FVB/N-Fance^{Tg(Tyr)2364E-2a2Ove}/Mmjax

Stock number: 017967
Strain synonyms: OVE2364E-2a2
Research areas: Hematological Research, Reproductive Biology Research

Description

The donating investigator reports that homozygous females are sterile, homozygous males are subfertile. The phenotype of the Fance mutants resembles the phenotype of other Fanconi anemia mutations in mice. Mouse model for Fanconi anemia. Studies of genome stability and DNA damage repair.

Detailed description

The donating investigator reports that homozygous females are sterile, homozygous males are subfertile. The phenotype of the Fance mutants resembles the phenotype of other Fanconi anemia mutations in mice. Mouse model for Fanconi anemia. Studies of genome stability and DNA damage repair.

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 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 OVE 2364E-2a2 exhibit a light grey coat color. Using inverse PCR analysis, the lentiviral integration site was identified in intron 8 of the Fanconi anemia E gene (Fance) on chromosome 17. The 3'-LTR is linked to the (+) strand of DNA at position 28,458,210 bp [NCB137/mm9; 3'-28,458,210(+)].

Allele

Symbol: Fance^{Tg(Tyr)2364E-2a2Ove}
Name: transgene insertion 2364E-2a2, Paul Overbeek
MGI accession ID: MGI:5467996
Generation method: Transgenic
Attributes: Null/Knockout; Inserted expressed sequence
Marker symbol: Fance
Marker name: Fanconi anemia, complementation group E
Chromosome: 17
Strain of origin: FVB/N
Expressed genes: Tyr,tyrosinase,mouse, laboratory
Molecular note: The Tyrosinase minigene is composed of the mouse Tyr enhancer region (623 bp), promoter region (657 bp), and 1566 bp cDNA sequence (including the stop codon). 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. Using inverse PCR analysis, the lentiviral integration site was identified in intron 8 of the Fanconi anemia E gene (Fance) on chromosome 17. The 3'-LTR is linked to the (+) strand of DNA at position 28,458,210 bp [NCB137/mm9; 3'-28,458,210(+)].