A novel engineered meganuclease induces homologous recombination in yeast and mammalian cells.
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.
Epinat
Jean-Charles
JC
CELLECTIS S.A., 28 rue du Dr Roux, 75724 Paris Cedex 15, France.
Arnould
Sylvain
S
Chames
Patrick
P
Rochaix
Pascal
P
Desfontaines
Dominique
D
Puzin
Clémence
C
Patin
Amélie
A
Zanghellini
Alexandre
A
Pâques
Frédéric
F
Lacroix
Emmanuel
E
D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D004247:DNA / Q000378:metabolism; D004262:DNA Restriction Enzymes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D015253:Deoxyribonucleases, Type I Site-Specific / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006358:Hot Temperature; D008958:Models, Molecular; D015202:Protein Engineering; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D011995:Recombination, Genetic; D015003:Yeasts / Q000235:genetics*
D011993:Recombinant Fusion Proteins; D004247:DNA; D004262:DNA Restriction Enzymes; C048901:endodeoxyribonuclease CreI; C081755:endodeoxyribonuclease I-Dmo I; D015253:Deoxyribonucleases, Type I Site-Specific