AUTHOR=Xiong Ting , Xu Zhongjin , Wan Qian , Chen Feng , Ye Yao , Wang Hong , Wu Chongjun 

TITLE=Identification of a novel ANK1 gene variant c.1504-9G>A and its mechanism of intron retention in hereditary spherocytosis

JOURNAL=Frontiers in Genetics

VOLUME=15

YEAR=2024

URL=https://www.frontiersin.org/journals/genetics/articles/10.3389/fgene.2024.1390924

DOI=10.3389/fgene.2024.1390924

ISSN=1664-8021

ABSTRACT=<p><bold>Objective:</bold> The objective of this study was to pinpoint pathogenic genes and assess the mutagenic pathogenicity in two pediatric patients with hereditary spherocytosis.</p><p><bold>Methods:</bold> We utilized whole-exome sequencing (WES) for individual analysis (case 1) and family-based trio analysis (case 2). The significance of the intronic mutation was validated through a Minigene splicing assay and supported by subsequent <italic>in vitro</italic> experiments.</p><p><bold>Results:</bold> Both probands received a diagnosis of hereditary spherocytosis. WES identified a novel <italic>ANK1</italic> c.1504-9G&gt;A mutation in both patients, causing the retention of seven nucleotides at the 5′ end of intron 13, as substantiated by the Minigene assay. This variant results in a premature stop codon and the production of a truncated protein. <italic>In vitro</italic> studies indicated a reduced expression of the <italic>ANK1</italic> gene.</p><p><bold>Conclusion:</bold> The novel <italic>ANK1</italic> c.1504-9G&gt;A variant is established as the causative factor for hereditary spherocytosis, with the c.1504-9G site functioning as a splicing receptor.</p>