Rapid and inexpensive screening of disease candidate genes in mice

描述（由申请人提供）：该资助旨在显著推进当前人类遗传疾病小鼠模型的生成技术。这涉及广泛的挑战领域（15）转化科学和具体挑战主题15- od (ORDR)-101：预防、早期发现和治疗罕见疾病的试点项目。尽管人类遗传技术的最新进展彻底改变了我们识别疾病候选基因的能力，但我们目前缺乏足够的高通量技术来经济有效地评估候选基因在小鼠体内后续研究中的相关性。我们提出了一种快速和低成本地生成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Conotruncal畸形）小鼠模型的新策略。我们在这项初步研究中建模的每种特定疾病的患病率在1/100,000到1/5,000之间，并且具有遗传异质性，因此这些疾病被称为罕见疾病。由于我们正在生成罕见的脑和心脏先天性畸形的小鼠模型，因此本建议对ODS， NINDS， NICHD和NHLBI有直接的兴趣。然而，值得注意的是，这项研究将对一般生物医学科学和公共卫生产生重大影响，并且可能引起多个NIH研究所的兴趣，因为提出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和其他正在支持大规模人类全基因组关联和CNV研究的机构特别相关，这些研究目前正在进行中，以确定人类疾病候选基因。该提案结合了RNAi技术、小鼠嵌合体生成和小鼠表型集中的最新进展，形成了一种新的策略，允许廉价和快速地生成高效的敲除小鼠模型。我们预测初始敲除表型可以在2个月内评估，每个基因大约2000美元。这大约是目前标准鼠标敲除的1/6时间和至少10%的成本。快速、高效和廉价的敲除小鼠的生产不会取代，而是补充正在进行的大规模NIH小鼠ENU和全基因组基因靶向计划。然而，这种新型基因敲除策略带来的时间和经济效益将彻底改变疾病候选基因分析。资金将使我们对人类疾病发病机制的理解迅速发展，这反过来将改善疾病的诊断和治疗，并进一步加速药物的发现。作为拨款的直接结果，将会新增1.5个全职职位，并保留2个全职职位。尽管人类基因组计划已经允许鉴定许多人类疾病基因，但目前在小鼠身上建立人类疾病模型的技术既耗时又昂贵，而且无法跟上目前人类基因发现的速度。在这个提议中描述了一种新的、廉价的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，改善疾病诊断，促进治疗和药物发现。

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.