The DMDgene analysed by field inversion gel electrophoresis

Dunnen, Johan T. den; Bakker, E.; Ommen, G.J.B. van; Pearson, P. L.

doi:10.1093/oxfordjournals.bmb.a072350

Cited by 18 publications

(7 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The presence of deletion hot spots in the hDMD gene is well established (1,42). The reason for this is presently unclear but may lie in the chromosomal context (e.g.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Generation and Characterization of Transgenic Mice with the Full-length Human DMD Gene

Hoen

Meijer

Boer

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

We report the generation of mice with an intact and functional copy of the 2.3-megabase human dystrophin gene (hDMD), the largest functional stretch of human DNA thus far integrated into a mouse chromosome. Yeast spheroplasts containing an artificial chromosome with the full-length hDMD gene were fused with mouse embryonic stem cells and were subsequently injected into mouse blastocysts to produce transgenic hDMD mice. Human-specific PCR, Southern blotting, and fluorescent in situ hybridization techniques demonstrated the intactness and stable chromosomal integration of the hDMD gene on mouse chromosome 5. Expression of the transgene was confirmed by RT-PCR and Western blotting. The tissue-specific expression pattern of the different DMD transcripts was maintained. However, the human Dp427p and Dp427m transcripts were expressed at 2-fold higher levels and human Dp427c and Dp260 transcripts were expressed at 2-and 4-fold lower levels than their endogenous counterparts. Ultimate functional proof of the hDMD transgene was obtained by crossing of hDMD mice with dystrophin-deficient mdx mice and dystrophin and utrophin-deficient mdx ؋ Utrn ؊/؊ mice. The hDMD transgene rescued the lethal dystrophic phenotype of the mdx ؋ Utrn ؊/؊ mice. All signs of muscular dystrophy disappeared in the rescued mice, as demonstrated by histological staining of muscle sections and gene expression profiling experiments. Currently, hDMD mice are extensively used for preclinical testing of sequence-specific therapeutics for the treatment of Duchenne muscular dystrophy. In addition, the hDMD mouse can be used to study the influence of the genomic context on deletion and recombination frequencies, genome stability, and gene expression regulation.

show abstract

“…The presence of deletion hot spots in the hDMD gene is well established (1,42). The reason for this is presently unclear but may lie in the chromosomal context (e.g.…”

Section: Discussionmentioning

confidence: 99%

“…It is located on the X chromosome (band Xp21.2), spans 2.3 Mb, 2 and contains 79 exons (1)(2)(3). Transcripts originating from seven different promoters throughout the gene have been described, whereas, in addition, extensive differential splicing of these transcripts has been acknowledged (details can be found on the Leiden Muscular Dystrophy Web pages).…”

mentioning

confidence: 99%

Generation and Characterization of Transgenic Mice with the Full-length Human DMD Gene

Hoen

Meijer

Boer

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…masked by the presence of the corresponding normal allele. Deletions therefore require either a quantitative method (such as MLPA or array CGH), or a qualitative method, such as pulsed field or field inversion gel electrophoresis [18].…”

Section: Carrier Diagnosis For a Known Familial Mutationmentioning

confidence: 99%

Best Practice Guidelines on molecular diagnostics in Duchenne/Becker muscular dystrophies

Abbs

Tuffery‐Giraud

Bakker

et al. 2010

Neuromuscular Disorders

View full text Add to dashboard Cite

“…[1][2][3][4][5]6 the DMD gene product, has been localised in the sarcolemma of the muscle cell.7'10 In muscle biopsies of Duchenne patients no dystrophin can be detected, whereas in BMD patients dystrophin of abnormal size or quantity is observed. 7 The combined results of protein and DNA studies4 [12][13][14][15] have confirmed the hypothesis'6 that the Duchenne phenotype might be caused by an 'out of frame' mutation of the gene, although some exceptions have been found. The mutations produce a prematurely terminated protein, which is apparently unstable and thus degraded.…”

mentioning

confidence: 79%

Immunohistochemical studies show truncated dystrophins in the myotubes of three fetuses at risk for Duchenne muscular dystrophy.

Ginjaar

Bakker

Paassen

et al. 1991

Journal of Medical Genetics

Self Cite

View full text Add to dashboard Cite

We have performed immunohistochemical studies on muscle tissue of three 12 week old fetuses at risk for DMD, using antisera directed against regions located NH2-proximally and centrally in the rod shaped spectrin-like domain and against the COOH-terminus of dystrophin. All three fetuses had a family history ofDMD. Truncateddystrophins were identified in all three cases by a positive reaction with the NH2-proximal antibody, different reactions with the central antibody, and a negative reaction with the COOH-terminal antibody. These data indicate that a panel of antibodies would, in principle, permit 'immunological' mapping of dystrophin mutations. This is diagnostically important in the 35% of families where no mutation is detectable at the DNA level. Secondly, by using this mapping technique it may also become possible to identify the at risk haplotype when DNA analysis is not informative. This may be of great value in DMD carrier detection.X linked Duchenne muscular dystrophy (DMD) is the most common of all severe muscular dystrophies

show abstract

The DMDgene analysed by field inversion gel electrophoresis

Cited by 18 publications

References 0 publications

Generation and Characterization of Transgenic Mice with the Full-length Human DMD Gene

Generation and Characterization of Transgenic Mice with the Full-length Human DMD Gene

Best Practice Guidelines on molecular diagnostics in Duchenne/Becker muscular dystrophies

Immunohistochemical studies show truncated dystrophins in the myotubes of three fetuses at risk for Duchenne muscular dystrophy.

Contact Info

Product

Resources

About