Rapid and inexpensive screening of disease candidate genes in mice

快速且廉价地筛选小鼠疾病候选基因

• 批准号: 7942828
• 负责人: Kathleen Joyce Millen
• 金额: $ 50万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7942828
• 关键词: Address; Area; Award; Behavioral; Biological; Birth; Brain; Candidate Disease Gene; Cell physiology; Cells; Cerebellum; Cerebrum; Chimera organism; Communities; Complement; Congenital Abnormality; Congenital Heart Defects; Dandy-Walker Syndrome; Data; Disease; Disease model; ES Cell Line; Early Diagnosis; Embryo; Employment; Ensure; Ethylnitrosourea; Follow-Up Studies; Funding; Gene Targeting; Generations; Genes; Genetic; Genomics; Gold; Grant; Heart; Heart Diseases; Hereditary Disease; Holt Oram syndrome; Human; Human Genetics; Human Genome; Human Genome Project; Institutes; Knock-out; Knockout Mice; Luciferases; Methods; MicroRNAs; Microcephaly; Modeling; Mus; Mutant Strains Mice; NIH Mouse; National Heart, Lung, and Blood Institute; National Human Genome Research Institute; National Institute of Child Health and Human Development; National Institute of Neurological Disorders and Stroke; Needles; Pathogenesis; Patients; Phenocopy; Phenotype; Pilot Projects; Polymerase; Positioning Attribute; Prevalence; Prevention; Production; Protocols documentation; Proxy; Public Health; Qualifying; RNA Interference; Rare Diseases; Reporter; Research; Science; Screening procedure; Specificity; Structure; Surgical incisions; Technology; Time; Translational Research; United States National Institutes of Health; Walkers; Zona Pellucida; abstracting; base; brain malformation; congenital heart disorder; cost; disease diagnosis; drug discovery; embryonic stem cell; expression vector; flexibility; gene function; gene interaction; genetic technology; genome wide association study; genome-wide; high throughput analysis; high throughput technology; human disease; improved; in vivo; interest; knock-down; loss of function; malformation; mouse model; new technology; novel; novel strategies; programs; promoter; small hairpin RNA

DESCRIPTION (provided by applicant): This grant proposes to significantly advance the current technology for generation of mouse models of human genetic disease. This addresses broad Challenge Area (15) Translational Science, and Specific Challenge Topic 15-OD(ORDR)-101: Pilot projects for prevention, early detection and treatment of rare diseases. Although recent advances in human genetic technologies have revolutionized our ability to identify disease candidate genes, we currently lack sufficient high-throughput technology to economically and efficiently assess the relevance of candidate genes in in vivo follow-up studies in mice. We propose a new strategy to rapidly and inexpensively generate mouse models of human structural birth defects of the cerebellum and cerebrum (Dandy-Walker malformation, microcephaly) and severe congenital heart defects (Holt-Oram Syndrome and Conotruncal malformations). Each specific disorder we are modeling in this pilot study has a prevalence of between 1/100,000 and 1/5,000 births and is genetically heterogeneous, qualifying these as Rare Disorders. Since we are generating mouse models of rare brain and heart congenital malformations, this proposal is of direct interest to ODS, NINDS, NICHD and NHLBI. Notably however, this research will have a high impact in general biomedical science and public health and is likely of interest to multiple NIH institutes, since the new technology proposed will be applicable to the generation of mouse models for any human genetic disorder. This technology is especially relevant to NHGRI and other institutes who are supporting large scale human Genome-wide Association and CNV studies currently underway to indentify human disease candidate genes. This proposal combines recent parallel advances in RNAi technology, mouse chimera generation and focused mouse phenotyping into a novel strategy allowing inexpensive and rapid generation of efficient knock-down mouse models. We predict that initial knock-down phenotypes can be assessed in 2 months for approximately $2,000 per gene. This is approximately 1/6 the time and at least 10% of the current cost of standard mouse knock-outs. Rapid, efficient and inexpensive production of knock-down mice will not replace, but rather complement ongoing large scale NIH mouse ENU and genome-wide gene targeting programs. The time and financial economies however, that result from this novel knock-down strategy will revolutionize disease candidate gene analysis. Funding will permit rapid advancement in our understanding the pathogenesis of human disease which in turn, will improve disease diagnosis and treatment and additionally accelerate drug discovery. As a direct result of funding 1.5 new full time employment (FTE) positions will be created and 2 FTE positions will be retained. Although the Human Genome Project has allowed identification of many human disease genes, current technology to model human diseases in mice is time consuming and expensive and cannot keep pace with the current rate of human genetic discoveries. Described in this proposal is a new, inexpensive and rapid strategy to generate mouse models of human disease. This technological advance will enable more rapid understanding of the biological basis of human disease, improve disease diagnosis and facilitate treatment and drug discovery.

描述(由申请者提供)：这项资助计划显著推进目前用于产生人类遗传病小鼠模型的技术。它涉及广泛的挑战领域(15)转化科学，以及具体的挑战主题15-OD(ORDR)-101：预防、早期发现和治疗罕见疾病的试点项目。尽管人类基因技术的最新进展使我们识别疾病候选基因的能力发生了革命性的变化，但我们目前缺乏足够的高通量技术来经济和有效地评估候选基因在小鼠体内后续研究中的相关性。我们提出了一种新的策略，以快速和廉价的方式建立人类小脑和大脑结构性出生缺陷(Dandy-Walker畸形，小头畸形)和严重先天性心脏缺陷(Holt-Oram综合征和圆锥干畸形)的小鼠模型。我们在这项初步研究中建模的每个特定疾病的患病率在1/100,000到1/5,000之间，并且在遗传上是不同的，这将这些疾病称为罕见疾病。由于我们正在建立罕见的大脑和心脏先天性畸形的小鼠模型，因此这项提议与ODS、NINDS、NICHD和NHLBI直接相关。然而，值得注意的是，这项研究将对普通生物医学科学和公共卫生产生高度影响，并可能引起多个NIH研究所的兴趣，因为拟议的新技术将适用于任何人类遗传疾病的小鼠模型的生成。这项技术与NHGRI和其他机构特别相关，这些机构正在支持目前正在进行的大规模人类基因组协会和CNV研究，以确定人类疾病候选基因。这一建议将RNAi技术、小鼠嵌合体生成和专注的小鼠表型的最新进展结合在一起，形成了一种新的策略，可以廉价而快速地生成高效的击倒小鼠模型。我们预测，最初的敲除表型可以在2个月内评估，每个基因大约2,000美元。这大约是标准鼠标淘汰的时间的六分之一，至少是目前成本的10%。快速、高效和廉价地生产击倒小鼠不会取代，而是对正在进行的大规模NIH小鼠ENU和全基因组基因靶向计划的补充。然而，这种新的击倒策略所产生的时间和金融经济将使疾病候选基因分析发生革命性变化。资金将使我们对人类疾病的发病机制的了解迅速进步，这反过来又将改善疾病的诊断和治疗，并进一步加速药物发现。作为供资的直接结果，将创造1.5个新的全职就业职位，并保留2个全职就业职位。尽管人类基因组计划已经能够识别许多人类疾病基因，但目前在小鼠身上模拟人类疾病的技术既耗时又昂贵，跟不上目前人类基因发现的速度。这项提案描述了一种新的、廉价和快速的策略来产生人类疾病的小鼠模型。这一技术进步将使人们能够更快地了解人类疾病的生物学基础，改进疾病诊断，促进治疗和药物发现。