High Thoughput FISH Mapping

为了实现我们的总体目标，即确定人类重要基因， 项目1（高重复率FISH作图）将绘制个体的染色体断裂点， 有明显的平衡重排和先天性异常我们假设染色体 这些个体中的重排是异常表型的原因， 单倍不足或一些其他遗传机制。将确定并招募研究受试者 通过我们不断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 将从知情同意的研究受试者中获得标本，并将淋巴细胞永生化， 为我们的研究提供持续的细胞材料来源。参与者可以选择 允许我们向NIGMS人类遗传细胞库提供这些样本的等分试样， 通过它，它们可以匿名地与整个科学界分享。强化前 断裂点定位，我们将使用阵列比较基因组杂交（aCGH）技术检测 可能混淆后续分析和进一步分析的隐蔽或亚显微变化 至于有重大删除或有证据证明有复杂重新安排的个案，则会押后处理。根据 我们现有的优先级标准，12例/年将被选择用于高分辨率断点映射， 使用完全或末端测序的BAG和fosmid克隆进行FISH。病例选择标准有利于具有以下特征的病例： 给定的染色体重排导致异常表型的可能性更高， 利用DGAP调查员在发展的不同领域的特殊专业知识 病理生物学断点映射将通过在裸DMA纤维上进行的FISH来增强（即，纤维 FISH），而不是中期染色体在选定的情况下，特别是那些与节段重复。 这种详细的绘图研究是确定和定性 发育重要基因（项目2），以及动物模型的产生（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。项目1将继续 通过互联网（http：//dqap.harvard.edu/）分享DGAP结果，以符合我们的核分型表型目标 全球临床遗传学家、细胞遗传学家和发育生物学家的资源。在 项目1将作为功能基因组学项目的切入点， 对人类发育至关重要的基因资源。 相关性（见说明）：发育基因组解剖学项目研究了一组患者， 卫生保健系统服务不足：那些由于染色体异常而先天性异常的人 重新安排我们的使命是发现在人类发育中被破坏的重要基因 通过这些染色体重排，基因很难通过更传统的人类遗传学鉴定， 战略，从而打开调查的障碍，他们所造成的。

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.