A novel strategy for identifying genes that regulate eye development

识别调节眼睛发育基因的新策略

基本信息

批准号：
8599120
负责人：
Iswar K. Hariharan
金额：
$ 22.61万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8599120
关键词：
Adhesives Adult Affect Affinity Age Anterior Award Biological Process Cell Communication Cell Proliferation Cell Proliferation Regulation Cell Survival Cell surface Cells Collection Complex Defect Development Drosophila eye Drosophila genus Epithelium Eye Eye Development Fertility Functional disorder Gene Targeting Generations Genes Genetic Genetic Screening Human Leg Maintenance Mediating Methods Morphology Mutation Organism Phenotype Photoreceptors Population Primordium Process Protein Kinase Protein phosphatase Proteins RNA Regulation Relative (related person)Research Personnel Research Project Grants Retina Retinal Retinal Degeneration Role Shapes Signal Transduction Staging Subgroup Tissues Vertebrates Wing base cell fate specification cell type design falls fitness fly gene function human disease imaginal disc in vivo method development mutant novel strategies preference public health relevance

项目摘要

DESCRIPTION (provided by applicant): The development and maintenance of a tissue such as the retina is contingent upon the proper regulation of cell proliferation, cell survival and appropriate adhesive interactions between cells and their neighbors. While great advances have been made in our understanding of the genetic regulation of many of these processes, studies in Drosophila imaginal discs have highlighted the existence of modes of cell-cell interactions that we still do not understand. For instance, studies of the phenomenon of cell competition have demonstrated that cells are capable of assessing the fitness of their neighbors and subsequently influencing their proliferation or survival. We also know that cells appear to preferentially adhere to other cells from the same portion of the imaginal disc while avoiding interactions with cells from other parts of the same primordium. This preference may be absolute such as the clear separation of anterior and posterior cells in leg and wing discs or relative, such as the tendency of clonal boundaries to fall along the equator in the developing Drosophila eye. While both cell competition and compartmentalization were discovered almost forty years ago, we still do not have a clear understanding of the mechanistic basis of either phenomenon. It is therefore likely that much remains to be discovered about the ways in which cells interact with the neighbors in tissues such as the developing retina. Mosaic screens in the Drosophila eye have been used to discover many genes that function in regulating cell proliferation, cell fate specification and photoreceptor differentiation. Importantly, the phenotypes elicited by mutations in some of these genes are only obvious in a situation where the disc is composed to two different types of cells: wild type and mutant. Under these conditions, changes in cell proliferation manifest as a change in the relative representation of the two populations of cells without necessarily affecting eye size and changes in cell-cell affinity can result in the formation of smooth, rather than irregular boundaries between the two populations. While screens based on RNA-mediated interference (RNAi) have been extremely effective in identifying genes in a number of biological processes, it is currently difficult to us this approach in the context of a mosaic screen. We have developed a new method, CoinFLP, which allows us to conduct mosaic screens in the Drosophila eye using RNAi and have validated this approach with a pilot screen that has already identified several genes not previously implicated in eye development. We propose to use this approach to identify and characterize genes that regulate cell proliferation and cell affinity in the developing Drosophila eye.

描述（由申请人提供）：诸如视网膜等组织的发育和维持取决于细胞增殖、细胞存活以及细胞与其邻近细胞之间适当的粘附相互作用的适当调节。虽然我们对许多这些过程的遗传调控的理解已经取得了巨大进展，但对果蝇成虫盘的研究强调了我们仍然不了解的细胞间相互作用模式的存在。例如，对细胞竞争现象的研究表明，细胞能够评估其邻居的适应性，并随后影响其增殖或存活。我们还知道，细胞似乎优先粘附于成虫盘同一部分的其他细胞，同时避免与同一原基其他部分的细胞相互作用。这种偏好可能是绝对的，例如腿和翼盘中前部和后部细胞的清晰分离，或者是相对的，例如发育中的果蝇眼睛中克隆边界沿着赤道下降的趋势。虽然细胞竞争和区室化都是在四十年前发现的，但我们仍然对这两种现象的机制基础没有清楚的了解。因此，关于细胞与组织（例如发育中的视网膜）中邻近细胞相互作用的方式，可能还有很多有待发现。果蝇眼睛中的马赛克筛选已被用来发现许多在调节细胞增殖、细胞命运规范和光感受器分化方面发挥作用的基因。重要的是，这些基因中的一些突变引起的表型仅在椎间盘由两种不同类型的细胞组成的情况下才明显：野生型和突变型。在这些条件下，细胞增殖的变化表现为两个细胞群的相对代表性的变化，而不一定影响眼睛的大小，并且细胞-细胞亲和力的变化可以导致两个细胞群之间形成平滑的边界，而不是不规则的边界。虽然基于 RNA 介导的干扰 (RNAi) 的筛选在识别许多生物过程中的基因方面非常有效，但目前很难在马赛克筛选的背景下使用这种方法。我们开发了一种新方法 CoinFLP，它允许我们使用 RNAi 在果蝇眼睛中进行马赛克筛选，并通过试点筛选验证了这种方法，该筛选已经识别出一些以前与眼睛发育无关的基因。我们建议使用这种方法来识别和表征在发育中的果蝇眼睛中调节细胞增殖和细胞亲和力的基因。