Overview

Also Known As:Dux4-fl, FLExDUX4

FLExDUX4 mice conditionally express human DUX4 following Cre-mediated recombination. This strain may be useful for studying facioscapulohumeral muscular dystrophy (FSHD).

Donating Investigator

Peter L. Jones, University of Nevada School of Medicine

Genetic overview

Genetic Background	Generation
	?+pN1F9 (2020-01-02 00:00:00)

Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}*

Allele Type	Gene Symbol	Gene Name
Targeted (Conditional ready (e.g. floxed), Inserted expressed sequence, Humanized sequence)	Gt(ROSA)26Sor	gene trap ROSA 26, Philippe Soriano

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Research Applications

Neurobiology Research
Research Tools

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Base Price

Starting at:

$270.00 Domestic price for female 4-week

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Details

Detailed Description

FLExDUX4 mice were created using a cre-dependent one-way genetic switch (FLEx) system. Homozygotes mice carrying this DUX4 conditional allele are viable and fertile. Two sets of incompatible outward facing recombination sites (loxP and lox511) flank an inverted human double homeobox 4 (DUX4) sequence, including exons 1-3 and both introns. The DUX4 gene encodes several alternative mRNA splicing variants. The hereditary muscle disorder, facioscapulohumeral muscular dystrophy (FSHD) is caused by the toxic gain-of-function of the DUX4-full-length (DUX4-fl) mRNA isoform. The DUX4-fl mRNA, which encodes a paired homeobox domain transcription factor, is typically not expressed in healthy muscle. However, in FSHD, the rare expression of DUX4-fl (in less than 1% of muscle fibers) initiates a pathogenic cascade of events including apoptosis, differentiation defects, muscle atrophy, and susceptibility to oxidative stress. Overall, FSHD is characterized by a slowly progressing muscular dystrophy that predominantly affects the skeletal muscles of the face, scapula, and upper arms but can affect muscles of the abdomen, hip girdle, and lower legs with ~20% of patients ultimately losing ambulation.

The DUX4 promoter drives expression of two short non-pathogenic isoforms (DUX4-s) and a longer cytotoxic isoform (DUX4-fl). This strain contains 4 point mutation in the 5’ splicing donor sites for the two DUX4-s mRNAs, abolishing expression of the short isoforms and only generating the pathogenic DUX4-fl mRNA isoform.

Because this construct was targeted for insertion into the Gt(ROSA)26Sor locus, DUX4-fl expression is determined by which tissue(s) express Cre recombinase. When bred to mice that express Cre recombinase, the resulting offspring will have the loxP or lox511 sites recombined, resulting in the inversion of the human DUX4-fl sequence, ending in a sense orientation.

Hemizygous and homozygous mice have low level DUX4-fl expression in the absence of Cre Recombinase. These mice exhibit alopecia, soft stool, inflammation, and muscle weakness. Homozygous are more affected, as are males compared to females.

When bred to STOCK Tg(ACTA1-cre/Esr1*)2Kesr/J mice (Stock No. 025750), resulting offspring express slightly more DUX4-fl than the FLExDUX4 hemizygous mice prior to tamoxifen induction (TMX). One TMX injection of 5 mg/kg leads to a useful mildly progressive FSHD model, while 2 injections of 10mg/kg leads to a more severe model.

Of note, the donating investigator reports that when bred to Sox2-Cre mice (Stock No. 008454), resulting offspring are embryonic lethal. When bred to ACTA1-rtTA, tetO-cre mice (Stock No. 012433), resulting offspring are embryonic lethal. When bred to ACTA1-Cre mice (Stock No. 006149), resulting offspring are still-born. When bred to UBC-CreERT2 mice (Stock No. 008085), without tamoxifen induction, offspring begin to show a severe neurological phenotype around 5 weeks of age, perhaps due to leaky DUX4 expression in the brain.

Development

The FLExDUX4 targeting vector was designed with (from 5’ to 3’) a loxP site, a frt-flanked neomycin resistance gene, a lox511 site, an inverted human double homeobox 4 (DUX4) gene with a polyadenylation sequence, and another loxP site and lox511 site, both in opposite orientation to the 5’ sites. The DUX4 gene contains 4 silent point mutations in exon 1 that prevent alternative splicing of a truncated isoform, DUX4-s, while maintaining the DUX4-fl ORF. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus via electroporation of C57BL/6-derived embryonic stem (ES) cells. Correctly targeted ES cells were injected into B6(Cg)-Tyr^c-2J/J blastocysts, and resulting chimeric males were bred to C57BL/6J females. Subsequently, mice were bred to mice expressing Flp recombinase on a C57BL/6N background to remove the neo cassette. Offspring were bred together to remove the Flp-expressing transgene and a homozygous colony of FLExDUX4 mice was created. Upon arrival at The Jackson Laboratory, sperm was cryopreserved. To establish our live colony, an aliquot of frozen sperm was used to fertilize C57BL/6J oocytes (Stock No. 000664).

A 48 SNP (single nucleotide polymorphism) panel analysis, with 43 markers covering all 19 chromosomes and the X chromosome, as well as 5 markers that distinguish between the C57BL/6J and C57BL/6N substrains, was performed on the rederived living colony at The Jackson Laboratory Repository upon arrival. While the 43 markers throughout the genome suggested a C57BL/6 genetic background, 5 of 5 markers that determine C57BL/6J from C57BL/6N were found to be segregating. These data suggest the mice sent to The Jackson Laboratory Repository were on a mixed C57BL/6N;C57BL/6J genetic background.

Expression Data

Expressed Gene	DUX4, double homeobox 4, human
Site of Expression

Control Suggestions

Approximate Controls

000664 C57BL/6J

Additional Information

Considerations for Choosing Controls

Selected References

Jones T; Jones PL. 2018. A cre-inducible DUX4 transgenic mouse model for investigating facioscapulohumeral muscular dystrophy. PLoS One 13(2):e0192657PubMed: 29415061 MGI: J:256652
Jones TI; Chew GL; Barraza-Flores P; Schreier S; Ramirez M; Wuebbles RD; Burkin DJ; Bradley RK; Jones PL. 2018. Transgenic mice expressing tunable levels of DUX4 develope characteristic facioscapulohumeral muscular dystrophy-like pathophysiology rangingin severity bioRxivMGI: J:266688

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Genetics

Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}*

Allele Symbol: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}*

Allele Name	targeted mutation 1.1, Peter Jones
Allele Type	Targeted (Conditional ready (e.g. floxed), Inserted expressed sequence, Humanized sequence)
Allele Synonym(s)	FLExDUX4
Gene Symbol and Name	Gt(ROSA)26Sor, gene trap ROSA 26, Philippe Soriano
Gene Synonym(s)
Expressed Gene	DUX4, double homeobox 4, human
Strain of Origin	C57BL/6
Chromosome	6
Molecular Note	The targeting vector was designed with (from 5 to 3) a loxP site, an FRT-flanked neomycin resistance gene, a lox511 site, an inverted human double homeobox 4 (DUX4) gene with a polyadenylation sequence, and another loxP site and lox511 site, both in opposite orientation to the 5 sites. The DUX4 gene contains 4 silent point mutations in exon 1 that prevent alternative splicing of a truncated isoform, DUX4-s, while maintaining the DUX4-fl ORF. This entire construct was inserted between exons 1 and 2 of the Gt(ROSA)26Sor locus. Flp-mediated recombination removed the FRT-flanked neo cassette.

Disease/Phenotype

Disease Terms

Model with phenotypic similarity to human disease where etiologies are distinct. Human genes are associated with this disease. Orthologs of these genes do not appear in the mouse genotype(s).

facioscapulohumeral muscular dystrophy

Research Areas By Phenotype

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

Neurobiology Research
- Muscular Dystrophy
- Cre-lox System
  - loxP-flanked Sequences
Research Tools
- Cre-lox System
  - loxP-flanked Sequences

Mammalian Phenotype Terms by Genotype

The following phenotype information is associated with a similar, but not exact match to this JAX^® Mice strain

Genotype: Edil3^{Tg(Sox2-cre)1Amc}/0 Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺
involves: C57BL/6 CBA

mortality/aging

embryonic lethality, complete penetrance

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Tg(ACTA1-cre)79Jme/0
involves: C57BL/6 SJL

growth/size/body region phenotype

decreased fetal size
- observed in E16.5 embryos

limbs/digits/tail phenotype

abnormal limb morphology
- E16.5 embryos exhibit deformed limbs

mortality/aging

perinatal lethality
- pups are stillborn and small
- pups develop until almost full term (E19.5)

nervous system phenotype

abnormal spinal cord morphology
- E16.5 embryos exhibit aberrant spinal columns

skeleton phenotype

small thoracic cage
- E16.5 embryos exhibit underdeveloped rib cages

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Ndor1^{Tg(UBC-cre/ERT2)1Ejb}/0
involves: 129S/SvEv C57BL/6

behavior/neurological phenotype

ataxia
- between 5-12 weeks of age pups develop progressive ataxia

impaired righting response
- pups lose the righting reflex between 5-12 weeks of age

seizures
- pups develop spontaneous seizures characterized by periods of wild and uncontrolled running by 12 weeks of age

tremors
- between 5-12 weeks of age pups develop tremors

skeleton phenotype

kyphosis
- between 5-12 weeks of age pups develop severe kyphosis

immune system phenotype

enlarged lymph nodes
- swollen subiliac lymph nodes

enlarged popliteal lymph nodes

integument phenotype

alopecia
- P10 pups exhibit mild alopecia by 3 weeks, progressing to severe alopecia by 12 weeks of age

retarded hair growth
- pups exhibit delayed fur growth at P10

nervous system phenotype

seizures
- pups develop spontaneous seizures characterized by periods of wild and uncontrolled running by 12 weeks of age

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Tg(ACTA1-cre/Esr1)2Kesr/0
involves: C3H * C57BL/6

behavior/neurological phenotype

ataxia
- mice exhibit ataxia within 7-8 days following IP injection of TMX

decreased grip strength
- mice exhibit loss of strength (hanging test) within 7-8 days following IP injection of TMX

cellular phenotype

skeletal muscle necrosis
- necrosis and phagocytosis are observed at time of euthanasia (9 days post TMX)

growth/size/body region phenotype

weight loss
- mice exhibit weight loss within 7-8 days following IP injection of TMX

immune system phenotype

increased acute inflammation
- skeletal muscle tissues exhibit large infiltrates of mononuclear cells and loss of membrane integrity at time of euthanasia (9 days post TMX)

muscle phenotype

centrally nucleated skeletal muscle fibers
- muscle from untreated mice has patches of centralized nuclei

decreased skeletal muscle fiber size
- muscle from untreated mice has small irregular fibers

dystrophic muscle

increased variability of skeletal muscle fiber size
- heterogeneous fiber size distribution at time of euthanasia (9 days post TMX)
- muscle from untreated mice has small irregular fibers

myopathy
- Tamoxifen (TMX)-treated mice exhibit progressive myopathy
- untreated mice exhibit a mild muscle phenotype

skeletal muscle fibrosis
- skeletal muscle tissues are mildly fibrotic at time of euthanasia (9 days post TMX)

skeletal muscle necrosis
- necrosis and phagocytosis are observed at time of euthanasia (9 days post TMX)

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Pax7^{tm1(cre/ERT2)Gaka}/?
involves: 129S6/SvEvTac C57BL/6 * C57BL/6NCrl

normal phenotype

no abnormal phenotype detected
- mice are viable and healthy in the absence of tamoxifen

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

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4*)Plj}/Gt(ROSA)26Sor⁺
B6(Cg)-Gt(ROSA)26Sor

growth/size/body region phenotype

decreased body weight
- adult size is slightly reduced
- female mice are 11% smaller than controls by 20 weeks of age
- male mice are 14% smaller than controls by 20 weeks of age

postnatal growth retardation
- females exhibit slower growth after 12 weeks of age
- males exhibit slower growth after 8 weeks of age

mammalian phenotype

muscle phenotype
- Normal - mice exhibit no overt myopathy

immune system phenotype

enlarged lymph nodes
- swollen subiliac lymph nodes

enlarged popliteal lymph nodes

integument phenotype

alopecia
- mild alopecia
- phenotype is more severe in females than males

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}
B6(Cg)-Gt(ROSA)26Sor

digestive/alimentary phenotype

abnormal defecation
- soft stools

rectal prolapse
- some mice develop rectal prolapse after 6 months of age

mammalian phenotype

muscle phenotype
- Normal - mice exhibit no overt myopathy

reproductive system phenotype

preputial gland inflammation
- males develop an inflamed preputial gland

endocrine/exocrine gland phenotype

preputial gland inflammation
- males develop an inflamed preputial gland

growth/size/body region phenotype

decreased body weight
- adult mice are smaller than controls; weight gain peaks at 9 weeks and plateaus at 21-23 grams
- adults are 43% smaller than wild-type and 33% smaller than heterozygotes
- males are smaller from birth

immune system phenotype

preputial gland inflammation
- males develop an inflamed preputial gland

integument phenotype

alopecia
- alopecia appears between 3-8 weeks of age
- alopecia is more severe than in hemizygous mutant mice

References

Jones T; Jones PL. 2018. A cre-inducible DUX4 transgenic mouse model for investigating facioscapulohumeral muscular dystrophy. PLoS One 13(2):e0192657PubMed: 29415061 MGI: J:256652
Jones TI; Chew GL; Barraza-Flores P; Schreier S; Ramirez M; Wuebbles RD; Burkin DJ; Bradley RK; Jones PL. 2018. Transgenic mice expressing tunable levels of DUX4 develope characteristic facioscapulohumeral muscular dystrophy-like pathophysiology rangingin severity bioRxivMGI: J:266688

Additional - Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}* related

Technical Support

CONTACT TECHNICAL SUPPORT

Genotyping Protocols
- Standard PCR:Gt(ROSA)26Sor
- Genotyping resources and troubleshooting
Dietary Information
- New Diet as of March 2015: Lab Diet® 5K0Q (6% fat)
Breeding Considerations

When maintaining a live colony, homozygous mice may be bred together.
- Additional Breeding and Husbandry Support
Mating System
- Homozygote x Homozygote
Citation
When using the FLExDUX4 mouse strain in a publication, please cite the originating article(s) and include JAX stock #028710 in your Materials and Methods section.

Animal Health Reports

Facility Barrier Level Descriptions

AX10 (Standard)

Pricing & Availability

Available

Domestic
International

Live Mouse

Age

Genotype

Price

weeks

Cryorecovery - Pricing

Service/Product	Description	Price
	Heterozygous for Gt(ROSA)26Sor<tm1.1(DUX4*)Plj>

Related Products and Services

Frozen Mouse Embryo	B6(Cg)-Gt(ROSA)26Sor<tm1.1(DUX4*)Plj>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6(Cg)-Gt(ROSA)26Sor<tm1.1(DUX4*)Plj>/J Frozen Embryo	$2595.00
Frozen Mouse Embryo	B6(Cg)-Gt(ROSA)26Sor<tm1.1(DUX4*)Plj>/J Frozen Embryo	$3373.50
Frozen Mouse Embryo	B6(Cg)-Gt(ROSA)26Sor<tm1.1(DUX4*)Plj>/J Frozen Embryo	$3373.50

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Also Known As:Dux4-fl, FLExDUX4

Donating Investigator

Genetic overview

Research Applications

Base Price

Detailed Description

Development

Expression Data

Control Suggestions

Approximate Controls

Additional Information

Selected References

Gt(ROSA)26Sortm1.1(DUX4*)Plj

Allele Symbol: Gt(ROSA)26Sortm1.1(DUX4*)Plj

Disease Terms

Model with phenotypic similarity to human disease where etiologies are distinct. Human genes are associated with this disease. Orthologs of these genes do not appear in the mouse genotype(s).

Research Areas By Phenotype

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

Mammalian Phenotype Terms by Genotype

Genotype: Edil3Tg(Sox2-cre)1Amc/0 Gt(ROSA)26Sortm1.1(DUX4*)Plj/Gt(ROSA)26Sor+involves: C57BL/6 * CBA

mortality/aging

Genotype: Gt(ROSA)26Sortm1.1(DUX4*)Plj/Gt(ROSA)26Sor+ Tg(ACTA1-cre)79Jme/0involves: C57BL/6 * SJL

growth/size/body region phenotype

limbs/digits/tail phenotype

mortality/aging

nervous system phenotype

skeleton phenotype

Genotype: Gt(ROSA)26Sortm1.1(DUX4*)Plj/Gt(ROSA)26Sor+ Ndor1Tg(UBC-cre/ERT2)1Ejb/0involves: 129S/SvEv * C57BL/6

behavior/neurological phenotype

skeleton phenotype

immune system phenotype

integument phenotype

nervous system phenotype

Genotype: Gt(ROSA)26Sortm1.1(DUX4*)Plj/Gt(ROSA)26Sor+ Tg(ACTA1-cre/Esr1*)2Kesr/0involves: C3H * C57BL/6

behavior/neurological phenotype

cellular phenotype

growth/size/body region phenotype

immune system phenotype

muscle phenotype

Genotype: Gt(ROSA)26Sortm1.1(DUX4*)Plj/Gt(ROSA)26Sor+ Pax7tm1(cre/ERT2)Gaka/?involves: 129S6/SvEvTac * C57BL/6 * C57BL/6NCrl

normal phenotype

Genotype: Gt(ROSA)26Sortm1.1(DUX4*)Plj/Gt(ROSA)26Sor+B6(Cg)-Gt(ROSA)26Sor

growth/size/body region phenotype

mammalian phenotype

immune system phenotype

integument phenotype

Genotype: Gt(ROSA)26Sortm1.1(DUX4*)Plj/Gt(ROSA)26Sortm1.1(DUX4*)PljB6(Cg)-Gt(ROSA)26Sor

digestive/alimentary phenotype

mammalian phenotype

reproductive system phenotype

endocrine/exocrine gland phenotype

growth/size/body region phenotype

immune system phenotype

integument phenotype

References

Additional - Gt(ROSA)26Sortm1.1(DUX4*)Plj related

Genotyping Protocols

Dietary Information

Breeding Considerations

Mating System

Citation

Animal Health Reports

Live Mouse

Cryorecovery - Pricing

Related Products and Services

Payment Terms and Conditions

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

Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}*

Allele Symbol: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}*

Genotype: Edil3^{Tg(Sox2-cre)1Amc}/0 Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺
involves: C57BL/6 CBA

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Tg(ACTA1-cre)79Jme/0
involves: C57BL/6 SJL

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Ndor1^{Tg(UBC-cre/ERT2)1Ejb}/0
involves: 129S/SvEv C57BL/6

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Tg(ACTA1-cre/Esr1)2Kesr/0
involves: C3H * C57BL/6

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor⁺ Pax7^{tm1(cre/ERT2)Gaka}/?
involves: 129S6/SvEvTac C57BL/6 * C57BL/6NCrl

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4*)Plj}/Gt(ROSA)26Sor⁺
B6(Cg)-Gt(ROSA)26Sor

Genotype: Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}/Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}
B6(Cg)-Gt(ROSA)26Sor

Additional - Gt(ROSA)26Sor^{tm1.1(DUX4)Plj}* related