Abstract
in English, Chinese N6-methyladenosine (m 6A) modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function. The m 6A reader YTHDF1 has been shown to enhance the translational efficiency of m 6A-modified mRNAs in the brain and is essential for learning and memory. However, its role in the mature retina remains unclear. Herein, we report a novel role of Ythdf1 in the maintenance of retinal function using a genetic knockout model. Loss of Ythdf1 resulted in impaired scotopic electroretinogram (ERG) responses and progressive retinal degeneration. Detailed analyses of rod photoreceptors confirmed substantial degenerative changes in the absence of ciliary defects. Single-cell RNA sequencing revealed comprehensive molecular alterations across all retinal cell types in Ythdf1-deficient retinas. Integrative analysis of methylated RNA immunoprecipitation (MeRIP) sequencing and RIP sequencing identified Tulp1 and Dhx38, two inheritable retinal degeneration disease-associated gene homologs, as direct targets of YTHDF1 in the retina. Specifically, YTHDF1 recognized and bound m 6A-modified Tulp1 and Dhx38 mRNA at the coding sequence (CDS), enhancing their translational efficiency without altering mRNA levels. Collectively, these findings highlight the essential role of YTHDF1 in preserving visual function and reveal a novel regulatory mechanism of m 6A reader proteins in retinal degeneration, identifying potential therapeutic targets for severe retinopathies. mRNA的N6-甲基腺苷（m6A）修饰作为一种重要的转录后修饰方式，在mRNA代谢调控中发挥关键作用。m6A阅读蛋白YTHDF1已被证实能增强大脑中被m6A修饰mRNA的翻译效率，从而在学习记忆过程中至关重要。然而，其在成熟视网膜和视觉中的作用仍不明确。该研究利用基因敲除模型揭示了YTHDF1在维持视网膜功能中的新作用：YTHDF1缺失导致暗适应视网膜电生理（ERG）反应受损，并伴随进行性视网膜变性。进一步深入分析发现，Ythdf1缺失主要引起视杆细胞病变丢失，但其纤毛功能未受影响。单细胞RNA测序（scRNA-seq）分析揭示了YTHDF1缺失导致视网膜中各类细胞的分子水平变化。通过结合甲基化RNA免疫共沉淀测序（MeRIP-seq）和RNA免疫共沉淀测序（RIP-seq），最终确定了遗传性视网膜变性疾病关键基因Tulp1和Dhx38作为YTHDF1在视网膜中的调控靶点。YTHDF1能够识别并结合 Tulp1和 Dhx38 mRNA编码区（CDS）上的m6A修饰位点，从而增强其翻译效率。该研究揭示了YTHDF1在维持视功能中的重要作用，阐明了m6A读取蛋白在视网膜变性中的新型调控机制，并为视网膜病变的治疗策略开发提供了潜在靶点。.