Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published March 26, 2023 | Submitted + Supplemental Material
Report Open

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

Supplemental Material - media-1.xlsx

Supplemental Material - media-10.xlsx

Supplemental Material - media-11.txt

Supplemental Material - media-12.txt

Supplemental Material - media-13.xlsx

Supplemental Material - media-14.xlsx

Supplemental Material - media-15.xlsx

Supplemental Material - media-16.xlsx

Supplemental Material - media-17.xlsx

Supplemental Material - media-18.xlsx

Supplemental Material - media-19.xlsx

Supplemental Material - media-2.xlsx

Supplemental Material - media-20.xlsx

Supplemental Material - media-21.xlsx

Supplemental Material - media-3.xlsx

Supplemental Material - media-4.xlsx

Supplemental Material - media-5.xlsx

Supplemental Material - media-6.xlsx

Supplemental Material - media-7.xlsx

Supplemental Material - media-8.xlsx

Supplemental Material - media-9.xlsx

Files

2022.08.30.505861v1.full.pdf
Files (20.4 MB)
Name Size Download all
md5:d4da2e84d49074764f206bc2ce1e0f1f
1.8 MB Download
md5:067745bf25bc5be966735649ba020c13
1.8 MB Download
md5:3a9dde82053d50f6fb5b539cd24c1830
5.6 kB Download
md5:1c8af4127474b95f3eb5099f51d3f223
7.0 kB Download
md5:bb776fd8b816abb20e55378f201fc65c
1.8 MB Download
md5:127311acdb73c08ff7eb0534f899d309
8.0 kB Download
md5:1465169d8e5825096f7e8e1e6c3b3a1d
27.4 kB Download
md5:60c66f2472d263b4394ef039053a6434
1.8 MB Download
md5:de00d53e895d138e6a54ba3dc12b4744
9.5 kB Download
md5:ff55d63e97dbb09b4ef7c422b7f7a8fb
7.0 kB Download
md5:5925200eabffe61e02a6a4dfd2502106
13.0 kB Download
md5:69474fffdfb5833124383046913baaf8
8.1 kB Download
md5:ea1f0c7f577e7d5ef87cec4dbfe2e845
1.8 MB Download
md5:36c3c9558fa36c2114c6707e5f1ef688
2.9 MB Download
md5:35955bdb8a4b21fa9b523acdd4ba2286
1.8 MB Download
md5:9cd0ff419348cfab753e3b3c9a33a3e2
37.0 kB Preview Download
md5:f57cd0c605f5cb3d9ebe0835d7a5a926
37.2 kB Preview Download
md5:68d92d822a8527fc5bce923c42f63249
6.6 kB Download
md5:48578b60bc1380731ad89f1aeec998e5
21.0 kB Download
md5:990593ef76d7c0eec003f7e4853998c8
59.3 kB Download
md5:4877dc65031bd1924d6c0dd15421cd2e
6.4 MB Preview Download
md5:b9d51b2bdd88fd932c53fb4d5831a419
9.7 kB Download

Additional details

Created:
August 20, 2023
Modified:
October 18, 2023