I am seeking to eliminate a double-stranded RNA virus from a host fungal pathogen using CRISPR-Cas technology. Currently, I am only able to gather that Cas13 (and related orthologs) cleaves ssRNA and not dsRNA.
Is there any CRISPR-Cas13 ortholog that has proven successful in cleaving dsRNA in vivo?
Could one attempt to disrupt the double-stranded RNA structure in vivo, creating two single-stranded RNA targets that could be cleaved by successive CRISPR-Cas13-mediated cleavages?
Double-stranded DNA (dsDNA) cleavage by Cas9 is a hallmark of type II CRISPR-Cas immune systems. Cas9–guide RNA complexes recognize 20-base-pair sequences in DNA and generate a site-specific double-strand break, a robust activity harnessed for genome editing Christopher Reynolds
Refer to the following article if this helps:
CRISPR-Csy4-Mediated Editing of Rotavirus Double-Stranded RNA Genome.
Papa et al. 2020
- PMCID: PMC7523552
- DOI: 10.1016/j.celrep.2020.108205
Cell Rep. 2020 Sep 29;32(13):108205.
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