Replacement of Fluorescence Imaging System

项目摘要 父母补助金的中心目标是开发一种全基因组配对鉴定的测定方法 同源DNA片段之间的相互作用。同源DNA序列的并置是 染色体结构、基因组稳定性和基因表达控制的决定因素。在体细胞中， 姐妹染色单体之间的配对确保了准确的染色体断裂修复和染色体断裂修复。 种族隔离在生殖细胞系中，同源染色体之间的配对确保了基因组的单倍体化 进行有性繁殖在发育过程中，同源染色体片段之间的相互作用 是建立单等位基因表达所必需的。配对缺陷与 染色体错误分离，基因组重排和异常基因剂量，导致出生 缺陷、癌症和过早衰老。开发一种检测方法，鉴定参与 同源配对是理解这一过程机理的重要步骤。在母 格兰特，我们正在开发同源配对捕获分析，以定量检测同源DNA 交互.虽然适用于大量的发展阶段和生物体，我们正在使用 芽殖酵母减数分裂作为分析开发的模型系统。可以诱导同步减数分裂 酵母培养物中的数百万个细胞，便于捕获处于全基因组同源阶段的细胞， 配对我们的许多方法涉及通过荧光监测减数分裂细胞周期进程 显微镜在这份补充提案中，我们要求提供资金， 荧光显微镜和一个灵敏的相机集成了最新的成像系统。我们 目前的系统已变得不可靠，其寿命即将结束。更新的荧光 显微镜将使我们能够保持和进一步加强高影响力的研究计划， 同源染色体配对的机制。

PROJECT SUMMARY The central aim of the parent grant is to develop an assay for genome-wide identification of pairing interactions between homologous DNA segments. Juxtaposition of homologous DNA sequences is a key determinant of chromosome structure, genome stability and gene expression control. In somatic cells, pairing between sister chromatids ensures accurate chromosome break repair and chromosome segregation. In the germ line, pairing between homologous chromosomes ensures genome haploidization for sexual reproduction. During development, interactions between homologous chromosome segments are required for establishing monoallelic gene expression. Pairing defects are associated with chromosome missegregation, genome rearrangements and aberrant gene dosage, resulting in birth defects, cancer, and premature aging. Developing an assay that identifies genome segments involved in homologous pairing is an essential step for a mechanistic understanding of this process. In the parent grant, we are developing the Homolog Pairing Capture assay, to quantitatively detect homologous DNA interactions. While applicable to a large number of developmental stages and organisms, we are using budding yeast meiosis as a model system for assay development. Synchronous meiosis can be induced in yeast cultures in millions of cells, facilitating the capture of cells at the stage of genome-wide homologous pairing. Many of our approaches involve monitoring of meiotic cell cycle progression via fluorescence microscopy. In this supplement proposal, we are requesting funds for a replacement for our aging routine fluorescence microscope and a sensitive camera integrated with an up-to-date imaging system. Our current system has become unreliable and is near the end of its lifetime. An updated fluorescence microscope will enable us to maintain and further enhance a high-impact research program into mechanisms of homologous chromosome pairing.