Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes

Xuechun Feng, Víctor López Del Amo, Enzo Mameli, Megan Lee, Alena L. Bishop, Norbert Perrimon and Valentino M. Gantz ()
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Xuechun Feng: University of California San Diego
Víctor López Del Amo: University of California San Diego
Enzo Mameli: Harvard Medical School
Megan Lee: University of California San Diego
Alena L. Bishop: University of California San Diego
Norbert Perrimon: Harvard Medical School
Valentino M. Gantz: University of California San Diego

Nature Communications, 2021, vol. 12, issue 1, 1-13

Abstract: 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.

Date: 2021
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DOI: 10.1038/s41467-021-23239-0

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