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

Anderson, M. A. E. et al. (2024) A multiplexed, confinable CRISPR/Cas9 gene drive can propagate in caged Aedes aegypti populations. Nature Communications, 15, 729. (doi: 10.1038/s41467-024-44956-2) (PMID:38272895)

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.

• Item Type: Articles
• Additional Information: Funding: M.A.E.A., E.G., J.X.D.A., L.S., K.N., S.A.N.V., and L.A. were funded through a Defense Advanced Research Projects Agency (DARPA) award [N66001-17-2-4054] to Kevin Esvelt at MIT. MPE and PTL were supported by the Wellcome Trust [110117/Z/15/Z]. L.A. and T.H.S. were funded by the UK Biotechnology and Biological Sciences Research Council [BBS/E/ I/00007033, BBS/E/I/00007038, and BBS/E/I/00007039 to The Pirbright Institute]. DKP’s PhD studentship was funded by The Pirbright Institute. Data availability: All raw reads from amplicon-Seq generated in this study were submitted to NCBI SRA with the accession number PRJNA741076. Addgene plasmid # 52891; http://n2t.net/addgene:52891; RRID:Addgene_52891. Plasmid sequences are available from NCBI OP728003 https://www.ncbi.nlm.nih.gov/nuccore/OP728003, OP728004, OP728005 https://www.ncbi.nlm.nih.gov/nuccore/ OP728005. The remaining data generated for this study is available in the Supplementary dataset 1 file. Source data are provided with this paper.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Purusothaman, Dr Deepak Kumar
• Authors: Anderson, M. A. E., Gonzalez, E., Edgington, M. P., Ang, J. X. D., Purusothaman, D.-K., Shackleford, L., Nevard, K., Verkuijl, S. A. N., Harvey-Samuel, T., Leftwich, P. T., Esvelt, K., and Alphey, L.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
• Journal Name: Nature Communications
• Publisher: Nature Research
• ISSN: 2041-1723
• ISSN (Online): 2041-1723
• Copyright Holders: Copyright: © 2024 The Author(s)
• First Published: First published in Nature Communications 15: 729
• Publisher Policy: Reproduced under a Creative Commons licence