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bioRxiv · 10.1101/846055

Programmable CRISPR interference for gene silencing using Cas13a in mosquitoes

Kulkarni, A.·Yu, W.·Moon, A. S.·Pandey, A.·Hanley, K. A.·Xu, J.
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Abstract

In the CRISPR-Cas systems, Cas13a is an RNA-guided RNA nuclease specifically targeting single strand RNA. We developed a Cas13a mediated CRISPR interference tool to target mRNA for gene silencing in mosquitoes. The machinery was tested in two mosquito species. A Cas13a expressing plasmid was delivered to mosquitoes by intrathoracic injection, and Cas13a transcripts were detectable at least10 days post-delivery. In Anopheles gambiae, vitellogenin gene was silenced by Vg-crRNA injection two hours post-blood meal, which was accompanied by a significant reduction in egg production. In Aedes aegypti, the - and {delta}-subunits of COPI genes were silenced by a post-blood meal crRNA injection, which resulted in mortality and fragile midguts, reproducing a phenotype reported previously. Co-silencing genes simultaneously is achievable when a cocktail of target crRNAs is given. No detectable collateral cleavages of non-target transcripts were observed in the study. This study adds a programmable CRISPR tool to manipulate RNA in mosquitoes.

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Kulkarni, A., Yu, W., Moon, A. S., Pandey, A., Hanley, K. A., Xu, J.. 2019-11-18. Programmable CRISPR interference for gene silencing using Cas13a in mosquitoes. https://doi.org/10.1101/846055

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