Gruzdev Artiom, Scott Greg J, Hagler Thomas B, Ray Manas K
Knockout Mouse Core, Reproductive and Developmental Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, NC, USA.
Methods Mol Biol. 2019;1960:1-21. doi: 10.1007/978-1-4939-9167-9_1.
The study of gene function in normal human physiology and pathophysiology is complicated by countless factors such as genetic diversity (~98 million SNPs identified in the human genome as of June 2015), environmental exposure, epigenetic imprinting, maternal/in utero exposure, diet, exercise, age, sex, socioeconomic factors, and many other variables. Inbred mouse lines have allowed researchers to control for many of the variables that define human diversity but complicate the study of the human genome, gene/protein function, cellular and molecular pathways, and countless other genetic diseases. Furthermore, genetically modified mouse models enable us to generate and study mice whose genomes differ by as little as a single point mutation while controlling for non-genomic variables. This allows researchers to elucidate the quintessential function of a gene, which will further not only our scientific understanding, but provide key insight into human health and disease. Recent advances in CRISPR/Cas9 genome editing have revolutionized scientific protocols for introducing mutations into the mammalian genome. The ensuing chapter provides an overview of CRISPR/Cas9 genome editing in murine embryonic stem cells for the generation of genetically modified mouse models.
在正常人体生理学和病理生理学中研究基因功能会受到无数因素的影响，如基因多样性（截至2015年6月，人类基因组中已鉴定出约9800万个单核苷酸多态性）、环境暴露、表观遗传印记、母体/子宫内暴露、饮食、运动、年龄、性别、社会经济因素以及许多其他变量。近交系小鼠使研究人员能够控制许多界定人类多样性但会使人类基因组、基因/蛋白质功能、细胞和分子途径以及无数其他遗传疾病研究变得复杂的变量。此外，基因编辑小鼠模型使我们能够生成并研究基因组差异小至单个点突变的小鼠，同时控制非基因组变量。这使研究人员能够阐明基因的基本功能，这不仅将增进我们的科学理解，还能为人类健康和疾病提供关键见解。CRISPR/Cas9基因组编辑技术的最新进展彻底改变了将突变引入哺乳动物基因组的科学方案。随后的章节概述了在小鼠胚胎干细胞中进行CRISPR/Cas9基因组编辑以生成基因编辑小鼠模型的方法。
