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Pre-mRNA splicing is a fundamental step in mRNA maturation and its discovery in 1977 revolutionized our understanding of gene expression. This collection includes reviews and research articles from across the Nature group of journals to showcase the latest advances in splicing research and the emerging understanding of how splicing regulates gene expression, and through it cell fate, development and physiology. The articles focus on topics that include alternative splicing, the architecture and function of spliceosome complexes, the coordination of splicing with other processes such as transcription and mRNA decay, and splicing-related diseases and therapeutics.