Identification of a Splice Site Mutation in the Cystathionine β-Synthase Gene Resulting in Variable and Novel Splicing Defects of Pre-mRNA

We used single-strand conformational polymorphism and direct nucleotide sequencing to identify a novel mutation in the cystathionine β-synthase (CBS) gene of two siblings with homocystinuria. Both patients are heterozygous carriers of the G₉₁₉A transition and the novel mutation which involves a G-to-A transition in the intron 12 splice donor site. Reverse transcription of RNA harvested from transformed lymphocytes followed by PCR showed a normal size product along with two shorter products involving the deletion of either exon 12 alone or both exons 11 and 12. To our knowledge, the skipping of more than one exon through a single base substitution at a splice-donor site has not been previously reported. The normal size splice product was found to have either a G or an A at nucleotide position 919, indicating that normal size mRNA was produced by both alleles.