CONCLUSION
In summary, we have carried out a large-scale analysis of exonic
variants in SLC12A3 related to GS through bioinformatics
predictions and minigene assays. The results indicated that six
previously presumed missense variants should be classified as splicing
variations, which induced the corresponding exon skipping. These
variants either break ESEs and
generate new ESSs, or interfere with the canonical splice sites, causing
significant pre-mRNA splicing alterations, probably associated with
their pathogenicity. This study highlights the important role of
assessing the effects of missense variants at the mRNA level in GS, as
well as the effectiveness of minigene splicing analysis.