Figure 4

Figure 4

Single molecule real time sequencing of f2^+/Δ14 mRNA demonstrates altered splicing in f2 following a deletion in exon 6. (top) Sanger sequencing of f2^+/+ and f2^Δ14/Δ14 cDNA reveals a canonical splice donor but alternative splice acceptor (red dashed lines indicate splice junctions). Allelic SNPs in exon 7 allowed the sorting of transcripts by their genomic allele in mRNA extracted from heterozygotes. Wild-type transcripts (middle) solely demonstrated canonical splicing. Mutant transcripts (bottom) primarily exhibited alternative splicing to a cryptic splice site with a resulting 45 bp inframe deletion, as well as low frequency canonical splicing with a resulting 14 bp out of frame deletion. gDNA, genomic DNA.