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

ABSTRACT: DNA variants affecting mRNA expression and processing in genetic diseases are often missed or poorly characterized. We previously reported a generic assay to identify variants that affect mRNA expression and splicing in Pompe disease, a monogenic disorder caused by deficiency of acid α-glucosidase (GAA). However, this assay could miss mRNA that is subjected to degradation. Here, we inhibited mRNA degradation using cycloheximide and performed unbiased splicing analysis of all GAA exons using exon flanking RT-PCR and exon internal RT-qPCR. In four patients that were suspected of harboring splicing variants but for which aberrant splicing could not be detected in normally growing cells, we detected a total of 10 novel splicing events in cells treated with cycloheximide. In addition, we found

SUBMITTER: Bergsma AJ 

PROVIDER: S-EPMC7940403 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature