High efficiency platform for rapid RNAi rat model development

摘要 药物开发的成本已经飙升到估计每种FDA批准的药物26亿美元，主要是 由于缺乏有效性或安全性而失败，这表明我们目前的临床前验证过程 在预测人类的治疗潜力和毒性方面还不够。动物模型是黄金 用于解剖疾病机制和评估体内新药靶点的标准;然而， 由于开发这些药物的准备时间很长，因此无法在药物发现过程中进行常规使用。随着近期 CRISPR/Cas9基因组编辑的发展，以及RNA干扰技术的进步，我们现在 有能力快速开发具有精确基因组修饰的动物模型， 病理学我们已经表明，RNAi作为基因缺失的快速替代物，也可以用于 基因工程小鼠模型，以评估新基因的治疗潜力和预测毒性 目标的这项提案的目标是将我们的能力扩展到mice之外，并开发一个平台， 并且在短短4个月内就能经济高效地生产出RNAi大鼠。尽管小鼠模型的实用性， 许多科学家认为，大鼠仍然是首选的啮齿动物，因为它们的体积更大，便于手术操作， 重复血液采样，以及他们的认知和生理特征，更接近于 人类比他们的老鼠同行。对于神经生物学、心脏生物学、免疫学和毒理学， 仍然是研究中的主要啮齿动物模型。我们目前的客户群中有近20%的人询问过利率 在过去的5年里，他们的化合物的大多数毒理学研究仍然是在大鼠中进行的， 第一阶段我们相信，大鼠将再次成为药物发现的首要啮齿动物模型 我们打算站在这一转变的前沿。CRISPR/Cas9基因组编辑现在提供了 的路径操纵大鼠基因组;然而，目前的方法使永久基因的衍生 敲除等位基因，但不允许时间和可逆的基因调控。我们的目标是从我们庞大的 建立小鼠模型的经验，并利用基于CRISPR的靶向效率来开发RNAi大鼠 模型，使诱导和可逆的基因沉默，以模拟治疗方案。这些RNAi大鼠 模型将通过评估潜在的药物反应来改变临床前验证过程， 研究体内的耐药机制，最终指导开发更安全、更有效的药物。

Abstract The cost of drug development has skyrocketed to an estimated $2.6B for every FDA approved drug primarily due to failures from lack of efficacy or safety, suggesting that our current preclinical validation process has been insufficient in predicting therapeutic potential and toxicity in humans. Animal models are the gold standard for dissecting disease mechanisms and evaluating novel drug targets in vivo; however, the cost and long lead time to develop them has prevented their routine use in the drug discovery process. With the recent developments in CRISPR/Cas9 genome editing, and advances in RNA interference technologies, we now have the ability to rapidly develop animal models with precise genomic modifications, and human-like disease pathologies. We have shown that RNAi serves as a fast alterative to gene deletion and can also be used within genetically engineered mouse models to assess the therapeutic potential and predict toxicities of novel gene targets. The goal of this proposal is to expand our capabilities beyond mice and develop a platform for rapid and cost-effective production of RNAi rats in as little as 4 months. Despite the utility of mouse models, for many scientists, the rat still remains the preferred rodent due to their larger size for surgical manipulation, repeat blood sampling, and their cognitive and physiological characteristics that more closely resemble humans than their mouse counterparts. For neurobiology, cardiobiology, immunology and toxicology, they are still the dominant rodent model in research. Nearly 20% of our current client base has inquired about rat models over the last 5 years, noting that most toxicology studies of their compounds are still done in rats prior to Phase I. We believe that rats will gain popularity once again as the premier rodent model in drug discovery and we intend to be at the forefront of this shifting paradigm. CRISPR/Cas9 genome editing now provides a path for manipulating the rat genome; however, current approaches enable the derivation of permanent gene knockout alleles, but do not allow temporal and reversible gene regulation. Our goal is to draw from our vast experience of mouse model creation and exploit the efficiency of CRISPR-based targeting to develop RNAi rat models that enable inducible and reversible gene silencing to simulate therapeutic regimes. These RNAi rat models will transform the preclinical validation process with assessment of potential drug response and resistance mechanisms in vivo, ultimately guiding the development of safer and more effective drugs.