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| These Six2-TGCtg mice express an EGFPCre fusion protein directed to nephron progenitor population cap mesenchyme from the onset of metanephric kidney development by the Six2 (sine oculis-related homeobox 2 homolog (Drosophila)) promoter/enhancer regions within the BAC transgene. Sequence updates published after this transgene was created revealed an upstream Six2 ATG start site that is in-frame with the tTA; thus an apparent and unintended fusion was created with the tTA (80 amino acids potentially added) that renders transgene expression unaffected by tetracycline/doxycycline administration. While not useful as a Tet-Off tool, these Six2-TGCtg mice may be useful as fluorescent or Cre-lox tools for lineage-tracing/marking Six2-expressing cells for studying multipotent nephron progenitor cell populations throughout kidney organogenesis. | |||||||||||||||||||
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Description
Strain Information
Former Names STOCK Tg(Six2-tTA,tetO-EGFP/cre)1Amc/J (Changed: 30-OCT-09 ) Type Mutant Stock; Transgenic; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Species laboratory mouse Donating Investigator Andrew McMahon, Harvard University Important Note
Sequence updates published after this transgene was created revealed an upstream Six2 ATG start site that is in-frame with the tTA; thus an apparent and unintended fusion was created with the tTA (80 amino acids potentially added) that renders transgene expression unaffected by tetracycline/doxycycline administration.Description
Hemizygous Six2-TGCtg mice are viable and fertile, harboring a BAC transgene with a Tet-off-eGFPCre under control of the Six2 promoter/enhancer regions within the BAC transgene. The Tet-off-eGFPCre contains both the tetracycline-controlled transactivator protein (tTA) as well as the tetracycline operator (tetO; also called tetracycline-responsive element [TRE] or tet-operator) upstream of an EGFPCre fusion protein. Sequence updates published after this transgene was created revealed an upstream Six2 ATG start site that is in-frame with the tTA; thus an apparent and unintended fusion was created with the tTA (80 amino acids potentially added) that renders transgene expression unaffected by tetracycline/doxycycline administration. It is not known whether the "mutant" tTA is resistant to tetracycline/doxycycline inactivation and still binds to TRE to activate EGFPCre expression or if the TRE acts as a minimal promoter adjacent to the Six2 promoter in the presence of a nonfunctional tTA. Either way, Tet-independent expression of the eGFPCre fusion protein (EGFP immunofluorescence and direct fluorescence/Cre recombinase activity) is directed to nephron progenitor population cap mesenchyme from the onset of metanephric kidney development. When bred with mice containing a loxP-flanked sequence of interest, Cre-mediated recombination will result in deletion of the floxed sequence(s) in such tissues of the offspring. While not useful as a Tet-Off tool, these Six2-TGCtg mice may be useful as fluorescent or Cre-lox tools for lineage-tracing/marking Six2-expressing cells for studying multipotent nephron progenitor cell populations throughout kidney organogenesis. The donating investigator reports they were unable to produce homozygous mice from hemizygous matings.Development
The ~181 kb C57BL/6J mouse bacterial artificial chromosome (BAC) RPCI23-311C1, containing the entire Six2 locus (and other genes), was modified by targeting a Tet-off-eGFPCre into the ATG start site of the Six2 locus. This inserted a Tet-off cassette (tetracycline-regulated transactivator [tTA], 2xpolyA signal, tetracycline-responsive element [TRE or teto with CMVmin promoter]) followed by an Enhanced Green Fluorescent Protein/Cre Recombinase (EGFP/Cre) fusion protein coding sequence, SV40 polyA signal, and frt-flanked kanamycin cassette. Sequence updates published after this transgene was created revealed an upstream Six2 ATG start site that is in-frame with the tTA; thus an apparent and unintended fusion was created with the tTA (80 amino acids potentially added) that renders transgene expression unaffected by tetracycline/doxycycline administration. The targeted BAC sequences were further modified using FLP recombination to remove the selection cassette and linearized to remove its original vector backbone. The resulting modified BAC was microinjected into CD-1 zygotes. Transgenic founders were obtained and bred to C57BL/6 mice to establish the Six2-TGCtg colony. These mice were subsequently maintained on CD-1;Swiss Webster;C57BL/6 mixed genetic background prior to arrival at The Jackson Laboratory. Upon arrival, transgenic mice were bred with C57BL/6J inbred mice (Stock No. 000664) for at least one generation to establish this colony.
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| Considerations for Choosing Controls | ||
Related Strains
Fluorescent Protein Strains
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009600 B6;129-Six2tm3(EGFP/cre/ERT2)Amc/J
007004 B6.Cg-Tg(Camk2a-tTA)1Mmay/DboJ 003767 B6.Cg-Tg(Eno2tTA)5021Nes/J 003763 B6.Cg-Tg(Eno2tTA)5030Nes/J 005964 B6.Cg-Tg(GFAP-tTA)110Pop/J 002618 B6.Cg-Tg(MMTVtTA)1Mam/J 008284 B6.Cg-Tg(Scg2-tTA)1Jt/J 006361 B6.Cg-Tg(Sp7-tTA,tetO-EGFP/cre)1Amc/J 003563 B6.Cg-Tg(tTALap)5Bjd/J 002709 B6;C3-Tg(TettTALuc)1Dgs/J 003010 B6;CBA-Tg(Camk2a-tTA)1Mmay/J 008344 B6;DBA-Tg(Fos-tTA,Fos-EGFP*)1Mmay Tg(tetO-lacZ,tTA*)1Mmay/J 010573 B6;SJL-Tg(Prl-tTA)6-5Jek/J 008082 B6;SJL-Tg(Tagln-tTA)1Mrab Tg(tetO-Mcpt1)1Mrab/J 005625 FVB-Tg(Pcp2-tTA)3Horr/J 003170 FVB.Cg-Tg(Myh6-tTA)6Smbf/J 006209 FVB.Cg-Tg(Tal1-tTA)19Dgt/J 005942 FVB/N-Tg(Pf4-tTA/VP16)42Kra/J 004937 NOD.Cg-Tg(Ins2-tTA)1Doi/DoiJ 006999 STOCK Dbttm1Geh Tg(tTALap)5Bjd Tg(tetO-DBT)A1Geh/J 003271 STOCK Tg(tTAhCMV)3Bjd/J 003275 STOCK Tg(tetL)1Bjd/J 003274 STOCK Tg(tetNZL)2Bjd/J
Additional Web Information
Fluorescent Proteins/lacZ Systems
Introduction to Cre-lox technology
Tet Expression Systems
Phenotype
Phenotype Information
assigned by genotype
Tg(Six2-EGFP/cre)1Amc/0
involves: C57BL/6J * CD-1 * Swiss Webster
- normal phenotype
- no abnormal phenotype detected (MGI Ref ID J:148455)
- mice are viable and fertile
This mouse can be used to support research in many areas including:
GFP relatedDevelopmental Biology Research
Internal/Organ Defects
kidney
Research Tools
Cre-lox System
Cre Recombinase Expression
Developmental Biology Research
Cre-lox System
Fluorescent Proteins
Genetics Research
Mutagenesis and Transgenesis: Cre-lox System
Tissue/Cell Markers: Cre-lox System
Tissue/Cell Markers: kidney specific marker
Tet Expression Systems
tTA/rtTA Expressing Strains
tTA/rtTA Responsive Strains
cre relatedResearch Tools
Fluorescent Proteins
Cre-lox System
Genetics Research
Mutagenesis and Transgenesis: Cre-lox System
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Tg(Six2-EGFP/cre)1Amc | ||
|---|---|---|---|
| Allele Name | transgene insertion 1, Andrew P McMahon | ||
| Allele Type | Transgenic (Cre/Flp) | ||
| Common Name(s) | Six2-TGC(tg); Six2-TGCtg; TGC BAC transgenic allele; Tet-off-eGFPCre BAC transgenic; Tg(Six2-tTA,tetO-EGFP/cre)1Amc; | ||
| Mutation Made By | Andrew McMahon, Harvard University | ||
| Strain of Origin | CD-1 | ||
| Expressed Gene | tTA, tetracycline-controlled transactivator, E. coli | ||
| The tetracycline-resistance gene (TetR), arose from chemically mutated Escherichia coli genome which was screened for tetracycline dependence (Gossen and Bujard, 1992). TetR was fused at the C-terminus with the viral co-activator, virion protein 16 of the herpes simplex virus (VP-16). The tetracycline-inhibitable transcription factor is a component of a bigenic system that allows doxycycline (a tetracycline analog) regulatable expression of genes that are under the direction of the tetracycline responsive promoter (TetOp)promoter. | |||
| 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. | |||
| Promoter | Six2, sine oculis-related homeobox 2 homolog (Drosophila), mouse, laboratory | ||
| Driver Note | Six2 | ||
| Inducible Note | induced by doxycycline | ||
| Molecular Note | The 181 kb C57BL/6J mouse bacterial artificial chromosome (BAC) RPCI23-311C1, containing the entire Six2 locus (and other genes), was modified by targeting a Tet-off-eGFPCre into the ATG start site of the Six2 locus. This inserted a Tet-off cassette (tetracycline-regulated transactivator (tTA), 2xpolyA signal, tetracycline-responsive element (TRE or tetO with CMV min promoter) followed by an Enhanced Greed Fluorescent Protein/Cre Recombinase (EGFP/Cre) fusion protein coding sequence, (SV40 polyA signal),and frt-flanked kanamycin cassette. The targeted BAC sequences were further modified using FLP recombination to remove the selection cassette and linearized to remove its original vector backbone. The resulting modified BAC was microinjected into CD-1 zygotes. Sequence updates published after this transgene was created revealed an upstream Six2 ATG start site that is in-frame with the tTA; thus an apparent and unintended fusion was created with the tTA (80 amino acids potentially added) that renders transgene expression unaffected by tetracycline/doxycycline administration. [MGI Ref ID J:148455] | ||
| Gene Symbol and Name | Tg(Six2-EGFP/cre)1Amc, transgene insertion 1, Andrew P McMahon | ||
| Chromosome | UN | ||
| Gene Common Name(s) | Six2-TGC(tg); Six2-TGCtg; TGC BAC transgenic allele; Tet-off-eGFPCre BAC transgenic; Tg(Six2-tTA,tetO-EGFP/cre)1Amc; | ||
Genotyping
Genotyping Information
This strain will not have a genotyping protocol or one is not currently available.
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
Kobayashi A; Valerius MT; Mugford JW; Carroll TJ; Self M; Oliver G; McMahon AP. 2008. Six2 defines and regulates a multipotent self-renewing nephron progenitor population throughout mammalian kidney development. Cell Stem Cell 3(2):169-81. [PubMed: 18682239] [MGI Ref ID J:148455]
Health & husbandry
Health & Colony Maintenance Information
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, hemizygous mice may be bred together or to wildtype (noncarrier) siblings. The donating investigator reports they were unable to produce homozygous mice from hemizygous matings.
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
This strain is currently Under Development for Production.
To register your interest in this strain go to the Strain Interest Form.
Estimated Available for Sale Date:
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Supply Details
| Standard Supply | Under Development for Distribution Colony |
|---|---|
| Supply Notes |
|
| Important Note | |
| Sequence updates published after this transgene was created revealed an upstream Six2 ATG start site that is in-frame with the tTA; thus an apparent and unintended fusion was created with the tTA (80 amino acids potentially added) that renders transgene expression unaffected by tetracycline/doxycycline administration. | |
Control Information
| Control | ||
|---|---|---|
| Noncarrier | ||
| 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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