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

Abstract RNA interference (RNAi) represents one of the most conserved pathways evolved by eukaryotic cells for regulating gene expression. RNAi utilises non-translatable double-stranded (dsRNA) molecules to sequester or degrade mRNA gene. In RNAi, specifically designed exogenous dsRNA delivered cell can silence a target gene, phenomenon that has been exploited in many functional studies and explored biopesticide applications. The search safe sustainable crop pest management options drives need offset effect inorganic pesticides on biodiversity. prospect replacing with spray is gaining popularity, enhanced its high-target specificity low environmental impact. However, reach pesticide market, it must be produced cost-effectively sustainably. this paper, we develop high-yield expression media generates up 15-fold yield compared existing utilising 1 mM IPTG. We also optimise low-cost purification method high-quality purified dsRNA. developed circumvents hazardous chemical reagents often found commercial kits nucleases eliminate contaminating DNA single-stranded (ssRNA) species. demonstrate production platform scalable, generating 6.29 mg from 259 wet E. coli pellet. results provide structural insights into heterogeneous species within microbial-derived pool. Finally, show ‘naked’ dsRNA, without prior formulation, induce insect toxicity under field conditions. This study provides novel, complete, process potential application industrial production. Graphical