These OVE2508G mice harbor a mutation created by random insertion of the SB-cHS4core-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2229). Using inverse PCR analysis, the transgene integration site was identified in intron 11 of the transmembrane anterior posterior transformation 1 gene (Tapt1) on chromosome 5. The donating investigator reports the phenotype of homozygous mice as: cleft palate, possible anemia.
Paul A Overbeek, Baylor College of Medicine
Genetic Background | Generation |
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Allele Type |
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Transgenic (Inserted expressed sequence) |
These OVE2508G (OVE#2508G) mice harbor a mutation created by random insertion of the SB-cHS4core-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2229). Using inverse PCR analysis, the lentiviral integration site was identified in intron 11 of the transmembrane anterior posterior transformation 1 gene (Tapt1) on chromosome 5. The 3'-LTR is linked to the (+) strand of DNA at position 44,574,289 bp [NCB137/mm9; 3'-44,574,289(+)]. The lentivirus is inserted in the antisense orientation relative to the disrupted mouse gene. The donating investigator reports the phenotype of homozygous mice as: cleft palate, possible anemia.
A lentiviral transgenic approach was used to generate these mice. The SB-cHS4core-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2229) was designed with a Sleeping Beauty (SB) transposon (inward-facing SB arms flanking a tandem pair of cHS4 core insulator elements), followed by a mouse tyrosinase minigene (Tyro) and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) in the FUGW self-inactivating HIV-based lentiviral vector backbone. The SB transposon and Tyro minigene replaced the ubiquitin-c promoter and EGFP sequences originally found in the FUGW lentiviral vector. The SB transposon used in this transgene has an inverted repeat/direct repeat sequence (IR/DR; the SB transposon recognition site), two copies of the 0.25 kb chicken beta-globin core insulator element (cHS4 core), and a second IR/DR sequence. The IR/DR sequences are inward-facing (pointed towards the cHS4core). The cHS4 core insulator element functions to terminate transcription of the upstream locus into which it integrates (and also enhance expression from the downstream Tyro minigene). The Tyro minigene is composed of the mouse Tyr enhancer region (623 bp), promoter region (657 bp), and 1566 bp cDNA sequence (including the stop codon); all in sense orientation relative to the FUGW backbone. There is no polyA site between the Tyro minigene and WPRE sequence. The WPRE sequence functions to enhance the mRNA transcript stability. This packaged lentiposon was injected subzonally (under the zona pellucida) into 1-8 cell stage FVB/N mouse embryos. Expression of the tyrosinase minigene results in melanin synthesis, so founder mice (F0) were identified by inspection for pigmentation. All F0 mice exhibited mosaic pigmentation. Most F0 mice had more than one lentiviral integration site. F0 mice were assigned an OVE number and then bred with FVB/N mice. Pigmented offspring (F1) with different coat colors were designated as subline A, B, C, etc., for each family. F1 mice were bred with FVB/N mice. F2 mice with different coat colors were designated as sublines A-1, A-2, etc., or B-1, B-2, etc. Breeding with FVB/N mice was continued until litters with only one or two different coat colors were obtained. Mice with identical coat colors were then inbred and assessed for the presence or absence of viable homozygotes. Hemizygous mice of line OVE2508G (OVE#2508G) exhibit medium brown coat color. Using inverse PCR analysis, the lentiviral integration site was identified in intron 11 of the transmembrane anterior posterior transformation 1 gene (Tapt1) on chromosome 5. The 3'-LTR is linked to the (+) strand of DNA at position 44,574,289 bp [NCB137/mm9; 3'-44,574,289(+)]. The lentivirus is inserted in the antisense orientation relative to the disrupted mouse gene. Transgenic males were sent to The Jackson Laboratory Repository Facebase collection. Upon arrival, these mice were bred to FVB/NJ inbred mice (Stock No. 001800) for at least one generation to establish the live colony.
Expressed Gene | Tyr, tyrosinase, mouse, laboratory |
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Site of Expression |
Allele Name | transgenic transposon lentiviral insertion 2508G, Paul A Overbeek |
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Allele Type | Transgenic (Inserted expressed sequence) |
Allele Synonym(s) | OVE#2508G; OVE2508G |
Gene Symbol and Name | Tapt1, transmembrane anterior posterior transformation 1 |
Gene Synonym(s) | |
Promoter | Tyr, tyrosinase, mouse, laboratory |
Expressed Gene | Tyr, tyrosinase, mouse, laboratory |
Strain of Origin | FVB/N |
Chromosome | 5 |
Molecular Note | The SB-cHS4core-SB-Tyro-WPRE-FUGW lentiposon transgene (LV2229) was designed with a Sleeping Beauty (SB) transposon (inward-facing SB arms flanking a tandem pair of cHS4 core insulator elements), followed by a mouse tyrosinase minigene (Tyro) and a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE) in the FUGW self-inactivating HIV-based lentiviral vector backbone. The SB transposon and Tyro minigene replaced the ubiquitin-c promoter and EGFP sequences originally found in the FUGW lentiviral vector. The SB transposon used in this transgene has an inverted repeat/direct repeat sequence (IR/DR; the SB transposon recognition site), two copies of the 0.25 kb chicken beta-globin core insulator element (cHS4 core), and a second IR/DR sequence. The IR/DR sequences are inward-facing (pointed towards the cHS4core). The cHS4 core insulator element functions to terminate transcription of the upstream locus into which it integrates (and also enhance expression from the downstream Tyro minigene). The Tyro minigene is composed of the mouse Tyr enhancer region (623 bp), promoter region (657 bp), and 1566 bp cDNA sequence (including the stop codon); all in sense orientation relative to the FUGW backbone. There is no polyA site between the Tyro minigene and WPRE sequence. The WPRE sequence functions to enhance the mRNA transcript stability. In line OVE2508G, a single copy of the lentiposon inserted into intron 11 [NCBI37/mm9; 3'-44,574,289(+)] in the antisense orientation. |
When maintaining a live colony, heterozygous mice may be bred together, bred with wildtype siblings, or bred with FVB/N inbred mice (Stock No. 001800).
When using the FVB/N-Tapt1TgTn(sb-cHS4,Tyr)2508GOve/J mouse strain in a publication, please cite the originating article(s) and include JAX stock #017436 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Hemizygous or Non carrier for Tapt1<TgTn(sb-cHS4,Tyr)2508GOve> |
Frozen Mouse Embryo | FVB/N-Tapt1<TgTn(sb-cHS4 Tyr)2508GOve>/J | $2595.00 |
Frozen Mouse Embryo | FVB/N-Tapt1<TgTn(sb-cHS4 Tyr)2508GOve>/J | $2595.00 |
Frozen Mouse Embryo | FVB/N-Tapt1<TgTn(sb-cHS4 Tyr)2508GOve>/J | $3373.50 |
Frozen Mouse Embryo | FVB/N-Tapt1<TgTn(sb-cHS4 Tyr)2508GOve>/J | $3373.50 |
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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. We do not guarantee breeding performance and therefore suggest that investigators order more than one breeding pair to avoid delays in their research.
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