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    A novel sustainable platform for scaled manufacturing of double‑stranded RNA biopesticides

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    Journal Article (2.158Mb)
    Date
    2022
    Author
    Nwokeoji, A.O.
    Nwokeoji, E.A.
    Chou, T.
    Togola, A.
    Type
    Journal Article
    Review Status
    Peer Review
    Target Audience
    Scientists
    Metadata
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    Abstract/Description
    RNA interference (RNAi) represents one of the most conserved pathways evolved by eukaryotic cells for regulating gene expression. RNAi utilises non-translatable double-stranded RNA (dsRNA) molecules to sequester or degrade mRNA molecules gene. In RNAi, specifically designed exogenous dsRNA delivered to the cell can silence a target gene, a phenomenon that has been exploited in many functional studies and explored in biopesticide applications. The search for safe and sustainable crop pest management options drives the need to offset the effect of inorganic pesticides on biodiversity. The prospect of replacing inorganic pesticides with dsRNA crop spray is gaining popularity, enhanced by its high-target specificity and low environmental impact. However, for dsRNA to reach the pesticide market, it must be produced cost-effectively and sustainably. In this paper, we develop a high-yield expression media that generates up to 15-fold dsRNA yield compared to existing expression media utilising 1 mM IPTG. We also optimise a low-cost purification method that generates high-quality and purified dsRNA. The developed method circumvents the need for hazardous chemical reagents often found in commercial kits or commercial nucleases to eliminate contaminating DNA or single-stranded RNA (ssRNA) species. We also demonstrate that the production platform is scalable, generating 6.29 mg dsRNA from 259 mg wet E. coli cell pellet. The results also provide structural insights into the heterogeneous dsRNA species within the microbial-derived dsRNA pool. Finally, we also show that the purified ‘naked’ dsRNA, without prior formulation, can induce insect toxicity under field conditions. This study provides a novel, complete, low-cost process dsRNA platform with potential for application in industrial dsRNA production.
    https://dx.doi.org/10.1186/s40643-022-00596-2
    Multi standard citation
    Permanent link to this item
    https://hdl.handle.net/20.500.12478/7962
    IITA Authors ORCID
    Abou TOGOLAhttps://orcid.org/0000-0001-6155-8292
    Digital Object Identifier (DOI)
    https://dx.doi.org/10.1186/s40643-022-00596-2
    Research Themes
    Plant Production and Health
    IITA Subjects
    Agronomy; Food Security; Pests of Plants
    Agrovoc Terms
    Pest Control; Biopesticides; Pest Management; Rna
    Hubs
    Headquarters and Western Africa Hub
    Journals
    Bioresources and Bioprocessing
    Collections
    • Journal and Journal Articles4836
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