Rapid and inexpensive screening of disease candidate genes in mice

描述（由申请人提供）：该资助旨在显着推进目前用于生成人类遗传疾病小鼠模型的技术。 这涉及广泛的挑战领域（15）转化科学和特定的挑战主题15-OD（ORR）-101：预防，早期发现和治疗罕见疾病的试点项目。 虽然人类遗传技术的最新进展已经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随访研究中的相关性。 我们提出了一种新的策略，以快速，廉价地产生小鼠模型的人类结构出生缺陷的小脑和大脑（Dandy-Walker畸形，小头畸形）和严重的先天性心脏病（Holt-Oram综合征和圆锥动脉干畸形）。 我们在这项试点研究中建模的每种特定疾病的患病率在1/100，000和1/5，000之间，并且具有遗传异质性，将其归类为罕见疾病。 由于我们正在生成罕见的大脑和心脏先天畸形的小鼠模型，因此该提议对ODS，NINDS，NICHD和NHLBI直接感兴趣。 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很大影响，并且可能会引起多个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和其他研究所特别相关，这些研究所正在支持目前正在进行的大规模人类全基因组关联和CNV研究，以识别人类疾病候选基因。 该提案将RNAi技术、小鼠嵌合体生成和集中的小鼠表型分析的最新平行进展结合到一种新策略中，从而允许廉价且快速地生成有效的敲除小鼠模型。 我们预测，最初的敲除表型可以在2个月内进行评估，每个基因大约2,000美元。 这大约是标准小鼠敲除的时间的1/6和当前成本的至少10%。 快速、高效和廉价地生产敲低小鼠不会取代，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，这种新的敲除策略所带来的时间和财政经济将彻底改变疾病候选基因分析。 资金将使我们能够快速推进对人类疾病发病机制的理解，从而改善疾病诊断和治疗，并加速药物发现。 作为资助的直接结果，将创建1.5个新的全职就业（FTE）职位，并保留2个FTE职位。 虽然人类基因组计划已经允许识别许多人类疾病基因，但目前在小鼠中建立人类疾病模型的技术既耗时又昂贵，并且无法跟上目前人类基因发现的速度。 该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.