D4Z4-2.5 transgenic mice recapitulate important epigenetic and DUX4 expression attributes seen in human FSHD1. D4Z4-2.5 mice, along with the control strain (D4Z4-12.5 transgenic mice carrying a normal sized, non-pathogenic allele; Stock No. 028012), may be useful in studying facioscapulohumeral dystrophy.
Silvere M van der Maarel, Leiden University Medical Center (LUMC)
Genetic Background | Generation |
---|---|
|
Allele Type |
---|
Transgenic (Inserted expressed sequence, Humanized sequence) |
Facioscapulohumeral dystrophy (FSHD) is an autosomal dominant muscular dystrophy predominantly affecting facial and upper extremity muscles. Hearing loss and retinopathy are frequently observed extramuscular features. FSHD1 is caused by contraction of the D4Z4 macrosatellite repeat to 10 or less D4Z4 units in the subtelomeric region of 4q on the 4A161 haplotype. DUX4 is more efficiently silenced with increasing D4Z4 copy number: human FSHD1 is associated with 1-10 D4Z4 repeat units, whereas the unaffected population has 11-100 D4Z4 repeat units. When expressed in skeletal muscle, the DUX4 transcription factor activates genes normally expressed in the germline (essentially inducing a stem cell program in the postmitotic muscle cell) and also suppresses several genes involved in the innate immune response.
The D4Z4-2.5 transgene contains a FSHD1-sized D4Z4 repeat derived from a human FSHD1 individual (haplotype 4A161; containing 2.5 D4Z4 repeat units followed by the pLAM DUX4 exon 3 with polyA signal). Hemizygous D4Z4-2.5 transgenic mice recapitulate important epigenetic and DUX4 expression attributes seen in human FSHD1; including high DUX4 expression levels in the germline, incomplete epigenetic repression of DUX4 in somatic tissue, and FSHD-specific variegated DUX4 expression in sporadic muscle nuclei associated with D4Z4 chromatin relaxation.
The phenotype listed below compares the D4Z4-2.5 transgenic mice carrying a D4Z4 genomic region from a contracted pathogenic FSHD1 allele (Stock No. 027991) with the D4Z4-12.5 transgenic mice carrying a normal sized, non-pathogenic allele (Stock No. 028012). Unless noted otherwise, hemizygous mice are described.
The fewer (2.5) D4Z4 repeat units in D4Z4-2.5 mice causes inefficient DUX4 repression in somatic tissue (bodywide hypomethylation of D4Z4) and manifests in features of human FSHD1. In contrast, D4Z4-12.5 mice have a D4Z4 repeat array (12.5) that maintains somatic tissue epigenetic silencing of DUX4 at a low enough level sufficient to preclude FSHD1 phenotype in humans.
D4Z4-2.5 mice have abundant levels of DUX4 mRNA in germ line tissues (most notably in testis), in ES cells and in early developmental stages. In D4Z4-12.5 mice, the pattern is similar but at lower levels.
In adult somatic tissue from D4Z4-2.5 mice, DUX4 mRNA expression is low and variegated in all analyzed skeletal muscles, and a variegated expression pattern of DUX4 protein in skeletal muscle nuclei is observed. In adult somatic tissue from D4Z4-12.5 mice, DUX4 transcripts are reproducibly detected only in the tibialis anterior and pectoralis muscles, whereas all other somatic tissues do not show reproducible DUX4 expression.
D4Z4-2.5 mice exhibit chromatin relaxation of the D4Z4 repeats compared to D4Z4-12.5 mice.
In both D4Z4-2.5 and D4Z4-12.5 mice, the overall morphology/histology of the limb and some head muscles appear normal. Hemizygous mice are viable and fertile. The donating investigator also reports that homozygous mice are viable and fertile, and that the number of D4Z4 repeats within each line stays the same through several generations of breeding (i.e., the D4Z4 repeat unit is stable).
Approximately half of the D4Z4-2.5 mice develop eye abnormalities (keratitis) around 8-12 weeks of age.
The D4Z4-2.5 transgene was designed by Dr. Silvere M. van der Maarel (Leiden University Medical Center, The Netherlands) with the human facioscapulohumeral dystrophy (FSHD)-permissive FSHD1 allele. Specifically, an EcoRI fragment derived from the lambda-42 clone of an FSHD1 allele (haplotype 4A161) was obtained. This allele contains two-and-a-half copies of the D4Z4 macrosatellite repeat units (each unit has exons 1-2 of the DUX4 retrogene; with exon 1 encoding a double homeobox transcription factor), the region telomeric to the last repeat (called pLAM) contains DUX4 exon 3 (harboring a polyA signal in haplotype 4A161) and lacks the more downstream DUX4 exons 4-7. This ~13.5 kbp transgene was microinjected into the pronuclei of fertilized oocytes of B6CBAF1/J mice (Charles River Laboratories), and then founder animals were bred to C57BL/6NCrl mice for germline transmission. A single founder line was generated, and found to have 4-6 copies of the transgene integrated onto chromosome 17. The D4Z4-2.5 transgenic colony was then bred with C57BL/6Jico (C57BL/6NCrl) wildtype mice for at least 20 generations prior to sending black male mice to The Jackson Laboratory Repository in 2015. Upon arrival, males were used to cryopreserve sperm. To establish the living D4Z4-2.5 mouse colony, an aliquot of the frozen sperm was used to fertilize oocytes from C57BL/6NJ inbred females (Stock No. 005304).
Expressed Gene | DUX4, double homeobox 4, human |
---|---|
Site of Expression | Reproducible levels of DUX4 mRNA were detected in all analyzed skeletal muscles of adult mice, including affected muscles of the limbs, trunk and head. |
Allele Name | transgene insertion 1, Silvere van der Maarel |
---|---|
Allele Type | Transgenic (Inserted expressed sequence, Humanized sequence) |
Allele Synonym(s) | D4Z4-2.5 |
Gene Symbol and Name | Tg(DUX4*)1Maar, transgene insertion 1, Silvere van der Maarel |
Gene Synonym(s) | |
Promoter | DUX4, double homeobox 4, human |
Expressed Gene | DUX4, double homeobox 4, human |
Site of Expression | Reproducible levels of DUX4 mRNA were detected in all analyzed skeletal muscles of adult mice, including affected muscles of the limbs, trunk and head. |
Strain of Origin | (C57BL/6J x CBA/J)F1 |
Chromosome | 17 |
Molecular Note | This transgene carries a 13.5 kb EcoRI fragment derived from a lambda-42 phage clone encompassing the partially deleted D4Z4 locus of a patient with facioscapulohumeral dystrophy. This allele contains the DUX4 pA signal, but lacks the more downstream exons 6 and 7 and contains 2.5 copies of the D4Z4 unit and flanking sequences. 4 copies of the fragment were integrated on chromosome 17. DUX4 mRNA was detected in germline tissues and variable expression is seen in analyzed somatic tissues including skeletal muscle and in the cerebellum. |
When maintaining a live colony, hemizygous mice may be bred to wildtype (noncarrier) siblings or to C57BL/6NJ inbred mice (Stock No. 005304). The donating investigator reports that homozygous mice are viable and fertile, and that the number of D4Z4 repeats stays the same through several generations of breeding (i.e., the D4Z4 repeat unit is stable).
When using the D4Z4-2.5 mouse strain in a publication, please cite the originating article(s) and include JAX stock #027991 in your Materials and Methods section.
Facility Barrier Level Descriptions
Service/Product | Description | Price |
---|---|---|
Hemizygous or Non Carrier for Tg(DUX4*)1Maar |
Frozen Mouse Embryo | B6N.Cg-Tg(DUX4*)1Maar/J | $2595.00 |
Frozen Mouse Embryo | B6N.Cg-Tg(DUX4*)1Maar/J | $2595.00 |
Frozen Mouse Embryo | B6N.Cg-Tg(DUX4*)1Maar/J | $3373.50 |
Frozen Mouse Embryo | B6N.Cg-Tg(DUX4*)1Maar/J | $3373.50 |
Terms are granted by individual review and stated on the customer invoice(s) and account statement. These transactions are payable in U.S. currency within the granted terms. Payment for services, products, shipping containers, and shipping costs that are rendered are expected within the payment terms indicated on the invoice or stated by contract. Invoices and account balances in arrears of stated terms may result in The Jackson Laboratory pursuing collection activities including but not limited to outside agencies and court filings.
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.
What information were you hoping to find through your search?
How easy was it to find what you were looking for?
We may wish to follow up with you. Enter your email if you are happy for us to connect and reachout to you with more questions.
Please Enter a Valid Email Address
Thank you for sharing your feedback! We are working on improving the JAX Mice search. Come back soon for exciting changes.