Administration of co-expressed Penaeus stylirostris densovirus-like particles and dsRNA-YHV-Pro provide protection against yellow head virus in shrimp

Sinnuengnong, Rapee; Attasart, Pongsopee; Smith, Duncan R.; Panyim, Sakol; Assavalapsakul, Wanchai

doi:10.1016/j.jbiotec.2018.01.002

Cited by 15 publications

(15 citation statements)

References 26 publications

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“…The potential for the use of virus like particles (VLPs) for dsRNA delivery to insect pests, and the challenges associated with this method, have been reviewed by Kolliopoulou et al (2017) and VLPs have been successfully used in order to protect against yellow head virus in shrimp ( Sinnuengnong et al, 2018 ). The technology utilises VLPs that co-assemble in the expression species, specifically non-infectious viral envelopes co-expressed in E. coli with the dsRNA and protect dsRNA from nucleases and harsh lysis steps, assisting in the isolation, purification and application of crop protecting agents.…”

Section: Strategies For Production Of Dsrna For Crop Protectionmentioning

confidence: 99%

Strategies for the production of dsRNA biocontrols as alternatives to chemical pesticides

Hough

Howard

Brown

et al. 2022

Front. Bioeng. Biotechnol.

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Current crop pest control strategies rely on insecticidal and fungicidal sprays, plant genetic resistance, transgenes and agricultural practices. However, many insects, plant viruses, and fungi have no current means of control or have developed resistance against traditional pesticides. dsRNA is emerging as a novel sustainable method of plant protection as an alternative to traditional chemical pesticides. The successful commercialisation of dsRNA based biocontrols for effective pest management strategies requires the economical production of large quantities of dsRNA combined with suitable delivery methods to ensure RNAi efficacy against the target pest. A number of methods exist for the production and delivery of dsRNA based biocontrols and here we review alternative methods currently employed and emerging new approaches for their production. Additionally, we highlight potential challenges that will need to be addressed prior to widespread adoption of dsRNA biocontrols as novel sustainable alternatives to traditional chemical pesticides.

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Section: Strategies For Production Of Dsrna For Crop Protectionmentioning

confidence: 99%

Strategies for the production of dsRNA biocontrols as alternatives to chemical pesticides

Hough

Howard

Brown

et al. 2022

Front. Bioeng. Biotechnol.

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“…Romo-Quiñonez et al ( Romo et al, 2020 ) were able to enhance the antiviral potential of Litopenaeus vannamei after using a silver nanoparticle, Argovit4, administered as feed. Sinnuengnong et al, (2018) co-expressed virus-like particles of pstDNV with dsRNA molecules of YHV-Pro to deliver dsRNA using viral proteins, which was also very effective. Thedcharoen et al, (2020b) used the pLVX-AcGFP1-N1 vector with a gene encoding long hairpin RNA, namely pLVX-lhRdRp, introduced under control of the CMV promoter, to inhibit the RdRp (RNA-dependent RNA polymerase) of YHV with significant effects.…”

Section: Preparation and Uptake Of Rnaimentioning

confidence: 99%

Potential dsRNAs can be delivered to aquatic for defense pathogens

Nie

Chen

Tang

et al. 2022

Front. Bioeng. Biotechnol.

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The use of antibiotics to facilitate resistance to pathogens in aquatic animals is a traditional method of pathogen control that is harmful to the environment and human health. RNAi is an emerging technology in which homologous small RNA molecules target specific genes for degradation, and it has already shown success in laboratory experiments. However, further research is needed before it can be applied in aquafarms. Many laboratories inject the dsRNA into aquatic animals for RNAi, which is obviously impractical and very time consuming in aquafarms. Therefore, to enable the use of RNAi on a large scale, the methods used to prepare dsRNA need to be continuously in order to be fast and efficient. At the same time, it is necessary to consider the issue of biological safety. This review summarizes the key harmful genes associated with aquatic pathogens (viruses, bacteria, and parasites) and provides potential targets for the preparation of dsRNA; it also lists some current examples where RNAi technology is used to control aquatic species, as well as how to deliver dsRNA to the target hydrobiont.

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“…Escherichia coli strain DH5α containing either a pET-17b plasmid containing two double-stranded RNAs which encode the ribonucleotide reductase small subunit (rr2) gene of white spot syndrome virus and the protease (pro) gene of yellow head virus (pET17b-dsrr2-dspro_one stem) [33], or a pET-28a(+) plasmid encoding the PstDNV capsid (pET28a-cpPstDNV) [32] was inoculated into Luria-Bertani (LB) broth supplemented with 100 µg/mL ampicillin or 50 µg/mL kanamycin, respectively. The bacteria were incubated at 37 • C overnight with vigorous shaking.…”

Section: Recombinant Bacteria and Plasmid Extractionmentioning

confidence: 99%

“…While different bacterial hosts are normally used for the expression of the dsRNA and the VLP, Wuthisathid and colleagues recently developed a novel E. coli host optimized to be able to express both molecules [31]. Interestingly, Sinnuengnong et al [32] reported that the co-expression of the PstDNV capsid protein (cpPstDNV) and dsRNA-YHV-pro in the same E. coli cell generated VLP that reduced YHV replication and shrimp mortality at a higher level than just administration of the dsRNA. However, two plasmids, encoding the viral capsid and dsRNA, are difficult to be continuously maintained in the same bacteria cell, as one of the two plasmids can be subsequently lost due to incompatibility.…”

Section: Introductionmentioning

confidence: 99%

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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Administration of co-expressed Penaeus stylirostris densovirus-like particles and dsRNA-YHV-Pro provide protection against yellow head virus in shrimp

Cited by 15 publications

References 26 publications

Strategies for the production of dsRNA biocontrols as alternatives to chemical pesticides

Strategies for the production of dsRNA biocontrols as alternatives to chemical pesticides

Potential dsRNAs can be delivered to aquatic for defense pathogens

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

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