A multiplexed, confinable CRISPR/Cas9 gene drive can propagate in caged Aedes aegypti populations

Anderson, Michelle A. E.; Gonzalez, Estela; Edgington, Matthew P.; Ang, Joshua X. D.; Purusothaman, Deepak-Kumar; Shackleford, Lewis; Nevard, Katherine; Verkuijl, Sebald A. N.; Harvey-Samuel, Timothy; Leftwich, Philip T.; Esvelt, Kevin; Alphey, Luke

Abstract

Aedes aegypti is the main vector of several major pathogens including dengue, Zika and chikungunya viruses. Classical mosquito control strategies utilizing insecticides are threatened by rising resistance. This has stimulated interest in new genetic systems such as gene drivesHere, we test the regulatory sequences from the Ae. aegypti benign gonial cell neoplasm (bgcn) homolog to express Cas9 and a separate multiplexing sgRNA-expressing cassette inserted into the Ae. aegypti kynurenine 3-monooxygenase (kmo) gene. When combined, these two elements provide highly effective germline cutting at the kmo locus and act as a gene drive. Our target genetic element drives through a cage trial population such that carrier frequency of the element increases from 50% to up to 89% of the population despite significant fitness costs to kmo insertions. Deep sequencing suggests that the multiplexing design could mitigate resistance allele formation in our gene drive system.

Citation

Anderson, M. A. E., Gonzalez, E., Edgington, M. P., Ang, J. X. D., Purusothaman, D.-K., Shackleford, L., …Alphey, L. (in press). A multiplexed, confinable CRISPR/Cas9 gene drive can propagate in caged Aedes aegypti populations. Nature communications, 15(1), Article 729. https://doi.org/10.1038/s41467-024-44956-2

• Journal Article Type: Article
• Acceptance Date: Jan 11, 2024
• Online Publication Date: Jan 25, 2024
• Deposit Date: Feb 24, 2025
• Journal: Nature Communications
• Print ISSN: 2041-1723
• Peer Reviewed: Peer Reviewed
• Volume: 15
• Issue: 1
• Article Number: 729
• DOI: https://doi.org/10.1038/s41467-024-44956-2
• Public URL: https://keele-repository.worktribe.com/output/1078998
• Publisher URL: https://www.nature.com/articles/s41467-024-44956-2
• Additional Information: Received: 9 August 2023; Accepted: 11 January 2024; First Online: 25 January 2024; : L.A. is an adviser to Synvect Inc and Biocentis Ltd, with equity and/or financial interest in those companies. The other authors declare that they have no competing interests.