Identification of eight exonic variants in the SLC4A1, ATP6V1B1 and
ATP6V0A4 gene that alter RNA splicing by minigene assay
Abstract
Primary distal renal tubular acidosis (dRTA) is a rare tubular disease
associated with variants in SLC4A1, ATP6V0A4, ATP6V1B1, FOXⅠ1 or WDR72
genes. Currently, there is growing evidence that all types of exonic
variants can alter splicing regulatory elements, affecting the pre-mRNA
splicing process. This study was to determine the consequences of
variants associated with dRTA on pre-mRNA splicing combined with
predictive bioinformatics tools and minigene assay. As a result, among
the 15 candidate variants, 8 variants distributed in SLC4A1
(c.1765C>T, p.Arg589Cys), ATP6V1B1( c.368G>T,
p.Gly123Val; c.370C>T, p.Arg124Trp; c.484G>T,
p.Glu162* and c.1102G>A, p.Glu368Lys) and ATP6V0A4 genes
(c.322C>T, p.Gln108*; c.1571C>T, p.Pro524Leu
and c.1572G>A, p.Pro524Pro) were identified to result in
whole or part of exon skipping by either disruption of ESEs and
generation of ESSs, or interference with the recognition of the classic
splicing site, or both. To our knowledge, this is the first study on
pre-mRNA splicing of exonic variants in the dRTA-related genes. These
results highlight the importance of assessing the effects of exonic
variants at the mRNA level and suggest that minigene analysis is an
effective tool for evaluating the effects of splicing on variants in
vitro