A confinable female-lethal population suppression system in the malaria vector, Anopheles gambiae
Abstract
Malaria is among the world's deadliest diseases, predominantly affecting sub-Saharan Africa, and killing over half a million people annually. Controlling the principal vector, the mosquito Anopheles gambiae, as well as other anophelines, is among the most effective methods to control disease spread. Here we develop an innovative genetic population suppression system termed Ifegenia (Inherited Female Elimination by Genetically Encoded Nucleases to Interrupt Alleles) in this deadly vector. In this bicomponent CRISPR-based approach, we disrupt a female-essential gene, femaleless (fle), demonstrating complete genetic sexing via heritable daughter gynecide. Moreover, we show that Ifegenia males remain reproductively viable, and can load both fle mutations and CRISPR machinery to induce fle mutations in subsequent generations, resulting in sustained population suppression. Through modeling, we demonstrate that iterative releases of non-biting Ifegenia males can act as an effective, confinable, controllable, and safe population suppression and elimination system.
Additional Information
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. We thank Judy Ishikawa and Michelle Bui for help with mosquito husbandry, Dhara Desai, Sanle Chen, Akshay Bharadwaj, and Martha L. Chow for laboratory assistance. Figures were Created with BioRender.com. This work was supported by funding from a DARPA Safe Genes Program Grant (HR0011-17-2-0047) and an NIH award (R01AI151004) awarded to O.S.A., and by funds from the Bill & Melinda Gates Foundation (INV-017683) awarded to J.M.M. The views, opinions, and/or findings expressed are those of the authors and should not be interpreted as representing the official views or policies of the U.S. government. Author contributions. O.S.A and A.L.S conceived and designed the experiments. J.P., R.A., A.L.S. and N.T. performed molecular and genetic experiments as well as mosquito crosses and husbandry I.A. performed the Oxford nanopore and RNA sequencing experiments and analysis. H.M.S.C., R.M.C., E.J.G. and J.M.M. performed mathematical modeling. All authors contributed to the writing, analyzed the data, and approved the final manuscript. Data availability. Complete sequence maps and plasmids are deposited at Addgene.org (#187238). All Illumina and Nanopore sequencing data has been deposited to the NCBI-SRA PRJNA862928, reviewer link: https://dataview.ncbi.nlm.nih.gov/object/PRJNA862928?reviewer=pbmvm5fb9in78ro2a1e7p227oc. All data used to generate figures are provided in the Supplementary Materials/Tables. Generated A. gambiae transgenic lines are available upon request to O.S.A. Ethical conduct of research. All animals were handled in accordance with the Guide for the Care and Use of Laboratory Animals as recommended by the National Institutes of Health and approved by the UCSD Institutional Animal Care and Use Committee (IACUC, Animal Use Protocol #S17187) and UCSD Biological Use Authorization (BUA #R2401). Competing Interest Statement. O.S.A is a founder of Agragene, Inc. and Synvect, Inc. with equity interest. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. A.L.S. is an author on a patent for related RNA-guided gene drive technology (WO2015105928A1). O.S.A, R.A., J.P., and A.S., have filed a provisional patent application on this technology. All other authors declare no competing interests.Attached Files
Submitted - 2022.08.30.505861v1.full.pdf
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Additional details
- Eprint ID
- 120352
- Resolver ID
- CaltechAUTHORS:20230322-368390000.38
- HR0011-17-2-0047
- Defense Advanced Research Project Agency (DARPA)
- R01AI151004
- NIH
- INV-017683
- Bill and Melinda Gates Foundation
- Created
-
2023-03-26Created from EPrint's datestamp field
- Updated
-
2023-08-13Created from EPrint's last_modified field