Shan Z.X.,Guangdong Academy of Medical science |
Shan Z.X.,Guangdong Provincial Cardiovascular Diseases Institute |
Shan Z.X.,Shantou University |
Lin Q.X.,Guangdong Academy of Medical science |
And 21 more authors.
Molecular Biology Reports | Year: 2010
Abstract MicroRNA-based short hairpin RNAs (shRNAs) are natural inducers of RNA interference and have been increasingly used in shRNA expression strategies. In thepresent study, we compared the efficiencies of exogenous green fluorescence protein (GFP) and endogenous glyceraldehyde- 3-phosphate dehydrogenase (GAPDH) knockdown and red fluorescent protein (RFP) indicator expression mediated by three differently designed plasmids. RFP was introduced either at the 50 end, at the 30 end of the human mir155- based target gene (TG) (e.g., GFP or GAPDH) shRNA expression cassette (EC), or at the 30 end of the chimeric intron-containing TG shRNA EC. Comparisons with the control vector showed an obvious reduction of GFP or GAPDH expression with the various shRNA expression plasmids (P\0.05).WhenRFPwas located at the 50 end or at the 30 end of the TG shRNA EC, RFP expression was low; whereas when RFP was connected with the chimeric introncontaining TG shRNA EC, RFP expression was high. Taken together, this study demonstrated an efficient plasmid design for both TG silencing induced by microRNA-based shRNA and indicator gene expression in vitro. © Springer Science+Business Media B.V. 2009.