2 transgene in the small intestine and colon (something which may happen often with genes that are expressed early during intestinal development). This variegated expression is advantageous for performing clonal lineage tracing and sorting intestinal stem cells, but may have limitations for more quantitative studies such as Lgr5-Cre driven knockout strategies. | |
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| USA, Canada and Mexico - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
| International - Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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