Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication

Tia S. Deas, Iwona Binduga-Gajewska, Mark Tilgner, Ping Ren, David A. Stein, Hong M. Moulton, Patrick L. Iversen, Elizabeth B. Kauffman, Laura D. Kramer, Pei-Yong Shi

Research output: Contribution to journalArticle

139 Citations (Scopus)

Abstract

RNA elements within flavivirus genomes are potential targets for antiviral therapy. A panel of phosphorodiamidate morpholino oligomers (PMOs), whose sequences are complementary to RNA elements located in the 5′- and 3′-termini of the West Nile (WN) virus genome, were designed to anneal to important cis-acting elements and potentially to inhibit WN infection. A novel Arg-rich peptide was conjugated to each PMO for efficient cellular delivery. These PMOs exhibited various degrees of antiviral activity upon incubation with a WN virus luciferase-replicon-containing cell line. Among them, PMOs targeting the 5′-terminal 20 nucleotides (5′End) or targeting the 3′-terminal element involved in a potential genome cyclizing interaction (3′CSI) exhibited the greatest potency. When cells infected with an epidemic strain of WN virus were treated with the 5′End or 3′CSI PMO, virus titers were reduced by approximately 5 to 6 logs at a 5 μM concentration without apparent cytotoxicity. The 3′CSI PMO also inhibited mosquito-borne flaviviruses other than WN virus, and the antiviral potency correlated with the conservation of the targeted 3′CSI sequences of specific viruses. Mode-of-action analyses showed that the 5′End and 3′CSI PMOs suppressed viral infection through two distinct mechanisms. The 5′End PMO inhibited viral translation, whereas the 3′CSI PMO did not significantly affect viral translation but suppressed RNA replication. The results suggest that antisense PMO-mediated blocking of cis-acting elements of flavivirus genomes can potentially be developed into an anti-flavivirus therapy. In addition, we report that although a full-length WN virus containing a luciferase reporter (engineered at the 3′ untranslated region of the genome) is not stable, an early passage of this reporting virus can be used to screen for inhibitors against any step of the virus life cycle.

Original languageEnglish (US)
Pages (from-to)4599-4609
Number of pages11
JournalJournal of Virology
Volume79
Issue number8
DOIs
StatePublished - Apr 2005
Externally publishedYes

Fingerprint

Flavivirus Infections
Flavivirus
Morpholinos
West Nile virus
Viral RNA
translation (genetics)
nucleotides
genome
infection
luciferase
viruses
Genome
Nucleotides
RNA
Flaviviridae
Antiviral Agents
replicon
therapeutics
3' untranslated regions
viral load

ASJC Scopus subject areas

  • Immunology

Cite this

Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication. / Deas, Tia S.; Binduga-Gajewska, Iwona; Tilgner, Mark; Ren, Ping; Stein, David A.; Moulton, Hong M.; Iversen, Patrick L.; Kauffman, Elizabeth B.; Kramer, Laura D.; Shi, Pei-Yong.

In: Journal of Virology, Vol. 79, No. 8, 04.2005, p. 4599-4609.

Research output: Contribution to journalArticle

Deas, TS, Binduga-Gajewska, I, Tilgner, M, Ren, P, Stein, DA, Moulton, HM, Iversen, PL, Kauffman, EB, Kramer, LD & Shi, P-Y 2005, 'Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication', Journal of Virology, vol. 79, no. 8, pp. 4599-4609. https://doi.org/10.1128/JVI.79.8.4599-4609.2005
Deas, Tia S. ; Binduga-Gajewska, Iwona ; Tilgner, Mark ; Ren, Ping ; Stein, David A. ; Moulton, Hong M. ; Iversen, Patrick L. ; Kauffman, Elizabeth B. ; Kramer, Laura D. ; Shi, Pei-Yong. / Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication. In: Journal of Virology. 2005 ; Vol. 79, No. 8. pp. 4599-4609.
@article{c3109541ad5f42dbaef6c59095d94395,
title = "Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication",
abstract = "RNA elements within flavivirus genomes are potential targets for antiviral therapy. A panel of phosphorodiamidate morpholino oligomers (PMOs), whose sequences are complementary to RNA elements located in the 5′- and 3′-termini of the West Nile (WN) virus genome, were designed to anneal to important cis-acting elements and potentially to inhibit WN infection. A novel Arg-rich peptide was conjugated to each PMO for efficient cellular delivery. These PMOs exhibited various degrees of antiviral activity upon incubation with a WN virus luciferase-replicon-containing cell line. Among them, PMOs targeting the 5′-terminal 20 nucleotides (5′End) or targeting the 3′-terminal element involved in a potential genome cyclizing interaction (3′CSI) exhibited the greatest potency. When cells infected with an epidemic strain of WN virus were treated with the 5′End or 3′CSI PMO, virus titers were reduced by approximately 5 to 6 logs at a 5 μM concentration without apparent cytotoxicity. The 3′CSI PMO also inhibited mosquito-borne flaviviruses other than WN virus, and the antiviral potency correlated with the conservation of the targeted 3′CSI sequences of specific viruses. Mode-of-action analyses showed that the 5′End and 3′CSI PMOs suppressed viral infection through two distinct mechanisms. The 5′End PMO inhibited viral translation, whereas the 3′CSI PMO did not significantly affect viral translation but suppressed RNA replication. The results suggest that antisense PMO-mediated blocking of cis-acting elements of flavivirus genomes can potentially be developed into an anti-flavivirus therapy. In addition, we report that although a full-length WN virus containing a luciferase reporter (engineered at the 3′ untranslated region of the genome) is not stable, an early passage of this reporting virus can be used to screen for inhibitors against any step of the virus life cycle.",
author = "Deas, {Tia S.} and Iwona Binduga-Gajewska and Mark Tilgner and Ping Ren and Stein, {David A.} and Moulton, {Hong M.} and Iversen, {Patrick L.} and Kauffman, {Elizabeth B.} and Kramer, {Laura D.} and Pei-Yong Shi",
year = "2005",
month = "4",
doi = "10.1128/JVI.79.8.4599-4609.2005",
language = "English (US)",
volume = "79",
pages = "4599--4609",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "8",

}

TY - JOUR

T1 - Inhibition of flavivirus infections by antisense oligomers specifically suppressing viral translation and RNA replication

AU - Deas, Tia S.

AU - Binduga-Gajewska, Iwona

AU - Tilgner, Mark

AU - Ren, Ping

AU - Stein, David A.

AU - Moulton, Hong M.

AU - Iversen, Patrick L.

AU - Kauffman, Elizabeth B.

AU - Kramer, Laura D.

AU - Shi, Pei-Yong

PY - 2005/4

Y1 - 2005/4

N2 - RNA elements within flavivirus genomes are potential targets for antiviral therapy. A panel of phosphorodiamidate morpholino oligomers (PMOs), whose sequences are complementary to RNA elements located in the 5′- and 3′-termini of the West Nile (WN) virus genome, were designed to anneal to important cis-acting elements and potentially to inhibit WN infection. A novel Arg-rich peptide was conjugated to each PMO for efficient cellular delivery. These PMOs exhibited various degrees of antiviral activity upon incubation with a WN virus luciferase-replicon-containing cell line. Among them, PMOs targeting the 5′-terminal 20 nucleotides (5′End) or targeting the 3′-terminal element involved in a potential genome cyclizing interaction (3′CSI) exhibited the greatest potency. When cells infected with an epidemic strain of WN virus were treated with the 5′End or 3′CSI PMO, virus titers were reduced by approximately 5 to 6 logs at a 5 μM concentration without apparent cytotoxicity. The 3′CSI PMO also inhibited mosquito-borne flaviviruses other than WN virus, and the antiviral potency correlated with the conservation of the targeted 3′CSI sequences of specific viruses. Mode-of-action analyses showed that the 5′End and 3′CSI PMOs suppressed viral infection through two distinct mechanisms. The 5′End PMO inhibited viral translation, whereas the 3′CSI PMO did not significantly affect viral translation but suppressed RNA replication. The results suggest that antisense PMO-mediated blocking of cis-acting elements of flavivirus genomes can potentially be developed into an anti-flavivirus therapy. In addition, we report that although a full-length WN virus containing a luciferase reporter (engineered at the 3′ untranslated region of the genome) is not stable, an early passage of this reporting virus can be used to screen for inhibitors against any step of the virus life cycle.

AB - RNA elements within flavivirus genomes are potential targets for antiviral therapy. A panel of phosphorodiamidate morpholino oligomers (PMOs), whose sequences are complementary to RNA elements located in the 5′- and 3′-termini of the West Nile (WN) virus genome, were designed to anneal to important cis-acting elements and potentially to inhibit WN infection. A novel Arg-rich peptide was conjugated to each PMO for efficient cellular delivery. These PMOs exhibited various degrees of antiviral activity upon incubation with a WN virus luciferase-replicon-containing cell line. Among them, PMOs targeting the 5′-terminal 20 nucleotides (5′End) or targeting the 3′-terminal element involved in a potential genome cyclizing interaction (3′CSI) exhibited the greatest potency. When cells infected with an epidemic strain of WN virus were treated with the 5′End or 3′CSI PMO, virus titers were reduced by approximately 5 to 6 logs at a 5 μM concentration without apparent cytotoxicity. The 3′CSI PMO also inhibited mosquito-borne flaviviruses other than WN virus, and the antiviral potency correlated with the conservation of the targeted 3′CSI sequences of specific viruses. Mode-of-action analyses showed that the 5′End and 3′CSI PMOs suppressed viral infection through two distinct mechanisms. The 5′End PMO inhibited viral translation, whereas the 3′CSI PMO did not significantly affect viral translation but suppressed RNA replication. The results suggest that antisense PMO-mediated blocking of cis-acting elements of flavivirus genomes can potentially be developed into an anti-flavivirus therapy. In addition, we report that although a full-length WN virus containing a luciferase reporter (engineered at the 3′ untranslated region of the genome) is not stable, an early passage of this reporting virus can be used to screen for inhibitors against any step of the virus life cycle.

UR - http://www.scopus.com/inward/record.url?scp=20144387945&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=20144387945&partnerID=8YFLogxK

U2 - 10.1128/JVI.79.8.4599-4609.2005

DO - 10.1128/JVI.79.8.4599-4609.2005

M3 - Article

VL - 79

SP - 4599

EP - 4609

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 8

ER -