Transgenic alternative-splicing reporters reveal tissue-specific expression profiles and regulation mechanisms in vivo
doi: 10.1038/nmeth944
pmid: 17060915
Transgenic alternative-splicing reporters reveal tissue-specific expression profiles and regulation mechanisms in vivo
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.
Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, Molecular Sequence Data, RNA-Binding Proteins, Exons, Receptors, Fibroblast Growth Factor, Alternative Splicing, Genes, Reporter, Mutation, Animals, Amino Acid Sequence, Transgenes, Caenorhabditis elegans, Caenorhabditis elegans Proteins
Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, Molecular Sequence Data, RNA-Binding Proteins, Exons, Receptors, Fibroblast Growth Factor, Alternative Splicing, Genes, Reporter, Mutation, Animals, Amino Acid Sequence, Transgenes, Caenorhabditis elegans, Caenorhabditis elegans Proteins
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