A novel and inexpensive application of RNAi technology to protect shrimp from viral disease

Saksmerprome, Vanvimon; Charoonnart, Patai; Gangnonngiw, Warachin; Withyachumnarnkul, Boonsirm

doi:10.1016/j.jviromet.2009.08.010

Cited by 60 publications

(50 citation statements)

References 17 publications

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“…It is not known to what extents the bacterial fixation and/or feed formulation processes, permeability of the shrimp gutwall barrier, dsRNA degradation by gut digestive nucleases and other possible biological obstacles contributed to the failure of the oral delivery approach to generate RNAi responses. Discounting this failure, however, it is important that such methods continue to be refined and explored, as approaches examined thus far have only offered non-specific or no or low-level specific protection against viral pathogens (Lauth et al 2010) even though many studies have now proven beyond doubt that virus-specific dsRNAs can be potent antiviral prophylactics when injected into shrimp (Robalino et al 2005, Tirasophon et al 2005, Yodmuang et al 2006, Saksmerprome et al 2009). Moreover, there remains great potential for using dsRNA muscle injection as a means, for example, of lessening loads or clearing viral infections from valuable broodstock prior to spawning to break vertical transmission cycles, particularly in regions where viruses such as WSSV continue to be a major impediment to shrimp aquaculture, or of interfering with biological pathways to promote ovary development and spawning in the absence of eye ablation that inevitably results in shrimp death from reproductive exhaustion.…”

Section: Discussionmentioning

confidence: 99%

“…In comparison, a study of RNAi responses to YHV infection in Penaeus monodon found that pre-injection of a dsRNA targeted to the 3CL-protease motif in the ORF1a gene afforded 95% protection against mortalities over a 10 d period post-challenge (Yodmuang et al 2006). More recently, a study of RNAi responses to YHV replication in L. vannamei found that injection of a dsRNA targeted to the RNA polymerase motif in the ORF1b gene afforded 87% protection against mortalities over a 14 d period post-challenge (Saksmerprome et al 2009). In the bioassays reported here, muscle injection of a pool of 5 GAV dsRNAs interfered strongly with GAV replication in juvenile P. monodon injected with a lethal dose of GAV that was selected to reliably generatẽ 50% accumulated mortality by 8 d p.i.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Penaeus monodon is protected against gill- associated virus by muscle injection but not oral delivery of bacterially expressed dsRNAs

Sellars

Rao²,

Arnold³

et al. 2011

Dis. Aquat. Org.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Penaeus monodon is protected against gill- associated virus by muscle injection but not oral delivery of bacterially expressed dsRNAs

Sellars

Rao²,

Arnold³

et al. 2011

Dis. Aquat. Org.

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“…In the past decade, RNAi-based technology has gained attention for its potential to control shrimp viral diseases that have caused significant losses in aquaculture. Inhibition of shrimp viruses by viral-specific dsRNA were demonstrated in insect cell lines (He et al, 2009;Theerawanitchpan et al, 2012) and in penaeid shrimp (Robalino et al, 2005;Yodmuang et al, 2006;Ongvarrasopone et al, 2008;Saksmerprome et al, 2009Saksmerprome et al, , 2013Thammasorn et al, 2013). Given that sequences of viral genes of interest are available, dsRNA targeting those genes can be synthesized by in vitro transcription and in vivo bacterial system.…”

Section: Introductionmentioning

confidence: 99%

Use of microalgae Chlamydomonas reinhardtii for production of double-stranded RNA against shrimp virus

Somchai

Jitrakorn

Thitamadee

et al. 2016

Aquaculture Reports

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a b s t r a c t RNA interference has been proposed to be a promising tool for combating shrimp viruses. Antiviral double-stranded (ds)RNA has been mostly produced in Escherichia coli-expression system because of its high efficiency and inexpensive operations. However, overusing the bacteria may raise concerns regarding public health and environmental contamination, and seeking for a new dsRNA production platform would be alternative for future molecular farming. In this study, we exploited the green microalgae Chlamydomonas reinhardtii to produce dsRNA targeting the lethal shrimp yellow head virus (YHV). The expression plasmid pSL18 for C. reinhardtii was constructed to contain YHV-specific hairpin RNA expression cassette, and the successful assembly of pSL18-YHV was confirmed by PCR and enzymatic digestions. Glass bead method was employed for transformation of C. reinhardtii nuclear genome with pSL18-YHV. Microalgal expression of dsRNA-YHV, approximately 45 ng from 100-mL culture, was detected by qRT-PCR. Oral feeding experiment on postlarval shrimp revealed that the formulated feed with C. reinhardtii expressing dsRNA-YHV, at the ratio of 1 × 10 8 transformants per gram feed, improved 22% survival rate after YHV challenge. The present study suggests that C. reinhardtii can be bioengineered to produce viralspecific dsRNA for shrimp viral disease control, and the developed qRT-PCR could detect microalgal dsRNA with detection limit of subpicogram.

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“…In lower vertebrates, RNAi can be delivered as long dsRNA, as in the example where L. vannamei shrimp were protected from White spot syndrome virus (Robalino et al 2005). Another study demonstrated protection from yellow head virus by injection of a dsRNA (Saksmerprome et al 2009). Penaeus japonicus (the kuruma prawn) was also protected from White spot syndrome virus after siRNA injection .…”

Section: Combating Infectious Diseasementioning

confidence: 99%

RNA interference-based technology: what role in animal agriculture?

et al. 2017

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Abstract. Animal agriculture faces a broad array of challenges, ranging from disease threats to adverse environmental conditions, while attempting to increase productivity using fewer resources. RNA interference (RNAi) is a biological phenomenon with the potential to provide novel solutions to some of these challenges. Discovered just 20 years ago, the mechanisms underlying RNAi are now well described in plants and animals. Intracellular double-stranded RNA triggers a conserved response that leads to cleavage and degradation of complementary mRNA strands, thereby preventing production of the corresponding protein product. RNAi can be naturally induced by expression of endogenous microRNA, which are critical in the regulation of protein synthesis, providing a mechanism for rapid adaptation of physiological function. This endogenous pathway can be co-opted for targeted RNAi either through delivery of exogenous small interfering RNA (siRNA) into target cells or by transgenic expression of short hairpin RNA (shRNA). Potentially valuable RNAi targets for livestock include endogenous genes such as developmental regulators, transcripts involved in adaptations to new physiological states, immune response mediators, and also exogenous genes such as those encoded by viruses. RNAi approaches have shown promise in cell culture and rodent models as well as some livestock studies, but technical and market barriers still need to be addressed before commercial applications of RNAi in animal agriculture can be realised. Key challenges for exogenous delivery of siRNA include appropriate formulation for physical delivery, internal transport and eventual cellular uptake of the siRNA; additionally, rigorous safety and residue studies in target species will be necessary for siRNA delivery nanoparticles currently under evaluation. However, genomic incorporation of shRNA can overcome these issues, but optimal promoters to drive shRNA expression are needed, and genetic engineering may attract more resistance from consumers than the use of exogenous siRNA. Despite these hurdles, the convergence of greater understanding of RNAi mechanisms, detailed descriptions of regulatory processes in animal development and disease, and breakthroughs in synthetic chemistry and genome engineering has created exciting possibilities for using RNAi to enhance the sustainability of animal agriculture.

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A novel and inexpensive application of RNAi technology to protect shrimp from viral disease

Cited by 60 publications

References 17 publications

Penaeus monodon is protected against gill- associated virus by muscle injection but not oral delivery of bacterially expressed dsRNAs

Penaeus monodon is protected against gill- associated virus by muscle injection but not oral delivery of bacterially expressed dsRNAs

Use of microalgae Chlamydomonas reinhardtii for production of double-stranded RNA against shrimp virus

RNA interference-based technology: what role in animal agriculture?

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