High Thoughput FISH Mapping

为了实现我们识别人类重要基因的总体目标 开发，项目1(高通量FISH图谱)将绘制个体的染色体断裂点 有明显的平衡重排和先天畸形。我们假设染色体 这些个体的重排是导致异常表型的原因，或者通过 单倍体不足或其他一些遗传机制(S)。研究对象将被确定和登记 通过我们日益增长的临床遗传学家、遗传咨询师和临床 细胞遗传学家和NIGMS人类遗传细胞库。外周淋巴细胞或成纤维细胞 样本将从同意的研究对象中获得，淋巴细胞将永生化以 为我们的研究提供持续的细胞材料来源。参与者可以选择授予 允许我们将这些样本的等量捐赠给NIGMS人类遗传细胞库， 通过它它们可以匿名地与整个科学界分享。在集约化之前 断点定位，我们将使用阵列比较基因组杂交(ACGH)技术来检测 隐晦的或亚微观的变化可能会扰乱后续的分析和进一步的分析 有重大删除或复杂重新安排证据的案件将被推迟。根据 我们现有的优先排序标准，每年12个案例将被选择用于高分辨率断点映射，通过 使用全序列或末端测序的袋状和梭状克隆的鱼。案例选择标准支持具有以下条件的案例 一个特定的染色体重排导致异常表型的可能性更高 利用DGAP调查人员在不同发展领域的特殊专业知识 病理生物学。在裸DMA光纤(即光纤)上执行FISH将增强断点映射 FISH)，而不是某些病例的中期染色体，特别是有节段性重复的病例。 这种详细的作图研究是识别和表征 发育重要基因(S)(项目2)，以及动物模型的生成(项目3)。 项目1实验室将领导基因发现、功能分析和候选基因的验证 涉及女性生殖道疾病和听力损失的DGAP病例。最后，项目1将继续 通过互联网分享DGAP结果(http://dqap.harvard.edu/)符合我们成为核型表型的目标 世界各地临床遗传学家、细胞遗传学家和发育生物学家的资源。在……里面 概述，项目1将作为功能基因组学项目的切入点，生成公共 对人类发展至关重要的基因资源。 相关性(参见说明)：发育基因组解剖项目研究了一组患者 卫生保健系统服务不足：因染色体而先天畸形的人 重新安排。我们的任务是发现人类发育过程中被破坏的重要基因 通过这些染色体重排，更传统的人类遗传学很难识别的基因 战略，从而开启对它们造成的疾病的调查。

To achieve our overall goal of identifying genes important in human development, Project 1 (High Throughput FISH Mapping) will map chromosomal breakpoints from individuals with apparently balanced rearrangements and congenital anomalies. We hypothesize that chromosomal rearrangements in these individuals are responsible for the abnormal phenotypes, either through haploinsufficiency or some other genetic mechanism(s). Study subjects will be identified and enrolled through our growing international network of clinical geneticists, genetic counselors, and clinical cytogeneticists, and from the NIGMS Human Genetic Cell Repository. Peripheral lymphocytes or fibroblast specimens will be obtained from consented research subjects, and lymphocytes will be immortalized to provide ongoing sources of cellular material for our studies. Participants have the option of granting permission for us to contribute aliquots of these samples to the NIGMS Human Genetic Cell Repository, through which they can be shared anonymously with the entire scientific community. Prior to intensive breakpoint mapping, we will use array comparative genomic hybridization (aCGH) technology to detect cryptic or submicroscopic changes that could potentially confound subsequent analyses, and further analysis of cases with significant deletions or evidence of complex rearrangements would be deferred. According to our existing prioritization criteria, 12 cases/year will be selected for high-resolution breakpoint mapping by FISH using fully or end-sequenced BAG and fosmid clones. The case selection criteria favor cases with a higher likelihood that a given chromosomal rearrangement is responsible for the abnormal phenotype and take advantage of the particular expertise of DGAP Investigators in diverse areas of developmental pathobiology. Breakpoint mapping will be enhanced by FISH performed on naked DMA fibers (i.e., fiber FISH) instead of metaphase chromosomes in selected cases, particularly those with segmental duplications. Such detailed mapping studies are the entry point for the identification and characterization of developmental^ important gene(s) (Project 2), as well as the generation of animal models (Project 3). Project 1 laboratories will lead the gene discovery, functional analysis, and validation of candidate genes in DGAP cases involving female genital tract disorders and hearing loss. Finally, Project 1 will continue to share DGAP results via the Internet (http://dqap.harvard.edu/) in keeping with our goal of being a karyotypephenotype resource for clinical geneticists, cytogeneticists, and developmental biologists worldwide. In summary, Project 1 will serve as the entry point for a functional genomics project generating a public resource of genes critical to human development. RELEVANCE (See instructions): The Developmental Genome Anatomy Project studies a group of patients underserved by the health care system: those with congenital abnormalities due to chromosome rearrangements. Our mission is to discover genes of importance in human development that are disrupted by these chromosomal rearrangements, genes that are difficult to identify by more traditional human genetic strategies, thereby opening investigation of the disorders that they cause.