The splicing of nuclear pre-mRNAs is a fundamental process required for the expression of most metazoan genes. The majority of the approximately 25,000 genes encoded by the human genome has been shown to produce more than one kind of transcripts through alternative splicing. Alternative splicing of pre-mRNAs can lead to the production of multiple protein isoforms from a single gene, significantly enriching the proteomic diversity of higher eukaryotic organisms. Because regulation of this process determines the timing and location that a particular protein isoform is produced, changes of alternative splicing patterns have the potential to modulate many cellular activities. Consequently, pre-mRNA splicing must occur with a high degree of specificity and fidelity to ensure the appropriate expression of functional mRNAs. Here we review recent progress made in understanding the extent of alternative splicing within the human genome with particular emphasis on splicing fidelity.