Genome Engineering with CRISPR-Cas9 in the Mosquito Aedes aegypti

The mosquito Aedes aegypti is a potent vector of the chikungunya, yellow fever, and dengue viruses, responsible for hundreds of millions of infections and over 50,000 human deaths per year. Mutagenesis in Ae. aegypti has been established with TALENs, ZFNs, and homing endonucleases, which require the engineering of DNA-binding protein domains to provide genomic target sequence specificity. Here, we describe the use of the CRISPR-Cas9 system to generate site-specific mutations in Ae. aegypti. This system relies on RNA-DNA base-pairing to generate targeting specificity, resulting in efficient and flexible genome-editing reagents. We investigate the efficiency of injection mix compositions, demonstrate the ability of CRISPR-Cas9 to generate different types of mutations via disparate repair mechanisms, and report stable germline mutations in several genomic loci. This work offers a detailed exploration into the use of CRISPR-Cas9 in Ae. aegypti that should be applicable to non-model organisms previously out of reach of genetic modification.

Abstract licence: CC BY-NC

• Non-retroviral Endogenous Viral Element Limits Cognate Virus Replication in Aedes aegypti Ovaries · 2020

• Development of a confinable gene drive system in the human disease vector Aedes aegypti · 2020

• The ion channel ppk301 controls freshwater egg-laying in the mosquito Aedes aegypti · 2019

• Targeted delivery of CRISPR-Cas9 ribonucleoprotein into arthropod ovaries for heritable germline gene editing · 2018

• CRISPR/Cas9 -mediated gene knockout of Anopheles gambiae FREP1 suppresses malaria parasite infection · 2018

• Progress and Prospects of CRISPR/Cas Systems in Insects and Other Arthropods · 2017

• The neurotranscriptome of the Aedes aegypti mosquito · 2016

A person counts once a day per paper. We keep no cookie and no address: a reader is a hash that changes every day. Any month under five reads shows no number.