Construction of Repulsive Guidance Molecule A-Specific RNA Interference Recombinant Adenovirus

Abstract

This study aims to construct an RGMa RNAi plasmid in preparation for a subsequent research, and three shRNAs were designed for collection. The designed shRNAs were taken into the target plasmid of pegensil-1 to silence the mRNA of RGMa. Sequencing and enzyme identification were performed, and PC12 cells were transfected with a shRNA-containing plasmid. Then, the effect of the silencing of the gene was tested by RT-PCR using PC12 cells and to choose the best one. The results showed that the first one was choose for the subsequent research among these three designed shRNAs, which had the highest transfection efficiency and a concentration of 2.5 × 109 pfu/ml. The concentration of Adv(–) was 2.1 × 109 pfu/ml for the blank control. The effect of the constructed RGMa RNAi plasmid for the subsequent research is stable and effective.