This pilot study investigates the potential pathogenic role of G-quadruplex (G4) structures in -associated retinal degeneration, starting from a case of suspected X-linked form affected family. We hypothesize that the stabilization of these structures might alter DNA replication and transcription, inducing genetic instability and influencing gene expression. We conducted whole genome amplification experiments and next-generation sequencing to detect the blockade of polymerase activity by G4 structures. Our specific focus was the gene, which hosts a high concentration of predicted G4-forming motifs and is implicated in most X-linked retinal degeneration cases. To understand the potential interference of G4 structures, we applied computational and 3D molecular modeling to visualize interferences in DNA replication and transcription regulation. Our data confirmed the obstruction of DNA polymerase enzymes by G4 structures, particularly when stabilized by the compound pyridostatin. This obstruction was evident in the reduced amplification of gene regions and a shift in the start/end sites of putative G4 motifs. Moreover, the modeling indicated a potential disruption of critical promoter elements and RNA polymerase binding, which could drastically alter gene expression. Our findings suggest that G4 formation in the gene could lead to genetic instability and affect the expression of RPGR, contributing to retinal dystrophy. Moreover, this study underscores the broader implications of G4 structures in other genetic disorders. Improved understanding of G4 structures could reveal novel therapeutic targets to combat genetic disorders, promoting the advancement of personalized medicine and precision health.

中文翻译：

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研究 RPGR 基因中的 G 四链体结构：对理解 X 连锁视网膜变性的意义

这项初步研究从一个疑似 X 连锁受影响家庭的病例开始，调查了 G 四联体 (G4) 结构在相关视网膜变性中的潜在致病作用。我们假设这些结构的稳定性可能会改变 DNA 复制和转录，从而引起遗传不稳定并影响基因表达。我们进行了全基因组扩增实验和新一代测序，以检测 G4 结构对聚合酶活性的阻断。我们特别关注的是该基因，该基因含有高浓度的预测 G4 形成基序，并且与大多数 X 连锁视网膜变性病例有关。为了了解 G4 结构的潜在干扰，我们应用计算和 3D 分子模型来可视化 DNA 复制和转录调控中的干扰。我们的数据证实了 G4 结构对 DNA 聚合酶的阻碍，特别是当被化合物吡啶他汀稳定时。这种阻碍在基因区域扩增的减少和假定的 G4 基序的起始/结束位点的变化中很明显。此外，模型表明关键启动子元件和 RNA 聚合酶结合可能受到破坏，这可能会极大地改变基因表达。我们的研究结果表明，该基因中 G4 的形成可能导致遗传不稳定并影响 RPGR 的表达，从而导致视网膜营养不良。此外，这项研究强调了 G4 结构在其他遗传性疾病中的更广泛的影响。提高对 G4 结构的理解可以揭示对抗遗传性疾病的新治疗靶点，促进个性化医疗和精准健康的进步。