Lentivirus-Mediated Expression Of Type I Collagen Shrna In Rat Mesangial Cells
Keywords
Collagen Type I, Rna Interference, Lentiviral Infection
Abstract
[Abstract Objective? To explore the expression of lentivirus-mediated collagen type I (ColI) short hairpin RNA (shRNA) in rat mesangial cells. method? ? Synthesis using three plasmids? How to synthesize lentiviral expression vector from expression plasmid, using infection enhancer (control group), empty lentiviral vector (pSC-GFP group) and lentiviral expression vector (pSC-GFP/Col¢ñ group) respectively? Infection of rat mesangial cells. Flow cytometry was used to detect the transfection efficiency of rat mesangial cells in each group, and RT-PCR and Western blotting methods were used to detect Col¢ñ? ? Expression of mRNA and Col¢ñ protein. result? ? Both pSC-GFP and pSC-GFP/Col¢ñ can efficiently infect rat mesangial cells, with the infection efficiency of pSC-GFP being 72.42%? The infection efficiency of pSC-GFP/Col¢ñ was as high as 75.42%. RT-PCR detection showed that the expression of Col¢ñmRNA in cells in the pSC-GFP/Col¢ñ group was significantly lower than that in the control group and pSC-GFP group? (P<0.05), the inhibition efficiency was 32.64%¡À0.12%, and there was no statistically significant difference in the expression of Col¢ñmRNA between the two groups. Westernblotting test showed that compared with the control group,? Compared with the pSC-GFP group, the protein expression of pSC-GFP/Col¢ñ group was significantly reduced (P<0.05), while the difference in Col¢ñ protein expression between the control group and the pSC-GFP group was not statistically significant. Knot? Argument? ? Lentiviral vectors can stably and efficiently transfect Col¢ñ shRNA into mesangial cells and significantly inhibit the expression of Col¢ñ, providing a theoretical basis for gene therapy of renal fibrosis. For further information of this article and research, feel free to contact our team for asssitance. Original research was done by Shan Yi, Zhou Chunhua
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