A generic assay for the identification of splicing variants that induce nonsense-mediated decay in Pompe disease.

摘要: DNA variants affecting mRNA expression and processing in genetic diseases are often missed or poorly characterized. We previously reported a generic assay to identify that affect splicing Pompe disease, monogenic disorder caused by deficiency of acid α-glucosidase (GAA). However, this could miss is subjected degradation. Here, we inhibited degradation using cycloheximide performed unbiased analysis all GAA exons exon flanking RT-PCR internal RT-qPCR. In four patients were suspected harboring but for which aberrant not be detected normally growing cells, total 10 novel events cells treated with cycloheximide. addition, found sequences of GAA introns 6 12 naturally included subset transcripts from healthy controls, indicating inefficient canonical splicing. Identification the common Asian variant c.546G>T allowed development an antisense oligonucleotide promoted pre-mRNA elevated enzymatic activity. Our results indicate this extended allows detection cases enable functional unknown targeted treatment options.