Methods to Interfere with Prokaryotic and Phage Translation and Noncoding RNA

Patent Status

Patent Pending

Brief Description

Classical methodologies for examining phage gene function, including UV/random mutagenesis and amber mutation, are difficult to assay efficiently on a genome-wide scale. Additionally, there are notable challenges in targeting phage genes with Cas9/12, such as epigenetic modifications, physical sequestration in the nucleus, absence of DNA genomes or intermediates in RNA phages, and efficient ligation/recombination processes. The limitation of current tools is also evident in failed attempts to apply transposon libraries in virulent phages, further underscoring the necessity for innovative approaches in phage functional genomics.

UC Berkeley researchers made the surprising discovery that catalytically inactivated Cas13 (dCas13) in complex with a guide RNA can bind to and modulate activity of viral target RNAs. Viruses have evolved numerous and diverse strategies to protect their genomes from host defenses, including encoding their genomes across several Baltimore classes (e.g., dsDNA, dsRNA, ssDNA, and ssRNA), employing diverse genome modification strategies, and employing advanced genome compartmentalization strategies. These protective strategies have severely limited the applicability and effectiveness of previously existing approaches. Thus, this invention provides methods and compositions for modulating the activity of a viral target RNA.

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