Water soluble lipopolymer can deliver siRNA targeting N-methy1-D-aspartate receptor 1 in vitro efficiently

Abstract

Objective To examine the potential application of a non-viral genecarrier,water soluble lipopolymer(WSLP)for delivering siRNA targeting N-methyl-D-aspartate receptor 1(NMDAR1)in vitro.Methods WSLP was complexed with siRNA designed to inhibit NR1 expression.Following serum stability and cytotoxicity observation,WSLP/siRNA(scrambled siRNA as a control)complexes were transfected in PC12 cells and then siRNA delivery efficiency of the complexes was evaluated by gene expression level assay using reverse transcriptive polymerase chain reaction(RT-PCR)and western-blot technique.Results WSLP protected siRNAs from enzymatic degradation in serum conditioned media,and the complexes of WSLP/siRNA had little cytotoxicity to cultured PC12 cell.NMDAR1 expression of PC12 cell was efficiently inhibited by WSLP/siRNA complexes,while complexes of WSLP with scrambled siRNA(0.64 ±0.13,4.32 ±1.09)did not show this inhibitory effect compared to unmodified siRNA (0.69±0.18,4.36± 1.02)neither by transcriptional level nor protein level.WSLP/siRNA complexes reduced NR 1 transcriptional level by 50％(0.35±0.21)and protein level by 55％(1.96±0.48)when compared to unmodified siRNA.Conclusion Our data suggest that Water soluble lipopolymer can deliver siRNA targeting NMDA receptor 1 in vitro efficiently and safely. Key words: NMDA receptor 1 ; Water soluble lipopolymer; siRNA