shRNA driven by Pol II/T7 dual-promoter system effectively induce cell-specific RNA interference in mammalian cells.
Although Pol III promoters synthesize shRNA and elicit RNAi efficiently, however, a major limitation is that they are constitutively expressed in all cell types. To circumvent this problem, in the present study, we described a novel shRNA vector based on Pol II/T7 dual-promoter couple system: the transcription of shRNA under the control of T7 promoter is dependent on the corresponding T7 RNA polymerase driven by Pol II promoter. Our results strongly demonstrated that such a dual-promoter system can efficiently mediate shRNA expression and specifically reduce the exogenous reporter gene expression in mammalian cells. Furthermore, when hepatoma specific AFP promoter was introduced to control T7 RNA polymerase expression, the RNA interference was permitted only in AFP-producing cells. To our knowledge, this is the first evidence that shRNA can be expressed in a cell-specific manner from Pol II/T7 dual-promoter system in mammalian cells.
Peng
Ying
Y
School of Life Science, Wenzhou Medical College, Wenzhou, Zhejiang 325025, People's Republic of China. ying7754@yahoo.com.cn
Lu
Jian-Xin
JX
Shen
Xin-Feng
XF
D004257:DNA Polymerase II / Q000235:genetics*; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D011401:Promoter Regions, Genetic / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*
D034741:RNA, Small Interfering; D004257:DNA Polymerase II