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1.发明公开公开(公告)号：US20240294864A1
公开(公告)日：2024-09-05
申请号：US18044891
申请日：2021-09-13
CPC分类号： C12N1/12 , A01N63/60 , A01N65/03 , A01P1/00 , C12N15/111 , C12N2310/14 , C12N2330/50
摘要： The invention relates to a novel method to produce small RNAs targeting virulence factors, essential genes and/or antimicrobial resistance genes of phytopathogens. 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. These EVs can be collected from the cell-free medium of Chlorella cultures, and further concentrated and purified for biocontrol applications. Importantly, Chlorella EVs protect small RNAs from ribonuclease-mediated digestion, indicating that these lipid-based particles not only act as natural vectors of small RNAs towards pathogenic cells, but also presumably limit their degradation in the environment. The invention can thus likely be used to reduce the pathogenicity and growth of a wide range of pathogens or, potentially, to enhance beneficial effects and growth of plant-associated symbiotic and commensal microbes. Furthermore, because the integrity of Chlorella EV-embedded antimicrobial siRNAs remains unaltered when produced in photobioreactors, and when stored frozen, this method has the potential to be further exploited for the industrialization and manufacturing of a novel generation of microalgae-based biologicals.
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2.发明公开公开(公告)号：US20240218387A1
公开(公告)日：2024-07-04
申请号：US18044868
申请日：2021-09-13
IPC分类号： C12N15/82 , A61K36/05 , C12N1/12 , C12N15/113 , C12R1/89
CPC分类号： C12N15/8257 , A61K36/05 , C12N1/125 , C12N15/113 , C12N2310/141 , C12R2001/89
摘要： 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.
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