Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos.
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.
Windbichler
Nikolai
N
Division of Cell and Molecular Biology, Imperial College London, Imperial College Road, London SW7 2AZ.
Papathanos
Philippos Aris
PA
Catteruccia
Flaminia
F
Ranson
Hilary
H
Burt
Austin
A
Crisanti
Andrea
A
D000818:Animals; D000852:Anopheles / Q000166:cytology / Q000196:embryology / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation; D004260:DNA Repair; D004275:DNA, Ribosomal / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D005785:Gene Conversion; D018390:Gene Targeting; D017930:Genes, Reporter; D007303:Insect Vectors / Q000166:cytology / Q000196:embryology / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D012339:RNA, Ribosomal, 28S / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA
D004275:DNA, Ribosomal; D012339:RNA, Ribosomal, 28S; D029701:Saccharomyces cerevisiae Proteins; D004706:Endodeoxyribonucleases; C064579:I-Ppo endonuclease; C037725:SCEI protein, S cerevisiae; D015252:Deoxyribonucleases, Type II Site-Specific