@article {1401902, title = {Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes}, journal = {Nat Commun}, volume = {12}, year = {2021}, month = {2021 05 20}, pages = {2960}, abstract = {Culex mosquitoes are a global vector for multiple human and animal diseases, including West Nile virus, lymphatic filariasis, and avian malaria, posing a constant threat to public health, livestock, companion animals, and endangered birds. While rising insecticide resistance has threatened the control of Culex mosquitoes, advances in CRISPR genome-editing tools have fostered the development of alternative genetic strategies such as gene drive systems to fight disease vectors. However, though gene-drive technology has quickly progressed in other mosquitoes, advances have been lacking in Culex. Here, we develop a Culex-specific Cas9/gRNA expression toolkit and use site-directed homology-based transgenesis to generate and validate a Culex quinquefasciatus Cas9-expressing line. We show that gRNA scaffold variants improve transgenesis efficiency in both Culex\ quinquefasciatus and Drosophila\ melanogaster and boost gene-drive performance in the fruit fly. These findings support future technology development to control Culex mosquitoes and provide valuable insight for improving these tools in other species.}, keywords = {Animals, Animals, Genetically Modified, CRISPR-Cas Systems, Culex, drosophila melanogaster, Female, Gene Drive Technology, Insecticide Resistance, Male, Mosquito Control, Mosquito Vectors, Mutagenesis, Site-Directed, RNA, Guide}, issn = {2041-1723}, doi = {10.1038/s41467-021-23239-0}, author = {Feng, Xuechun and L{\'o}pez Del Amo, V{\'\i}ctor and Mameli, Enzo and Lee, Megan and Bishop, Alena L and Perrimon, Norbert and Gantz, Valentino M} }