Minigene splicing assays reveal new insights into exonic variants of the SLC12A3 gene in Gitelman syndrome. Issue 4 (3rd January 2023)

• Record Type: Journal Article
• Authors: Shi, Xiaomeng; Wang, Hong; Zhang, Ruixiao; Liu, Zhiying; Guo, Wencong; Wang, Sai; Liu, Xuyan; Lang, Yanhua; Bottillo, Irene; Dong, Bingzi; Shao, Leping
• Is Part Of: Molecular genetics & genomic medicine. Volume 11:Issue 4(2023)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2023-01-03
• Subjects: exon splicing -- exonic variant -- Gitelman syndrome -- minigene assay -- SLC12A3; Medical genetics -- Periodicals; Genomics -- Periodicals; 616.042
• DOI: 10.1002/mgg3.2128
• Languages: English
• ISSNs: 2324-9269

Abstract

Abstract: Background: Gitelman syndrome (GS) is a type of salt‐losing tubular disease, most of which is caused by SLC12A3 gene variants, and missense variants account for the majority. Recently, the phenomenon of exon skipping, in which variants disrupt normal pre‐mRNA splicing, has been related to a variety of diseases. Therefore, we hypothesize that a certain proportion of SLC12A3 variants can result in disease via interfering with the normal splicing process. Methods: We analyzed 342 previously presumed SLC12A3 missense variants using bioinformatics programs and identified candidate variants that may alter the splicing of pre‐mRNA through minigene assays. Results: Our study revealed that, among ten candidate variants, six variants (c.602G>A, c.602G>T, c.1667C>T, c.1925G>A, c.2548G>C, and c.2549G>C) led to complete or incomplete exon skipping by affecting exonic splicing regulatory elements and/or disturbing canonical splice sites. Conclusion: It is worth mentioning that this is the largest study on pre‐mRNA splicing of SLC12A3 exonic variants. In addition, our study emphasizes the importance of detecting splicing function at the mRNA level in GS and indicates that minigene analysis is a valuable tool for splicing functional assays of variants in vitro. Abstract : The purpose of this study was to identify the effect of previously presumed missense SLC12A3 variants on pre‐mRNA splicing using bioinformatics tools and minigenes. The results revealed that, among ten candidate variants, six variants (c.602G>A, c.602G>T, c.1667C>T, c.1925G>A, c.2548G>C, and c.2549G>C) led to complete or incomplete exon skipping by affecting exonic splicing regulatory elements and/or disturbing canonical splice sites. It is worth mentioning that this is the largest study on pre‐mRNA splicing of SLC12A3 exonic variants, and our study emphasizes the importance of detecting splicing function at the mRNA level in GS and indicates that minigene analysis is a valuable tool for splicing functional assays of variants in vitro.