Cloning zebrafish visual system mutants by whole-genome sequencing & SNP mapping

通过全基因组测序克隆斑马鱼视觉系统突变体

• 批准号: 8518344
• 负责人: Jeffrey Gross
• 金额: $ 18.34万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8518344
• 关键词: Affect; Age related macular degeneration; Anatomy; Animal Model; Animals; Anterior; Biochemical; Biological Assay; Biological Models; Birth; Blindness; Caenorhabditis elegans; Cataract; Cell physiology; Cells; Childhood; Chromosome Mapping; Clinical; Cloning; Collection; Coloboma; Congenital Disorders; Data; Defect; Development; Developmental Biology; Disease; Disease model; Ethylnitrosourea; Etiology; Eye Abnormalities; Genetic; Genetic Screening; Genome; Glaucoma; Grant; Hereditary Disease; Human; Human Genetics; Image; Laboratories; Lead; Maps; Mediating; Methods; Modeling; Molecular; Mutate; Mutation; Ocular Albinism; Oculocutaneous Albinism; Organ; Patients; Phenotype; Physiology; Positioning Attribute; Process; Proteins; Reporting; Research; Research Personnel; Resolution; Retinal Degeneration; Retinitis Pigmentosa; Single Nucleotide Polymorphism Map; System; Techniques; Testing; Time; Tissues; Validation; Visual impairment; Visual system structure; Zebrafish; age related; base; blind; cell type; congenital cataract; cost; gene discovery; genome sequencing; human disease; in vivo; in vivo Model; innovation; interest; lens; malformation; mutant; novel; positional cloning; protein function; research study; simple sequence length polymorphism

DESCRIPTION (provided by applicant): Visual impairments affect over 160 million people worldwide, and of these, roughly 37 million are blind (1). Major causes of blindness include cataracts, glaucoma, retinitis pigmentosa and age-related macular degeneration and the development of effective, low-cost therapies for these disorders is of the highest priority. Model organisms with similar anatomy and physiology to humans are vital to understand the molecular and cellular mechanisms underlying these diseases and in which to develop and test potential therapies. The zebrafish, Danio rerio, is an ideal system for modeling human disease, and studies utilizing zebrafish to elucidate the molecular and cellular underpinnings of congenital ocular disorders have made a significant impact in the field. Indeed, the combination of forward genetic screens to identify zebrafish mutants with ocular defects, and the rapid pace of technological advancement in analyzing the mutant phenotypes has positioned the zebrafish system at the forefront of those that contribute to our knowledge of the mechanistic underpinnings of human congenital eye diseases. Research in this grant will utilize an innovative whole-genome sequencing and SNP mapping approach to rapidly and affordably clone recessive zebrafish visual system mutants identified from a recently completed forward genetic screen in our lab. 23 mutants were identified in our screen that presented with congenital cataracts, anterior segment dysgenesis, colobomas, oculocutaneous albinism, retinal degeneration and other developmental defects. Our novel mapping technique will enable us to clone the affected loci in most, if not all, of these mutants and then perform targeted, hypothesis-driven experiments to determine the underlying molecular and cellular mechanisms that lead to ocular defects. We will focus these further research efforts on the congenital cataract mutants. Congenital cataracts occur in ~40 per 100,000 human births and represent the most common cause of childhood blindness in the developed world. Cataracts are also a common clinical feature in nearly 200 different human genetic diseases making them a frequent component of an otherwise heterogeneous collection of disorders. While much is known about age-related and environmentally induced cataracts, less is known about the etiology of congenital cataracts. Thus, the studies proposed here will have significant scientific merit as they will identify gene products required for normal lens development and they will provide animal congenital cataract models through which an understanding of disease mechanism can be can be advanced and a model through which novel therapies can be developed and tested.

描述（由申请人提供）：全球有超过1.6亿人受到视力障碍的影响，其中约有3700万人失明(1)。失明的主要原因包括白内障、青光眼、视网膜色素变性和老年性黄斑变性，开发有效、低成本的治疗这些疾病的方法是当务之急。与人类解剖学和生理学相似的模式生物对于了解这些疾病的分子和细胞机制以及开发和测试潜在疗法至关重要。斑马鱼（Danio rerio）是模拟人类疾病的理想系统，利用斑马鱼阐明先天性眼部疾病的分子和细胞基础的研究在该领域产生了重大影响。事实上，识别斑马鱼眼部缺陷突变体的前向基因筛选，以及分析突变表型的快速技术进步，已经将斑马鱼系统定位在那些有助于我们了解人类先天性眼病机制基础的系统的前沿。这项资助的研究将利用创新的全基因组测序和SNP定位方法，快速、经济地克隆从我们实验室最近完成的前向遗传筛选中鉴定出的隐性斑马鱼视觉系统突变体。在我们的筛查中发现了23个突变体，这些突变体表现为先天性白内障、前段发育不良、结肠瘤、眼皮肤白化、视网膜变性和其他发育缺陷。我们的新定位技术将使我们能够克隆大多数（如果不是全部的话）这些突变的受影响基因座，然后进行有针对性的、假设驱动的实验，以确定导致眼部缺陷的潜在分子和细胞机制。我们将把这些进一步的研究重点放在先天性白内障突变体上。先天性白内障发生率约为10万分之40，是发达国家儿童失明的最常见原因。白内障也是近200种不同的人类遗传疾病的常见临床特征，使其成为其他异质性疾病的常见组成部分。虽然对年龄相关和环境诱发的白内障了解很多，但对先天性白内障的病因了解较少。因此，这里提出的研究将具有重要的科学价值，因为它们将确定正常晶状体发育所需的基因产物，并将提供动物先天性白内障模型，通过该模型可以进一步了解疾病机制，并通过该模型可以开发和测试新的治疗方法。