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
© 2023 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY). We thank J. Ishikawa and M. Bui for help with mosquito husbandry; D. Desai, S. Chen, A. Bharadwaj, and M. L. Chow for laboratory assistance; and K. Dickerson for assistance in CRISPR QC analysis. Funding: This work was supported by funding from NIH awards (R01AI151004, R01GM132825, RO1AI148300, RO1AI175152, and DP2AI152071), EPA STAR award (RD84020401), and an Open Philanthropy award (309937-0001) 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. Ethics statement: 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 University of California San Diego (UCSD) Institutional Animal Care and Use Committee (animal use protocol no. S17187) and UCSD Biological Use Authorization (no. R2401). Author contributions: O.S.A. and A.L.S. conceived and designed the system and experiments. J.J.P., R.A.A., A.L.S., and N.T. performed molecular and genetic experiments. 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. Competing interests: 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. O.S.A., R.A.A., J.J.P., and A.L.S. have filed a provisional patent application on Ifegenia titled "Confinable population suppression system," University of California San Diego, 26 May 2022, 63/346,207. The other authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper and/or the Supplementary Materials. Complete sequence maps and plasmids are deposited at Addgene.org (no. 187238). All Illumina and Nanopore sequencing data have been deposited to the NCBI-SRA PRJNA862928. Generated A. gambiae transgenic lines are available upon request to O.S.A. Figures were created with BioRender.com.Attached Files
Published - sciadv.ade8903.pdf
Supplemental Material - sciadv.ade8903_sm.pdf
Supplemental Material - sciadv.ade8903_tables_s1_to_s21.zip
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Additional details
- PMCID
- PMC10321730
- Eprint ID
- 122510
- Resolver ID
- CaltechAUTHORS:20230725-705875000.5
- R01AI151004
- NIH
- R01GM132825
- NIH
- RO1AI148300
- NIH
- RO1AI175152
- NIH
- DP2AI152071
- NIH
- RD84020401
- Environmental Protection Agency (EPA)
- 309937-0001
- Open Philanthropy
- INV-017683
- Bill and Melinda Gates Foundation
- Created
-
2023-08-13Created from EPrint's datestamp field
- Updated
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2023-08-14Created from EPrint's last_modified field