White spot syndrome virus VP28 specific double-stranded RNA provides protection through a highly focused siRNA population

Nilsen, Pål; Karlsen, Marius; Sritunyalucksana, Kallaya; Thitamadee, Siripong

doi:10.1038/s41598-017-01181-w

Cited by 38 publications

(12 citation statements)

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“…The maximum dose used in the present work was 3.0 µg/shrimp with 95% protection at 504 hpi (21 days). These evaluated doses are lower than those previously reported from 5.5 μg doses [66] up to 31 μg of dsRNA/shrimp [23]. This work has demonstrated the efficacy of the dose, and the sequence of the dsRNA used.…”

Section: Discussionmentioning

confidence: 53%

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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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Section: Discussionmentioning

confidence: 53%

“…The RNAi trials using an intramuscular injection (IM) have shown that VP28 glycoprotein is the target of choice to block WSSV infection in shrimp [14,23,24]. However, RNAi intramuscular (IM) administration is limited to lab-scale experiments since its use is not yet viable for applications on a large scale, as found in salmon farms [25].…”

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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“…RNA interference (RNAi) is an endogenous gene-silencing mechanism that can be repurposed to combat viral infection through sequence-specific recognition and degradation of viral RNA [ 10 ]. As one of the most infamous shrimp viruses, RNAi applications against White Spot Syndrome Virus (WSSV) have been extensively investigated [ 11 , 12 ] and dsRNA targeting the VP28 mRNA, which encodes the virus envelope protein that has been implicated in host entry [ 13 , 14 ], has become one of the most effective targets [ 15 ]. Despite the promising results at a laboratory level, the implementation of the RNAi technology at a farm level is still at its infantile stage due to the lack of effective delivery approaches.…”

Section: Introductionmentioning

confidence: 99%

Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain

et al. 2021

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Background Viruses cause significant economic losses to shrimp aquaculture worldwide. In severe cases, they can lead to 100% mortality within a matter of days, hence the aquaculture industry requires antiviral strategies to minimize economic impacts. Currently, a double-stranded RNA (dsRNA)-based platform has been proven effective at a laboratory scale. The bottleneck for its industrialization is the lack of low-cost, efficient and practical delivery approaches. In an effort to bridge the gap between laboratory and farm applications, virus-like particles (VLP) have been used as nanocarriers of dsRNA. However, the implementation of this approach still suffers from high costs and a lengthy procedure, co-expression of subunits of VLP or capsid proteins (CPs) and dsRNA can be the solution for the problem. CP and dsRNA are traditionally expressed in two different E. coli hosts: protease-deficient and RNase III-deficient strains. To condense the manufacturing of dsRNA-containing VLP, this study constructed a novel E. coli strain that is able to co-express viral capsid proteins and dsRNA in the same E. coli cell. Results A novel bacterial strain DualX-B15(DE3) was engineered to be both protease- and RNase III-deficiency via P1 phage transduction. The results revealed that it could simultaneously express recombinant proteins and dsRNA. Conclusion Co-expression of viral capsid proteins and dsRNA in the same cell has been shown to be feasible. Not only could this platform serve as a basis for future cost-effective and streamlined production of shrimp antiviral therapeutics, it may be applicable for other applications that requires co-expression of recombinant proteins and dsRNA.

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“…Moreover, this envelop protein plays significant role in initiating the WSSV infection in shrimp [22]. The VP28 gene of the WSSV is suggestively involved in endosome escape through its interaction with host Rab7 [24], and has been identified as a potent target for dsRNA treatment in comparative studies [25]. Previous analyses of strain variability reported that competitive fitness depends on the size of the genome [26,27], however, these studies are far from enough to illustrate the mechanisms of WSSV pathogenesis.…”

Section: Introductionmentioning

confidence: 99%

Circulating Phylotypes of White Spot Syndrome Virus in Bangladesh and Their Virulence

Hasan¹,

Hoque²,

Ahmed³

et al. 2021

Preprint

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White Spot Syndrome Virus (WSSV) has emerged as one of the most prevalent and lethal viruses globally, and infects both shrimps and crabs in the aquatic environment. This study aimed to investigate the occurrence of WSSV in different ghers of Bangladesh and the virulence of the circulating phylotypes. We collected 360 shrimp (Penaeus monodon) and 120 crab (Scylla sp.) samples from the South-East (Cox’s Bazar) and South-West (Satkhira) coastal regions of Bangladesh. The VP28 gene-specific PCR assays and sequencing revealed statistically significant (p < 0.05, Kruskal Wallis test) differences in the prevalence of WSSV in shrimps and crabs between the study areas (Cox’s Bazar and Satkhira), and over the study periods (2017-2019). The mean Log load of WSSV varied from 8.40 (Cox’s Bazar) to 10.48 (Satkhira) per gram of tissue. The mean values for salinity, dissolved oxygen, temperature and pH were 14.71±0.76 ppt, 3.7±0.1 ppm, 34.11±0.38˚C and 8.23±0.38, respectively in the WSSV-positive ghers. The VP28 gene-based phylogenetic analysis showed an amino-acid substitution (E→G) at 167th position in the isolates from Cox’s Bazar (referred to as phylotype BD2) compared to the globally circulating one (BD1). Shrimp PL artificially challenged with BD1 and BD2 phylotypes with filtrates of tissue containing 0.423 X 109 copies of WSSV per mL resulted a median LT50 value of 73 hrs and 75 hrs, respectively. The in-vivo trial showed higher mean Log WSSV copies (6.47±2.07 per mg tissue) in BD1 challenged shrimp PL compared to BD2 (4.75±0.35 per mg tissue). Crabs infected with BD1 and BD2 showed 100% mortality within 48 hrs and 62 hrs of challenge, respectively with mean Log WSSV copies of 12.06±0.48 and 9.95±0.37 per gram tissue, respectively. Moreover, shrimp antimicrobial peptides (AMPs) penaeidin and lysozyme expression was lower in BD1 challenged group compared to BD2 challenged shrimps. These results collectively demonstrated that relative virulence properties of WSSV based on mortality rate, viral load and expression of host immune genes in artificially infected shrimp PL could be affected by single aa substitution in VP28.

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White spot syndrome virus VP28 specific double-stranded RNA provides protection through a highly focused siRNA population

Cited by 38 publications

References 38 publications

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

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

Co-expression of double-stranded RNA and viral capsid protein in the novel engineered Escherichia coli DualX-B15(DE3) strain

Circulating Phylotypes of White Spot Syndrome Virus in Bangladesh and Their Virulence

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