Specific inhibition of GFP expression by small hairpin RNA in colon cancer LoVo cell

Objective To construct a expression vector carrying small hairpin RNA (SHi pU6 GFP) for GFP gene.To study the specific inhibition of GFP gene using SHi pU6 GFP in LoVo cell.Methods The reporter gene plasmid pCX GFP (5 510 bp) and pU6 (3.3 kb) were used in this study.Plasmid pCX GFP,pU6 and SHi pU6 GFP carrying hairpin siRNA for GFP gene were transfected in K562 and LoVo cell by using Lipofectamine TM 2000 according to the manufacturer's instructions.Results The constructed SHi pU6 GFP carrying hairpin RNA for GFP gene and pU6 were proven to be the same as designed by restriction endonuclease analysis.The expression of GFP gene were specifically inhibited by SHi pU6 GFP in LoVo cell.Conclusion There is RNA interference phenomenon in LoVo cell.Short hairpin RNA (shRNA) could induce sequence specific GFP gene silencing in LoVo cell.The specific shRNAs could silence GFP gene.shRNA induced silencing may shed light on the application of RNA interference in colorectal cancer gene therapy.