Chlorella-based production of extracellular vesicle-embedded small RNAs for prophylactic or therapeutic applications

Abstract

The invention relates to a novel method to produce small RNAs targeting pathogenicity factors, essential genes and/or antimicrobial resistance genes of animal pathogens. This method also includes the production of small RNAs directed against host susceptibility factors, whose silencing, inactivation, or deletion, is known to enhance resistance towards the targeted pathogen(s). More specifically, the invention involves the expression of exogenous RNA interference (RNAi) precursor(s) in Chlorella cells, which in turn express and release Extracellular Vesicle (EV)-embedded antimicrobial small RNAs. Importantly, Chlorella EVs protect antimicrobial small RNAs from ribonuclease-mediated digestion. They are also rapidly and efficiently internalized by human alveolar epithelial cells, highlighting their potential for delivering antimicrobial small RNAs in these cells, and for controlling respiratory infections. The invention can thus be used for prophylactic or therapeutic treatments, to reduce various infectious diseases in animals, including humans. Furthermore, because the integrity and functionality of Chlorella EVs remain unaltered when produced in photobioreactors, and when stored frozen, this novel method has the potential to be further exploited for the industrialization of EV-based anti-infective products.

Dependencies are inferred from the claim text.