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用于电穿孔和分离小鼠(亚)室管膜区细胞的方案,用于单细胞组学。

Protocol for electroporating and isolating murine (sub)ventricular zone cells for single-nuclei omics.

机构信息

Department of Biological Sciences, Clemson University, Clemson, SC 29631, USA.

Department of Biological Sciences, Clemson University, Clemson, SC 29631, USA.

出版信息

STAR Protoc. 2024 Jun 21;5(2):103095. doi: 10.1016/j.xpro.2024.103095. Epub 2024 May 31.

Abstract

In vivo genetic modification of neural stem cells is necessary to model the origins and pathogenesis of neurological disorders. Electroporation is a technique that applies a transient electrical field to direct charged molecules into living cells to genetically modify the mouse brain. Here, we provide a protocol to electroporate the neural stem cells surrounding the neonatal ventricles. We describe subsequent steps to isolate and prepare nuclei from the cells and their cellular progeny for single-nuclei omics. For complete details on the use and execution of this protocol, please refer to Riley et al..

摘要

在体神经干细胞遗传修饰对于模拟神经疾病的起源和发病机制是必要的。电穿孔是一种将短暂的电场施加于带电荷的分子以将其导入活细胞从而修饰小鼠大脑的技术。在这里,我们提供了一种在新生鼠脑室周围电穿孔神经干细胞的方案。我们还描述了随后的步骤,包括从细胞及其细胞后代中分离和制备细胞核,以进行单细胞组学分析。有关此方案的使用和执行的完整详细信息,请参见 Riley 等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/167c/11179414/35ceefe848a7/fx1.jpg

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