Yuan Celvie L, Hu Yueh-Chiang
Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, 3333 Burnet Avenue, MLC7007, Cincinnati, OH, 45229, USA.
Adv Exp Med Biol. 2017;1016:75-90. doi: 10.1007/978-3-319-63904-8_4.
The use of animal models, particularly rodents, has been immensely important to nearly all aspects of biomedical research, from basic science exploration to translational discoveries into clinical applications. The transgenic core facility that provides animal model production, preservation, and recovery services has been fundamental to the success of research efforts using animals. Recent advances in genome editing technologies, especially the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) enzyme system, have transformed the tedious animal model production into a simple and effective procedure. We, as a transgenic core facility established in 1993, adopted the CRISPR/Cas9 technology in early 2014 and have experienced the dramatic shift in the practice of animal model production, from the conventional embryonic stem cell approach to the direct genomic editing in rodent embryos. In this chapter, we describe the lessons that we learned from more than 200 genome editing projects performed in this core facility within the past 3 years. We also provide the practical guidelines for efficient generation of animal models using this technology and the insights into where new technologies lead us.
动物模型的使用，尤其是啮齿动物模型，对于生物医学研究的几乎所有方面都极为重要，从基础科学探索到转化为临床应用的研究发现。提供动物模型生产、保存和恢复服务的转基因核心设施对于利用动物的研究工作取得成功至关重要。基因组编辑技术的最新进展，特别是成簇规律间隔短回文重复序列（CRISPR）/CRISPR相关（Cas）酶系统，已将繁琐的动物模型生产转变为简单有效的程序。我们作为1993年成立的转基因核心设施，于2014年初采用了CRISPR/Cas9技术，并经历了动物模型生产实践中的巨大转变，从传统的胚胎干细胞方法转变为在啮齿动物胚胎中直接进行基因组编辑。在本章中，我们描述了过去3年在该核心设施中进行的200多个基因组编辑项目所学到的经验教训。我们还提供了使用该技术高效生成动物模型的实用指南以及对新技术将引领我们走向何方的见解。
