A novel strategy for identifying genes that regulate eye development

描述（由申请人提供）：组织（如视网膜）的发育和维持取决于细胞增殖、细胞存活和细胞与其相邻细胞之间适当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很大的进展，但对果蝇成虫盘的研究突出了我们仍然不了解的细胞-细胞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，细胞似乎优先粘附到其他细胞从同一部分的成虫盘，而避免与细胞的相互作用，从其他部分相同的原基。这种偏好可能是绝对的，如腿和翅盘中前后细胞的明显分离，也可能是相对的，如果蝇眼睛发育中克隆边界沿着赤道下降的趋势。虽然细胞竞争和区室化在近40年前就被发现了，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（如发育中的视网膜）中的邻居相互作用的方式，可能还有很多东西有待发现。 果蝇眼睛中的镶嵌屏幕已经被用来发现许多在调节细胞增殖、细胞命运指定和感光细胞分化中起作用的基因。重要的是，由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是明显的：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体的相对代表性的变化，而不一定影响眼睛大小，并且细胞-细胞亲和力的变化可以导致两个群体之间形成平滑而不是不规则的边界。虽然基于RNA介导的干扰（RNAi）的筛选在许多生物过程中识别基因方面非常有效，但目前很难在镶嵌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，并通过一个试验性筛选验证了这种方法，该筛选已经确定了几个以前与眼睛发育无关的基因。我们建议使用这种方法来确定和表征基因，调节细胞增殖和细胞亲和力在发展中的果蝇眼睛。

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.