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Comparison of transfection efficiency of rat dorsal root ganglion satellite glial cells with different transfection methods

         

摘要

In order to compare the transfection efficiency of lncRNA malat1 plasmid and lentivirus vectors in primary cultured rat DRG-derived satelite glial cells(SGCs).FISH was employed to detect the subcellular localization IncRNA-malat1l,qPCR was used to detect the expression of IncRNA-malat1 before and after transfection of the interference vector.It was found that malat1 was not silenced by IncRNA Smart silencers plasmid.qPCR results showed no significant difference in expression level of malat1 mRNA before and after transfection smart silencers plasmid of malat1 at 24 h,48h,3 d and7 d.The subcellular localization analysis found that malat1 was mainly located in the nucleus.It is speculated that it is difficult to achieve silencing of lncRNA located in the nucleus by applying interference technology of siRNA plasmid vector.qPCR results showed that the expression level of malat1 mRNA before and after malat1 transfection was significantly decreased at 24 h and 3 d compared with the normal group and the no-load group(P<0.05).In conclusion,it was suggested that the SGCs of DRG belonged to primary cells.

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