Human Molecular Genetics Advance Access originally published online on October 28, 2008
Human Molecular Genetics 2009 18(1):202-211; 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, Children's Hospital Boston, Boston, MA, USA
* To whom correspondence should be addressed at: Program in Genomics Children's Hospital Boston Enders Rm 570 300 Longwood Ave, Boston, MA 02115, USA. Tel: +1 6173558200; Fax: +1 6173557588; Email: kunkel{at}enders.tch.harvard.edu
Received June 16, 2008; Accepted October 8, 2008
Sapje-like (sapcl100) was one of eight 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.
Present address: Auke Bay Laboratories, National Marine Fisheries Service, Ted Stevens Marine Research Institute, Juneau, AK, USA.