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Alternative splicing of pre-mRNAs greatly contributes to the spatiotemporal diversity of gene expression in metazoans. However, the molecular basis of developmental regulation and the precise sequence of alternative pre-mRNA processing in vivo are poorly understood. In the present study, we focus on the developmental switching of the mutually exclusive alternative splicing of the  let-2  gene of  Caenorhabditis elegans  from the exon 9 form in embryos to the exon 10 form in adults. By visualizing the usage of the  let-2  mutually exclusive exons through differential expression of green fluorescent protein (GFP) and red fluorescent protein (RFP), we isolated several switching-defective mutants and identified the  alternative splicing defective-2  ( asd-2 ) gene, encoding a novel member of the evolutionarily conserved STAR (signal transduction activators of RNA) family of RNA-binding proteins. Comparison of the amounts of partially spliced  let-2  RNAs in synchronized wild-type and  asd-2  mutant worms suggested that either of the introns downstream from the  let-2  mutually exclusive exons is removed prior to the removal of the upstream ones, and that  asd-2  promotes biased excision of intron 10 in the late larval stages. We propose that the developmental switching between alternative sequences of intron removal determines the ratio between the mature  let-2  mRNA isoforms.

STAR family RNA-binding protein ASD-2 regulates developmental switching of mutually exclusive alternative splicing in vivo