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Programmable CRISPR interference for gene silencing using Cas13a in mosquitoes

biorxiv. 2019-12; 
Aditi Kulkarni, Wanqin Yu, Alex Moon, Ashmita Pandey, Kathryn A. Hanley, Jiannong Xu
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摘要

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 α- and δ-subunits of COPI genes were silenced by a post-blood meal crRNA injection, which resulted in mortality... More

關鍵詞

CRISPR-Cas13a, RNA interference, Anopheles gambiae, Aedes aegypti, gene silencing, CRISPRi
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