Abstract
Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy are caused in most cases by deletions of the DMD gene. These rearrangements are detectable in affected boys and men by a simple multiplex polymerase chain reaction approach. However, this technique is not able to disclose DMD deletions in heterozygous female carriers, and different approaches must be used in these cases. Here, we describe an approach based on the combined use of primed in situ labeling and fluorescence in situ hybridization techniques for the detection of single DMD exons in fixed metaphase chromosomes and interphase nuclei of both male and female subjects, suggesting the usefulness of this tool in the detection of small intragenic deletions of the DMD gene.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Sellner, L. N. and Taylor, G. R. (2004) MLPA and MAPH: new techniques for detection of gene deletions. Hum. Mutat. 23, 413–419.
Muntoni, F., Torelli, S., and Ferlini, A. (2003) Dystrophin and mutations: one gene, several proteins, multiple phenotypes. Lancet Neurol. 2, 731–740.
Mandel, J. L. (1989). Dystrophin. The gene and its product. Nature 339, 584–586
Chamberlain, J. S., Gibbs, R. A., Ranier, J. E., Nguyen, P. N., and Caskey, C. T. (1988) Deletion screening of the Duchenne muscular dystrophy locus via multiplex DNA amplification. Nucleic Acids Res. 16, 11,141–11,156.
Den Dunnen, J. T., Grootscholten, P. M., Bakker, E., Blonden, L., Ginjaar, H. B., Wapenaar, M. C., et al. (1989). Topography of the Duchenne muscular dystrophy (DMD) gene: FIGE and cDNA analysis of 194 cases reveals 115 deletions and 13 duplications. Am. J. Hum. Genet. 45, 835–847.
Beggs, A. H., Koenig, M., Boyce, F. M., and Kunkel, L. M. (1990). Detection of 98% of DMD/BMD gene deletions by polymerase chain reaction. Hum. Genet. 86, 45–48.
Koenig, M., Monaco, A. P., and Kunkel, L. M. (1988). The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein. Cell 53, 219–226.
Muntoni, F. and Strong, P. N. (1989) Transcription of the dystrophin gene in Duchenne muscular dystrophy muscle. FEBS Lett. 252, 95–98.
Acknowledgment
We thank Dr. Carmela Trimarchi for technical assistance and her for the helpful and constructive comments. This work was supported by CNR grant.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Stuppia, L., La Sala, D., Cinti, C. (2006). Combined Fluorescence In Situ Hybridization and PRINS for the Analysis of the Dystrophin Gene. In: Pellestor, F. (eds) PRINS and In Situ PCR Protocols. Methods in Molecular Biology™, vol 334. Humana Press. https://doi.org/10.1385/1-59745-068-5:115
Download citation
DOI: https://doi.org/10.1385/1-59745-068-5:115
Publisher Name: Humana Press
Print ISBN: 978-1-58829-549-1
Online ISBN: 978-1-59745-068-3
eBook Packages: Springer Protocols