CRE rat for Psychiatric Disorders

描述（由申请人提供）：据估计，成瘾及其相关成本：犯罪、家庭暴力和虐待儿童、医疗保健成本、失业和家庭结构的损失，每年超过5000亿美元。与毒瘾作斗争不是意志力的问题，而是一场了解药物如何影响大脑并永久改变其功能的战斗。了解成瘾等复杂疾病的细胞和遗传基础的关键限制之一是缺乏良好的模型来测试与细胞和基因及其功能相关的假设。实验室大鼠是一种具有巨大价值的模型，因为它的生理、生化和行为特征被广泛研究，而且它们的基因组序列与人类相似。超过2000种疾病相关性状的遗传决定因素已经在大鼠中被初步描述，然而，由于操纵大鼠基因组的技术的限制，很难精确地将特定基因与这些性状联系起来。在过去的4年里，我们一直在开发新的工具来缩小这一技术差距。我们已经开发了新的和有效的方法，使转基因大鼠使用转座元件是高度可复制的，我们是第一个应用锌指核酸酶（ZFN）技术靶向和破坏，或敲除，在大鼠的特定基因。这是两项非常重要的技术，可以让我们做很多事情，但还需要做更多的工作。很多时候，敲除整个动物体内的一个基因会妨碍研究它在特定细胞或组织中的作用，因为该基因对早期胚胎发育至关重要，因此敲除它会导致动物死亡。在其他情况下，敲除整个动物的基因并不能让人们区分基因在哪个器官（或大脑的一部分，例如）起作用导致疾病。在这项试点/可行性研究中，我们建议朝着这一目标发展下一个关键技术步骤——能够在特定时间特异性地破坏特定细胞或组织中的基因。我们将培育在特定组织中表达可诱导的CRE/loxP重组酶系统的转基因大鼠。这个系统将允许我们创造条件敲除大鼠，我们可以控制基因功能从细胞中移除的地点和时间。我们将重点关注已知对研究成瘾和行为很重要的脑神经系统——多巴胺能和血清素能神经元。为了证明这个系统是有效的，我们将敲除产生血清素的基因，这种激素在大脑的奖励系统中很重要，在成瘾和其他精神疾病中起着不可或缺的作用，特别是在后脑的神经元中。如果成功，这种方法将改变我们在实验室大鼠模型中研究基因、细胞和疾病的方式，从而影响成瘾和药物滥用的社会经济负担。

DESCRIPTION (provided by applicant): It is estimated that addiction and its associated costs: crime, domestic violence and child abuse, health care costs, and loss of employment and family structure, exceed half of a trillion dollars per year. Fighting addiction is not a matter of willpower, it is a battle to understand how drugs affect the brain and can permanently alter its function. One of the key limitations to understanding the cellular and genetic basis of a complex disease like addiction is the availability of good models to test hypotheses related to cells and genes and their functions. The laboratory rat is one model which has tremendous value because of its intensely studied physiological, biochemical, and behavioral characteristics and their genome sequence similarities to humans. More than 2000 genetic determinants of disease-related traits have been initially described in the rat, however, it has been difficult to precisely correlate specific genes with these traits because of limitations in technology for manipulating the rat genome. In the past 4 years, we have been developing new tools to close this technology gap. We have developed new and efficient ways of making transgenic rats using transposable elements which are highly reproducible and we were the first to apply zinc-finger nuclease (ZFN) technology to target and disrupt, or knock out, specific genes in the rat. These are two very important technologies that allow us to do many things, but more work is needed. Many times, knocking out a gene in the whole animal precludes studying its role in a particular cell or tissue because the gene is essential for early embryo development and so knocking it out causes the animal to die. In other cases, knocking out the gene in the whole animal doesn't allow one to distinguish what organ (or part of the brain, for example) the gene is functioning in to cause the disease. In this pilot/feasibility study, we propose to develop the next key technological step toward this goal - to be able to specifically disrupt genes in a particular cell or tissue at a particular time. We will develop transgenic rats which express an inducible CRE/loxP recombinase system in specific tissues. This system will allow us to create conditional knockout rats where we can control where and when a gene function is removed from a cell. We will focus brain neuron systems known to be important for studying addiction and behavior - the dopaminergic and serotonergic neurons. As a proof of principal that the system is working, we will knock out the gene that produces serotonin, a hormone that is important in the reward system of the brain and an integral player in addiction and other psychiatric disorders, specifically in neurons of the hindbrain. If successful, this approach will change the way we can approach genes, cells and diseases in the laboratory rat model to impact the socioeconomic burdens of addiction and drug abuse. PUBLIC HEALTH RELEVANCE: The key to developing effective therapies for the treatment of addiction and other psychiatric disorders is the identification of genes and the cells they work in to target new drugs and approaches. Lab rats are the preferred model for studying addiction by many resources, but the technology to study a particular gene by knocking out its function in a particular cell or tissue has not been demonstrated. This pilot and feasibility study aims to develop that technology in the lab rat so we can enable studies of specific genes and their roles in addictive behavior and other diseases.