FVB/N-Cnpy4^{Tg(Tyr)2356C-2a1Ove}/Mmjax

Stock number: 017418

Description

These OVE2356C-2a1 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: fertile.

Detailed description

These OVE2356C-2a1 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 intron 3 of the canopy 4 homolog [zebrafish] gene (Cnpy4) on chromosome 5. The 5'-LTR is linked to the (-) strand of DNA at position 138,632,847 bp [NCB137/mm9; 5'-138,632,847(-)]. The lentivirus is inserted in the sense orientation relative to the disrupted mouse gene. The donating investigator reports the phenotype of homozygous mice as: fertile.

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 OVE2356C-2a1 (OVE#2356C-2a1) exhibit light grey mottle coat color. Using inverse PCR analysis, the lentiviral integration site was identified in intron 3 of the canopy 4 homolog [zebrafish] gene (Cnpy4) on chromosome 5. The 5'-LTR is linked to the (-) strand of DNA at position 138,632,847 bp [NCB137/mm9; 5'-138,632,847(-)]. The lentivirus is inserted in the sense orientation relative to the disrupted mouse gene. 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: Cnpy4^{Tg(Tyr)2356C-2a1Ove}
Name: transgene lentiviral insertion 2356C-2a1, Paul A Overbeek
MGI accession ID: MGI:5286685
Generation method: Transgenic
Attributes: Reporter
Synonyms: OVE#2356C-2a1; OVE2356C-2a1
Marker symbol: Cnpy4
Marker name: canopy FGF signaling regulator 4
Marker synonyms: 2610019P18Rik; Cnpy; PRAT4B; RGD1307636
Chromosome: 5
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 2356C-2a1, the lentiviral transgene inserted into intron 3 (NCBI37/mm9; 5'-138,632,847(-)) in sense orientation relative to the disrupted gene.