The development of sequencing techniques identified numerous genetic variants, and accurate evaluation of the clinical significance of these variants facilitates the diagnosis of Mendelian diseases. In the present study, 549 rare single- nucleotide variants of uncertain significance were extracted from the ADPKD and ClinVar databases. MaxEntScan scoresplice is an in silico splicing prediction tool that was used to analyze rare PKD1 and PKD2 variants of unknown significance. An in vitro minigene splicing assay was used to verify 37 splicing-altering candidates that were located within seven residues of the splice donor sequence excluding canonical GT dinucleotides or within 21 residues of the acceptor sequence excluding canonical AG dinucleotides of PKD1 and PKD2. We demonstrated that eight PKD1 variants alter RNA splicing and were predicted to be pathogenic.

测序技术的发展确定了许多遗传变异，准确评估这些变异的临床意义有助于孟德尔疾病的诊断。在本研究中，从 ADPKD 和 ClinVar 数据库中提取了 549 个意义不确定的罕见单核苷酸变异。MaxEntScan scoresplice 是一种计算机剪接预测工具，用于分析未知意义的罕见PKD1和PKD2变体。体外小基因剪接试验用于验证 37 个剪接改变候选者，这些候选者位于剪接供体序列的 7 个残基内，不包括经典 GT 二核苷酸，或位于受体序列的 21 个残基内，不包括PKD1的规范 AG 二核苷酸和公钥簿2。我们证明了八种PKD1变体改变了 RNA 剪接并被预测为致病性。