FVB/N-Zfp553^{Tg(Tyr)2412B-1Ove}/Mmjax

Stock number: 017409
Research areas: Developmental Biology Research

Description

These OVE2412B-1 mice harbor a mutation created by random insertion of the Tyro-IRES-sd-loxP-FUGW lentiviral transgene (LV2187). The donating investigator reports the phenotype of homozygous mice as: embryonic day (E)11 lethal.

Detailed description

These OVE2412B-1 mice harbor a mutation created by random insertion of the Tyro-IRES-sd-loxP-FUGW lentiviral transgene (LV2187). Using inverse PCR analysis, the lentiviral integration site was identified in an intron of the putative zinc finger protein 553 (Zfp553) gene on chromosome 7. The 3'-LTR is linked to position 134,689,198 bp [NCB137/mm9;]. The donating investigator reports the phenotype of homozygous mice as: embryonic day (E)11 lethal.

Development

A lentiviral transgenic approach was used to generate these mice. The Tyro-IRES-sd-loxP-FUGW lentiviral transgene (LV2187) was designed with a mouse tyrosinase minigene (Tyro) followed by an IRES::3xATG::splice-donor::loxP sequence in the FUGW self-inactivating HIV-based lentiviral vector backbone. The Tyro-IRES-sd-loxP replaced the ubiquitin-c promoter, EGFP, and WPRE sequences originally found in the FUGW lentiviral vector. 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 antisense orientation relative to the FUGW backbone. The Tyro minigene is followed by an internal ribosome entry site (IRES) ending with ATGs in all three reading frames (3xATG), an artificial splice donor sequence, and a loxP site in this construct. This packaged lentivirus 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 OVE2412B-1 (OVE#2412B-1) exhibit tan coat color. Using inverse PCR analysis, the lentiviral integration site was identified in an intron of the putative zinc finger protein 553 (Zfp553) gene on chromosome 7. The 3'-LTR is linked to position 134,689,198 bp [NCB137/mm9;]. Transgenic males were sent to The Jackson Laboratory Repository node of the Mutant Mouse Regional Resource Center (MMRRC-JAX). Upon arrival, these mice were bred to FVB/NJ inbred mice for at least one generation to establish the live colony.

Allele

Symbol: Zfp553^{Tg(Tyr)2412B-1Ove}
Name: transgene lentiviral insertion 2412B-1, Paul A Overbeek
MGI accession ID: MGI:5286668
Generation method: Transgenic
Attributes: Reporter
Marker symbol: Zfp553
Marker name: zinc finger protein 553
Chromosome: 7
Strain of origin: FVB/N
Expressed genes: Tyr,tyrosinase,mouse, laboratory
Molecular note: The Tyro-IRES-sd-loxP-FUGW lentiviral transgene (LV2187) was designed with a mouse tyrosinase minigene (Tyro) followed by an IRES::3xATG::splice-donor::loxP sequence in the FUGW self-inactivating HIV-based lentiviral vector backbone. The Tyro-IRES-sd-loxP replaced the ubiquitin-c promoter, EGFP, and WPRE sequences originally found in the FUGW lentiviral vector. 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 antisense orientation relative to the FUGW backbone. The Tyro minigene is followed by an internal ribosome entry site (IRES) ending with ATGs in all three reading frames (3xATG), an artificial splice donor sequence, and a loxP site in this construct. In line 2412B-1, the lentiviral transgene inserted into an intron (NCBI37/mm9; 134,689,198 bp(+)) in sense orientation relative to the disrupted gene.