Human Molecular Genetics Advance Access published online on October 28, 2008
Human Molecular Genetics, doi:10.1093/hmg/ddn337
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Genetic isolation and characterization of a splicing mutant of zebrafish dystrophin
1 Division of Genetics, Children's Hospital, Boston, MA, USA 2 Department of Genetics, Harvard Medical School, Boston, MA, USA 3 Howard Hughes Medical Institute, Boston, MA, USA, Children's Hospital Boston, MA, USA
* To whom correspondence should be addressed at: Louis M. Kunkel, Ph.D., Director, Program in Genomics, Children's Hospital Boston, Enders Rm 570, 300 Longwood Ave, Boston, MA 02115, Tel: 617-355-8200, Fax: 617-355-7588, Email: kunkel{at}enders.tch.harvard.edu
Received June 16, 2008; Revised October 8, 2008; Accepted October 8, 2008
Sapje-like sapcl100) was one of 8 potential zebrafish muscle mutants isolated as part of an early-pressure screen of 500 families. This mutant shows a muscle tearing phenotype similar to sapje (dys -/-) and both mutants fail to genetically complement suggesting they have a mutation in the same gene. Protein analysis confirms a lack of dystrophin in developing sapje-like embryos. Sequence analysis of the sapje-like dystrophin mRNA shows that exon 62 is missing in the dystrophin transcript causing exon 63 to be translated out of frame terminating translation at a premature stop codon at the end of exon 63. Sequence analysis of sapje-like genomic DNA identified a mutation in the donor splice junction at the end of dystrophin exon 62. This mutation is similar to splicing mutations associated with human forms of Duchenne Muscular Dystrophy. Sapje-like is the first zebrafish dystrophin splicing mutant identified to date and represents a novel disease model which can be used in future studies to identify therapeutic compounds for treating diseases caused by splicing defects.
Current Address: Auke Bay Laboratories, National Marine Fisheries Service, Ted Stevens Marine Research Institute, Juneau, AK