MOLECULAR MECHANISMS IN RETINAL DEGENERATIONS

视网膜变性的分子机制

• 批准号: 2162158
• 负责人: DEBORA B FARBER
• 金额: $ 15.89万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2162158
• 关键词: DNA; RNA; chromosome aberrations; chromosome disorders; congenital vision disorder; disease /disorder model; gene expression; gene mutation; genetic library; genetic transcription; genetic translation; in situ hybridization; laboratory mouse; linkage mapping; messenger RNA; molecular cloning; molecular pathology; nucleic acid probes; nucleic acid sequence; protein sequence; retina degeneration; retinitis pigmentosa; transposon /insertion element; visual photoreceptor; western blottings

The long-term objective of this proposal is to characterize, chromosome map and isolate genes that may be involved in inherited retinal degenerations affecting animals and humans. Knowledge about the genes that are responsible for these diseases may increase our understanding of the process of pathogenesis and in addition, it may help to develop studies on how to control or even arrest the expression of the abnormal genes. The rd mouse model of retinal degeneration will be studied first. the basic strategy involves the development of a novel probe consisting of photoreceptor-enriched cDNAs which will allow selection of low abundance photoreceptor-specific clones from a normal mouse retinal cDNA library. These clones will be re-screened with probes consisting of rd adult retinal cDNAs and rd 9-11 day-old retinal cDNAs to pinpoint rd candidate genes. After Northern analysis of developing rd and normal mouse retinal RNAs with the rd candidate cDNAs to determine any differences between the rd and normal mRNAs, the mouse chromosome location of each potential rd cDNA will be determined and, if on chromosome 5, its specific position within this chromosome will be established. the nucleotide and reduced amino acid sequences as well as the genomic organization of the rd candidate genes and their in vitro transcription and translation will then be determined. Final proof of the identify of the rd gene with cloned gene will require correcting the rd phenotype. Experiments involving transgenic mice will be planned for the future but are not within the scope of this application. Isolated photoreceptor-specific cDNAs will be used in human studies to search for alterations in the patterns of RFLPs in families affected with any of the different types of retinitis pigmentosa. To carry out the proposed work, methods currently used in Molecular Biology will be employed. These studies will add to the knowledge of the molecular properties of normal and abnormal retinas since in addition to the rd gene other photoreceptor-specific genes may be characterized.

该提议的长期目标是表征染色体 可能涉及遗传视网膜的地图和分离基因 影响动物和人类的退化。 关于基因的知识 负责这些疾病的原因可能会增加我们对 发病机理的过程以及此外，它可能有助于发展 关于如何控制甚至阻止异常表达的研究 基因。 将首先研究视网膜变性的RD小鼠模型。 基本策略涉及开发新的探针 富含光感受器的cDNA，该cDNA将允许选择低 来自正常小鼠视网膜cDNA的丰度光感受器特异性克隆 图书馆。 这些克隆将通过由RD组成的探针重新筛选 成人视网膜CDNA和RD 9-11天大的视网膜cDNA，以查明RD 候选基因。 在北部分析了RD和正常状态之后 带有RD候选cDNA的小鼠视网膜RNA确定任何 RD和正常mRNA之间的差异，小鼠染色体 将确定每个电势RD cDNA的位置，如果在 5号染色体，其在该染色体中的特定位置将是 已确立的。 核苷酸和还原氨基酸序列以及 RD候选基因的基因组组织及其体外 然后将确定转录和翻译。 最终证明 用克隆基因识别RD基因将需要纠正 RD表型。 将计划涉及转基因小鼠的实验 未来，但不在本应用程序的范围内。 分离的光感受器特异性cDNA将在人类研究中使用 在受影响的家庭中寻找RFLP模式的变化 任何不同类型的视网膜炎色素。 为了进行拟议的工作，目前用于分子的方法 生物学将被使用。 这些研究将增加 正常和异常视网膜的分子特性，因为 RD基因可以表征其他感光特异性基因。