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

Viral challenge in mice: Mice, in groups of eleven, were anesthetized with dry ice and inoculated intracerebrally or intramuscularly (in masseter muscle) with the rAAV (2 by ITRs. An siRNA transcript was produced by the U6 promoter, and a cGFP transcript was produced by the CMV promoter. CMV IE was the enhancer for the U6 promoter and CMV promoter. × 10 12 vg/kg). After 48hr, the mice were infected with 10 LD 50 of RAbV CVS-11 for intracerebral challenge and 20 LD 50 of RAbV CVS-11 for intramuscular challenge at the same inoculation site as the rAAV inoculation site. The third group of mice was inoculated intracerebrally with the rAAV and then challenged with 20 LD 50 of RAbV CVS-11 intramuscularly (in masseter muscle) more than 48 hr after inoculation with rAAV (2 × 10 12 vg/kg). The forth group of mice was infected with RAbV intramuscularly and inoculated with rAAV-N796 or rAAV-Neg intracerebrally and intramuscularly after infection for 24 hr. At 6 days post infection with RAbV CVS-11, three mice from each group were anesthetized, and their brains were collected for assay of virus titer; the other mice were observed for 21 days for survival. All animal researches were conducted under the guidance of the CDC's Institutional Animal Care and Use Committee and in an Association for Assessment and Accreditation of Laboratory Animal Care International-accredited facility.