Sequence-optimized and targeted double-stranded RNA as a therapeutic antiviral treatment against infectious myonecrosis virus in Litopenaeus vannamei

Loy, John Dustin; Loy, Duan S.; Mogler, Mark; Janke, Bruce H.; Kamrud, Kurt I.; Harris, D. L.

doi:10.3354/dao02600

Cited by 13 publications

(7 citation statements)

References 25 publications

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“…It is thus not possible to predict, without undertaking longer-term challenge trials, to what level and over what time period shrimp would remain protected against developing acute IMNV infection and associated disease. Nonetheless, the promising results reported here and elsewhere (Tirasophon et al 2007, Loy et al 2012, 2013 clearly justify follow-up studies to examine whether RNAi approaches can be applied in breeding programs, hatcheries, or farms to effectively suppress IMNV infection over the life of L. vannamei, thus minimizing risks of IMN production losses occurring during culture.…”

Section: Discussionmentioning

confidence: 72%

“…The RNAi mechanism exists in shrimp, where it can be mobilized as a component of the antiviral defense response. Synthetic dsRNAs targeted to various viral and host gene sequences have thus been investigated for their ability to activate prophylactic RNAi responses against viruses including the Taura syndrome virus (TSV, Robalino et al 2004), white spot syndrome virus (WSSV, Robalino et al 2005), yellow head virus (YHV, Yodmuang et al 2006, Tirasophon et al 2007, and IMNV (Loy et al 2013). To activate the RNAi mechanism in shrimp effectively, long dsRNA molecules have been delivered by injection.…”

Section: Introductionmentioning

confidence: 99%

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RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei

Feijó

Maggioni²,

Martins³

et al. 2015

Dis. Aquat. Org.

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Disease in Pacific white shrimp Litopenaeus vannamei caused by the infectious myonecrosis virus (IMNV) causes significant socioeconomic impacts in infection-prone shrimp aquaculture regions. The use of synthetic dsRNA to activate an RNA interference (RNAi) response is being explored as a means of disease prophylaxis in farmed shrimp. Here, survival was tracked in L. vannamei injected with long synthetic dsRNAs targeted to IMNV open reading frame (ORF) 1a, ORF1b, and ORF2 genome regions prior to injection challenge with IMNV, and real-time RT-PCR was used to track the progress of IMNV infection and mRNA expression levels of the host genes sid1, dicer2, and argonaute2. Injection of dsRNAs targeting the ORF1a and ORF1b genes but not the ORF2 gene strongly inhibited IMNV replication over a 3 wk period following IMNV challenge, and resulted in 90 and 83% shrimp survival, respectively. Host gene mRNA expression data indicated that the Sid1 protein, which forms a transmembrane channel involved in cellular import/export of dsRNA, increased in abundance most significantly in shrimp groups that were most highly protected by virus-specific dsRNA injection. Subclinical IMNV infections present in the experimental L. vannamei used increased markedly in the 2 d between injection of any of the 4 virus-specific or non-specific dsRNAs tested and IMNV challenge. While handling and injection stress are implicated in increasing IMNV replication levels, the underlying molecular factors that may have been involved remain to be elucidated.

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Section: Discussionmentioning

confidence: 72%

Section: Introductionmentioning

confidence: 99%

RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei

Feijó

Maggioni²,

Martins³

et al. 2015

Dis. Aquat. Org.

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“…The injection dose depends on the size and the RNAi sensitivity of the target species and tissues and usually ranges from about 5 to 1000 ng per injection. Haemolymph injection (Yodmuang et al 2006;Ongvarrasopone et al 2011) and intramuscular injection (Kim et al 2007;Loy et al 2013;Hou et al 2014) are used to deliver dsRNA into bigger shrimp species (Table 2).…”

Section: Molecular Mechanismmentioning

confidence: 99%

RNA interference in shrimp and potential applications in aquaculture

Nguyen

Christiaens

Bossier

et al. 2016

Reviews in Aquaculture

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As the RNA interference mechanism was discovered, researchers have made huge strides in unravelling biological mechanisms at the gene level in a wide range of organisms. RNAi is a simple and rapid method that allows silencing gene expression at the post-transcriptional level and is triggered by the introduction of double-stranded RNA into the cell, leading to the degradation of the corresponding mRNA. In this review, we will provide an overview of successful RNAi experiments to date in Crustacea, and shrimp in particular, with special focus on various potential applications of RNA interference in research and aquaculture. An important aspect of RNAi in any species is the delivery of the double-stranded RNA and the barriers that have to be overcome to get the dsRNA into the target cells. Different options, experimental procedures, success rates and challenges in using RNAi are discussed in this review

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“…As the major challenge regarding the use of RNAi as antiviral treatment in mammals has been the delivery method 7 , 8 , this has been more promising in invertebrates due to the systemic nature of RNAi in many of these species. Trials in farmed shrimps have shown that muscular injection of dsRNA can be used therapeutically against yellow head virus and infectious myonecrosis virus, at least to decrease the viral load and increase survival rate of infected animals 9 – 11 . In insects such as the honeybee, oral treatment with dsRNA simultaneously with virus inoculation has been shown to induce resistance to the israeli acute paralysis virus (IAPV) 12 , and even large scale application of dsRNA against this virus has been attempted 13 .…”

Section: Introductionmentioning

confidence: 99%

RNAi-mediated treatment of two vertically transmitted rhabdovirus infecting the salmon louse (Lepeophtheirus salmonis)

Øvergård

Hamre

Kongshaug

et al. 2017

Sci Rep

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Rhabdoviruses are a family of enveloped negative-sense single-stranded RNA viruses infecting a variety of hosts. Recently, two vertically transmitted salmon louse (Lepeophtheirus salmonis) rhabdoviruses (LsRV) have been identified. The prevalence of these viruses was measured along the Norwegian coast and found to be close to 100%, and with the present lack of suitable cell lines to propagate these viruses, it is challenging to obtain material to study their host impact and infection routes. Thus, virus free lice strains were established from virus infected lice carrying one or both LsRVs by treating them with N protein dsRNA twice during development. The viral replication of the N protein was specifically down-regulated following introduction of virus-specific dsRNA, and virus-free lice strains were maintained for several generations. A preliminary study on infection routes suggested that the LsRV-No9 is maternally transmitted, and that the virus transmits from males to females horizontally. The ability to produce virus free strains allows for further studies on transmission modes and how these viruses influences on the L.salmonis interaction with its salmonid host. Moreover, this study provides a general fundament for future studies on how vertically transmitted rhabdoviruses influence the biology of their arthropod hosts.

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Sequence-optimized and targeted double-stranded RNA as a therapeutic antiviral treatment against infectious myonecrosis virus in Litopenaeus vannamei

Cited by 13 publications

References 25 publications

RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei

RNAi-based inhibition of infectious myonecrosis virus replication in Pacific white shrimp Litopenaeus vannamei

RNA interference in shrimp and potential applications in aquaculture

RNAi-mediated treatment of two vertically transmitted rhabdovirus infecting the salmon louse (Lepeophtheirus salmonis)

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