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

The antiviral activity of rAAV-N796 in NA cells depended on the titer of rAAV-N796. In the in vivo experiment, rAAV-N796 pretreatment did not achieve complete protection, which may be associated with the inoculation dose of rAAV-N796. The biggest rAAV titer achieved in our lab through co-transfection in HEK293 cells is 4 × 10 11 vg/ml. So, a better method to produce rAAV particles needs to be established. The production of rAAV vectors in insect cells via the use of recombinant baculovirus technology has proven to be an efficient and scalable means of rAAV production. The yield of genome-containing particles produced per Sf9 cell approached 5 × 10 4 ; thus, 1,000 ml of cultured Sf9 cells produced the equivalent of between 500 to 1,000 × 175 cm 2 flasks of 293 cells. This robust system provides a simple, cost-effective method for AAV vector production [18] . However, to achieve complete protection, other siRNAs targeting the RAbV gene, the rAAV titer and the timing of therapeutic administration should be assessed to generate an optimal treatment strategy [20] .