The role of microRNAs in the development and differentiation of mouse photorecept

描述(申请人提供)：视网膜色素变性(RP)是最常见的遗传性视网膜变性[1]。RP可由多种不同基因的突变引起，其中许多基因是光感受器特有的[1]。因此，阐明光感受器基因调控网络是弄清光感受器发育和退化机制的基础。小RNA加工所需的一种关键酶--DICER的视网膜缺失导致的表型已经确立了microRNAs(MiRNAs)，它是视网膜发育和退化的关键因素[2，3]。MiRNAs在转录后水平上改变基因表达[4]。尽管如此，关于它们在光感受器发育和退化中的作用还知之甚少。本研究的目的是研究miRNAs在光感受器发育、分化和退化过程中的作用。为了实现这一目标，人们设计了两个目标来阐明调控光感受器miRNA表达的转录网络和了解视网膜中表达最高的miRNA miR-124的功能。这些目标将通过下一代测序、生物信息学分析、原位杂交、RT-PCR、免疫组织化学、克隆技术和组织电穿孔来实现。 公共卫生相关性：了解视网膜神经变性背后的细胞机制对于治疗导致失明的疾病至关重要，如视网膜色素变性。MicroRNAs在转录后调节细胞中的基因表达。阐明视网膜中microRNAs的功能可能有助于开发治疗和治疗视网膜色素变性的新方法。

DESCRIPTION (provided by applicant): Retinitis pigmentosa (RP) is the most common inherited form of retinal degeneration [1]. RP can be caused by mutations in an assortment of different genes, many of them photoreceptor-specific [1]. Therefore, elucidation of the photoreceptor gene regulatory network is fundamental to discerning the mechanism behind photoreceptor development and degeneration. Phenotypes resulting from the retinal deletion of Dicer, a critical enzyme required for small RNA processing, have established microRNAs (miRNAs), as key factors in retinal development and degeneration [2, 3]. MiRNAs modify gene expression at the post-transcriptional level [4]. Nonetheless, little is known about their role within photoreceptor development and degeneration. The goal of this proposal is to study the function of miRNAs in photoreceptor development, differentiation and degeneration. To achieve this goal, two aims have been designed to elucidate the transcriptional network regulating miRNA expression in photoreceptors and to understand the function of miR-124, the most highly expressed miRNA in retina. These aims will be accomplished using next generation sequencing, bioinformatic analysis, in situ hybridization, RT-PCR, immunohistochemistry, cloning techniques and tissue electroporation. PUBLIC HEALTH RELEVANCE: Understanding the cellular mechanism behind retinal neurodegeneration is essential for treating diseases that result in blindness, such as retinitis pigmentosa. MicroRNAs act post-transcriptionally to modify gene expression in cells. Elucidating the function of microRNAs in the retina may yield in the development of novel methods for therapy and treatment of retinitis pigmentosa.