Peptide-conjugated morpholino oligomers inhibit porcine reproductive and respiratory syndrome virus replication.
Porcine reproductive and respiratory syndrome (PRRS) has been devastating the global swine industry for more than a decade, and current strategies to control PRRS are inadequate. In this study we characterized the inhibition of PRRS virus (PRRSV) replication by antisense phosphorodiamidate morpholino oligomers (PMO). Of 12 peptide-conjugated PMO (PPMO), four were found to be highly effective at inhibiting PRRSV replication in cell culture in a dose-dependant and sequence-specific manner. PPMO 5UP2 and 5HP are complementary to sequence in the 5' end of the PRRSV genome, and 6P1 and 7P1 to sequence in the translation initiation regions of ORF6 and ORF7, respectively. Treatment of cells with 5UP2 or 5HP caused a 4.5log(10) reduction in PRRSV yield, compared to a control PPMO. Combination of 6P1 and 7P1 led to higher level reduction than 6P1 or 7P1 alone. 5UP2, 5HP, and a combination of 6P1 and 7P1 inhibited PRRSV replication in porcine alveolar macrophages and protected the cells from PRRSV-induced cytopathic effect. Northern blot and real-time RT-PCR results demonstrated that the effective PPMO led to a reduction of PRRSV RNA level. 5UP2 and 5HP inhibited virus replication of 10 other strains of PRRSV. Results from this study suggest potential applications of PPMO for PRRS control.
Patel
Deendayal
D
Molecular Virology Laboratory, VA-MD Regional College of Veterinary Medicine, University of Maryland, College Park, MD 20742, USA.
Opriessnig
Tanja
T
Stein
David A
DA
Halbur
Patrick G
PG
Meng
Xiang-Jin
XJ
Iversen
Patrick L
PL
Zhang
Yan-Jin
YJ
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D001483:Base Sequence; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002474:Cell-Free System; D019084:Fluorescent Antibody Technique, Indirect; D017930:Genes, Reporter; D006801:Humans; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000821:virology; D051379:Mice; D009025:Morpholines / Q000138:chemical synthesis / Q000494:pharmacology*; D060172:Morpholinos; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010455:Peptides / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D019316:Porcine respiratory and reproductive syndrome virus / Q000187:drug effects*; D014176:Protein Biosynthesis / Q000187:drug effects; D012367:RNA, Viral / Q000096:biosynthesis; D016415:Sequence Alignment; D013552:Swine; D014779:Virus Replication / Q000187:drug effects*
D000998:Antiviral Agents; D009025:Morpholines; D060172:Morpholinos; D016376:Oligonucleotides, Antisense; D010455:Peptides; D012367:RNA, Viral