背景:Gitelman综合征(GS)是一种脱盐肾小管疾病,其中大部分是由SLC12A3基因变异引起的,错觉变体占大多数。最近,外显子跳跃的现象,其中变体破坏正常的pre-mRNA剪接,与多种疾病有关。因此,我们假设一定比例的SLC12A3变异体可通过干扰正常剪接过程导致疾病.
方法:我们使用生物信息学程序分析了342个先前假定的SLC12A3错义变体,并通过小基因测定鉴定了可能改变前mRNA剪接的候选变体。
结果:我们的研究表明,在十个候选变体中,六个变种(c.602G>A,c.602G>T,c.1667C>T,c.1925G>A,c.2548G>C,和c.2549G>C)通过影响外显子剪接调控元件和/或干扰规范剪接位点而导致完整或不完整的外显子跳跃。
结论:值得一提的是,这是关于SLC12A3外显子变体的前mRNA剪接的最大研究。此外,我们的研究强调了在GSmRNA水平检测剪接功能的重要性,并表明小基因分析是体外变异体剪接功能分析的有价值的工具.
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.
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.
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.
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.