Antisense morpholino-oligomers directed against the 5' end of the genome inhibit coronavirus proliferation and growth.
Conjugation of a peptide related to the human immunodeficiency virus type 1 Tat represents a novel method for delivery of antisense morpholino-oligomers. Conjugated and unconjugated oligomers were tested to determine sequence-specific antiviral efficacy against a member of the Coronaviridae, Mouse hepatitis virus (MHV). Specific antisense activity designed to block translation of the viral replicase polyprotein was first confirmed by reduction of luciferase expression from a target sequence-containing reporter construct in both cell-free and transfected cell culture assays. Peptide-conjugated morpholino-oligomers exhibited low toxicity in DBT astrocytoma cells used for culturing MHV. Oligomer administered at micromolar concentrations was delivered to >80% of cells and inhibited virus titers 10- to 100-fold in a sequence-specific and dose-responsive manner. In addition, targeted viral protein synthesis, plaque diameter, and cytopathic effect were significantly reduced. Inhibition of virus infectivity by peptide-conjugated morpholino was comparable to the antiviral activity of the aminoglycoside hygromycin B used at a concentration fivefold higher than the oligomer. These results suggest that this composition of antisense compound has therapeutic potential for control of coronavirus infection.
Neuman
Benjamin W
BW
The Scripps Research Institute, Department of Neuropharmacology, Division of Virology, 10550 North Torrey Pines Rd., La Jolla, CA 92037, USA. bneuman@scripps.edu
Stein
David A
DA
Kroeker
Andrew D
AD
Paulino
Amy D
AD
Moulton
Hong M
HM
Iversen
Patrick L
PL
Buchmeier
Michael J
MJ
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D016679:Genome, Viral; D051379:Mice; D008969:Molecular Sequence Data; D009025:Morpholines / Q000494:pharmacology*; D060172:Morpholinos; D006517:Murine hepatitis virus / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D014764:Viral Proteins / Q000096:biosynthesis
D009025:Morpholines; D060172:Morpholinos; D016376:Oligonucleotides, Antisense; D014764:Viral Proteins