Interference of EGFP RNA in Human NT-2/D1 Cell Lines Using Human U6 Promoter-based siRNA PCR Products

  • Kwak, Young-Don (Burnett College of Biomedical Sciences, Biomolecular Science Center, University of Central Florida) ;
  • Sugaya, Kiminobu (Burnett College of Biomedical Sciences, Biomolecular Science Center, University of Central Florida)
  • Published : 2006.06.30


RNA interference (RNAi), a process of sequence-specific gene suppression, has been known as a natural gene regulatory mechanism in a wide range of lower organisms. Recently, we have reported that a transfection of human U6 promoter (hU6) driven hairpin small-interference RNA (siRNA) plasmid specifically knocks down the target gene by post-transcriptional gene silencing in mammalian cells. Here we report that transfection of polymerase chain reaction (PCR) products, containing human U6 promoter with hairpin siRNA, knocks down the target gene expression in human teratocarcinoma NT-2/D1 cells. Moreover, we showed 3' end termination sequence, 5 Ts, is not critical elements for knocking down in PCR-based siRNA system. Therefore, the PCR-based siRNA system is a promising tool not only for the screening but also to temporally regulate gene expression in the human progenitor cells.


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