title: Whole genome sequencing reveals a 7 base-pair deletion in DMD exon 42 in a dog with muscular dystrophy.
abstract: Dystrophin is a key cytoskeletal protein coded by the Duchenne muscular dystrophy (DMD) gene located on the X-chromosome. Truncating mutations in the DMD gene cause loss of dystrophin and the classical DMD clinical syndrome. Spontaneous DMD gene mutations and associated phenotypes occur in several other species. The mdx mouse model and the golden retriever muscular dystrophy (GRMD) canine model have been used extensively to study DMD disease pathogenesis and show efficacy and side effects of putative treatments. Certain DMD gene mutations in high-risk, the so-called hot spot areas can be particularly helpful in modeling molecular therapies. Identification of specific mutations has been greatly enhanced by new genomic methods. Whole genome, next generation sequencing (WGS) has been recently used to define DMD patient mutations, but has not been used in dystrophic dogs. A dystrophin-deficient Cavalier King Charles Spaniel (CKCS) dog was evaluated at the functional, histopathological, biochemical, and molecular level. The affected dog's phenotype was compared to the previously reported canine dystrophinopathies. WGS was then used to detect a 7 base pair deletion in DMD exon 42 (c.6051-6057delTCTCAAT mRNA), predicting a frameshift in gene transcription and truncation of dystrophin protein translation. The deletion was confirmed with conventional PCR and Sanger sequencing. This mutation is in a secondary DMD gene hotspot area distinct from the one identified earlier at the 5' donor splice site of intron 50 in the CKCS breed.
authors.affiliation: Department of Veterinary Integrative Biosciences (Mail Stop 4458), College of Veterinary Medicine, Texas A&M University, College Station, TX, 77843-4458, USA. pnghiem@cvm.tamu.edu.
authors.forename: Peter P
authors.identifier: 0000-0002-8796-8123
authors.initials: PP
authors.lastname: Nghiem
authors.affiliation: Department of Neurosciences, University of Padova, Via Giustiniani 5, 35128, Padova, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Bello
authors.affiliation: Department of Veterinary Integrative Biosciences (Mail Stop 4458), College of Veterinary Medicine, Texas A&M University, College Station, TX, 77843-4458, USA.
authors.forename: Cindy
authors.initials: C
authors.lastname: Balog-Alvarez
authors.affiliation: Department of Veterinary Integrative Biosciences (Mail Stop 4458), College of Veterinary Medicine, Texas A&M University, College Station, TX, 77843-4458, USA.
authors.forename: Sara Mata
authors.initials: SM
authors.lastname: López
authors.affiliation: Department of Veterinary Integrative Biosciences (Mail Stop 4458), College of Veterinary Medicine, Texas A&M University, College Station, TX, 77843-4458, USA.
authors.forename: Amanda
authors.initials: A
authors.lastname: Bettis
authors.affiliation: Department of Veterinary Integrative Biosciences (Mail Stop 4458), College of Veterinary Medicine, Texas A&M University, College Station, TX, 77843-4458, USA.
authors.forename: Heather
authors.initials: H
authors.lastname: Barnett
authors.affiliation: Department of Veterinary Integrative Biosciences (Mail Stop 4458), College of Veterinary Medicine, Texas A&M University, College Station, TX, 77843-4458, USA.
authors.forename: Briana
authors.initials: B
authors.lastname: Hernandez
authors.affiliation: Veterinary Emergency and Specialty Center of Santa Fe, 2001 Vivigen Way, Santa Fe, NM, 87505, USA.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Schatzberg
authors.affiliation: Department of Clinical Sciences and Services, Royal Veterinary College, London, UK.
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Piercy
authors.affiliation: Department of Veterinary Integrative Biosciences (Mail Stop 4458), College of Veterinary Medicine, Texas A&M University, College Station, TX, 77843-4458, USA.
authors.forename: Joe N
authors.initials: JN
authors.lastname: Kornegay
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D004195:Disease Models, Animal; D004285:Dogs; D016189:Dystrophin / Q000235:genetics*; D005091:Exons / Q000235:genetics; D006801:Humans; D007438:Introns / Q000235:genetics; D051379:Mice; D018101:Mice, Inbred mdx / Q000235:genetics; D009136:Muscular Dystrophies / Q000235:genetics*; D009154:Mutation; D012333:RNA, Messenger; D017384:Sequence Deletion / Q000235:genetics*; D000073336:Whole Genome Sequencing / Q000379:methods*
chemical_list: D016189:Dystrophin; D012333:RNA, Messenger