Lentivirus (LV) construction and infection

PJ Pengyue Jiang
SY Shengyu Ye
XF Xiaobin Fan
YT Yini Tian
DZ Dongmei Zhang
WP Weiqing Pan
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The expression plasmid for Malat1 was constructed by PCR and inserted into the pCDH-CMV-EF1-Puro vector. LV-Malat1 or LV-Scramble was produced by co-transfecting HEK293T cells cultured in 10-cm dishes with pCDH-CMV-EF1-Puro-Malat1 or empty vectors and packaging plasmids (psPAX.2 and pMD2.G) at a ratio of 4:3:1. After 48 h of transfection, the lentivirus supernatant was filtered through a 0.45-μm filter, followed by infection of cells with 10 μg/ml polybrene. One day after infection, the medium was replaced with fresh medium and cultured for 48 h before cells were collected for further analysis.

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