Infectious hypodermal and hematopoietic necrosis virus-like particles encapsulating VP28 double-stranded RNA protect shrimp from white spot syndrome virus

Jariyapong, Pitchanee; Chotwiwatthanakun, Charoonroj; Pooljun, Chettupon; Weerachatyanukul, Wattana

doi:10.1016/j.aquaculture.2019.02.001

Cited by 11 publications

(6 citation statements)

References 31 publications

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“…However, when shrimp were treated with Linked PstDNV VLPs-dspro and other constructs followed by YHV challenge, shrimp receiving Linked PstDNV VLPs-dspro showed the lowest cumulative mortality amongst those treated with CoEx PstDNV VLPs/dspro, PstDNV alone, dspro alone, or NaCl, suggesting that the Linked PstDNV VLPs-dspro could effectively shield dsRNA from host nuclease and immune response, thus assisting in endocytosis and successful delivery. The protection toward therapeutic nucleotides of PstDNV VLP and MrNV VLP have been already demonstrated both in vitro and in vivo in shrimp [27][28][29][30]32,[44][45][46]. Dspro alone could reduce shrimp's mortality from YHV infection, but at a comparatively lower rate.…”

Section: Discussionmentioning

confidence: 97%

Simultaneous Production of a Virus-Like Particle Linked to dsRNA to Enhance dsRNA Delivery for Yellow Head Virus Inhibition

Worawittayatada

Angsujinda

Sinnuengnong

et al. 2022

Viruses

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A co-expressed Penaeus stylirostris densovirus (PstDNV) capsid and dsRNA specific to the yellow head virus (YHV) protease (CoEx cpPstDNV/dspro) has been shown to suppress YHV replication in the Pacific white-legged shrimp (Litopenaeus vannamei). However, maintaining two plasmids in a single bacterial cell is not desirable; therefore, a single plasmid harboring both the PstDNV capsid and the dsRNA-YHV-pro gene was constructed under the regulation of a single T7 promoter, designated pET28a-Linked cpPstDNV-dspro. Following induction, this novel construct expressed an approximately 37-kDa recombinant protein associated with a roughly 400-bp dsRNA (Linked cpPstDNV-dspro). Under a transmission electron microscope, the virus-like particles (VLP; Linked PstDNV VLPs-dspro) obtained were seen to be monodispersed, similar to the native PstDNV virion. A nuclease digestion assay indicated dsRNA molecules were both encapsulated and present outside the Linked PstDNV VLPs-dspro. In addition, the amount of dsRNA produced from this strategy was higher than that obtained with a co-expression strategy. In a YHV infection challenge, the Linked PstDNV VLPs-dspro was more effective in delaying and reducing mortality than other constructs tested. Lastly, the linked construct provides protection for the dsRNA cargo from nucleolytic enzymes present in the shrimp hemolymph. This is the first report of a VLP carrying virus-inhibiting dsRNA that could be produced without disassembly and reassembly to control virus infection in shrimp.

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Section: Discussionmentioning

confidence: 97%

Simultaneous Production of a Virus-Like Particle Linked to dsRNA to Enhance dsRNA Delivery for Yellow Head Virus Inhibition

Worawittayatada

Angsujinda

Sinnuengnong

et al. 2022

Viruses

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“…In both cases, the VLPs were synthesized from viruses that infect shrimp. One was with the macrobrachium rosenbergii nodavirus (MrNv) [66], whereas the second was with the infectious hypodermal and hematopoietic necrosis virus (IHHNV) [31]; both studies showed a survival rate of 44.5 and 40% by IM injection (6 μg of dsRNAvp28 per shrimp), respectively. Here, we were able to obtain similar results when VLP-dsRNAvp28 was administered per os.…”

Section: Discussionmentioning

confidence: 99%

“…One solution is a nanocarrier [11,27,29] to protect, stabilize and maintain the integrity of the RNAi in the environment [14]. Recently, dsRNA has been integrated into nanovehicles such as non-virulent capsids or virus-like particles (VLPs) [30][31][32] lacking the viral genome. Their small size (20-140 nm), allows them to permeate the cell membranes without causing toxicity or immune response in the treated organisms [30,[32][33][34][35][36].…”

Section: Introductionmentioning

confidence: 99%

Antiviral therapy in shrimp through plant virus VLP containing VP28 dsRNA against WSSV

Ramos-Carreño¹,

Giffard‐Mena²,

Zamudio-Ocadiz³

et al. 2021

Beilstein J. Org. Chem.

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The white spot syndrome virus (WSSV), currently affecting cultured shrimp, causes substantial economic losses to the worldwide shrimp industry. An antiviral therapy using double-stranded RNA interference (dsRNAi) by intramuscular injection (IM) has proven the most effective shrimp protection against WSSV. However, IM treatment is still not viable for shrimp farms. The challenge is to develop an efficient oral delivery system that manages to avoid the degradation of antiviral RNA molecules. The present work demonstrates that VLPs (virus-like particles) allow efficient delivery of dsRNAi as antiviral therapy in shrimp. In particular, VLPs derived from a virus that infects plants, such as cowpea chlorotic mottle virus (CCMV), in which the capsid protein (CP) encapsidates the dsRNA of 563 bp, are shown to silence the WSSV glycoprotein VP28 (dsRNAvp28). In experimental challenges in vivo, the VLPs- dsRNAvp28 protect shrimp against WSSV up to 40% by oral administration and 100% by IM. The novel research demonstrates that plant VLPs, which avoid zoonosis, can be applied to pathogen control in shrimp and also other organisms, widening the application window in nanomedicine.

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“…The VP28 has been identified as a viral surface protein that plays a key role in the initial steps of systemic WSSV infection in shrimp [27]. Several reports have experimentally tested dsRNAs targeting the structural gene VP28 for their efficacy to confer prevention of WSSV infection in P. monodon and L. vannamei [28,29]. Thus, RNAi technology is now applicable as a prevention strategy against WSSV infection in shrimp.…”

Section: Discussionmentioning

confidence: 99%

Bacillus subtilis expressing dsVP28 improved shrimp survival from WSSV challenge

Saelim¹,

Loprasert²,

Phongdara³

2020

ScienceAsia

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White spot syndrome virus (WSSV) is a potent shrimp viral pathogen responsible for significant economic losses to shrimp aquaculture all over the world. Several studies have demonstrated efficient RNAi-based approaches for suppressing viral genes and reducing shrimp mortality. However, the application has been difficult in real practical use. The present work aimed to develop an efficient approach for the delivery of VP28 specific-siRNA using Bacillus subtilis, a novel live oral vaccine vehicle, to protect shrimp against WSSV infection. A plasmid that can function in both bacteria and eukaryotic cells was developed by combining pBE-sDNA and the pCMV promoter from the pcDNA4 plasmid to obtain pBE:cDNA4. Subsequently, to evaluate the efficacy of pBE:cDNA4 in vivo, dsGFP-expressing pBE:cDNA4 (pBE:cDNA4-dsGFP) and dsVP28-expressing pBE:cDNA4 (pBE:cDNA4-dsVP28) were used to knockdown target transcripts in WSSV-infected shrimp. A high survival rate was shown for WSSV-infected shrimp injected with pBE:cDNA4-dsVP28 at 7-14 days post-infection (dpi). The pBE:cDNA4-dsVP28 plasmid was transformed into B. subtilis, and B. subtilis was used as a delivery vehicle during oral feeding. WSSV-infected shrimp fed with B. subtilis carrying pBE:cDNA4-dsVP28 showed 91.67% survival compared to the control, which showed only 28.57% survival. The results here demonstrate the efficient delivery of specific siRNAs by using bactofection with B. subtilis.

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Infectious hypodermal and hematopoietic necrosis virus-like particles encapsulating VP28 double-stranded RNA protect shrimp from white spot syndrome virus

Cited by 11 publications

References 31 publications

Simultaneous Production of a Virus-Like Particle Linked to dsRNA to Enhance dsRNA Delivery for Yellow Head Virus Inhibition

Simultaneous Production of a Virus-Like Particle Linked to dsRNA to Enhance dsRNA Delivery for Yellow Head Virus Inhibition

Antiviral therapy in shrimp through plant virus VLP containing VP28 dsRNA against WSSV

Bacillus subtilis expressing dsVP28 improved shrimp survival from WSSV challenge

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