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

Autor: Macias VM; Department of Entomology, The Pennsylvania State University, University Park, PA 16802., McKeand S; Department of Entomology, The Pennsylvania State University, University Park, PA 16802., Chaverra-Rodriguez D; Department of Entomology, The Pennsylvania State University, University Park, PA 16802., Hughes GL; Department of Entomology, The Pennsylvania State University, University Park, PA 16802., Fazekas A; Department of Molecular Biology and Biochemistry, University of California Irvine, CA 92697., Pujhari S; Department of Entomology, The Pennsylvania State University, University Park, PA 16802., Jasinskiene N; Department of Molecular Biology and Biochemistry, University of California Irvine, CA 92697., James AA; Department of Molecular Biology and Biochemistry, University of California Irvine, CA 92697.; Department of Microbiology and Molecular Genetics School of Medicine, University of California Irvine, CA 92697., Rasgon JL; Department of Entomology, The Pennsylvania State University, University Park, PA 16802 jlr54@psu.edu.; Center for Infection Disease Dynamics, Huck Institutes for the Life Sciences, The Pennsylvania State University, University Park, PA 16802.
Jazyk: angličtina
Zdroj: G3 (Bethesda, Md.) [G3 (Bethesda)] 2020 Apr 09; Vol. 10 (4), pp. 1353-1360. Date of Electronic Publication: 2020 Apr 09.
DOI: 10.1534/g3.120.401133
Abstrakt: Innovative tools are essential for advancing malaria control and depend on an understanding of molecular mechanisms governing transmission of malaria parasites by Anopheles 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 Anopheles ) are difficult and inefficient, particularly for non-specialist laboratories. Here, we adapted the ReMOT Control ( Re ceptor- m ediated O vary T ransduction of C argo) technique to deliver Cas9 ribonucleoprotein complex to adult mosquito ovaries, generating targeted and heritable mutations in the malaria vector Anopheles stephensi without injecting embryos. In Anopheles , ReMOT Control gene editing was as efficient as standard embryo injections. The application of ReMOT Control to Anopheles 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.
(Copyright © 2020 Macias et al.)
Databáze: MEDLINE
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