Overview

These OVE2372E-3a3 mice harbor a mutation created by random insertion of the Tyro-IRES-sd-loxP-FUGW lentiviral transgene (LV2187). Hemizygous mice of line OVE2372E-3a3 exhibit very light tan/grey coat color and red eyes. The donating investigator reports the phenotype of homozygous mice as: fertile.

Donating Investigator

Paul A Overbeek, Baylor College of Medicine

Genetic overview

Genetic Background	Generation

Klhl20^{Tg(Tyr)2372E-3a3Ove}

Allele Type
Transgenic (Inserted expressed sequence)

View Genetics

Research Applications

View All Research Applications

Details

Detailed Description

These OVE2372E-3a3 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 last intron (12) of the kelch-like 20 gene (Klhl20) on chromosome 1. The 3'-LTR is linked to the (-) strand of DNA at position 163,021,329 bp [NCB137/mm9; 3'-163,021,329(-)]. The lentivirus is inserted in the sense orientation relative to the disrupted mouse gene. Hemizygous mice of line OVE2372E-3a3 exhibit very light tan/grey coat color and red eyes. 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 OVE2372E-3a3 exhibit very light tan/grey coat color and red eyes. Using inverse PCR analysis, the lentiviral integration site was identified in last intron (12) of the kelch-like 20 gene (Klhl20) on chromosome 1. The 3'-LTR is linked to the (-) strand of DNA at position 163,021,329 bp [NCB137/mm9; 3'-163,021,329(-)]. The lentivirus is inserted in the sense orientation relative to the disrupted mouse gene. Transgenic males were later found to exhibit coat color differences. Further investigation revealed a second integration site in intron 2 of the activating transcription factor 7 gene (Atf7) on chromosome 15. After additional breeding, mice harboring only the Klhl20 integration site 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.

Expression Data

Expressed Gene	Tyr, tyrosinase, mouse, laboratory
Site of Expression

Control Suggestions

Wild-type from the colony
001800 FVB/NJ

Additional Information

Considerations for Choosing Controls

Genetics

Klhl20^{Tg(Tyr)2372E-3a3Ove}

Allele Symbol: Klhl20^{Tg(Tyr)2372E-3a3Ove}

Allele Name	transgene lentiviral insertion 2372E-3a3, Paul A Overbeek
Allele Type	Transgenic (Inserted expressed sequence)
Allele Synonym(s)	OVE#2372E-3a3; OVE2372E-3a3
Gene Symbol and Name	Klhl20, kelch-like 20
Gene Synonym(s)
Promoter	Tyr, tyrosinase, mouse, laboratory
Expressed Gene	Tyr, tyrosinase, mouse, laboratory
Strain of Origin	FVB/N
Chromosome	1
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 2372E-3a3, the lentiviral transgene inserted into last intron (12; NCBI37/mm9; 3'-163,021,329(-)) in sense orientation relative to the disrupted gene.
Mutations Made By	Paul Overbeek, Baylor College of Medicine

Disease/Phenotype

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

The following phenotype relates to a compound genotype created using this strain

Genotype: Klhl20^{Tg(Tyr)2372E-3a3Ove}/Klhl20^{Tg(Tyr)2372E-3a3Ove}
FVB/N-Klhl20

reproductive system phenotype

abnormal fertility/fecundity
- males may be sterile

References

Additional - Klhl20^{Tg(Tyr)2372E-3a3Ove} related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Separated PCR:Klhl20-3'
- Genotyping resources and troubleshooting
Breeding Considerations

When maintaining a live colony, heterozygous mice may be bred together, bred with wildtype siblings, or bred with FVB/N inbred mice.
- Additional Breeding and Husbandry Support
Citation
When using the FVB/N-Klhl20^{Tg(Tyr)2372E-3a3Ove}/Mmjax mouse strain in a publication, please cite the originating article(s) and include MMRRC stock #36274 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

The Jackson Laboratory's Genotype Promise

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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Donating Investigator

Genetic overview

Research Applications

Detailed Description

Development

Expression Data

Control Suggestions

Additional Information

Klhl20Tg(Tyr)2372E-3a3Ove

Allele Symbol: Klhl20Tg(Tyr)2372E-3a3Ove

Disease Terms

Research Areas By Phenotype

This mouse can be used to support research in many areas including:

Mammalian Phenotype Terms by Genotype

Genotype: Klhl20Tg(Tyr)2372E-3a3Ove/Klhl20Tg(Tyr)2372E-3a3OveFVB/N-Klhl20

reproductive system phenotype

References

Additional - Klhl20Tg(Tyr)2372E-3a3Ove related

Genotyping Protocols

Breeding Considerations

Citation

Animal Health Reports

Production of mice from cryopreserved embryos or sperm occurs in a maximum barrier room, G200

The Jackson Laboratory's Genotype Promise

Terms Of Use

Licensing Information

JAX® Mice, Products & Services Conditions of Use

No Warranty

Credit for PRODUCTS or SERVICES

Animal Care and Use for SERVICES

No Liability

Klhl20^{Tg(Tyr)2372E-3a3Ove}

Allele Symbol: Klhl20^{Tg(Tyr)2372E-3a3Ove}

Genotype: Klhl20^{Tg(Tyr)2372E-3a3Ove}/Klhl20^{Tg(Tyr)2372E-3a3Ove}
FVB/N-Klhl20

Additional - Klhl20^{Tg(Tyr)2372E-3a3Ove} related