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Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes
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Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes
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Journal Article
Optimized CRISPR tools and site-directed transgenesis towards gene drive development in Culex quinquefasciatus mosquitoes
2021
Overview
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.
Culex
mosquitoes are a global vector for insect-borne diseases, though progress with genetic tools lags behind other mosquito species. Here the authors present a Cas9-based toolkit and methods that could support future gene drive development in these mosquitoes.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 13/109
/ 38/44
/ 42
/ 42/35
/ 64
/ Animals
/ Animals, Genetically Modified
/ CRISPR
/ CRISPR-Cas Systems - genetics
/ Culex
/ Drosophila melanogaster - genetics
/ Female
/ Gene Drive Technology - methods
/ Genomes
/ gRNA
/ Homology
/ Humanities and Social Sciences
/ Insects
/ Malaria
/ Male
/ Mutagenesis, Site-Directed - methods
/ RNA, Guide, CRISPR-Cas Systems - genetics
/ Science
/ Toolkits
/ Vectors
