非典型溶血尿毒综合征致病基因CFI突变位点的RNA剪接分析
RNA Splicing Analysis of the CFI Mutation Site in the Causative Gene for Atypical Hemolytic Uremic Syndrome
DOI: 10.12677/acm.2025.1551472, PDF,   
作者: 徐 宁, 张译尹, 张 然, 刘绪言, 刘晓淼：青岛大学青岛医学院，山东 青岛；王 至, 张炳莹：山东第二医科大学临床医学院，山东 青岛；张 艳：北大医疗鲁中医院肾内科，山东 淄博；尤青青：青岛市市立医院肾内科，山东 青岛；邵乐平：厦门大学附属第一医院肾内科，福建 厦门
关键词: 补体因子I；CFI；基因突变；迷你基因；mRNA剪接；Complement Factor I； CFI； Gene Mutation； Minigene； mRNA Splicing
摘要: 目的：CFI基因编码的补体因子I (FI)是补体系统调节过程中重要的丝氨酸蛋白酶，其功能缺陷与多种疾病相关，然而许多突变的致病机制并未完全阐明。本实验旨在通过迷你基因实验验证CFI基因突变对mRNA剪接的影响，为解释致病机制、发现治疗靶点提供新思路。方法：本研究首先通过生物信息学软件，从人类基因数据库中筛选出目标突变，并将包含该突变位点的基因片段导入载体中。随后将构建好的含有野生型和突变型的质粒分别转染至人胚肾细胞(HEK293T)中，并利用RT-PCR和测序技术分析其mRNA产物的变化。结果：与野生型5号外显子相比，c.772G>A突变导致外显子完全跳变，产生了异常剪接产物。这一异常剪接可能导致产生的FI蛋白功能域缺陷，从而影响其蛋白酶活性或与其他补体因子的结合。结论：本研究成功构建了CFI基因的迷你基因实验体系，证实了c.772G>A突变可导致mRNA剪接异常。这为深入理解CFI基因突变的致病机制提供了重要的实验依据，也为aHUS疾病的基因诊断和治疗提供了新的靶点。
Abstract: Objective: Complement factor I (FI), encoded by the CFI gene, is an important serine protease regulating the complement system, and its functional defects have been associated with a wide range of diseases. However, the pathogenic mechanisms of many mutations have not been fully elucidated. This experiment aims to verify the effect of CFI gene mutation on mRNA splicing through minigene experiments, which will provide new ideas to explain the pathogenic mechanism and discover therapeutic targets. Methods: In this study, the target mutation was first screened out from the human gene database by bioinformatics software, and the gene fragment containing the mutation site was introduced into the vector. Subsequently, the constructed plasmids containing wild-type and mutant were transfected into human embryonic kidney cells (HEK293T) respectively, and the changes in their mRNA products were analyzed by RT-PCR and sequencing. Results: Compared with wild-type exon 5, the c.772G>A mutation resulted in a complete exon skipping, producing an aberrant splicing product. This aberrant splicing may lead to defects in the functional domains of the resulting FI proteins, thus affecting their protease activity or binding to other complement factors. Conclusion: In this study, we successfully constructed a minigene experimental system for the CFI gene and confirmed that the c.772G>A mutation could lead to abnormal mRNA splicing. This provides an essential experimental basis for the in-depth understanding of the pathogenic mechanism of CFI gene mutation and also provides a new target for the genetic diagnosis and treatment of aHUS disease.
文章引用：徐宁, 王至, 张译尹, 张艳, 张然, 张炳莹, 尤青青, 刘绪言, 刘晓淼, 邵乐平. 非典型溶血尿毒综合征致病基因CFI突变位点的RNA剪接分析[J]. 临床医学进展, 2025, 15(5): 1109-1120. https://doi.org/10.12677/acm.2025.1551472

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