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Abstract
Cas9-mediated gene editing is a powerful tool for addressing research questions in arthropods. Current approaches rely upon delivering Cas9 ribonucleoprotein (RNP) complex by embryonic microinjection, which is challenging, is limited to a small number of species, and is inefficient even in optimized taxa. Here we develop a technology termed Receptor-Mediated Ovary Transduction of Cargo (ReMOT Control) to deliver Cas9 RNP to the arthropod germline by injection into adult female mosquitoes. We identify a peptide (P2C) that mediates transduction of Cas9 RNP from the female hemolymph to the developing mosquito oocytes, resulting in heritable gene editing of the offspring with efficiency as high as 0.3 mutants per injected mosquito. We demonstrate that P2C functions in six mosquito species. Identification of taxa-specific ovary-specific ligand–receptor pairs may further extend the use of ReMOT Control for gene editing in novel species.
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1 Department of Entomology, The Huck Institutes of the Life Sciences, and the Center for infectious Disease Dynamics, The Pennsylvania State University, University Park, PA, USA
2 Department of Entomology, The Huck Institutes of the Life Sciences, and the Center for infectious Disease Dynamics, The Pennsylvania State University, University Park, PA, USA; Department of Pathology, Institute for Human Infections and Immunity, Center for Tropical Diseases, Center for Biodefense and Emerging Infectious Disease, University of Texas Medical Branch, Galveston, TX, USA
3 Department of Entomology, The Huck Institutes of the Life Sciences, and the Center for infectious Disease Dynamics, The Pennsylvania State University, University Park, PA, USA; Institut Pasteur, Viruses and RNA Interference Unit, CNRS Unité Mixte de Recherche, Paris, France