Gavin Herbert Eye Institute-Center for Translational Vision Research, Department of Ophthalmology, University of California, Irvine, California, USA; Department of Physiology and Biophysics, University of California, Irvine, California, USA.
Department of Ophthalmology and UCLA Stein Eye Institute, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California, USA.
J Biol Chem. 2024 Jun;300(6):107344. doi: 10.1016/j.jbc.2024.107344. Epub 2024 May 4.
MicroRNAs (miRs) are short, evolutionarily conserved noncoding RNAs that canonically downregulate expression of target genes. The miR family composed of miR-204 and miR-211 is among the most highly expressed miRs in the retinal pigment epithelium (RPE) in both mouse and human and also retains high sequence identity. To assess the role of this miR family in the developed mouse eye, we generated two floxed conditional KO mouse lines crossed to the RPE65-ERT2-Cre driver mouse line to perform an RPE-specific conditional KO of this miR family in adult mice. After Cre-mediated deletion, we observed retinal structural changes by optical coherence tomography; dysfunction and loss of photoreceptors by retinal imaging; and retinal inflammation marked by subretinal infiltration of immune cells by imaging and immunostaining. Single-cell RNA sequencing of diseased RPE and retinas showed potential miR-regulated target genes, as well as changes in noncoding RNAs in the RPE, rod photoreceptors, and Müller glia. This work thus highlights the role of miR-204 and miR-211 in maintaining RPE function and how the loss of miRs in the RPE exerts effects on the neural retina, leading to inflammation and retinal degeneration.
微小 RNA（miRs）是短的、进化上保守的非编码 RNA，通常下调靶基因的表达。miR-204 和 miR-211 组成的 miR 家族在人和鼠的视网膜色素上皮（RPE）中是表达最高的 miR 之一，并且具有高度的序列同一性。为了评估这个 miR 家族在发育中的鼠眼的作用，我们生成了两个 floxed 条件性 KO 小鼠品系，与 RPE65-ERT2-Cre 驱动小鼠品系杂交，在成年小鼠中进行该 miR 家族的 RPE 特异性条件性 KO。在 Cre 介导的缺失后，我们通过光学相干断层扫描观察视网膜的结构变化；通过视网膜成像观察感光细胞的功能障碍和损失；通过成像和免疫染色观察免疫细胞在视网膜下的浸润来观察视网膜炎症。患病的 RPE 和视网膜的单细胞 RNA 测序显示了潜在的 miR 调节靶基因，以及 RPE、杆状光感受器和 Muller 胶质细胞中非编码 RNA 的变化。因此，这项工作强调了 miR-204 和 miR-211 在维持 RPE 功能中的作用，以及 RPE 中 miR 的缺失如何对神经视网膜产生影响，导致炎症和视网膜变性。
