Advanced genetic engineering technology development

先进基因工程技术开发

• 批准号: 8145528
• 负责人: Aron M Geurts
• 金额: $ 229.5万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-30 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8145528
• 关键词: Achievement; Address; Affect; American; Animal Model; Animals; Antihypertensive Agents; Award; Biological Models; Cardiovascular Diseases; Cells; Developmental Process; Disease; Embryo; Engineering; Genes; Genetic Engineering; Genetic Research; Genome; Genome engineering; Gold; Human; Hypertension; Kidney Diseases; Kidney Failure; Knock-in Mouse; Knock-out; Laboratories; Laboratory Rat; Laboratory mice; Methods; Modeling; Mus; Mutagenesis; Names; Physiology; Process; Rattus; Research; Research Personnel; Role; Scientist; Stem cells; Technology; Testing; Tissues; United States National Institutes of Health; Work; Zinc Fingers; abstracting; gene replacement; human disease; innovation; interest; knockout animal; knockout gene; novel strategies; nuclease; public health relevance; research study; technology development; vertebrate genome

DESCRIPTION (Provided by the applicant) Abstract: The NIH created the Director's New Innovator Award to ""support exceptionally creative new investiga- tors who propose highly innovative projects that have the potential for unusually high impact."" I have been working for more than ten years on new and innovative approaches for manipulating vertebrate genomes. Having the ability to manipulate specific genes in laboratory model systems has been the gold standard of genetic research for many years, but the most useful strategies for specifically knocking out (disrupting) or knocking in (adding) specific genes precisely into the genome has been limited to mice. However, laboratory mice are not always the best model system for understanding human physiology and disease. If we can expand these genome engineering approaches to other species it will have a tremendous impact on human disease research because we will finally be able to study specific genes in the most relevant animal models. Recently, my colleagues and I were the first in the world to target and knock out genes in the laboratory rat, the most widely studied research animal model by physiologists, behavioralists, neuroscientists, nutritionalists, and by researchers in- terested in cardiovascular and kidney diseases. This technological achievement was named by The Scientist as one of the 'Top 10 Innovations' in 2009. By applying a Zinc-Finger Nuclease (ZFN) technology to the rat embryo, generating knockout animal models for specific genes becomes very rapid because it does not require the labor of engineering stem cells and then turning those stem cells into a whole animal. We have been very successful with this approach and have now knocked out 57 genes related to hypertension and renal failure in humans in the past year - something that was not possible even 2 years ago. Using ZFNs for gene knockout, however, is just the beginning of what can be done with this technology. Specifically, we now know that gene knockin approaches in the rat embryo are now possible , but much remains to be explored. We propose to understand the capabilities and limitations of ZFNs of knockin engineering to enable rapid and reproducible methods for addressing hypotheses related to genes involved in any vertebrate model system for virtually any disease or developmental process. For us, the model system is the rat and the disease we study is hypertension, but the innovative approaches developed by this proposal will serve as a blueprint for any model system where embryos can be isolated. Specifically, we will use ZFN knockin technology to enable conditional mutagenesis and whole gene replacement approaches in the rat to develop new and valuable models to address hypotheses related to hypertension and evaluate new anti- hypertensive therapies and make an impact on the estimated 28% of Americans and more than one billion people world-wide affected by this disease. Public Health Relevance: The key to developing effective targeted therapies for the treatment of human disease such is the understanding of how genes, cells and tissues are involved in specific disease processes. Genetic engineering in mice to understand these roles has been the gold standard for many years, but we have developed new approaches which are potentially applicable to any research animal model, including rats. The proposed studies aim to establish highly innovative and widely applicable approaches for studying specific genes in cells and tissues in any model system and to develop better animal models for testing therapies for diseases like hypertension.

描述（申请人提供） 摘要：NIH创建了导演的新创新者奖，以“支持”具有异常创新的新调查，他们提出了具有异常高影响力的高度创新的项目。多年来，具有在实验室模型系统中操纵特定基因的能力一直是遗传研究的黄金标准，但是专门敲除（破坏）或敲入（添加）特定基因的最有用的策略精确地限于小鼠。但是，实验室小鼠并不总是理解人类生理和疾病的最佳模型系统。如果我们可以将这些基因组工程方法扩展到其他物种，它将对人类疾病的研究产生巨大影响，因为我们最终将能够研究最相关的动物模型中的特定基因。最近，我和我的同事是世界上第一个靶向和淘汰实验室大鼠中基因的人，这是生理学家，行为主义者，神经科学家，营养学家，以及在心血管疾病和肾脏疾病中遭受高估的研究人员。这项技术成就由科学家命名为2009年的“十大创新”之一。通过将锌指核酸酶（ZFN）技术应用于大鼠胚胎，为特定基因产生敲除动物模型变得非常迅速，因为它不需要工程干细胞的劳动，然后将这些干细胞变成整个动物。在这种方法中，我们一直非常成功，现在已经淘汰了过去一年与人类高血压和肾衰竭有关的57个基因 - 即使在两年前，这也是不可能的。但是，将ZFN用于基因敲除只是该技术可以做的事情的开始。具体来说，我们现在知道，现在可以使用大鼠胚胎中的基因敲击蛋白方法，但仍有许多待探索。我们建议了解敲蛋白工程的ZFN的能力和局限性，以实现与任何脊椎动物模型系统有关的基因相关的假设，以实现快速且可重复的方法，用于几乎任何疾病或发育过程。对我们来说，模型系统是大鼠，我们研究的疾病是高血压，但是该提案开发的创新方法将成为任何可以隔离胚胎的模型系统的蓝图。具体而言，我们将使用ZFN敲蛋白技术来实现大鼠中有条件的诱变和整个基因替代方法，以开发新的和有价值的模型，以解决与高血压有关的假设，并评估新的抗高血压疗法，并对估计的28％的美国人和该疾病全球范围内的十亿多人产生影响。 公共卫生相关性：开发有效的靶向疗法以治疗人类疾病的关键是对基因，细胞和组织如何参与特定疾病过程的理解。多年来，了解这些角色的小鼠基因工程一直是黄金标准，但是我们开发了新方法，这些方法可能适用于包括大鼠在内的任何研究动物模型。拟议的研究旨在建立高度创新的和广泛适用的方法，用于在任何模型系统中研究细胞和组织中的特定基因，并开发更好的动物模型，用于测试高血压等疾病的疗法。