Inhibition of foot-and-mouth disease virus replication by small interfering RNA.
Foot-and-mouth disease, caused by foot-and-mouth disease virus (FMDV), is one of the most dangerous diseases of cloven-hoofed animals and is a constant threat to the dairy and beef industries in the Middle East and other regions of the world, despite intensive vaccination programmes. In this work, the ability of specific small interfering (si)RNAs to inhibit virus replication in BHK-21 cells was examined. By using bioinformatic computer programs, all FMDV sequences in public-domain databases were analysed. The analysis revealed three regions of at least 22 bp with 100 % identity in all FMDV entries. From these sequences, three specific siRNA molecules were prepared and used to test the ability of siRNAs to inhibit virus replication. By using real-time quantitative PCR to measure the amount of viral RNA in infected cells, it was shown that virus replication was inhibited in cells that were transfected with siRNAs. When viral titres were examined, 100 % inhibition of growth could be demonstrated in cells transfected with a mixture of all three anti-FMDV siRNAs, compared with control cells transfected with anti-LacZ siRNA.
Virology Division, Kimron Veterinary Institute, PO Box 12, Beit-Dagan 50250, Israel.
Ronen
R
Kahana
Virology Division, Kimron Veterinary Institute, PO Box 12, Beit-Dagan 50250, Israel.
Larisa
L
Kuznetzova
Virology Division, Kimron Veterinary Institute, PO Box 12, Beit-Dagan 50250, Israel.
Arie
A
Rogel
Pathology Division, Kimron Veterinary Institute, PO Box 12, Beit-Dagan 50250, Israel.
Mordechai
M
Shemesh
Virology Division, Kimron Veterinary Institute, PO Box 12, Beit-Dagan 50250, Israel.
Dalia
D
Hai
Virology Division, Kimron Veterinary Institute, PO Box 12, Beit-Dagan 50250, Israel.
Hagai
H
Yadin
Virology Division, Kimron Veterinary Institute, PO Box 12, Beit-Dagan 50250, Israel.
Yehuda
Y
Stram
D000818:Animals; D002460:Cell Line; D029544:Foot-and-Mouth Disease Virus / Q000235:genetics / Q000502:physiology*; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000138:chemical synthesis / Q000494:pharmacology / Q000502:physiology*; D012367:RNA, Viral / Q000032:analysis / Q000037:antagonists & inhibitors; D013997:Time Factors; D014162:Transfection; D014779:Virus Replication
D034741:RNA, Small Interfering; D012367:RNA, Viral