circRNA Cdr1as 调控视网膜变性的机理研究

circRNA is a kind of special non-coding RNAs. It is highly conserved with robust expression and not easily degraded. Recently it has been demonstrated to be closely related to a variety of diseases. Cdr1as was firstly approved to be associated with brain function and neurological diseases. Our previous study discovered that circRNAs were highly expressed in mice retina and differently expressed at developmental stages. As one of the most abundant circRNAs, Cdr1as express abnormally in retinal degeneration mice model, and can sponge miR-204, which is related to retinal disease. Based on our previous researches, at first, we will study the circRNAs in developmental stages and ceRNA regulatory network. Then, we will dissect the function of Cdr1as in retina through Cdr1as conditional knockout mice. Last, we will rescue or relieve Cdr1as-relevant retinal diseases via sub-retinal injection AAV-Cdr1as in mice. This is the first time to elucidate the regulation of Cdr1as in retina, which would enrich the non-coding RNA regulatory network and provide new direction for study mechanism and treatment of retinal diseases.

circRNA是一类特殊的非编码RNA分子，在进化上高度保守，表达稳定且不易被降解，近年来已被证实与多种疾病紧密相关。其中Cdr1as首次被证实与大脑功能存在关联且与神经性疾病相关。我们前期研究发现，小鼠视网膜中也存在大量高丰度circRNA，而且不同发育阶段中circRNA表达迥异。表达量位于前三的Cdr1as在视网膜变性疾病模型小鼠中表达异常，并且能够调控与视网膜疾病相关的miRNA。本项目拟研究与视网膜发育相关的circRNA及ceRNA调控网络；利用Cdr1as敲除小鼠模型深入剖析其在视网膜中的重要调控作用及内在机制；通过AAV介导circRNA表达的方式rescue或缓解与Cdr1as相关的视网膜变性疾病模型。本课题首次阐明Cdr1as在视网膜中的调控作用及机制，在丰富视网膜中非编码RNA调控网络的同时，为视网膜相关疾病的机理与治疗研究提供了新的思路。

环状RNA (circRNAs)是一类新型的非编码RNA，具有组织和阶段特异性表达模式，它们比线性转录本更稳定。它们是构成了基因调控网络的重要组成部分。到目前为止，circRNA在视网膜中的表达模式和调控潜力并未被发掘。本课题对小鼠视网膜发育5个阶段（E18.5，P1，P7，P14和P30)）的RNA进行高通量测序后，得到9029个circRNAs。分析了不同发育阶段circRNA及其亲本基因的表达模式及特点；研究了与视网膜发育相关的circRNA及其ceRNA调控网络；通过AAV介导circRNA表达的方式将circTulp4在小鼠视网膜内干扰，研究了circTulp4在视网膜中的重要调控作用及其机制；通过成功构建的Cdr1as敲除小鼠模型深入阐明了Cdr1as在视网膜中的重要功能及调控机制。作为miRNA和circRNA行使功能中的重要调控因子，Ago2在视网膜中的调控作用也通过AAV干扰和过表达的方式得到了进一步的阐述。本课题丰富了视网膜中RNA的调控网络，为视网膜相关疾病的机理研究与干预治疗提供了新的思路。