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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 tamoxifen 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). | |||||||||||||||||||
- Description
- Phenotype
- Genes & alleles
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Description
Strain Information
Former Names B6.129P2-Lgr5tm1(cre/ESR1)Cle/J (Changed: 30-MAR-10 ) 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 Generation N11+N1F3 (31-JAN-11)
Generation DefinitionsDonating 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 tamoxifen 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 donating investigator reported that the resulting Lgr5-EGFP-IRES-creERT2 mice (floxed-neo removed) were then backcrossed to C57BL/6J (see SNP note below) for at least 4 generations. Next, Lgr5-EGFP-IRES-creERT2 mice were bred with Rosa26-lacZ mice (C57BL/6 genetic background; see Stock No. 003474). Mice with both mutations were further backcrossed for more than 6 generations. Upon arrival at The Jackson Laboratory, the mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish the colony. Mice were then selectively bred to remove the Rosa26-lacZ allele.A 32 SNP (single nucleotide polymorphism) panel analysis, with 27 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository. While the 27 markers throughout the genome suggested a C57BL/6 genetic background, 2 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6J ; C57BL/6N genetic background.
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
View Fluorescent Protein Strains (345 strains)
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/ERT2)Cle/Lgr5tm1(cre/ERT2)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:
GFP relatedCancer Research
Other
tumor metastasis
Internal/Organ Research
Gastrointestinal Defects
Research Tools
Cancer Research
Cre-lox System
Cre Recombinase Expression
Cre Recombinase Expression: Inducible
Developmental Biology Research
Cre-lox System
transplantation marker for embryonic and adult tissue
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers
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
cre relatedResearch Tools
Fluorescent Proteins
Cre-lox System
Genetics Research
Mutagenesis and Transgenesis
Mutagenesis and Transgenesis: Cre-lox System
Genes & Alleles
Gene & Allele Information provided by MGI
| Allele Symbol | Lgr5tm1(cre/ERT2)Cle | ||
|---|---|---|---|
| Allele Name | targeted mutation 1, Hans Clevers | ||
| Allele Type | Targeted (knock-in) | ||
| Common Name(s) | Lgr5 EGFP-IRES-creERT2; Lgr5-EGFP; Lgr5-EGFP-IRES-creERT2; Lgr5-cre; Lgr5tm1(cre/ESR1)Cle; | ||
| Mutation Made By | Nick Barker, Hubrecht Institute | ||
| Strain of Origin | 129P2/OlaHsd | ||
| ES Cell Line Name | IB10/E14IB10 | ||
| ES Cell Line Strain | 129P2/OlaHsd | ||
| Site of Expression | Variegated EGFP fluorescence is observed in crypt base columnar cells in small intestine (stem cells of the small intestine) and colon. | ||
| Expressed Gene | cre, cre recombinase, bacteriophage P1 | ||
| Cre recombinase is an enzyme derived from the bacteriophage P1 that specifically recognizes loxP sites. Cre has been shown to effectively mediate the excision of DNA located between loxP sites. After the excision event, the DNA ends recombine leaving a single loxP site in place of the intervening sequence. | |||
| Expressed Gene | GFP, Green Fluorescent Protein, jellyfish | ||
| Green Fluorescent Protein (GFP), derived from the jellyfish Aequorea victoria, is a versatile reporter molecule which has found use in many biological applications. In some constructs the original molecule has been modified in order to enhance its fluorescence intensity (EGFP, enhanced GFP). When utilized in a transgenic construct, tissue expressing sufficient amounts of GFP will fluoresce when exposed to a 488 nm light source. | |||
| Driver Note | Lgr5 | ||
| Inducible Note | induced by tamoxifen | ||
| Molecular Note | An EGFP-IRES-creERT2 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 tamoxifen treatment. [MGI Ref ID J:127123] | ||
| 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; | ||
Genotyping
Genotyping Information
Genotyping Protocols
Lgr5tm1(cre/ERT2)Cle, Melt Curve Analysis
Lgr5tm1(cre/ESR1)Cle DI's Assay, Standard PCR
Lgr5tm1(cre/ESR1)Cle STD PCR, Standard PCR
Helpful Links
Genotyping resources and troubleshooting
References
References provided by MGI
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/ERT2)Cle relatedArnold K; Sarkar A; Yram MA; Polo JM; Bronson R; Sengupta S; Seandel M; Geijsen N; Hochedlinger K. 2011. Sox2(+) adult stem and progenitor cells are important for tissue regeneration and survival of mice. Cell Stem Cell 9(4):317-29. [PubMed: 21982232] [MGI Ref ID J:177655]
Barker N; Huch M; Kujala P; van de Wetering M; Snippert HJ; van Es JH; Sato T; Stange DE; Begthel H; van den Born M; Danenberg E; van den Brink S; Korving J; Abo A; Peters PJ; Wright N; Poulsom R; Clevers H. 2010. Lgr5(+ve) stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro. Cell Stem Cell 6(1):25-36. [PubMed: 20085740] [MGI Ref ID J:157075]
Barker 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]
Chai R; Xia A; Wang T; Jan TA; Hayashi T; Bermingham-McDonogh O; Cheng AG. 2011. Dynamic expression of lgr5, a wnt target gene, in the developing and mature mouse cochlea. J Assoc Res Otolaryngol 12(4):455-69. [PubMed: 21472479] [MGI Ref ID J:173579]
Gerbe F; van Es JH; Makrini L; Brulin B; Mellitzer G; Robine S; Romagnolo B; Shroyer NF; Bourgaux JF; Pignodel C; Clevers H; Jay P. 2011. Distinct ATOH1 and Neurog3 requirements define tuft cells as a new secretory cell type in the intestinal epithelium. J Cell Biol 192(5):767-80. [PubMed: 21383077] [MGI Ref ID J:170320]
Grachtchouk M; Pero J; Yang SH; Ermilov AN; Michael LE; Wang A; Wilbert D; Patel RM; Ferris J; Diener J; Allen M; Lim S; Syu LJ; Verhaegen M; Dlugosz AA. 2011. Basal cell carcinomas in mice arise from hair follicle stem cells and multiple epithelial progenitor populations. J Clin Invest 121(5):1768-81. [PubMed: 21519145] [MGI Ref ID J:173930]
Itzkovitz S; Lyubimova A; Blat IC; Maynard M; van Es J; Lees J; Jacks T; Clevers H; van Oudenaarden A. 2011. Single-molecule transcript counting of stem-cell markers in the mouse intestine. Nat Cell Biol 14(1):106-14. [PubMed: 22119784] [MGI Ref ID J:178925]
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]
Kasper M; Jaks V; Are A; Bergstrom A; Schwager A; Barker N; Toftgard R. 2011. Wounding enhances epidermal tumorigenesis by recruiting hair follicle keratinocytes. Proc Natl Acad Sci U S A 108(10):4099-104. [PubMed: 21321199] [MGI Ref ID J:170330]
Kim TH; Shivdasani RA. 2011. Notch signaling in stomach epithelial stem cell homeostasis. J Exp Med 208(4):677-88. [PubMed: 21402740] [MGI Ref ID J:176833]
Montgomery RK; Carlone DL; Richmond CA; Farilla L; Kranendonk ME; Henderson DE; Baffour-Awuah NY; Ambruzs DM; Fogli LK; Algra S; Breault DT. 2011. Mouse telomerase reverse transcriptase (mTert) expression marks slowly cycling intestinal stem cells. Proc Natl Acad Sci U S A 108(1):179-84. [PubMed: 21173232] [MGI Ref ID J:169012]
Qiu W; Wang X; Leibowitz B; Liu H; Barker N; Okada H; Oue N; Yasui W; Clevers H; Schoen RE; Yu J; Zhang L. 2010. Chemoprevention by nonsteroidal anti-inflammatory drugs eliminates oncogenic intestinal stem cells via SMAC-dependent apoptosis. Proc Natl Acad Sci U S A 107(46):20027-32. [PubMed: 21041628] [MGI Ref ID J:166608]
Sato T; van Es JH; Snippert HJ; Stange DE; Vries RG; van den Born M; Barker N; Shroyer NF; van de Wetering M; Clevers H. 2011. Paneth cells constitute the niche for Lgr5 stem cells in intestinal crypts. Nature 469(7330):415-8. [PubMed: 21113151] [MGI Ref ID J:168728]
Sei Y; Lu X; Liou A; Zhao X; Wank SA. 2011. A stem cell marker-expressing subset of enteroendocrine cells resides at the crypt base in the small intestine. Am J Physiol Gastrointest Liver Physiol 300(2):G345-56. [PubMed: 21088235] [MGI Ref ID J:168494]
Snippert HJ; Haegebarth A; Kasper M; Jaks V; van Es JH; Barker N; van de Wetering M; van den Born M; Begthel H; Vries RG; Stange DE; Toftgard R; Clevers H. 2010. Lgr6 marks stem cells in the hair follicle that generate all cell lineages of the skin. Science 327(5971):1385-9. [PubMed: 20223988] [MGI Ref ID J:157895]
Snippert HJ; van der Flier LG; Sato T; van Es JH; van den Born M; Kroon-Veenboer C; Barker N; Klein AM; van Rheenen J; Simons BD; Clevers H. 2010. Intestinal crypt homeostasis results from neutral competition between symmetrically dividing Lgr5 stem cells. Cell 143(1):134-44. [PubMed: 20887898] [MGI Ref ID J:164644]
Takeda N; Jain R; Leboeuf MR; Wang Q; Lu MM; Epstein JA. 2011. Interconversion Between Intestinal Stem Cell Populations in Distinct Niches. Science :. [PubMed: 22075725] [MGI Ref ID J:178296]
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):189-93. [PubMed: 21983963] [MGI Ref ID J:180048]
Vandussen KL; Carulli AJ; Keeley TM; Patel SR; Puthoff BJ; Magness ST; Tran IT; Maillard I; Siebel C; Kolterud A; Grosse AS; Gumucio DL; Ernst SA; Tsai YH; Dempsey PJ; Samuelson LC. 2012. Notch signaling modulates proliferation and differentiation of intestinal crypt base columnar stem cells. Development 139(3):488-97. [PubMed: 22190634] [MGI Ref ID J:179656]
Yan KS; Chia LA; Li X; Ootani A; Su J; Lee JY; Su N; Luo Y; Heilshorn SC; Amieva MR; Sangiorgi E; Capecchi MR; Kuo CJ. 2012. The intestinal stem cell markers Bmi1 and Lgr5 identify two functionally distinct populations. Proc Natl Acad Sci U S A 109(2):466-71. [PubMed: 22190486] [MGI Ref ID J:179994]
de Lau W; Barker N; Low TY; Koo BK; Li VS; Teunissen H; Kujala P; Haegebarth A; Peters PJ; van de Wetering M; Stange DE; van Es JE; Guardavaccaro D; Schasfoort RB; Mohri Y; Nishimori K; Mohammed S; Heck AJ; Clevers H. 2011. Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling. Nature 476(7360):293-7. [PubMed: 21727895] [MGI Ref ID J:174842]
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
| Pricing for USA, Canada and Mexico shipping destinations |
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Live Mice
Price (US dollars $) Gender Genotypes Provided Individual Mouse $261.00 Female or Male Heterozygous for Lgr5tm1(cre/ERT2)Cle
Pairs /Price (US dollars $) Pair Genotype $321.00 Heterozygous for Lgr5tm1(cre/ERT2)Cle x Wild-type for Lgr5tm1(cre/ERT2)Cle $321.00 Wild-type for Lgr5tm1(cre/ERT2)Cle x Heterozygous for Lgr5tm1(cre/ERT2)Cle Standard Supply
Repository-Live. The Repository Strains represent an exclusive set of over 1500 unique mouse models maintained at The Jackson Laboratory to support a vast array of research areas. The breeding colonies for Repository Strains provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. We treat orders for these strains as custom orders. Within 2 business days, we respond to each availability inquiry or order with various delivery options. Repository Strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping.
| Pricing for International shipping destinations |
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Live Mice
Price (US dollars $) Gender Genotypes Provided Individual Mouse $339.30 Female or Male Heterozygous for Lgr5tm1(cre/ERT2)Cle
Pairs /Price (US dollars $) Pair Genotype $417.30 Heterozygous for Lgr5tm1(cre/ERT2)Cle x Wild-type for Lgr5tm1(cre/ERT2)Cle $417.30 Wild-type for Lgr5tm1(cre/ERT2)Cle x Heterozygous for Lgr5tm1(cre/ERT2)Cle Standard Supply
Repository-Live. The Repository Strains represent an exclusive set of over 1500 unique mouse models maintained at The Jackson Laboratory to support a vast array of research areas. The breeding colonies for Repository Strains provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. We treat orders for these strains as custom orders. Within 2 business days, we respond to each availability inquiry or order with various delivery options. Repository Strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping.
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Standard Supply
Repository-Live. The Repository Strains represent an exclusive set of over 1500 unique mouse models maintained at The Jackson Laboratory to support a vast array of research areas. The breeding colonies for Repository Strains provide mice for both large and small orders and fluctuate in size depending on current demand for each strain. We treat orders for these strains as custom orders. Within 2 business days, we respond to each availability inquiry or order with various delivery options. Repository Strains typically are delivered at 4 to 8 weeks of age and will not exceed 12 weeks of age on the day of shipping.
General Supply Notes
- This strain is included in the Induced Mutant Resource Colony collection.
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 000664 C57BL/6J | ||
| Considerations for Choosing Controls | ||
| Control Pricing Information for Genetically Engineered Mutant Strains. | ||
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.
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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.Ordering Information
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