RT Journal Article SR Electronic T1 Genome-engineering with CRISPR-Cas9 in the mosquito Aedes aegypti JF bioRxiv FD Cold Spring Harbor Laboratory SP 013276 DO 10.1101/013276 A1 Kathryn E. Kistler A1 Leslie B. Vosshall A1 Benjamin J. Matthews YR 2014 UL http://biorxiv.org/content/early/2014/12/27/013276.abstract AB The mosquito Aedes aegypti is a potent vector of the Chikungunya, yellow fever, and Dengue viruses, which result in hundreds of millions of infections and over 50,000 human deaths per year. Loss-of-function mutagenesis in Ae. aegypti has been established with TALENs, ZFNs, and homing endonucleases, which require the engineering of DNA-binding protein domains to generate target specificity for a particular stretch of genomic DNA. Here, we describe the first use of the CRISPR-Cas9 system to generate targeted, site-specific mutations in Ae. aegypti. CRISPR-Cas9 relies on RNA-DNA base pairing to generate targeting specificity, resulting in cheaper, faster, and more flexible genome-editing reagents. We investigate the efficiency of reagent concentrations and compositions, demonstrate the ability of CRISPR-Cas9 to generate several different types of mutations via disparate repair mechanisms, and show that stable germ-line mutations can be readily generated at the vast majority of genomic locitested. This work offers a detailed exploration into the optimal use of CRISPR-Cas9 in Ae. aegypti that should be applicable to non-model organisms previously out of reach of genetic modification.