Averina O A, Vysokikh M Y, Permyakov O A, Sergiev P V
Institute of functional genomics, Lomonosov Moscow State University, Moscow, 119991 Russia.
Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, 119991 Russia.
Acta Naturae. 2020 Jan-Mar;12(1):42-50. doi: 10.32607/actanaturae.10937.
The generation of transgenic model organisms (primarily mice) is an integral part of modern fundamental and applied research. Simple techniques based on the biology of these laboratory rodents can often increase efficiency when generating genome-edited mouse strains. In this study, we share our three years of experience in the optimization of mouse genome editing based on microinjection of CRISPR/Cas9 components into ca. 10,000 zygotes. We tested a number of techniques meant to improve efficiency in generating knockout mice, such as optimization of the superovulation method and choosing the optimal mouse strains to be used as zygote donors and foster mothers. The presented results might be useful to laboratories aiming to quickly and efficiently create new mouse strains with tailored genome editing.
转基因模式生物（主要是小鼠）的产生是现代基础研究和应用研究不可或缺的一部分。基于这些实验啮齿动物生物学特性的简单技术，在生成基因组编辑小鼠品系时通常可以提高效率。在本研究中，我们分享了基于将CRISPR/Cas9组件显微注射到约10000个受精卵中对小鼠基因组编辑进行优化的三年经验。我们测试了许多旨在提高敲除小鼠生成效率的技术，比如优化超排卵方法，选择用作受精卵供体和代孕母鼠的最佳小鼠品系。所展示的结果可能对旨在快速高效地创建具有定制基因组编辑的新小鼠品系的实验室有用。
