German Center for Neurodegenerative Diseases, Tatzberg 41, Germany and Center for Regenerative Therapies Dresden, Fetscherstraße 101, 01307 Dresden, Germany.
STAR Protoc. 2021 Apr 16;2(2):100472. doi: 10.1016/j.xpro.2021.100472. eCollection 2021 Jun 18.
Genetic manipulation of neural precursor cells is an important tool to study mechanisms underlying proliferation, fate specification, and neuron formation. The CRISPR/Cas9 system enables efficient genome editing but requires the clonal expansion of cells containing the desired mutation. Here, we describe a protocol for the effective generation of clonal mouse hippocampal neural precursor lines with CRISPR/Cas9-based gene knockouts. Edited cell lines can be used to investigate gene regulatory networks driving neuronal differentiation and for modeling of diseases that involve hippocampal neurogenesis. For complete details on the use and execution of this protocol, please refer to Pötzsch et al. (2021).
神经前体细胞的遗传操作是研究增殖、命运特化和神经元形成的重要工具。CRISPR/Cas9 系统可实现高效的基因组编辑，但需要克隆扩增含有所需突变的细胞。在此，我们描述了一种利用 CRISPR/Cas9 进行基因敲除的方法，可有效地生成具有克隆性的小鼠海马神经前体细胞系。编辑后的细胞系可用于研究驱动神经元分化的基因调控网络，并用于建模涉及海马神经发生的疾病。有关该方案的使用和执行的完整详细信息，请参阅 Pötzsch 等人（2021 年）。
