Interference of shRNA on Keap1 in prostate cancer PC3 cell mediated by lentivirus

Abstract

Abstract: AIM: To construct the shRNA interference lentivirus vector for Keap1 and evaluate the interference effects in human prostate cancer PC3 cell mediated by lentivirus. METHODS: Bioinformatics methods were used to design RNAi sequences for Keap1. A lentiviral vector was applied for construction of Keap1 shRNA vectors, which was expressed in E. coli packaged by 293T cells. Transfection efficiency and titer were measured by dilution method according to the green fluorescent protein (GFP) tracer. Real-time fluorescence quantitative method was applied to compare interfere effects of target sequences.RESULTS: Four Keap1 targeting sequences were constructed completely and the corresponding Keap1 shRNA lentiviral vectors were screened for efficiency. One shRNA lentiviral vector with best efficiency was screened by transient transfection (interference efficiency reached 80%) and the working condition was established as well.CONCLUSION: shRNA lentiviral vectors for Keap1 was constructed and screened successfully in PC3 cells, which effectively inhibit the expression of Keap1.

Key words: Keap1, shRNA, lentivirus, PC3 cell

CLC Number:

WANG Tan-heng, WU Xin-li, LIU Xiao-pin. Interference of shRNA on Keap1 in prostate cancer PC3 cell mediated by lentivirus[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2014, 19(8): 861-865.

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