Homing Endonuclease Mediated Gene Targeting in Anopheles Gambiae Cells and Embryos

Nucleic Acids Res. 2007;35(17):5922-33. doi: 10.1093/nar/gkm632. Epub 2007 Aug 28.


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anopheles / cytology
  • Anopheles / embryology
  • Anopheles / genetics*
  • Cell Line
  • Cell Proliferation
  • DNA Repair
  • DNA, Ribosomal / metabolism
  • Deoxyribonucleases, Type II Site-Specific / genetics
  • Deoxyribonucleases, Type II Site-Specific / metabolism*
  • Embryo, Nonmammalian / metabolism
  • Endodeoxyribonucleases / genetics
  • Endodeoxyribonucleases / metabolism*
  • Gene Conversion
  • Gene Targeting*
  • Genes, Reporter
  • Insect Vectors / cytology
  • Insect Vectors / embryology
  • Insect Vectors / genetics*
  • Plasmids / genetics
  • RNA, Ribosomal, 28S / genetics
  • Repetitive Sequences, Nucleic Acid
  • Saccharomyces cerevisiae Proteins
  • Sequence Analysis, DNA


  • DNA, Ribosomal
  • RNA, Ribosomal, 28S
  • Saccharomyces cerevisiae Proteins
  • Endodeoxyribonucleases
  • I-Ppo endonuclease
  • SCEI protein, S cerevisiae
  • Deoxyribonucleases, Type II Site-Specific