Construction and Verification of RPL11 Gene Stable Knockdown Cell Lines
LIU Wei1, GUO Peidong1, JIA Nannan2, SUN Yingjie2, TAN Lei2, LIU Weiwei2, QIU Xusheng2, LIAO Ying2, SONG Cuiping2, DING Chan1,2*, MENG Chunchun2*
In order to construct stable knockdown cell lines of RPL11, three different shRNA target sequences of RPL11 gene were designed and ligated to PrNAT-U6.1 with green fluorescent label to obtain recombinant plasmids. The recombinant plasmids were transfected into HeLa cells, and G418 was added for pressure screening. The positive cells were subcloned and purified by limited dilution method. Western blot and qPCR were used to verify the expanded culture of monoclone cells, and RPL11 gene stable knockdown cell line was obtained. CCK-8 cell viability test kit and puromycin labeling method were used to determine the growth activity and new protein synthesis ability of RPL11 stable knockdown cell lines. The results showed that RPL11 stable knockdown did not affect the cell viability and protein synthesis level, which laid a foundation for the follow-up study of RPL11 non ribosomal function.
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