These OVE#2323C 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 3 of the PHD finger protein 5A gene (Phf5a) on chromosome 15 (specifically at the 5'-81,697,556(+) bp position in sense orientation). The donating investigator reports the phenotype of homozygous mice as: may be lethal at embryonic day (E)6.
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 OVE#2323C 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 3 of the PHD finger protein 5A gene (Phf5a) on chromosome 15 (specifically at the 5'-81,697,556(+) bp position in sense orientation). The donating investigator reports the phenotype of homozygous mice as: may be lethal at embryonic day (E)6.
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 (containing a cHS4 core insulator element), 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), a 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 (620 bp), promoter region (500 bp), and 1.7 kbp 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 construct was injected subzonally (under the zona pellucida) into 1-8 cell stage FVB/N mouse embryos. Because of the random and multiple lentiviral integration sites of a transgene containing tyrosinase, founder mice (F0) may be identified by mosaic pigmentation. Founder mice from line OVE#2323C were bred with FVB/N mice, and pigmented offspring (F1) were bred to FVB/N mice. The resulting offspring (F2) with identical coat colors (dark grey) were subsequently inbred. Using inverse PCR analysis, the transgene integration site was identified in intron 3 of the PHD finger protein 5A gene (Phf5a) on chromosome 15 (specifically at the 5'-81,697,556(+) bp position in sense orientation). Transgenic males were sent to The Jackson Laboratory Repository (MMRRC-JAX). Upon arrival, these mice were bred to FVB/NJ inbred mice 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 2323C, Paul A Overbeek |
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Allele Type | Transgenic (Inserted expressed sequence) |
Allele Synonym(s) | OVE#2323C; OVE2323C |
Gene Symbol and Name | Phf5a, PHD finger protein 5A |
Gene Synonym(s) | |
Promoter | Tyr, tyrosinase, mouse, laboratory |
Expressed Gene | Tyr, tyrosinase, mouse, laboratory |
Strain of Origin | FVB/N |
Chromosome | 15 |
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 OVE2323S, a single copy of the lentiposon inserted into intron 3 [NCBI37/mm9; 5'-81,697,556(+)] in the sense orientation. |
When maintaining a live colony, heterozygous mice may be bred together, bred with wildtype siblings, or bred with FVB/N inbred mice.
When using the FVB/N-Phf5aTgTn(sb-cHS4,Tyr)2323COve/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #36228 in your Materials and Methods section.
Facility Barrier Level Descriptions
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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