Nature Methods
- 3, 909 - 915 (2006)
Published online: 23 October 2006; | doi:10.1038/nmeth944
Transgenic alternative-splicing reporters reveal tissue-specific expression profiles and regulation mechanisms in vivoHidehito Kuroyanagi1, 2, Tetsuo Kobayashi3, 4, Shohei Mitani3, 4 & Masatoshi Hagiwara1, 21
School of Biomedical Science, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan. 2
Medical Research Institute, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, Tokyo 113-8510, Japan. 3
Department of Physiology, Tokyo Women's Medical University School of Medicine, Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. 4
CREST, JST, Hon-cho, Kawaguchi, Saitama 332-0012, Japan.
Correspondence should be addressed to Hidehito Kuroyanagi kuroyana.end@tmd.ac.jp or Masatoshi Hagiwara m.hagiwara.end@mri.tmd.ac.jp Alternative splicing of pre-mRNAs allows multicellular organisms to create a huge diversity of proteomes from a finite number of genes. But extensive studies in vitro or in cultured cells have not fully explained the regulation mechanisms of tissue-specific or developmentally regulated alternative splicing in living organisms. Here we report a transgenic reporter system that allows visualization of expression profiles of mutually exclusive exons in Caenorhabditis elegans. Reporters for egl-15 exons 5A and 5B showed tissue-specific profiles, and we isolated mutants defective in the tissue specificity. We identified alternative-splicing defective-1 (asd-1), encoding a new RNA-binding protein of the evolutionarily conserved Fox-1 family, as a regulator of the egl-15 reporter. Furthermore, an asd-1;
fox-1 double mutant was defective in the expression of endogenous egl-15 (5A) and phenocopied egl-15 (5A) mutant. This transgenic reporter system can be a powerful experimental tool for the comprehensive study of expression profiles and regulation mechanisms of alternative splicing in metazoans.
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