Nucleic Acids Research Advance Access originally published online on August 28, 2007
Nucleic Acids Research 2007 35(17):5922-5933; doi:10.1093/nar/gkm632
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Nucleic Acids Research, 2007, Vol. 35, No. 17 5922-5933
© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Molecular Biology |
Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos
1Division of Cell and Molecular Biology, Imperial College London, Imperial College Road, London SW7 2AZ and 2Division of Biology and NERC Centre for Population Biology, Imperial College London, Silwood Park, Ascot, Berks SL5 7PY, UK
*To whom correspondence should be addressed. Tel: +44 (0)20 75945426; Fax: +44 (0)20 75945439; Email: acrs{at}icex.imperial.ac.uk
Received June 29, 2007. Revised July 31, 2007. Accepted July 31, 2007.
Homing endonuclease genes (HEGs) are ‘selfish’ genetic elements that combine the capability to selectively disrupt specific gene sequences with the ability to rapidly spread from a few individuals to an entire population through homologous recombination repair events. Because of these properties, HEGs are regarded as promising candidates to transfer genetic modifications from engineered laboratory mosquitoes to wild-type populations including Anopheles gambiae the vector of human malaria. Here we show that I-SceI and I-PpoI homing endonucleases cleave their recognition sites with high efficiency in A. gambiae cells and embryos and we demonstrate HEG-induced homologous and non-homologous repair events in a variety of functional assays. We also propose a gene drive system for mosquitoes that is based on our finding that I-PpoI cuts genomic rDNA located on the X chromosome in A. gambiae, which could be used to selectively incapacitate X-carrying spermatozoa thereby imposing a severe male-biased sex ratio.
The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors
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