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- Description
- Phenotype
- Genes & alleles
- Genotyping
- Health & husbandry
- References
- Purchasing information
- Terms of Use
Description
Strain Information
Former Names STOCK Dbttm1Geh Tg(tTALap)5Bjd Tg(tetO-DBT*)A1Geh/J (Changed: 11-MAY-07 ) Type Mutant Stock; Targeted Mutation; Transgenic; Additional information on Genetically Engineered and Mutant Mice. Visit our online Nomenclature tutorial. Mating System See Colony Maintenance Species laboratory mouse Generation ?+F3 (15-JAN-09) Donating Investigator Gregg Homanics, University of Pittsburgh Description
Mice homozygous for the Dbt (E2) targeted mutation and carrying both the LAP-tTA and TRE-E2 transgenes are viable and fertile. The E2-targeted mutation leads to absence of branched-chain keto acid dehydrogenase (BCKDH) activity and E2 protein in liver tissue, but this absence is rescued by the two transgenes: liver-directed expression of the modified human BCKDH E2 subunit from the complimentary "Tet-off" transgenes abrogates the severity of Maple Syrup Urine Disease (MSUD) phenotype observed in E2-deficient single mutant mice. Triple mutant mice are a model for intermediate MSUD (iMSUD); BCKDH activity is only 5-6% of that found in wildtype mice. This low level of BCKDH activity is sufficient to allow survival, but insufficient to normalize circulating branched chain amino acids levels. Because these mice have near normal amounts of E2 protein, but only 5-6% of normal BCKDH enzyme activity, it is probable that the c-myc tag at the carboxy-terminus of the human E2 transgene interferes with enzymatic activity. The donating investigator reports that addition of the tetracycline analog doxycycline does not affect MSUD phenotype rescue. These mutant mice may be useful in studying metabolic disease, biochemistry, and both the complete/classic and intermediate forms of Maple Syrup Urine Disease.Development
The mutant allele of branched-chain keto acid dehydrogenase E2 subunit (Dbt gene) was created by Dr. Gregg Homanics (University of Pittsburgh) using a targeting vector designed to replace part of exon 4 and all of exon 5 with a PGKneo cassette. The construct was electroporated into (129X1/SvJ x 129S1/Sv)F1-derived R1 embryonic stem (ES) cells. Correctly targeted ES cells were microinjected into C57BL/6J blastocysts. Chimeric mice were bred to C57BL/6J to generate heterozygous mice. Heterozygotes were then bred to LAP-tTA transgenic mice on a mixed (NMRI outbred;FVB;C57BL/6J) background. These mice were created by Dr. Hermann Bujard (Universitat Heidelberg) to express tetracycline-controlled transactivator (tTA) under control of the rat liver-enriched activator protein promoter. The TRE-E2 (or tetO-DBT*) transgene, also created by Dr. Gregg Homanics, contains a tetracycline response element and minimal human CMV promoter controlling a human E2 subunit (DBT gene) cDNA which has been modified to contain a 4x alanine linker followed by a c-myc epitope tag at the carboxy terminus. After injection of the transgene into C57BL/6J embryos, founder line A mice were bred with E2-deficient, LAP-tTA double mutant mice to generate this triple mutant strain.
Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| Considerations for Choosing Controls | ||
Related Strains
Strains carrying Tg(tTALap)5Bjd allele
003563 B6.Cg-Tg(tTALap)5Bjd/J View Strains carrying Tg(tTALap)5Bjd (1 strain)
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 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 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 003271 STOCK Tg(tTAhCMV)3Bjd/J 003275 STOCK Tg(tetL)1Bjd/J 003274 STOCK Tg(tetNZL)2Bjd/J
Phenotype
Phenotype Information
Maple Syrup Urine Disease - Models with phenotypic similarity to human disease where etiologies involve orthologs.1
1 Human genes are associated with this disease. Orthologs of those genes appear in the mouse genotype(s).
assigned by genotype
Dbttm1Geh/Dbttm1Geh Tg(tetO-DBT)A1Geh/0 Tg(tTALap)5Bjd/0
involves: 129S1/Sv * 129X1/SvJ * C57BL/6J * FVB * NMRI
- lethality-postnatal
- postnatal lethality (MGI Ref ID J:119973)
- approximately 4% of pups surviving to weaning have this genotype, compared to a theoretical survival rate or 14%, indicating partial rescue of the Dbttm1Geh phenotype
- life span-post-weaning/aging
- premature death (MGI Ref ID J:119973)
- transgenic mice survival is only ~16 weeks, when mice become moribund and are sacrificed
- homeostasis/metabolism phenotype
- abnormal circulating amino acid level (MGI Ref ID J:119973)
- blood levels of alanine, glutamate, and glutamine are reduced compared to controls, but greater than in Dbttm1Geh homozygous mice
- BCAA/alanine (BCAA -branched-chain amino acids - leucine + isoleucine + valine) values are intermediate between controls and Dbt-null mice between 4-7 weeks of age
- abnormal enzyme/coenzyme activity (MGI Ref ID J:119973)
- levels of branched-chain keto acid dehydrogenase (BCKDH) activity are only 5-6% of controls levels, despite nearly equal protein levels, indicating suboptimal BCKDH activity of protein assembled with human E2
Genes & Alleles
Gene & Allele Information
| Allele Symbol | Dbttm1Geh | ||
|---|---|---|---|
| Allele Name | targeted mutation 1, Gregg E Homanics | ||
| Allele Type | Targeted (knock-out) | ||
| Common Name(s) | E2 KO; | ||
| Strain of Origin | (129X1/SvJ x 129S1/Sv)F1-Kitl<+> | ||
| ES Cell Line Name | R1 | ||
| ES Cell Line Strain | (129X1/SvJ x 129S1/Sv)F1-Kitl<+> | ||
| Gene Symbol and Name | Dbt, dihydrolipoamide branched chain transacylase E2 | ||
| Chromosome | 3 | ||
| Gene Common Name(s) | BCATE2; BCKAD E2; D3Wsu60e; DNA segment, Chr 3, Wayne State University 60, expressed; E2; E2B; MGC9061; dihydrolipoyl transacylase; dihydrolipoyllysine-residue (2-methylpropanoyl)transferase; | ||
| Molecular Note | A targeting vector was used to replace part of exon 4 and all of exon 5 with a PGKneo cassette.Southern blot and immunohistochemical analyses verify the absence of wild-type E2 transcripts and lack of E2 protein detection in homozygotes. [MGI Ref ID J:119973] | ||
| Allele Symbol | Tg(tTALap)5Bjd | ||
| Allele Name | transgene insertion 5, Hermann Bujard | ||
| Allele Type | Transgenic (random, expressed) | ||
| Common Name(s) | LAP-tTA; Tg(tTALap)5Uh; | ||
| Mutation Made By | Hermann Bujard, Universität Heidelberg | ||
| Strain of Origin | NMRI | ||
| Site of Expression | Expresses tTA at high levels in the liver; see Stock No. 003563 | ||
| 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. | |||
| Promoter | Cebpb, CCAAT/enhancer binding protein (C/EBP), beta, rat | ||
| General Note | Transgenic mice on a C57BL/6J background are viable and fertile, and express tTA in the liver. | ||
| Molecular Note | This transgene contains the gene encoding the tetracycline regulated transactivator protein (tTA) driven by the rat Cebpb promoter (previously called liver-enriched activator protein). [MGI Ref ID J:93000] | ||
| Allele Symbol | Tg(tetO-DBT)A1Geh | ||
| Allele Name | transgene insertion A, Gregg E Homanics | ||
| Allele Type | Transgenic (random, expressed) | ||
| Common Name(s) | TRE-E2 (line A); | ||
| Strain of Origin | C57BL/6J | ||
| Site of Expression | Expresses tTA at high levels in the liver; see Stock No. 003563 | ||
| 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 | DBT, dihydrolipoamide branched chain transacylase E2, human | ||
| Promoter | tetO, tet operator, | ||
| Molecular Note | The TRE-E2 (or tetO-DBT) transgene contains a tetracycline response element and minimal human CMV promoter controlling a human E2 subunit (DBT gene) cDNA which has been modified to contain a 4x alanine linker followed by a c-myc epitope tag at the carboxy terminus to facilitate detection. Levels of the human E2 protein are approximately equal to mouse E2 protein in livers of control mice. However, when crossed to animals that express a transgene encoding the tetracycline regulated transactivator protein (Tg(tTALap)5Bjd) and are homozygous for a knockout allele of mouse E2 (Dbttm1Geh) are treated with doxycycline, the E2 protein levels correlate with only ~5-6% of normal branched-chain keto acid dehydrogenase (BCKDH) activity. It is one hypothesis that addition of the c-myc tag interferes with BCKDH subunit assembly (ie. interaction the human E2 subunit with mouse E1 and E3 complexes), thus reducing enzymatic activity. [MGI Ref ID J:119973] | ||
Genotyping
Genotyping Information
Genotyping Protocols
Tg(tetO-DBT)A1Geh, QPCR, vers. 2
Dbttm1Geh, SEP PCR, vers. 1
Dbttm1Geh, STD PCR, vers. 2
Tg(tTA), MCA, vers. 4
Tg(tTA), QPCR, vers. 3
Tg(tTA), STD PCR, vers. 2
Tg(tTALap)5Bjd, SEP PCR, vers. 1
Tg(tetO-DBT)A1Geh, STD PCR, vers. 1
Helpful Links
Genotyping resources and troubleshooting
References
References
Selected Reference(s)
Homanics GE; Skvorak K; Ferguson C; Watkins S; Paul HS. 2006. Production and characterization of murine models of classic and intermediate maple syrup urine disease. BMC Med Genet 7:33. [PubMed: 16579849] [MGI Ref ID J:119973]
Additional References
Tg(tTALap)5Bjd relatedBeer S; Komatsubara K; Bellovin DI; Kurobe M; Sylvester K; Felsher DW. 2008. Hepatotoxin-induced changes in the adult murine liver promote MYC-induced tumorigenesis. PLoS ONE 3(6):e2493. [PubMed: 18560566] [MGI Ref ID J:139884]
Beer S; Zetterberg A; Ihrie RA; McTaggart RA; Yang Q; Bradon N; Arvanitis C; Attardi LD; Feng S; Ruebner B; Cardiff RD; Felsher DW. 2004. Developmental Context Determines Latency of MYC-Induced Tumorigenesis. PLoS Biol 2(11):E332. [PubMed: 15455033] [MGI Ref ID J:93372]
Cairo S; Armengol C; De Reynies A; Wei Y; Thomas E; Renard CA; Goga A; Balakrishnan A; Semeraro M; Gresh L; Pontoglio M; Strick-Marchand H; Levillayer F; Nouet Y; Rickman D; Gauthier F; Branchereau S; Brugieres L; Laithier V; Bouvier R; Boman F; Basso G;Michiels JF; Hofman P; Arbez-Gindre F; Jouan H; Rousselet-Chapeau MC; Berrebi D; Marcellin L; Plenat F; Zachar D; Joubert M; Selves J; Pasquier D; Bioulac-Sage P; Grotzer M; Childs M; Fabre M; Buendia MA. 2008. Hepatic stem-like phenotype and interplay of Wnt/beta-catenin and Myc signaling in aggressive childhood liver cancer. Cancer Cell 14(6):471-84. [PubMed: 19061838] [MGI Ref ID J:142029]
Carpenter B; Lin Y; Stoll S; Raffai RL; McCuskey R; Wang R. 2005. VEGF is crucial for the hepatic vascular development required for lipoprotein uptake. Development 132(14):3293-303. [PubMed: 15944181] [MGI Ref ID J:100427]
Goga A; Yang D; Tward AD; Morgan DO; Bishop JM. 2007. Inhibition of CDK1 as a potential therapy for tumors over-expressing MYC. Nat Med 13(7):820-7. [PubMed: 17589519] [MGI Ref ID J:125802]
Hasan MT; Schonig K; Berger S; Graewe W; Bujard H. 2001. Long-term, noninvasive imaging of regulated gene expression in living mice. Genesis 29(3):116-22. [PubMed: 11252052] [MGI Ref ID J:127660]
Kistner A; Gossen M; Zimmermann F; Jerecic J; Ullmer C; Lubbert H; Bujard H. 1996. Doxycycline-mediated quantitative and tissue-specific control of gene expression in transgenic mice. Proc Natl Acad Sci U S A 93(20):10933-8. [PubMed: 8855286] [MGI Ref ID J:93000]
Kornmann B; Schaad O; Bujard H; Takahashi JS; Schibler U. 2007. System-driven and oscillator-dependent circadian transcription in mice with a conditionally active liver clock. PLoS Biol 5(2):e34. [PubMed: 17298173] [MGI Ref ID J:141590]
Roy CN; Mak HH; Akpan I; Losyev G; Zurakowski D; Andrews NC. 2007. Hepcidin antimicrobial peptide transgenic mice exhibit features of the anemia of inflammation. Blood 109(9):4038-44. [PubMed: 17218383] [MGI Ref ID J:145326]
Shachaf CM; Kopelman AM; Arvanitis C; Karlsson A; Beer S; Mandl S; Bachmann MH; Borowsky AD; Ruebner B; Cardiff RD; Yang Q; Bishop JM; Contag CH; Felsher DW. 2004. MYC inactivation uncovers pluripotent differentiation and tumour dormancy in hepatocellular cancer. Nature 431(7012):1112-7. [PubMed: 15475948] [MGI Ref ID J:93899]
Sun Y; Quinn B; Xu YH; Leonova T; Witte DP; Grabowski GA. 2006. Conditional expression of human acid beta-glucosidase improves the visceral phenotype in a Gaucher disease mouse model. J Lipid Res 47(10):2161-70. [PubMed: 16861620] [MGI Ref ID J:116525]
Tilli MT; Furth PA. 2003. Conditional mouse models demonstrate oncogene-dependent differences in tumor maintenance and recurrence. Breast Cancer Res 5(4):202-5. [PubMed: 12817992] [MGI Ref ID J:84503]
Tward AD; Jones KD; Yant S; Cheung ST; Fan ST; Chen X; Kay MA; Wang R; Bishop JM. 2007. Distinct pathways of genomic progression to benign and malignant tumors of the liver. Proc Natl Acad Sci U S A 104(37):14771-14776. [PubMed: 17785413] [MGI Ref ID J:124960]
Wang R; Ferrell LD; Faouzi S; Maher JJ; Bishop JM. 2001. Activation of the met receptor by cell attachment induces and sustains hepatocellular carcinomas in transgenic mice. J Cell Biol 153(5):1023-34. [PubMed: 11381087] [MGI Ref ID J:69731]
Yang YA; Zhang GM; Feigenbaum L; Zhang YE. 2006. Smad3 reduces susceptibility to hepatocarcinoma by sensitizing hepatocytes to apoptosis through downregulation of Bcl-2. Cancer Cell 9(6):445-57. [PubMed: 16766264] [MGI Ref ID J:110133]
Health & husbandry
Health & Colony Maintenance Information
Animal Health Reports
Room Number AX11
Colony Maintenance
Breeding & Husbandry When maintaining a live colony, mice that are heterozygous for the targeted mutation and carriers of both transgenes can be bred together. Loss of one or both of the transgenes results in perinatal lethality (and Maple Syrup Urine Disease or MSUD phenotype) for mice homozygous for the targeted mutation. Mating System See above Diet Information LabDiet® 5K52/5K67
Purchasing information
Pricing, Supply Level & Notes, Controls, General Terms & Conditions
Pricing
| Pricing for USA, Canada and Mexico shipping destinations |
|
Weeks of Age Price (US dollars $) Gender Genotypes Provided Individual Mouse $243.50 Female or Male Heterozygous for Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh $300.70 Female or Male Homozygous for Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh
Pairs /Price (US dollars $) Pair Genotype $487.00 Heterozygous for Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh x Heterozygous for Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh
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 Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh $391.00 Female or Male Homozygous for Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh
Pairs /Price (US dollars $) Pair Genotype $633.10 Heterozygous for Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh x Heterozygous for Dbttm1Geh, Homozygous for Tg(tTALap)5Bjd, Homozygous for Tg(tetO-DBT)A1Geh
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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Control Information
| Control | ||
|---|---|---|
| Wild-type from the colony | ||
| 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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