Construction and identification of shRNA expressing vectors targeting RhoA gene

AIM:To construct and identify shRNA interference vector targeting RhoA gene.METHODS:Two pairs of single stranded oligonueleotides encoding RhoA siRNA sequence were designed and synthesized.After annealing,its were inserted into vector pGPU6/GFP/Neo,constructed recombinant vectors and then were identified by restrictive digestion and DNA sequencing.LoVo cells were transfected with the recombinant DNA samples by the liposome complex method,and then fluo-rescence photographs were taken.RESULTS:Enzyme digestion and DNA sequencing showed that the oligonucleotide fragmentswere correctly inserted into pGPU6/GFP/Neo vector.CONCLUSION:The RhoA gene-targeted siRNA and its vector are successfully constructed.