2005•Junshi Yixue Kexueyuan yuankanRequires access
RNA interference suppresses expression of extraneous {\sl PC--1} gene in NIH3T3 cells
Liquan Zhou, Hui Zhang, Xuesong Gao, Wang Jian, Ruixia Liang, Hong Baofa, Jianguang Zhou
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Abstract
Recombinant plasmids expressing short hairpin RNA targeting PC-1 mRNA were constructed using DNA recombinant technology and transfected into NIH3T3 cells with PC-1-EGFP plasmid respectively. The most effective RNA interferencing construct was selected from 5 candidates by observing the expression of EGFP using fluorescence microscope 48 hours after transfection. The effect of RNA interference was further confirmed by reverse transcriptase polymerase chain reaction and Western-blotting analysis. PC-1 gene expression was surpressed by the most effective RNAi target. This strategy is adapted to select the most effective target from many candidates. The work provides basis for researching the impact of suppression of the endogenous PC-1 gene expression on the biological behavior of prostate cancer cell.