Zhang Lichen, Li Wushan, Liu Zhuangzhuang, Liu Yang, Liu Zhilong, Gu Yanrong, He Le, Zhou Binhui, Li Tianhan, Chao Tianzhu, Liang Yinming, Lu Liaoxun
Laboratory of Genetic Regulators in the Immune System, Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang, China.
Henan Key Laboratory of Immunology and Targeted Therapy, Xinxiang Medical University, Xinxiang, China.
Front Cell Dev Biol. 2022 Jan 18;9:769673. doi: 10.3389/fcell.2021.769673. eCollection 2021.
Functional genomics in a mammalian model such as mice is fundamental for understanding human biology. The CRISPR/Cas9 system dramatically changed the tempo of obtaining genetic mouse models due to high efficiency. However, experimental evidence for the establishment of sgRNA knock-in animals and analyses of their value in functional genomics are still not sufficient, particularly in mammalian models. In this study, we demonstrate that the establishment of sgRNA knock-in mice is feasible, and more importantly, crosses between sgRNA knock-in mice and the Cas9 constitutively expressing mice result in complete deletion of the target gene. Such sgRNA knock-in provides an alternative approach for genetic modification and can be useful in multiple circumstances, such as maintenance of genetically modified animals, which are difficult to breed as homozygotes, and cross of such mice to diverse genomic backgrounds to obtain genetically modified animals.
在诸如小鼠这样的哺乳动物模型中进行功能基因组学研究对于理解人类生物学至关重要。由于效率高，CRISPR/Cas9系统极大地改变了获取基因小鼠模型的速度。然而，关于建立sgRNA敲入动物及其在功能基因组学中的价值分析的实验证据仍然不足，尤其是在哺乳动物模型中。在本研究中，我们证明建立sgRNA敲入小鼠是可行的，更重要的是，sgRNA敲入小鼠与组成型表达Cas9的小鼠杂交会导致靶基因完全缺失。这种sgRNA敲入为基因修饰提供了一种替代方法，并且在多种情况下都可能有用，例如维持难以纯合繁殖的基因修饰动物，以及将此类小鼠与不同基因组背景杂交以获得基因修饰动物。
