Inhibition of norovirus replication by morpholino oligomers targeting the 5′-end of the genome

Bok, Karin; Cavanaugh, Victoria J.; Matson, David O.; González-Molleda, Lorenzo; Chang, Kyeong‐Ok; Zintz, Carmelann; Smith, Alvin W.; Iversen, Patrick L.; Green, Kim Y.; Campbell, Ann E.

doi:10.1016/j.virol.2008.08.007

Cited by 22 publications

(23 citation statements)

References 35 publications

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Section: Introductionmentioning

confidence: 99%

“…The identification of new antiviral approaches has been limited due to the inability of human noroviruses to complete a productive infection in cell culture 3 . The recent isolation of a murine norovirus (MNV), closely related to human norovirus 4 but which can be propagated in cells 5 has opened new avenues for the investigation of these pathogens 6,7 .MNV replication results in the synthesis of new positive sense genomic and subgenomic RNA molecules, the latter of which corresponds to the last third of the viral genome (Figure 1). MNV contains four different open reading frames (ORFs), of which ORF1 occupies most of the genome and encodes seven non-structural proteins (NS1-7) released from a polyprotein precursor.…”

mentioning

confidence: 99%

“…The recent isolation of a murine norovirus (MNV), closely related to human norovirus 4 but which can be propagated in cells 5 has opened new avenues for the investigation of these pathogens 6,7 .…”

mentioning

confidence: 99%

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Reverse Genetics Mediated Recovery of Infectious Murine Norovirus

Arias

Ureña

Thorne

et al. 2012

JoVE

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Human noroviruses are responsible for most cases of human gastroenteritis (GE) worldwide and are recurrent problem in environments where close person-to-person contact cannot be avoided 1,2 . During the last few years an increase in the incidence of outbreaks in hospitals has been reported, causing significant disruptions to their operational capacity as well as large economic losses. The identification of new antiviral approaches has been limited due to the inability of human noroviruses to complete a productive infection in cell culture 3 . The recent isolation of a murine norovirus (MNV), closely related to human norovirus 4 but which can be propagated in cells 5 has opened new avenues for the investigation of these pathogens 6,7 .MNV replication results in the synthesis of new positive sense genomic and subgenomic RNA molecules, the latter of which corresponds to the last third of the viral genome (Figure 1). MNV contains four different open reading frames (ORFs), of which ORF1 occupies most of the genome and encodes seven non-structural proteins (NS1-7) released from a polyprotein precursor. ORF2 and ORF3 are contained within the subgenomic RNA region and encode the capsid proteins (VP1 and VP2, respectively) ( Figure 1). Recently, we have identified that additional ORF4 overlapping ORF2 but in a different reading frame is functional and encodes for a mitochondrial localised virulence factor (VF1) 8 .Replication for positive sense RNA viruses, including noroviruses, takes place in the cytoplasm resulting in the synthesis of new uncapped RNA genomes. To promote viral translation, viruses exploit different strategies aimed at recruiting the cellular protein synthesis machinery [9][10][11] . Interestingly, norovirus translation is driven by the multifunctional viral protein-primer VPg covalently linked to the 5' end of both genomic and subgenomic RNAs [12][13][14] . This sophisticated mechanism of translation is likely to be a major factor in the limited efficiency of viral recovery by conventional reverse genetics approaches.Here we report two different strategies based on the generation of murine norovirus-1 (referred to as MNV herewith) transcripts capped at the 5' end. One of the methods involves both in vitro synthesis and capping of viral RNA, whereas the second approach entails the transcription of MNV cDNA in cells expressing T7 RNA polymerase. The availability of these reverse genetics systems for the study of MNV and a small animal model has provided an unprecedented ability to dissect the role of viral sequences in replication and pathogenesis [15][16][17] . Video LinkThe video component of this article can be found at https://www.jove.com/video/4145/ Protocol RNA Transcription and Capping for The Recovery of Infectious MNVThis protocol is designed to allow the efficient recovery of infectious MNV from cDNA via in vitro transcription and its subsequent in vitro capping (section 1.1). The resulting capped transcripts are then transfected into cells to recover infectious MNV (sections 1.2 and ...

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Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Reverse Genetics Mediated Recovery of Infectious Murine Norovirus

Arias

Ureña

Thorne

et al. 2012

JoVE

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“…A panel of peptide-conjugated phosphorodiamidiate morpholino oligomers (PPMOs) specific for the 5′-UTR of MNV proved to be effective in inhibiting its replication in vitro (Bok et al, 2008). Also, a consensus PPMO (designated Noro 1.1), designed to target the corresponding region of several diverse human norovirus genotypes, inhibited Norwalk virus protein expression in replicon-bearing cells (Bok et al, 2008). Moreover, PMOs targeting the 5′-UTR of the FCV genome were used successfully in three clinical trials during FCV outbreaks in kittens .…”

Section: Wwwintechopencommentioning

confidence: 99%

Targeting Norovirus: Strategies for the Discovery of New Antiviral Drugs

Rocha‐Pereira¹,

Nascimento²

2012

Antiviral Drugs - Aspects of Clinical Use and Recent Advances

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“…In the Caliciviridae, peptide-linked phosphorodiamidiate morpholino oligomers (PPMOs) targeting a sequence near the 5 -UTR of the murine norovirus genome were effective in reducing translation of viral proteins in vitro (Bok et al, 2008). Our preliminary investigations have made use of the PPMO-approach to inhibit multiplication of the feline calicivirus in vitro.…”

Section: The Antisense Approachmentioning

confidence: 99%

Antiviral strategies to control calicivirus infections

et al. 2010

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Caliciviridae are human or non-human pathogenic viruses with a high diversity. Some members of the Caliciviridae, i.e. human pathogenic norovirus or rabbit hemorrhagic disease virus (RHDV), are worldwide emerging pathogens. The norovirus is the major cause of viral gastroenteritis worldwide, accounting for about 85% of the outbreaks in Europe between 1995 and 2000. In the United States, 25 million cases of infection are reported each year. Since its emergence in 1984 as an agent of fatal hemorrhagic diseases in rabbits, RHDV has killed millions of rabbits and has been dispersed to all of the inhabitable continents. In view of their successful and apparently increasing emergence, the development of antiviral strategies to control infections due to these viral pathogens has now become an important issue in medicine and veterinary medicine. Antiviral strategies have to be based on an understanding of the epidemiology, transmission, clinical symptoms, viral replication and immunity to infection resulting from infection by these viruses. Here, we provide an overview of the mechanisms underlying calicivirus infection, focusing on the molecular aspects of replication in the host cell. Recent experimental data generated through an international collaboration on structural biology, virology and drug design within the European consortium VIZIER is also presented. Based on this analysis, we propose antiviral strategies that may significantly impact on the epidemiological characteristics of these highly successful viral pathogens.

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Inhibition of norovirus replication by morpholino oligomers targeting the 5′-end of the genome

Cited by 22 publications

References 35 publications

Reverse Genetics Mediated Recovery of Infectious Murine Norovirus

Reverse Genetics Mediated Recovery of Infectious Murine Norovirus

Targeting Norovirus: Strategies for the Discovery of New Antiviral Drugs

Antiviral strategies to control calicivirus infections

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