Cas9-Mediated Gene-Editing in the Malaria Mosquito Anopheles stephensi by ReMOT Control

• Published in: G3 : genes - genomes - genetics Vol. 10; no. 4; pp. 1353 - 1360
• Main Authors: Macias, Vanessa M, McKeand, Sage, Chaverra-Rodriguez, Duverney, Hughes, Grant L, Fazekas, Aniko, Pujhari, Sujit, Jasinskiene, Nijole, James, Anthony A, Rasgon, Jason L
• Format: Journal Article
• Language: English
• Published: United States Genetics Society of America 01.04.2020; Oxford University Press
• Subjects: crispr/cas9; Investigations; ovary translocation; reverse genetics; CRISPR/Cas9; reverse genetics; ovary translocation
• ISSN: 2160-1836; 2160-1836
• DOI: 10.1534/g3.120.401133

Innovative tools are essential for advancing malaria control and depend on an understanding of molecular mechanisms governing transmission of malaria parasites by mosquitoes. CRISPR/Cas9-based gene disruption is a powerful method to uncover underlying biology of vector-pathogen interactions and can itself form the basis of mosquito control strategies. However, embryo injection methods used to genetically manipulate mosquitoes (especially ) are difficult and inefficient, particularly for non-specialist laboratories. Here, we adapted the ReMOT Control ( ceptor- ediated vary ransduction of argo) technique to deliver Cas9 ribonucleoprotein complex to adult mosquito ovaries, generating targeted and heritable mutations in the malaria vector without injecting embryos. In , ReMOT Control gene editing was as efficient as standard embryo injections. The application of ReMOT Control to opens the power of CRISPR/Cas9 methods to malaria laboratories that lack the equipment or expertise to perform embryo injections and establishes the flexibility of ReMOT Control for diverse mosquito species.