Transgenic alternative-splicing reporters reveal tissue-specific expression profiles and regulation mechanisms in vivo

• Published in: Nature methods Vol. 3; no. 11; pp. 909 - 915
• Main Authors: Kuroyanagi, Hidehito, Hagiwara, Masatoshi, Kobayashi, Tetsuo, Mitani, Shohei
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 01.11.2006
• Subjects: Alternative Splicing; Amino Acid Sequence; Analysis; Animals; Caenorhabditis elegans; Caenorhabditis elegans - genetics; Caenorhabditis elegans - metabolism; Caenorhabditis elegans Proteins - genetics; Caenorhabditis elegans Proteins - metabolism; Cell culture; DNA microarrays; Exons; Gene expression; Gene Expression Profiling; Genes, Reporter - genetics; Genetic engineering; Metazoa; Methods; Molecular Sequence Data; Mutants; Mutation; Proteins; Receptors, Fibroblast Growth Factor - genetics; Receptors, Fibroblast Growth Factor - metabolism; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleic acid; RNA; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Tissues; Transgenes - genetics; Japan
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth944

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.