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 specificity. 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 vector. The translation of these achievements into control measures requires an effective technology to spread a genetic modification from laboratory mosquitoes to field populations. We have suggested previously that homing endonuclease genes (HEGs), a class of simple selfish genetic elements, could be exploited for this purpose. Here we demonstrate that a synthetic genetic element, consisting of mosquito regulatory regions and the homing endonuclease gene I-SceI, 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.
MeSH Terms
Animals
Animals, Genetically Modified
Anopheles/genetics
Deoxyribonucleases, Type II Site-Specific/genetics
Female
Genes, Reporter/genetics
Genetic Engineering
Genotype
Insect Vectors/genetics
Male
Molecular Sequence Data
Mosquito Control/methods
Saccharomyces cerevisiae Proteins/genetics
Chemicals
Saccharomyces cerevisiae Proteins
AI4 protein, S cerevisiae
Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Windbichler Nikolai
Imperial College London, Department of Life Sciences, South Kensington Campus, London, SW7 2AZ, UK.
Menichelli Miriam
Papathanos Philippos Aris
Thyme Summer B
Li Hui
Ulge Umut Y
Hovde Blake T
Baker David
Monnat Raymond J
Burt Austin
Crisanti Andrea
References (30)
30 references, click to expand
-
Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos.
Nucleic Acids Res. 2007;35(17):5922-33
PMID: 17726053
-
Computational reprogramming of homing endonuclease specificity at multiple adjacent base pairs.
Nucleic Acids Res. 2010 Sep;38(16):5601-8
PMID: 20435674
-
Exploitation of binding energy for catalysis and design.
Nature. 2009 Oct 29;461(7268):1300-4
PMID: 19865174
-
Possible use of translocations to fix desirable genes in insect pest populations.
Nature. 1968 Apr 27;218(5139):368-9
PMID: 5649682
-
An Anopheles transgenic sexing strain for vector control.
Nat Biotechnol. 2005 Nov;23(11):1414-7
PMID: 16244659
-
Molecular basis of engineered meganuclease targeting of the endogenous human RAG1 locus.
Nucleic Acids Res. 2011 Jan;39(2):729-43
PMID: 20846960
-
Heritable targeted mutagenesis in maize using a designed endonuclease.
Plant J. 2010 Jan;61(1):176-87
PMID: 19811621
-
Homing endonuclease I-CreI derivatives with novel DNA target specificities.
Nucleic Acids Res. 2006;34(17):4791-800
PMID: 16971456
-
Extraordinary sex ratios. A sex-ratio theory for sex linkage and inbreeding has new implications in cytogenetics and entomology.
Science. 1967 Apr 28;156(3774):477-88
PMID: 6021675
-
Outcrossed sex allows a selfish gene to invade yeast populations.
Proc Biol Sci. 2001 Dec 22;268(1485):2537-42
PMID: 11749707
-
Molecular basis of xeroderma pigmentosum group C DNA recognition by engineered meganucleases.
Nature. 2008 Nov 6;456(7218):107-11
PMID: 18987743
-
Transgenic anopheline mosquitoes impaired in transmission of a malaria parasite.
Nature. 2002 May 23;417(6887):452-5
PMID: 12024215
-
Generation of single-chain LAGLIDADG homing endonucleases from native homodimeric precursor proteins.
Nucleic Acids Res. 2009 Apr;37(5):1650-62
PMID: 19153140
-
Homing endonuclease genes: the rise and fall and rise again of a selfish element.
Curr Opin Genet Dev. 2004 Dec;14(6):609-15
PMID: 15531154
-
An intron-encoded protein is active in a gene conversion process that spreads an intron into a mitochondrial gene.
Cell. 1985 Jun;41(2):383-94
PMID: 3886163
-
Activation of Akt signaling reduces the prevalence and intensity of malaria parasite infection and lifespan in Anopheles stephensi mosquitoes.
PLoS Pathog. 2010 Jul 15;6(7):e1001003
PMID: 20664791
-
An anti-Chitinase malaria transmission-blocking single-chain antibody as an effector molecule for creating a Plasmodium falciparum-refractory mosquito.
J Infect Dis. 2005 Sep 1;192(5):878-87
PMID: 16088838
-
Gene drive systems for insect disease vectors.
Nat Rev Genet. 2006 Jun;7(6):427-35
PMID: 16682981
-
A synthetic maternal-effect selfish genetic element drives population replacement in Drosophila.
Science. 2007 Apr 27;316(5824):597-600
PMID: 17395794
-
Site-specific selfish genes as tools for the control and genetic engineering of natural populations.
Proc Biol Sci. 2003 May 7;270(1518):921-8
PMID: 12803906
-
Stable introduction of a life-shortening Wolbachia infection into the mosquito Aedes aegypti.
Science. 2009 Jan 2;323(5910):141-4
PMID: 19119237
-
Site-specific integration and expression of an anti-malarial gene in transgenic Anopheles gambiae significantly reduces Plasmodium infections.
PLoS One. 2011 Jan 25;6(1):e14587
PMID: 21283619
-
Computational redesign of endonuclease DNA binding and cleavage specificity.
Nature. 2006 Jun 1;441(7093):656-9
PMID: 16738662
-
Efficient targeting of a SCID gene by an engineered single-chain homing endonuclease.
Nucleic Acids Res. 2009 Sep;37(16):5405-19
PMID: 19584299
-
High-resolution profiling of homing endonuclease binding and catalytic specificity using yeast surface display.
Nucleic Acids Res. 2009 Nov;37(20):6871-80
PMID: 19740766
-
I-SceI endonuclease, a new tool for studying DNA double-strand break repair mechanisms in Drosophila.
Genetics. 1999 Jul;152(3):1037-44
PMID: 10388822
-
Engineered I-CreI derivatives cleaving sequences from the human XPC gene can induce highly efficient gene correction in mammalian cells.
J Mol Biol. 2007 Aug 3;371(1):49-65
PMID: 17561112
-
Homing endonuclease structure and function.
Q Rev Biophys. 2005 Feb;38(1):49-95
PMID: 16336743
-
Bee venom phospholipase inhibits malaria parasite development in transgenic mosquitoes.
J Biol Chem. 2002 Oct 25;277(43):40839-43
PMID: 12167627
-
Malaria control with genetically manipulated insect vectors.
Science. 2002 Oct 4;298(5591):119-21
PMID: 12364786