Abstract
Genetic methods of manipulating or eradicating disease vector populations have long been discussed as an attractive alternative to existing control measures because of their potential advantages in terms of effectiveness and species specificity1,2,3. The development of genetically engineered malaria-resistant mosquitoes has shown, as a proof of principle, the possibility of targeting the mosquito’s ability to serve as a disease vector4,5,6,7. The translation of these achievements into control measures requires an effective technology to spread a genetic modification from laboratory mosquitoes to field populations8. We have suggested previously that homing endonuclease genes (HEGs), a class of simple selfish genetic elements, could be exploited for this purpose9. Here we demonstrate that a synthetic genetic element, consisting of mosquito regulatory regions10 and the homing endonuclease gene I-SceI11,12,13, can substantially increase its transmission to the progeny in transgenic mosquitoes of the human malaria vector Anopheles gambiae. We show that the I-SceI element is able to invade receptive mosquito cage populations rapidly, validating mathematical models for the transmission dynamics of HEGs. Molecular analyses confirm that expression of I-SceI in the male germline induces high rates of site-specific chromosomal cleavage and gene conversion, which results in the gain of the I-SceI gene, and underlies the observed genetic drive. These findings demonstrate a new mechanism by which genetic control measures can be implemented. Our results also show in principle how sequence-specific genetic drive elements like HEGs could be used to take the step from the genetic engineering of individuals to the genetic engineering of populations.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 51 print issues and online access
$199.00 per year
only $3.90 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Curtis, C. F. Possible use of translocations to fix desirable genes in insect pest populations. Nature 218, 368–369 (1968)
Hamilton, W. D. Extraordinary sex ratios. A sex-ratio theory for sex linkage and inbreeding has new implications in cytogenetics and entomology. Science 156, 477–488 (1967)
Alphey, L. et al. Malaria control with genetically manipulated insect vectors. Science 298, 119–121 (2002)
Corby-Harris, V. et al. Activation of Akt signaling reduces the prevalence and intensity of malaria parasite infection and lifespan in Anopheles stephensi mosquitoes. PLoS Pathog. 6, e1001003 (2010)
Ito, J., Ghosh, A., Moreira, L. A., Wimmer, E. A. & Jacobs-Lorena, M. Transgenic anopheline mosquitoes impaired in transmission of a malaria parasite. Nature 417, 452–455 (2002)
Moreira, L. A. et al. Bee venom phospholipase inhibits malaria parasite development in transgenic mosquitoes. J. Biol. Chem. 277, 40839–40843 (2002)
Li, F., Patra, K. P. & Vinetz, J. M. An anti-chitinase malaria transmission-blocking single-chain antibody as an effector molecule for creating a Plasmodium falciparum-refractory mosquito. J. Infect. Dis. 192, 878–887 (2005)
Sinkins, S. P. & Gould, F. Gene drive systems for insect disease vectors. Nature Rev. Genet. 7, 427–435 (2006)
Burt, A. Site-specific selfish genes as tools for the control and genetic engineering of natural populations. Proc. R. Soc. Lond. B 270, 921–928 (2003)
Catteruccia, F., Benton, J. P. & Crisanti, A. An Anopheles transgenic sexing strain for vector control. Nature Biotechnol. 23, 1414–1417 (2005)
Jacquier, A. & Dujon, B. An intron-encoded protein is active in a gene conversion process that spreads an intron into a mitochondrial gene. Cell 41, 383–394 (1985)
Bellaiche, Y., Mogila, V. & Perrimon, N. I-SceI endonuclease, a new tool for studying DNA double-strand break repair mechanisms in Drosophila . Genetics 152, 1037–1044 (1999)
Windbichler, N. et al. Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos. Nucleic Acids Res. 35, 5922–5933 (2007)
Stoddard, B. L. Homing endonuclease structure and function. Q. Rev. Biophys. 38, 49–95 (2005)
Goddard, M. R., Greig, D. & Burt, A. Outcrossed sex allows a selfish gene to invade yeast populations. Proc. R. Soc. Lond. B 268, 2537–2542 (2001)
Meredith, J. M. et al. Site-specific integration and expression of an anti-malarial gene in transgenic Anopheles gambiae significantly reduces Plasmodium infections. PLoS ONE 6, e14587 (2011)
Burt, A. & Koufopanou, V. Homing endonuclease genes: the rise and fall and rise again of a selfish element. Curr. Opin. Genet. Dev. 14, 609–615 (2004)
Chen, C. H. et al. A synthetic maternal-effect selfish genetic element drives population replacement in Drosophila . Science 316, 597–600 (2007)
McMeniman, C. J. et al. Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti . Science 323, 141–144 (2009)
Ashworth, J. et al. Computational redesign of endonuclease DNA binding and cleavage specificity. Nature 441, 656–659 (2006)
Jarjour, J. et al. High-resolution profiling of homing endonuclease binding and catalytic specificity using yeast surface display. Nucleic Acids Res. 37, 6871–6880 (2009)
Ashworth, J. et al. Computational reprogramming of homing endonuclease specificity at multiple adjacent base pairs. Nucleic Acids Res. 38, 5601–5608 (2010)
Thyme, S. B. et al. Exploitation of binding energy for catalysis and design. Nature 461, 1300–1304 (2009)
Gao, H. et al. Heritable targeted mutagenesis in maize using a designed endonuclease. Plant J. 61, 176–187 (2010)
Grizot, S. et al. Efficient targeting of a SCID gene by an engineered single-chain homing endonuclease. Nucleic Acids Res. 37, 5405–5419 (2009)
Munoz, I. G. et al. Molecular basis of engineered meganuclease targeting of the endogenous human RAG1 locus. Nucleic Acids Res. 39, 729–743 (2010)
Redondo, P. et al. Molecular basis of xeroderma pigmentosum group C DNA recognition by engineered meganucleases. Nature 456, 107–111 (2008)
Arnould, S. et al. Engineered I-CreI derivatives cleaving sequences from the human XPC gene can induce highly efficient gene correction in mammalian cells. J. Mol. Biol. 371, 49–65 (2007)
Rosen, L. E. et al. Homing endonuclease I-CreI derivatives with novel DNA target specificities. Nucleic Acids Res. 34, 4791–4800 (2006)
Li, H., Pellenz, S., Ulge, U., Stoddard, B. L. & Monnat, R. J., Jr Generation of single-chain LAGLIDADG homing endonucleases from native homodimeric precursor proteins. Nucleic Acids Res. 37, 1650–1662 (2009)
Sheng, G., Thouvenot, E., Schmucker, D., Wilson, D. S. & Desplan, C. Direct regulation of rhodopsin 1 by Pax-6/eyeless in Drosophila: evidence for a conserved function in photoreceptors. Genes Dev. 11, 1122–1131 (1997)
Lobo, N. F., Clayton, J. R., Fraser, M. J., Kafatos, F. C. & Collins, F. H. High efficiency germ-line transformation of mosquitoes. Nature Protocols 1, 1312–1317 (2006)
Catteruccia, F., Godfray, H. C. & Crisanti, A. Impact of genetic manipulation on the fitness of Anopheles stephensi mosquitoes. Science 299, 1225–1227 (2003)
Scalley-Kim, M., McConnell-Smith, A. & Stoddard, B. L. Coevolution of a homing endonuclease and its host target sequence. J. Mol. Biol. 372, 1305–1319 (2007)
Thyme, S. B. et al. Exploitation of binding energy for catalysis and design. Nature 461, 1300–1304 (2009)
Doyon, J. B., Pattanayak, V., Meyer, C. B. & Liu, D. R. Directed evolution and substrate specificity profile of homing endonuclease I-SceI. J. Am. Chem. Soc. 128, 2477–2484 (2006)
Argast, G. M., Stephens, K. M., Emond, M. J. & Monnat, R. J., Jr I-PpoI and I-CreI homing site sequence degeneracy determined by random mutagenesis and sequential in vitro enrichment. J. Mol. Biol. 280, 345–353 (1998)
Ulge, U. Y., Baker, D. A. & Monnat, R. J. Comprehensive computational design of mCreI homing endonuclease cleavage specificity for genome engineering. Nucleic Acids Res. 10.1093/nar/gkr022 (1 February 2011)
McConnell Smith, A. et al. Generation of a nicking enzyme that stimulates site-specific gene conversion from the I-AniI LAGLIDADG homing endonuclease. Proc. Natl Acad. Sci. USA 106, 5099–5104 (2009)
Das, R. & Baker, D. Macromolecular modeling with Rosetta. Annu. Rev. Biochem. 77, 363–382 (2008)
Studier, F. W. Protein production by auto-induction in high density shaking cultures. Protein Expr. Purif. 41, 207–234 (2005)
Acknowledgements
We thank M. Ashburner, S. Russell, D. Huen and S. Chan for comments, assistance and for plasmids. We thank M. P. Calos for providing the pET11phiC31polyA plasmid. We thank M. J. Fraser Jr for providing the pBSII-IFP2-orf plasmid. We thank J. Meredith and P. Eggleston for providing the docking strain. We thank A. Hall, T. Nolan, K. Magnusson, D. Rogers and S. Fuchs for assistance. We thank S. Arshiya Quadri and M. Szeto for experimental support and the members of the laboratories of D. Baker, R. Monnat, A. Scharenberg and B. Stoddard for their collective support of HEG engineering. A. F. M. Hackmann provided graphics support. Funded by a grant from the Foundation for the National Institutes of Health through the Vector-Based Control of Transmission: Discovery Research (VCTR) program of the Grand Challenges in Global Health initiative and by NIH RL1 awards GM084433 to D.B. and CA133831 to R.J.M.
Author information
Authors and Affiliations
Contributions
N.W. designed the experiments. N.W., M.M. and P.A.P. performed the experiments. N.W. and P.A.P. generated the transgenic lines. M.M. maintained mosquito populations. N.W. analysed the data. A.B. and N.W. generated the population dynamic models. A.C. and A.B. inspired the work and wrote the paper together with N.W. HEG redesign and target site cleavage analyses were performed by S.B.T., H.L., U.Y.U. (contributed equally) and B.T.H. with guidance from D.B. and R.J.M. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Information
This file contains Supplementary Figures 1-6 with legends and Supplementary Table 1. (PDF 2210 kb)
PowerPoint slides
Rights and permissions
About this article
Cite this article
Windbichler, N., Menichelli, M., Papathanos, P. et al. A synthetic homing endonuclease-based gene drive system in the human malaria mosquito. Nature 473, 212–215 (2011). https://doi.org/10.1038/nature09937
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nature09937
This article is cited by
-
Field investigation combined with modeling uncovers the ecological heterogeneity of Aedes albopictus habitats for strategically improving systematic management during urbanization
Parasites & Vectors (2023)
-
Evaluation of anti-malaria potency of wild and genetically modified Enterobacter cloacae expressing effector proteins in Anopheles stephensi
Parasites & Vectors (2022)
-
Versioning biological cells for trustworthy cell engineering
Nature Communications (2022)
-
A CRISPR endonuclease gene drive reveals distinct mechanisms of inheritance bias
Nature Communications (2022)
-
Gene drives gaining speed
Nature Reviews Genetics (2022)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.