Section of Cell and Developmental Biology, University of California San Diego, La Jolla, CA, USA.
Department of Genetics, Blavatnik Institute, Harvard Medical School, Boston, MA, USA.
Nat Commun. 2021 May 20;12(1):2960. doi: 10.1038/s41467-021-23239-0.
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.
库蚊是多种人类和动物疾病的全球性传播媒介，包括西尼罗河病毒、淋巴丝虫病和禽疟，对公共卫生、牲畜、伴侣动物和濒危鸟类构成持续威胁。尽管杀虫剂耐药性的上升已经威胁到了库蚊的控制，但 CRISPR 基因组编辑工具的进步促进了替代遗传策略的发展，如基因驱动系统来对抗病媒。然而，尽管基因驱动技术在其他蚊子中迅速发展，但在库蚊中却缺乏进展。在这里，我们开发了一个库蚊特异性的 Cas9/gRNA 表达工具包，并使用基于靶向同源性的转基因技术生成和验证了一个表达 Cas9 的斑翅果蝇品系。我们表明，gRNA 支架变体提高了在库蚊和果蝇中的转基因效率，并提高了果蝇中的基因驱动性能。这些发现支持了未来控制库蚊的技术发展，并为在其他物种中改进这些工具提供了有价值的见解。
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