Abstract
Objective To construct a lentiviral vector of RNA interference of chemokine CC motif ligand-5 (CCLS) gene and identify its knockout effectiveness in Hela cells. Methods The pGCSIL-GFP plasmids which expressed four RNAi sequences ( a1, a2, a3, a4) respectively for human CCL5 gene togeth-er with pHelper 1.0 and Helper 2.0 plasmids were transfected into 293T cells in order to construct tentivi-ral vectors of RNA interference of CCL5 gene. Then the lentiviral vectors were transfected into Hela cells, and RT-PCR was used to evaluate its knockout effectiveness for CCL5. Results The knockout effective-ness of a1 ,a2 or a3 was beyond 95% excepting a4 ,whose knockout effectiveness was beyond 90% in Hela cells. Conclusion The knockout effectiveness of the constructed lentviral vectors for human CCL5 was satisfactory. Our results demonstrated that RNAi is able to silence the potential key gene CCL5 in human cancer cells. Key words: CCL5 gene ; RNA interference; Lentiviral vector; Construction; Knockout effectiveness
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