Cas13b-dependent and Cas13b-independent RNA knockdown of viral sequences in mosquito cells following guide RNA expression

Tng, P Y L; Carabajal Paladino, L; Verkuijl, S A N; Purcell, J; Merits, A; Leftwich, P T; Fragkoudis, R; Noad, R; Alphey, L

Abstract

Aedes aegypti and Aedes albopictus mosquitoes are vectors of the RNA viruses chikungunya (CHIKV) and dengue that currently have no specific therapeutic treatments. The
development of new methods to generate virus-refractory mosquitoes would be beneficial.
Cas13b is an enzyme that uses RNA guides to target and cleave RNA molecules and has been
reported to suppress RNA viruses in mammalian and plant cells. We investigated the
potential use of the Prevotella sp. P5-125 Cas13b system to provide viral refractoriness in
mosquito cells, using a virus-derived reporter and a CHIKV split replication system. Cas13b in
combination with suitable guide RNAs could induce strong suppression of virus-derived
reporter RNAs in insect cells. Surprisingly, the RNA guides alone (without Cas13b) also gave
substantial suppression. Our study provides support for the potential use of Cas13b in
mosquitoes, but also caution in interpreting CRISPR/Cas data as we show that guide RNAs
can have Cas-independent effects.

Citation

Tng, P. Y. L., Carabajal Paladino, L., Verkuijl, S. A. N., Purcell, J., Merits, A., Leftwich, P. T., …Alphey, L. (2020). Cas13b-dependent and Cas13b-independent RNA knockdown of viral sequences in mosquito cells following guide RNA expression. Communications Biology, 3(1), https://doi.org/10.1038/s42003-020-01142-6

• Journal Article Type: Article
• Acceptance Date: Jul 9, 2020
• Publication Date: Jul 31, 2020
• Deposit Date: Aug 6, 2020
• Publicly Available Date: Aug 6, 2020
• Journal: Communications Biology
• Publisher: Nature Research
• Peer Reviewed: Peer Reviewed
• Volume: 3
• Issue: 1
• DOI: https://doi.org/10.1038/s42003-020-01142-6
• Public URL: https://rvc-repository.worktribe.com/output/1376451
• Publisher URL: https://doi.org/10.1038/s42003-020-01142-6