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Construction of Lentivirus-mediated HPV16-E7 Gene RNAi Cell Model |
ABUDUKADEER Abida1, HASIMU Axiangu2, AINIWAER Saimire2, ABULIZI Abudula3, SHENG Lei3 |
1. The First Afiliated Hospital, Xinjiang Medical University, Urumqi 830011, China;2. College of Basic Medicine, Xinjiang Medical University, Urumqi 830011, China;3. Xinjiang Key Laboratory of Molecular Biology and Endemic Diseases, Xinjiang Medical University, Urumqi 830011, China |
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Abstract: The HPV16-E7 ggene RNAi cell model was constructed. A recombinant lentiviral siRNA expression vector was constructed using a HPV16-E7 oncogene specific siRNA fragment, and an RNAi cell model stably expressing the HPV16-E7-siRNA was established by transfection of 293FT virus-packiging cells with the viral vector followed by infection of HPV16-positive SiHa carcinoma cells with recombinant virus, antibiotica selection and molecular biological characterization. RNAi cell model was successfully established, stably transcribing the HPV16-E7-siRNA fragment, and the inhitory effect on target gene (E7) was also characterized by western blotting and RT-PCR. The RNAi cell model expressing the HPV16-E7 gene-specific siRNA fragment using lentiviral vector was stable, effective and feasible, which lays the foundation for the role study and carcinogenic mechanism of E7 protein.
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Received: 28 June 2013
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