Resources for Comparative Mendelian Disease Genomics

比较孟德尔疾病基因组学资源

基本信息

批准号：
8998309
负责人：
LAURA G REINHOLDT
金额：
$ 85.39万
依托单位：
JACKSON LABORATORY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8998309
关键词：
Achondroplasia Address Affect Albers-Schonberg disease Ally American Bone Diseases Breeding CRISPR/Cas technology Clinic Clinical Code Copy Number Polymorphism DNA Sequence Analysis Data Databases Detection Diagnosis Disease Disease model Etiology Family Gene Transfer Techniques Genes Genetic Genetic Engineering Genetic Heterogeneity Genome Genomics Goals Hereditary Malignant Neoplasm High-Throughput DNA Sequencing Human Human Genetics Human Genome Inbred Strain Inherited Lesion Libraries Maps Mendelian disorder Methods Modeling Molecular Molecular Diagnosis Mouse Strains Mus Mutation Mutation Detection Neonatal Osteochondromatosis Other Genetics Partner in relationship Patients Perinatal Phenotype Population Process Proteins Public Health Research Resources Source Source Code Techniques Technology Testing Translating Untranslated RNA Validation Variant accurate diagnosis base comparative congenital heart disorder consanguineous family cost effective design exome exome sequencing experience gene discovery genome editing genome sequencing improved in vivo mouse genome mutant neonate nervous system disorder new technology public health relevance research study reverse genetics segregation skin disorder success tool transcriptome sequencing whole genome

项目摘要

DESCRIPTION (provided by applicant): Approximately 20-30 million Americans are affected by Mendelian genetic disorders with broad clinical consequences including congenital heart disease, congenital bone diseases, inherited skin diseases, hereditary neurological disorders, hereditary cancers, and others. Over the last several years, high-throughput, whole-exome sequencing has been used for molecular diagnosis and as a research tool for discovery of new disease-causative gene(s). Since the first successful application of this technology six years ago, the fundamental genetic bases for over 100 Mendelian diseases have been identified. Despite these advances, the overall success rate for human Mendelian disease gene discovery by whole-exome sequencing remains at slightly less than 50 percent. In contrast to these clinical cases, the discovery of Mendelian disease genes in mice is powered by genetically defined inbred strain backgrounds, large consanguineous pedigrees for segregation analysis, and disease modeling through the use of exciting new CRISPR/Cas9 approaches and more traditional genetic engineering techniques. With these allied technologies, the application of whole-exome sequencing in recent years has increased the rate of mutation discovery in mouse by nearly ten fold. Still, as in humans, the success rate for Mendelian disease gene discovery in the mouse is only slightly better than 50 percent. Possible limitations of whole-exome sequencing for disease gene discovery in both the mouse and human genomes include shortcomings of variant calling tools, insufficient resources describing `normal' genome variation, and the likely existence of regulatory mutations that, residing outside of protein-codin regions, escape detection by exome-sequencing. With the promise of exploring and surmounting these limitations, our long-term goal is to develop genomic resources that facilitate functionalization of both the coding and non-coding portions of the mouse genome through forward genetic discovery approaches and reverse genetic validation. More specifically, the objectives of this proposal are to expand the scope of gene discovery beyond exome-imposed limitations, and to use these data to develop a mouse genome variation database, and other optimized resources, that will deliver improved mutation discovery success rates. Moreover, we will apply these new technologies to explore de novo mutations and perinatal lethal phenotypes in a large population of mouse neonates, as well as identifying the heritable molecular lesions in established, genetically defined Mendelian disease models. In both settings, we will validate and model the mutations using modern CRISPR/Cas9 and other genetic engineering approaches.

描述（由适用提供）：大约有20-30万美国人受到孟德尔遗传疾病的影响，包括先天性心脏病，先天性骨疾病，遗传性皮肤病，遗传性神经系统疾病，遗传性癌症等。在过去的几年中，高通量，全象征测序已用于分子诊断，并作为发现新型疾病疾病基因的研究工具。自六年前该技术的第一次成功应用以来，已经确定了100多种孟德尔疾病的基本遗传基础。尽管取得了这些进步，但全异位测序发现人类孟德尔疾病基因的总体成功率仍然低于50％。与这些临床病例相反，小鼠中孟德尔疾病基因的发现由基因定义的近交菌株背景，大的近亲血统，用于隔离分析的大阴谋和疾病建模，通过使用令人兴奋的新CRISPR/CAS9方法和更传统的基因工程技术。借助这些相关技术，近年来全异位测序的应用使小鼠突变发现率提高了近十倍。尽管如此，就像人类一样，小鼠中孟德尔病基因发现的成功率仅略高于50％。小鼠和人类基因组中疾病基因发现的全异位体测序的可能局限性包括变异通话工具的缺点，描述“正常”基因组变异的资源不足以及调节突变的可能性存在，这些突变的可能性存在，这些突变的可能性存在于蛋白质 - 二烷区域之外的逃逸，并通过蛋白质 - 毒素区域之外的逃生来检测。有了探索和克服这些局限性的承诺，我们的长期目标是开发基因组资源，通过远期遗传发现方法和反向遗传验证，促进小鼠基因组的编码和非编码部分的功能化。更具体地说，该提案的目标是将基因发现的范围扩展到外显型限制之外，并使用这些数据来开发鼠标基因组变异数据库以及其他优化资源，这些资源将提供改进的突变发现成功率。此外，我们将应用这些新技术来探索大量小鼠新生儿中的从头突变和围产期致死表型，并确定已建立的遗传定义的孟德尔氏病模型中可遗传的分子病变。在这两种情况下，我们都将使用现代CRISPR/CAS9和其他基因工程方法来验证和建模突变。