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Generation of stable CD19 knockout (CD19KO) cell lines

GL Guangchao Li
QZ Qing Zhang
ZL Zhi Liu
HS Huijuan Shen
YZ Yangmin Zhu
ZZ Zhao Zhou
WD Wen Ding
SH Siqi Han
JZ Jie Zhou
RO Ruiming Ou
ML Min Luo
SL Shuang Liu
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Stable CD19KO cell lines were generated through the CD19-CRISPR-Cas9 technique, according to the manufacturer’s instructions for the LentiCRISPRv2 vector. Briefly, the sgRNA against CD19 (SEQ ID NO: 11: 5’-CAGTCCTATGAGGATATGAG-3) was cloned into the LentiCRISPRv2 vector, which was digested using BsmBI enzyme. The linkage system included 1 µL (50 ng) of linear LentiCRISPRv2 vector, 1 µL of double-stranded DNA corresponding to sgRNA, 1 µL of T4 DNA ligase, 5 µL of 2 × Quick linkage buffer, and 3 µL of ddH2O. Raji was then transfected with lentivirus packaged using LentiCRISPRv2-CD19-sgRNA, and the stably transfected Raji was then sorted with 5 µg/mL puromycin.

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