Department of Biomedicine, Basel University Hospital and University of Basel, Basel, Switzerland.
Transplantation Immunology & Nephrology, Basel University Hospital, Basel, Switzerland.
Methods Mol Biol. 2021;2285:255-264. doi: 10.1007/978-1-0716-1311-5_20.
The CRISPR/Cas technology allows for genome editing in primary T cells. We herein describe the activation of primary murine CD4 or CD8 T cells, followed by electroporation with plasmid or ribonucleoproteins (RNP) for gene modification. Gene edited T cells can subsequently be transferred to host mice for in vivo studies or cultured in vitro for further characterization. This protocol enables sophisticated genetic analysis of T cells using commonly available virus-free reagents.
CRISPR/Cas 技术可实现原代 T 细胞的基因组编辑。本文描述了原代小鼠 CD4 或 CD8 T 细胞的激活，随后通过质粒或核糖核蛋白（RNP）电穿孔进行基因修饰。经基因编辑的 T 细胞可随后转移至宿主小鼠进行体内研究，或在体外培养进行进一步鉴定。本方案使用常用的无病毒试剂实现了 T 细胞的复杂遗传分析。
