用狂犬病毒核蛋白基因小干扰RNA库抑制狂犬病毒复制和感染的研究

Inhibition of Rabies Virus Replication and Infection by Rabies Virus Nucleoprotein Gene siRNA Cocktail

采用RNaseⅢ消化长片段双链RNA的方法,制备了狂犬病毒N基因、P基因和G基因小干扰RNA库(siRNACocktail)。将siRNA转染BSR和MNA细胞单层后,采用直接免疫荧光法观察发现,N基因siRNACocktail能够对RV的复制和感染产生明显和稳定的抑制作用,并且通过RT-PCR在转录水平探测到mRNA产量的降低;而P基因或G基因siRNACocktail对RV的复制和感染不产生或仅产生弱的抑制作用。这一结果为RNA干扰在RV研究中靶基因的选择及进一步的应用提供了依据。

Small interference RNA mixtures （siRNA Cocktails ） of Rabies virus （RV） N gene, P gene and G gene were prepared by digesting long double-stranded RNA （dsRNA） with RNase Ⅲ. These siRNA cocktails were transfecteing into BSR or MNA cell monolayers and the direct immunofluorescence assay （FA） was used to measure the replication and infection of RV on the cell monolayers. We found that the siRNA Cocktails of N gene can obviously and stably inhibit the replication and infection of RV. A descent of mRNA of N gene was also detected by RT-PCR. Under the same situation, the siRNA Cocktails of P gene or G gene showed no or weak inhibition to the replication and infection of RV. All of these results provide a basis for selecting target gene of RV in RNA itreatment and its further application.