CRISPR-Mediated Genome Engineering in Aedes aegypti

R. Sun, M. Li, C. J. McMeniman and O. S. Akbari,  piRNA: Methods and Protocols,  2022.

CRISPR-mediated genome engineering technologies have been adapted to a wide variety of organisms with high efficiency and specificity. The yellow fever mosquito, Aedes aegyptiAedes aegypti, is one such organism. It is also responsible for transmitting a wide variety of deadly viruses including Dengue, Zika, Yellow fever, and Chikungunya. The key to successful CRISPR-mediated gene editingGene editing applications is the delivery of both Cas9 ribonuclease and single-guide RNA (sgRNASingle guide RNA (sgRNA)) to the nucleus of desired cells. Various methods have been developed for supplying the Cas9 endonuclease, sgRNASingle guide RNA (sgRNA), and donor DNA to Ae. aegypti. In this chapter, we focus on methods of direct embryoEmbryosdelivery of editing components, presenting detailed step-by-step CRISPR/Cas9-based genome-editing protocols for inducing desired heritable edits in mosquitoes as well as insights into successful application of these protocols. We also highlight potential opportunities for customizing these protocols to manipulate the mosquito genome for innovative in vivoIn vivo gene function studies.

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