Optimization of Plasmid Electroporation of Jurkat Cells for Delivery of CRISPR/Cas9 System
Zhao Ri1,2, Meng Lu3, Feng Jianqiong2, Liu Pan3, Jia Ting1, Zheng Wuyan4, Zhang Yue5, Liu Jiahui4, Zhang Tao2, Zou Qiang3, Li Hua2*
CRISPR/Cas9, a highly efficient gene editing technology, can be used for gene modification in T cells to enhance their specific immune response in adaptive cellular therapy of tumor. However, the delivery methods and efficiency of CRISPR/Cas9 system affects the efficiency of gene editing. Electroporation is a safe, simple, economical and reproducible transfection strategy and could be used to deliver CRISPR/Cas9 system into the cells. In this study, Jurkat cells were used as a T cell model and the plasmid with green fluorescent protein gene was used as a detective marker to observe the transfection efficiency and optimize the parameters in plasmid electroporation of Jurkat cells.Furthermore, using these optimized parameters, the CRISPR/Cas9 system was successfully delivered into Jurkat cells and the β2M gene was effectively knocked out to become HLA-I− Jurkat cells. This study provided an optimal system of plasmid electroporation of Jurkat cells suitable for delivery of CRISPR/Cas9 system, laid a foundation for subsequent application of CRISPR/Cas9 system in adaptive cellular therapy of tumor.
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