Adeno-Associated Viruses Serotype 2-Mediated RNA Interference Efficiently Inhibits Rabies Virus Replication In Vitro and In Vivo

WU, Hong-Xia; WANG, Hua-Lei; GUO, Xiao-Feng; YANG, Yu-Jiao; MA, Jin-Zhu; WANG, Tie-Cheng; GAO, Yu-Wei; ZHAO, Yong-Kun; YANG, Song-Tao; XIA, Xian-Zhu

To evaluate the antiviral effects of rAAV-N796 targeting the conserved sequences of the CVS11-N gene, the rAAV-N796 and a negative control (rAAV-Neg) were respectively inoculated into NA cells with 10 vg/cell. The transduction efficiency of rAAV in NA cells was more than 90% (result not shown). After transduction for 24 hr, the cells were infected with 0.01 MOI of the RAbV CVS-11 virus. At the different time points, the cells supernatants were collected for analysis of RAbV titer. As shown in Fig. 2A , the virus titer in the rAAV-N796 group was significantly lower (P<0.05) than that in the rAAV-Neg group at each time point. At 72 hr post infection, the virus titer (TCID 50 /ml of 3.5 ± 0.65) in the rAAV-N796 pretreatment group was reduced by nearly 10 4 times (P<0.01) compared with the rAAV-Neg group (TCID 50 /ml of 7.8 ± 0.82). When RAbV infection was before rAAV transduction, no significant difference was observed at 24 and 48 hr post infection. Only at 72 and 96 hr post infection (Fig. 2b) was the RABV titer reduced significantly (P<0.05). These results indicated that the inoculation of rAAV-N796 before RAbV infection could inhibit the virus replication well, but that inoculation of rAAV-N796 after RAbV infection inhibited virus replication only at later time points.