Identification and characterisation of human dysferlin transcript variants: implications for dysferlin mutational screening and isoforms

• Published in: Human genetics Vol. 125; no. 4; pp. 413 - 420
• Main Authors: Pramono, Zacharias Aloysius Dwi, Tan, Chin Lai, Seah, Irene Ai Lian, See, Joseph Shean Long, Kam, Siok Yuen, Lai, Poh San, Yee, Woon Chee
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01.05.2009; Springer-Verlag; Springer; Springer Nature B.V
• Subjects: Alternative Splicing; Base Sequence; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Classical genetics, quantitative genetics, hybrids; Distal Myopathies - genetics; DNA Mutational Analysis; DNA Primers - genetics; Dysferlin; Exons; Fundamental and applied biological sciences. Psychology; Gene Function; Genetic aspects; Genetic Variation; Genetics of eukaryotes. Biological and molecular evolution; Human; Human Genetics; Humans; Introns; Membrane Proteins - genetics; Messenger RNA; Metabolic Diseases; Molecular Medicine; Muscle Proteins - genetics; Muscle, Skeletal - metabolism; Muscles; Muscular Dystrophies, Limb-Girdle - genetics; Musculoskeletal system; Mutation; Neurosciences; Original Investigation; Protein Isoforms - genetics; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - blood; RNA, Messenger - genetics; RNA, Messenger - metabolism; Singapore; Alternative Promoter; Alternative Splice; Skeletal Muscle; Limb Girdle Muscular Dystrophy; Transcript Variant; Human; Variant; Screening; Molecular form; Genetics; Identification; Mutation; Gene expression
• ISSN: 0340-6717; 1432-1203
• DOI: 10.1007/s00439-009-0632-y

In conducting dysferlin mutational screening using blood mRNA instead of genomic DNA, we identified the occurrence of alternative splicing involving novel dysferlin exons, i.e. exons 5a and 40a, in addition to previously reported alternative splicing of exon 17. Further study employing long range RT-PCR and subcloning revealed a total of fourteen dysferlin transcripts with maintained dysferlin reading frame. The study also characterised the differences in relative frequencies of the dysferlin transcripts in skeletal muscle and blood. The findings have potential implications for molecular diagnosis of dysferlinopathy and the identification of dysferlin isoforms.