1. A novel engineered meganuclease induces homologous recombination in yeast and mammalian cells
- Author
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Alexandre Zanghellini, Amélie Patin, Sylvain Arnould, Emmanuel Lacroix, Dominique Desfontaines, Patrick Chames, Frédéric Pâques, Pascal Rochaix, Clémence Puzin, and Jean-Charles Epinat
- Subjects
Models, Molecular ,Protein Folding ,I-CreI ,Hot Temperature ,Recombinant Fusion Proteins ,Computational biology ,Protein Engineering ,Homing endonuclease ,Genome engineering ,Endonuclease ,Yeasts ,Genetics ,Animals ,Recombination, Genetic ,biology ,Base Sequence ,Deoxyribonucleases, Type I Site-Specific ,Gene targeting ,DNA ,DNA Restriction Enzymes ,Articles ,Reverse genetics ,Protein Structure, Tertiary ,Meganuclease ,COS Cells ,biology.protein ,Homologous recombination - Abstract
Homologous gene targeting is the ultimate tool for reverse genetics, but its use is often limited by low efficiency. In a number of recent studies, site- specific DNA double-strand breaks (DSBs) have been used to induce efficient gene targeting. Engineering highly specific, dedicated DNA endonucleases is the key to a wider usage of this technology. In this study, we present two novel, chimeric meganucleases, derived from homing endonucleases. The first one is able to induce recombination in yeast and mammalian cells, whereas the second cleaves a novel (chosen) DNA target site. These results are a first step toward the generation of custom endonucleases for the purpose of targeted genome engineering.
- Published
- 2003