Inhibition of foot-and-mouth disease virus replication in vitro and in vivo by small interfering RNA.
By using bioinformatics computer programs, all foot-and-mouth disease virus (FMDV) genome sequences in public-domain databases were analyzed. Based on the results of homology analysis, 2 specific small interfering RNA (siRNA) targeting homogenous 3D and 2B1 regions of 7 serotypes of FMDV were prepared and 2 siRNA-expression vectors, pSi-FMD2 and pSi-FMD3, were constructed. The siRNA-expressing vectors were used to test the ability of siRNAs to inhibit virus replication in baby hamster kidney (BHK-21) cells and suckling mice, a commonly used small animal model. The results demonstrated that transfection of BHK-21 cells with siRNA-expressing plasmids significantly weakened the cytopathic effect (CPE). Moreover, BHK-21 cells transiently transfected with short hairpin RNA (shRNA)-expressing plasmids were specifically resistant to the infection of the FMDV serotypes A, O, and Asia I and this the antiviral effects persisted for almost 48 hours. We measured the viral titers, the 50% tissue culture infective dose (TCID50) in cells transfected with anti-FMDV siRNAs was found to be lower than that of the control cells. Furthermore, subcutaneous injection of siRNA-expressing plasmids in the neck of the suckling mice made them less susceptible to infection with O, and Asia I serotypes of FMDV.
Pengyan
Wang
W
College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang 832003, PR China. wwwpy_322@163.com
Yan
Ren
R
Zhiru
Guo
G
Chuangfu
Chen
C
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D003588:Cytopathogenic Effect, Viral; D005536:Foot-and-Mouth Disease / Q000401:mortality / Q000628:therapy / Q000821:virology*; D029544:Foot-and-Mouth Disease Virus / Q000145:classification / Q000235:genetics / Q000502:physiology*; D051379:Mice; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics* / Q000494:pharmacology; D014779:Virus Replication
D034741:RNA, Small Interfering