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

Nat Methods. 2006 Nov;3(11):909-15. doi: 10.1038/nmeth944.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alternative Splicing*
  • Amino Acid Sequence
  • Animals
  • Caenorhabditis elegans / genetics*
  • Caenorhabditis elegans / metabolism*
  • Caenorhabditis elegans Proteins / genetics*
  • Caenorhabditis elegans Proteins / metabolism
  • Exons
  • Gene Expression Profiling*
  • Genes, Reporter / genetics*
  • Molecular Sequence Data
  • Mutation
  • RNA-Binding Proteins / genetics*
  • RNA-Binding Proteins / metabolism
  • Receptors, Fibroblast Growth Factor / genetics*
  • Receptors, Fibroblast Growth Factor / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Transgenes / genetics*

Substances

  • Asd-1 protein, C elegans
  • Caenorhabditis elegans Proteins
  • EGL-15 protein, C elegans
  • FOX-1 protein, C elegans
  • RNA-Binding Proteins
  • Receptors, Fibroblast Growth Factor

Associated data

  • GENBANK/DQ485531