Inhibition of rabies virus replication by multiple artificial microRNAs

Israsena, Nipan; Supavonwong, Pornpan; Ratanasetyuth, Nitipol; Khawplod, Pakamatz; Hemachudha, Thiravat

doi:10.1016/j.antiviral.2009.07.012

Cited by 56 publications

(51 citation statements)

References 39 publications

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“…For this reason we advocate the use of multimeric (polycistronic) amiRNA, where multiple sequences in the pathogen are targeted simultaneously, greatly reducing the opportunity for natural diversity or mutation in the pathogen to escape surveillance and degradation ( 30,59 ) . In 2009, a similar rationale was followed in the work of Israsena and colleagues ( 27 ) in which they designed precursor genes encoding three amiRNA against rabies virus and tested them in cell culture. Likewise, multiple siRNAs were introduced in the backbone of a multiplex miRNA, directed against HIV in cultured cells ( 56,60 ) .…”

Section: Introductionmentioning

confidence: 91%

“…Transgenic lines expressing these two amiRNAs speci fi cally conferred resistance to TYMV or TuMV ( 30 ) . Since then, a similar strategy has been successfully applied to other virus-plant systems using a number of different miRNA precursors as backbones for delivery of amiRNAs ( 27,(30)(31)(32)(33)(34)(35)(36) .…”

Section: Introductionmentioning

confidence: 98%

“…It is possible to alter the sequence of the 21-nt mature miRNAs within the natural miRNA precursor, without affecting miRNA biogenesis and maturation, as long as the secondary structure of the pre-miRNA is maintained. This opens up the prospect of creating amiRNAs with new targets by design ( 23,(27)(28)(29) .…”

Section: Introductionmentioning

confidence: 98%

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Designing Effective amiRNA and Multimeric amiRNA Against Plant Viruses

Fahim¹,

Larkin

2012

Methods in Molecular Biology

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RNA-mediated virus resistance is increasingly becoming a method of choice for antiviral defense in plants when effective natural resistance is unavailable. In this chapter we discuss the design principles of artificial micro RNA (amiRNA), in which a natural miRNA precursor gene is modified to target a different species of RNA, in particular viral RNA. In addition, we explore the advantages and effectiveness of multiple amiRNAs within one polycistronic amiRNA precursor against a virus, as illustrated with Wheat streak mosaic virus, WSMV. The judicious selection of amiRNAs, which are sequences of short length as compared to other related methodologies of RNA interference, greatly assists in avoiding unintended off-targets in the host plant. The viral sequences targeted can be genomic or replicative and should be derived from conserved regions of the published WSMV genome. In short, using published folding and miRNA selection rules and algorithms, candidate miRNA sequences are selected from conserved regions between a number of WSMV genomes, and are BLASTed against wheat TIGR ESTs. Five miRNAs are selected that are least likely to interfere with the expression of transcripts from the wheat host. Then, the natural miRNA in each of the five arms of the polycistronic rice miR395 is replaced in silico with the chosen artificial miRNAs. This artificial precursor is transformed into wheat behind a ubiquitin promoter, and its integration into transformed wheat plants is confirmed by PCR and Southern blot analysis. We have demonstrated the effectiveness of this methodology using an amiRNA precursor that we have termed Fanguard. The processing of amiRNAs in transgenic leaves is verified through splinted ligation assay, and the functionality of the transgene in preventing viral replication is verified by virus bioassay. Resistance is confirmed using mechanical virus inoculation over two subsequent generations. This example demonstrates the potential of polycistronic amiRNA to achieve stable immunity to economically important viruses.

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Section: Introductionmentioning

confidence: 91%

Section: Introductionmentioning

confidence: 98%

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Designing Effective amiRNA and Multimeric amiRNA Against Plant Viruses

Fahim¹,

Larkin

2012

Methods in Molecular Biology

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“…By this viewpoint, considering the conservation of the aminoacidic sequence of the target region and the codon usage constraint, few alternative version of a siRNA aiming to these targets may protect from the majority of influenza strains infecting a species. In this perspective, the option of simultaneously using more than one siRNA sequence is realistic option both in the use of synthetic siRNA as drugs or in making transgenic animals with shRNA vectors (Wang et al, 2006;Pu et al, 2011;Israsena et al, 2009). Further in vivo validation of this approach will be required, but the extent of the effects set good premises towards the development of transgenic animals and effective antiviral therapies.…”

Section: Discussionmentioning

confidence: 97%

Expression of a single siRNA against a conserved region of NP gene strongly inhibits in vitro replication of different Influenza A virus strains of avian and swine origin

et al. 2015

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“…Além disso, um trabalho publicado sobre RNAi contra a raiva utilizando plasmídeos com sequências de pré-microRNAs transfectado com lipofetamina dirigidos também ao RNAm da nucleoproteína em cultivo de células N2A demonstrou também eficiência deste sistema e deste alvo para o decréscimo do nível de RNA genômico viral em relação à amostra CVS de RABV (ISRASENA et al, 2009).…”

Section: Imunofluorescência Direta Das Amostras De Snc Dos Camundongounclassified

Inibição da replicação do virus da raiva in vitro e in vivo por meio de interferência por RNA

Ono¹

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Inhibition of rabies virus replication by multiple artificial microRNAs

Cited by 56 publications

References 39 publications

Designing Effective amiRNA and Multimeric amiRNA Against Plant Viruses

Designing Effective amiRNA and Multimeric amiRNA Against Plant Viruses

Expression of a single siRNA against a conserved region of NP gene strongly inhibits in vitro replication of different Influenza A virus strains of avian and swine origin

Inibição da replicação do virus da raiva in vitro e in vivo por meio de interferência por RNA

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