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目的:分析一个X连锁显性遗传Alport综合征家系n COL4A5基因的剪接突变位点，探索外显子特异性U1核小RNA（small nuclear RNA，snRNA）基因治疗的可能性。n 方法:收集Alport综合征先证者及其家族成员临床资料，高通量测序技术检测先证者肾病系列基因全外显子组基因突变。用在线软件分析n COL4A5 c.546+5G>A变异引起的剪接位点改变及其致病性。用minigene实验验证和分析Alport综合征家系先证者n COL4A5基因突变位点c.546+5G>A及瞬时转染导入修饰过的U1 snRNA纠正剪接突变的效果。n 结果:基因测序结果显示，先证者和其同母异父的兄长均存在n COL4A5基因半合子变异，变异位点为c.546+5G>A。在线软件分析剪接变异致病性的结果显示，突变后原有的Donor剪切位点检测不到，提示影响剪切可能性很大。杂交小基因检测结果验证了n COL4A5基因c.546+5G>A突变的异常剪接模式——外显子9缺失。经过修饰的U1 snRNA可部分纠正该异常的剪接突变，ExSpeU1（MT）、ExSpeU1（E9+1）、ExSpeU1（E9+9）、ExSpeU1（E9+11）对c.546+5G>A引发的外显子9缺失的纠正比例分别为0、43.81%、52.09%、48.12%。n 结论:COL4A5新发剪接突变具有致病性，修饰过的U1 snRNA可部分纠正异常剪接。n “,”Objective:To analyze the splicing mutation site of n COL4A5 gene in a family with X-linked dominant Alport syndrome and explore the possibility of exon specific U1 small nuclear RNA (snRNA) gene therapy.n Methods:The clinical data of the proband and family members of Alport syndrome were collected, and the gene mutations in the whole exon of a series of nephropathy genes in the proband were detected by high-throughput sequencing. The splice site changes and pathogenicity caused by n COL4A5 c.546+5G>A mutation were analyzed by online software. Minigene experiment was used to verify and analyze the effect ofn COL4A5 gene mutation site c.546+5G>A in the proband of Alport syndrome family, and transient transfection and introduction of modified U1 snRNA to correct splicing mutation.n Results:The results of gene sequencing showed that there was a hemizygous variation of n COL4A5 gene in the proband and his half brother, and the variation site was c.546+5G>A. The results of online software for analyzing the pathogenicity of splice variation showed that the original donor splicing site could not be detected after mutation, suggesting that there was a great possibility of affecting splicing. The abnormal splicing mode ofn COL4A5 gene with c.546+5G>A mutation—deletion of exon 9 was verified by hybridized small gene detection. The abnormal splicing mutation could be partially corrected by the modified U1 snRNA. The correction ratios of ExSpeU1 (MT), ExSpeU1(E9+1), ExSpeU1(E9+9) and ExSpeU1(E9+11) to exon 9 deletion caused by c.546+5G>A were 0, 43.81%, 52.09% and 48.12%, respectively.n Conclusions:The pathogenicity of the new splicing mutation of n COL4A5 is verified, and the modified U1 snRNA can partially correct the abnormal splicing.n